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Year : 2022  |  Volume : 65  |  Issue : 6  |  Page : 4-92
Oral paper abstracts

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Date of Web Publication28-Nov-2022

How to cite this article:
. Oral paper abstracts. Indian J Pathol Microbiol 2022;65, Suppl S2:4-92

How to cite this URL:
. Oral paper abstracts. Indian J Pathol Microbiol [serial online] 2022 [cited 2023 Jan 30];65, Suppl S2:4-92. Available from:

   AOP1 Top

   Autopsy Pathology Top

Histopathological evaluation of fetal cardia – An autopsy study

Jahnvi Vijay, B Deepak Kumar, V Srinivasa Murthy

ESIC Medical College and PGIMSR, Bengaluru, Karnataka, India

Introduction: Cardiac anomalies form a significant proportion of fetal congenital anomalies which can lead to non-viable pregnancy. The incidence of congenital heart defects is 6 to 7 per 1000 live births with much higher rates detected during fetal autopsy. The study was undertaken as cardiac autopsy places high demands on pathologists with regards to proper morphological assessment. Objectives: To study the spectrum and prevalence of fetal cardiac anomalies based on morphological assessment and to analyze the association of the same with other congenital anomalies. Materials and Methods: Fetal cardia were retrospectively analysed over a period of 5 years and examined morphologically for any anomalies and were categorised as cardiac and extracardiac. Results: A total of 231 fetal cardia were evaluated. The prevalence rate of cardiac anomalies was 12.5%. The cases were divided as those with exclusive cardiac anomalies (6.5%); with combined cardiac and extracardiac anomalies (6%); with normal cardia and exclusive organ specific anomalies (15.6%); with normal cardia and multiorgan extracardiac anomalies (10.8%). 61% of cases showed no anomalies. ASD (1.3%), 2 chambered heart (0.86%), anencephaly (5.2%) and craniorachischisis (2.6%) were the commonest anomalies in the above categories respectively. In addition, the above categories showed more than ten different types of anomalies in each of them. Conclusion: A detailed fetal cardiac morphological evaluation helps to identify cardiac anomalies which form a significant proportion of fetal congenital anomalies. Recognition of these with associated extracardiac anomalies helps in understanding the prevailing pathology and also guides in parental counselling for future pregnancies.

Keywords: Anomalies, cardia, fetus

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   Autopsy Pathology Top

Single umbilical artery and associated birth defects in perinatal autopsies – Need for prenatal diagnosis and management

Manushree Saxena, Bhagyashri R Hungund

Jawaharlal Nehru Medical College, Belagavi, Karnataka, India

Introduction: The umbilical artery develops between 13 and 38 days after conception and normally consists of two umbilical arteries and one umbilical vein. However, in 0.5-1% of fetuses, one umbilical artery is either obliterated or does not form giving rise to single umbilical artery (SUA). The incidence is higher (0.3-7%) among twins, fetal deaths, abortuses and autopsies. SUA is more often found in association with a wide range of other malformations than as an isolated defect. Hence there is need for detailed evaluation of fetus to look for other anomalies when SUA is discovered during routine ultrasound. Objectives: To estimate association between Single Umbilical Artery and congenital anomalies in perinatal autopsies. Materials and Methods: This is a retrospective study done at Jawaharlal Nehru Medical College and Research Centre, Belagavi, India. The data was obtained from hospital pathology laboratory records from year 2013 to 2022. We evaluated 1277 fetuses sent for autopsy. The birth defects were grouped by organ/system for analysis including cardiovascular, urinary tract, nervous system, GIT, musculoskeletal, lung anomalies and others. Results: Of 1277 autopsies, 110 (8.61%) had single umbilical artery with gestational age ranging from 13 to 40 weeks. Among these, 66 (60%) were associated with other congenital anomalies with predominant involvement of the urinary tract. Conclusion: We found single umbilical artery in 8.61% of the autopsies. It is more often found in association with other congenital malformations than as an isolated defect with predominant involvement of the urinary system.

Keywords: Birth-defects, perinatal autopsy, single umbilical artery

   AOP3 Top

   Autopsy Pathology Top

Congenital anomalies in fetal autopsy

Aiswarya George, K Nagarekha, C Bharath

Vijayanagar Institute of Medical Sciences, Bellary, Karnataka, India

Introduction: The common indications of fetal autopsies are termination of pregnancy for congenital anomalies, intrauterine fetal death and inevitable abortion. Perinatal autopsy either confirms or refutes ultrasound diagnosis and complements ultrasound examination in making precise diagnosis of the cause of intrauterine fetal death. Objectives: To evaluate the causes of intrauterine fetal death and avoid recurrence in future pregnancies. Materials and Methods: Retrospective study was conducted in Department of Pathology, VIMS Bellary. A total of 100 cases of intrauterine fetal death specimens were received and were grossly analyzed. Results: Out of 100 cases studied, 78 were normal by autopsy findings. Rest 22 were abnormal. Among these, the most common anomaly detected was anencephaly (3 out of 22 cases, 13.6%) Other anomalies included mermaid syndrome, meningocele, bilateral club foot, solitary kidney, anophthalmia, ambiguous genitalia and hydrops fetalis. Conclusion: This study confirms the utility of fetal autopsy in providing valuable information in identifying cause of fetal loss. Even though prenatal ultrasonogram reasonably predicts malformations, fetal autopsy gives significant additional information about malformations, hence plays an important role in genetic counselling and management of future pregnancies.

Keywords: Congenital anomalies, fetal autopsy, intrauterine death

   AOP4 Top

   Autopsy Pathology Top

Correlation between prenatal ultrasound and morphological findings of fetal autopsy in congenital anomalies

Ankita Madinkar, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Ultrasound examination during pregnancy is considered to be an important part of prenatal care, as it may reveal fetal developmental anomalies. Postmortem examination of aborted fetuses is an important tool for assessing the quality and accuracy of prenatal ultrasound report. Moreover the examination may provide additional information to that obtained by prenatal ultrasonography. Objectives: To Evaluate discrepancies between ultrasonographic and autopsy findings following the intrauterine death of fetus and to categorise them into different categories. Materials and Methods: This is a Prospective unicentre study of total 50 cases of fetal anomalies over a period of 1.5 years. For Prospective study data including prenatal usg report was collected from case sheet. A standard protocol of autopsy procedure was followed, which included external examination, internal examination and histopathological examination. Results: The findings of Prenatal Ultrasound were compared with Autopsy findings in 50 Fetuses received in the department of Pathology. The Primary diagnosis showed 32 CNS cases, 1 case of RS, 6 cases of CVS, 1 case of Genitourinary system, 2 cases of skeletal malformation, 2 cases GI system, 6 cases with other abnormalities. 24% of total anomalies had multiple malformation. In 29 cases (58%) prenatal ultrasound was confirmed with autopsy findings, In 15 cases (30%) USG findings was confirmed with additional findings, In 6 cases (12%) USG findings was Unconfirmed with autopsy findings. Conclusion: Even though Prenatal ultrasonographic examination detects the fetal anomalies, postmortem examination is still necessary to verify or improve the prenatal diagnosis and may influence future counseling.

Keywords: Congenital anomalies, fetal autopsy, prenatal ultrasonography

   AOP5 Top

   Autopsy Pathology Top

Spectrum of histopathological findings in the heart: An autopsy study in a tertiary care hospital

Manpreet Kaur, R K Sharma, Permeet Kaur Bagga, Jaspreet Singh

GMC, Amritsar, Punjab, India

Introduction: Sudden death in adults without any past history of chronic illness are increasing world over and is an issue of concern for medical professionals. Even in this modernized medical world, diagnostic tools lack precision to find the clinical cause of death in comparison to autopsy cause of death. The main aim of autopsy is to find out the most possible cause of death. World health organization defines sudden death as “death within 24 hours from the onset of symptoms”. This study was conducted to analyze histopathologically the possible cardiovascular causes of sudden death. Objectives: This study was planned to observe various histomorphological cardiac changes in patients presenting with sudden death of cardiac etiology. Materials and Methods: This prospective study was carried out in the Department of Pathology, Government Medical College, Amritsar from January 2021 to October 2022. A total of 100 cases were studied during this period. Results: Of the total 100 cases studied MI was the major cause of death (38%) followed by myocarditis (4%) cardiomyopathy (2%). AP in coronary arteries were noted in 55% of cases. Conclusion: Sudden death in adults is an issue of concern and a meticulous postmortem and HPE is necesaary to ascertain its cause.

Keywords: AP, MI, HPE

   AOP6 Top

   Autopsy Pathology Top

The book-keeping of make every mother and child count: An experience at tertiary care hospital

Prasad Charpe, Lalita Y Patil, Shruti Lande, Kusum Jashnani

TNMC and B.Y.L Nair Ch. Hospital, Mumbai, Maharashtra, India

Introduction: Maternal mortality is a major public health challenge despite numerous strategies. Pathological autopsy is an important tool in investigation of maternal deaths as it provides critical information about pathophysiological changes in various organs important in the sequence of events leading to death. Objectives: To find out various causes leading to maternal deaths and contribution of different etiologies responsible for same. Materials and Methods: Retrospective study executed at a tertiary care centre from January 2014 to December 2021. Total 311 maternal mortality autopsies were performed and histopathological and clinicopathological correlation was studied in detail. The medicolegal cases were excluded. Results: On evaluation, maximum maternal mortality was due to indirect causes-222 (71.39%) of which hepatic cause was most common-67 (30.18%). The most common direct cause was pregnancy induced hypertension-18 (20.22%). Maximum deaths were between age group 21-25 years-132 (42.44%). Maximum deaths occured in post partum period- 190 (61.09%) and in multiparous females- 107 (56.31%). Conclusion: Autopsy based study provides detailed analysis of various causes (direct and indirect) leading to maternal deaths, in addition detalied gross and microscopic examination of each organ can be done. The various risk factors which contribute to maternal deaths and multiple parameters like age, gravida, parity, duration of hospital stay, trimester can be studied in collaboration. The loop holes regarding health of mother and child in our health system can be identified and necessary steps advised by WHO to reduce mother and child loss and upliftment of health system can be taken on priority bases.

Keywords: Histopathology, maternal mortality, pathological autopsy

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   Autopsy Pathology Top

Histopathological spectrum of traumatic brain injury

Prem Kalagi, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Traumatic brain injury is an umbrella term for a range of focal and diffuse pathologies which can be associated with trauma. It encompasses blunt, non-missile head injury, especially involving motor vehicle accidents, and penetrating, missile injury. Head injuries are classified into 2 major categories of brain damage; primary and secondary damages. Primary damages include scalp lacerations, skull fractures, contusions and lacerations of the brain, diffuse axonal injury, and the intracranial hemorrhages. Secondary damages arise from the primary damages and include the lesions produced by increased intracranial pressure, ischemia, brain swelling, and infection. Objectives: To determine Spectrum of Brain Pathology in Traumatic Brain Injuries. Materials and Methods: All medico-legal autopsy subjects irrespective of age, sex & cause of death, satisfying inclusion and exclusion criteria, included in the study. A prospective study was done for 18 months. The brain specimen was examined grossly and microscopically. Special stains like LFB, GFAP, NFP, BetaAPP were used wherever required. All the cases were analyzed by descriptive statistics. Results: A total of 230 patients were studied during 18 months period, and majority (81.7%) were males. 118/230 (51.3%) were having intraparenchymal hemorrhage, 83/230 (36.1%) had subarachnoid hemorrhage, 16/230 (6.9%) had subdural hemorrhage, 13/230 (5.6%) were suspicious of diffuse axonal injury. Conclusion: The most common cause of traumatic brain injury in our study was found to be road traffic accidents. Thus, it can be inferred that morbidity and mortality from traumatic brain injuries can be reduced by taking steps to prevent road traffic accidents.

Keywords: Autopsy, intracranial hemorrhage, road traffic accidents

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   Autopsy Pathology Top

Alarm! Coronary atherosclerosis in non-cardiac deaths: An autopsy study

Vivek Mannammanavar, S Rukmini, U T Kanchana, C Prabhashree

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Heart disease has remained the first leading cause of death at the global level for last 20 yrs. To evaluate the prevalence of atherosclerosis, an autopsy-based study conducted on subjects who died of non cardiac causes can be a valuable tool. Objectives: Age and sex distribution and importance of early surveillance of atherosclerosis in deaths due to non-cardiac causes. Assessment of pattern of involvement coronaries including grading of severity in atherosclerosis. Materials and Methods: This study was conducted from January to December 2021 at Department of Pathology, KIMS Hubballi. Patients who died of noncardiac causes underwent autopsy, heart specimens were sent in 10% formalin for histopathological analysis. Right and Left coronary arteries were dissected and examined for any atherosclerotic plaques and associated pathological lesions and graded according to the classification given by American Heart Association. Results: A total of 215 autopsy cases were evaluated. Amongst them 210 were deaths due to noncardiac causes, of which 131 (62.38%) subjects had evidence of atherosclerosis. In the study, 120 (91.61%) were males and 11 (8.39%) were females. Left coronary artery (64%) was the most commonly involved coronary artery. Right and left vessel disease was found in 22% of subjects. Conclusion: The study showed alarmingly high prevalence of atherosclerosis. The pathogenesis of coronary atherosclerosis begins at a younger age in Indian population. Though the incidence of atherosclerosis is more common in males compared to females; coronary atherosclerosis is an important risk factor for IHDs in both sexes and screening for the same should begin at an early age.

Keywords: Atherosclerosis, autopsy, coronary

   AOP9 Top

   Autopsy Pathology Top

Postmortem pathological changes in lungs, liver, heart and kidney in burn cases

Alokabandhu Nayak, Pranati Mohanty, Soumya Ranjan Nayak, Lity Mohanty, Asranti Kar

Department of Pathology, SCB Medical College, Cuttack, Odisha, India

Introduction: A severe burn injury is the most devastating injury a person can sustain and yet hope to survive. Every year more than 2 million people sustained burns in India. Thermal burns are most common incidents which stands next to road traffic accidents. There is lack of information on the cause of death due to burns and histopathological changes hence this study was planned to study on the cause of death due to burns and its histopathological changes. Objectives: To study whether there is any correlation between survival period and pathological changes in vital organs in burn cases. Materials and Methods: All the autopsy samples (kidney, liver, heart and lungs) of burn patients were fixed with 10% neutral buffered formalin. Grossing was given from the representative areas, tissue processing and embedding in paraffin wax was done according to standard protocol. Staining was done with H &E and sections examined under low and high power. Results: 40 cases were studied. In lungs 30 cases (75%) showed congestion and 17 cases (42.5%) showed edema. In kidney 24 cases (60%) showed congestion, 15 cases (37.5%) showed necrosis. In liver 20 cases (50%) showed congestion, 18 cases (45%) showed necrosis. Conclusion: From the study we concluded that the most common histopathological change associated with visceral organs in burns is congestion followed by necrosis.

Keywords: Autopsy, burn, congestion, grossing

   AOP10 Top

   Autopsy Pathology Top

Histopathological study of medicolegal autopsies with incidental and interesting cases

Swati Ghanghurde, Vaibhav Bari, Anuja Yadav

Rajiv Gandhi Medical College, Thane, Mumbai, Maharashtra, India

Introduction: Autopsy is a medical practice in which thorough body examination done after death. The internal organs are studied grossly and histopathologically to determine cause or manner of death. This study spotlights the various histopathological findings in autopsies for academic and research purpose. Objectives: 1. To study histopathological spectrum of lesions encountered in medicolegal autopsy. 2. To highlight various incidental and interesting microscopic findings. Materials and Methods: It was a retrospective study of medicolegal autopsies for two years, from Jan 2018-Dec 2019. All medicolegal autopsies for these two years were included & clinical autopsies were excluded from the study. Clinical history, gross and microscopic findings were noted. Results: 262 autopsies were sent for histopathological examination. Majority of patients were seen in 21-30 years of age and majority of them were males. Out of 262 cases, most of them were brought dead followed by others, GIT complications & Poisoning. Most common histopathological finding was Pulmonary Oedema 199 cases (76%), followed by Pulmonary haemorrhage 133 cases (51%), Fatty change liver 121 cases (46%), Atherosclerosis 108 cases (41%) etc. Some of the incidental findings were RCC, Membranous nephropathy, Sex-cord stromal tumour, Embolism (Amniotic fluid, bone marrow embolism), Fungal Abscess in lungs & Sickle cell disease. Conclusion: This study provides an insight into the true prevalence of disease or lesions in the society. This study highlights the various incidental cases, which are imperative in academic and research purposes.

Keywords: Autopsy, histopathology, medicolegal

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   Autopsy Pathology Top

Spectrum of cardiac anomalies in foetal and perinatal autopsies: An experience of a decade in a tertiary care hospital

Basavaraj Yamakanamardi, Hephzibah Rani, Ravikala V Rao

Department of Pathology, SDM College of Medical Sciences and Hospital, Dharwad, Karnataka, India

Introduction: Congenital cardiac anomalies are the most common defects in newborns with global incidence of 3.3 to 8.1 per 1000 live births. Identifying the cause of death & the associated anomaly is important to determine the risk of recurrence. Objectives: 1. To evaluate the spectrum of cardiac anomalies in foetal and perinatal autopsies 2. To analyze its association with extra cardiac malformations. Materials and Methods: It is a 10 year retrospective study. Fetuses diagnosed with cardiac abnormalities over the last decade were included in the study. The clinical & radiological details were collected from the medical records. Autopsy photographs & histopathological slides were retrieved from laboratory archives. Results: Total of 48 fetal autopsies were included in this study. Of these, isolated cardiac defect was seen in 25 cases and complex cardiac anomalies in 23 cases. The most common isolated cardiac anomaly was Septal defects (36%). Truncus arteriosus was the most common complex cardiac anomaly observed in 23 cases (32%). Association with extra cardiac malformations was seen in 16 cases, with renal being the most common (31.25%). Antenatal USG identified cardiac anomalies in 35 cases, and was missed in the remaining 13 cases. Conclusion: Septal defects are the most common isolated cardiac anomalies and truncus arteriosus is the most identified defect among the complex cardiac anomalies. A systematic fetal autopsy still remains the gold standard for the diagnosis of fetal anomalies.

Keywords: Cardiac anomalies, septal defects, truncus arteriosus

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   Bone/Joints/Soft Tissue Pathology Top

Pleomorphic Rhabdomyosarcoma – Clinicopathologic study of 15 rare tumors

Puja Thakur, Bharat Rekhi, Mukta Ramadwar

Tata Memorial Hospital, Mumbai, Maharashtra, India

Introduction: Primary pleomorphic rhabdomyosarcoma (RMS) constitutes a relatively uncommon, high-grade sarcoma, invariably observed in adult age-group with few reported case series, but none from our country. Objectives: To study clinicopathological features of diagnosed cases of pleomorphic RMS at our Institution from 2018 to 2022 (5 years). Materials and Methods: Slides of diagnosed cases of pleomorphic RMS, at our Institution from 2018 to 2022 (5 years) were retrieved and reviewed. Results: 15 cases were analyzed. The mean age was 53 years (range 36-74 years), including 12 males and 3 females. There were 5 biopsies, 2 resections and 8 biopsies with corresponding resections. Most common site was lower limb, especially thigh. T-size varied from 3.5 - 19 cm (mean 10.9). Histologically, tumor cells were arranged in sheets and fascicles showing pleomorphic bizzare cells, rhabdomyoblasts (n=11), frequent mitotic figures, including atypical forms and necrosis (n=10). Immunohistochemically, 13/15 (86.7%) tumors were diffusely positive for desmin while 2 showed patchy positivity. 8 of 15 tumors were focally positive for myogenin and 14/15 (93.3%) tumors were positive for MyoD1 (mostly diffuse or patchy). Treatment wise (n=10), 10 patients underwent surgery out of which 1 patient received adjuvant chemotherapy. 3 patients received neoadjuvant chemotherapy and 3 received adjuvant chemotherapy+radiation. With regards to outcomes (n=5), a single developed recurrence. 1 patient succumbed to the disease. A single patient developed metastasis. Conclusion: Pleomorphic RMS is a rare high- grade RMS associated with an aggressive outcome. Differential diagnoses will be presented.

Keywords: Desmin, MyoD1, pleomorphic rhabdomyosarcoma

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   Bone/Joints/Soft Tissue Pathology Top

Myxoid sarcomas: Histopathological spectrum with varied vascular patterns

R Saba Afreen, T N Chandrashekhar, Priyadarshini Berguman, B Kusumanjali

Shimoga Institute of Medical Sciences, Shivamogga, Karnataka, India

Introduction: Myxoid soft tissue tumors encompass a complex group of mesenchymal neoplasms characterized by the production of abundant extracellular myxoid matrix. Myxoid tumors can be benign or frankly malignant. Myxoid sarcomas include Myxoid liposarcoma, myxofibrosarcoma, low-grade fibromyxoid sarcoma and extraskeletal myxoid chondrosarcoma. Objectives: To describe histomorphological spectrum and vascular pattern of various myxoid soft tissue tumors. Materials and Methods: Retrospective and prospective observational study of 224 cases of myxoid histo-morphology with special emphasis on vascular pattern. Results: Majority of lesions are benign (73.21%), (19.19%) malignant cases and (7.58%) intermediate lesions. Lesions have male preponderance and arise predominantly from extremities. Most common benign myxoid lesions were of neural origin. Myxoid neurofibroma constituting (29. 01%) cases, followed by schwannoma (16.9%). Myxoid Dermatofibrosarcoma protuberans was the most common intermediate lesion. Tumors of adipocytic differentiation were predominant among the malignant lesions. Conclusion: The histomorphological patterns with varied vascular patterns are helpful for baseline differentiation of tumors pave way for effective diagnosis with ancillary techniques. Vascular pattern in the myxoid lesions are crucial in arriving at a histo-morphological diagnosis. Further studies correlating the vascular pattern with the genetic profile of these tumours can help arriving at a histo-morphological diagnosis of myxoid lesions.

Keywords: Histo-morphology, myxoid, soft tissue, vascular pattern

   AOP14 Top

   Bone/Joints/Soft Tissue Pathology Top

Solitary fibrous tumor: A clinicopathological study of 14 cases and metastatic risk stratification assessment

Pooja H Upase, G Champaka, S Balu

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Solitary fibrous tumor occurring in pleura and extra pleural sites represents a spectrum of mesenchymal neoplasms, which are of intermediate (locally aggressive and rarely metastasizing) and malignant categories (WHO Soft tissue & bone tumors 2020). These are characterized by NAB2-STAT6 gene rearrangement. They were previously termed as hemangiopericytoma which is no longer recommended. Objectives: To assess the metastatic risk stratification of solitary fibrous tumors diagnosed in our institute. Materials and Methods: All cases of Solitary fibrous tumors diagnosed in our institute Pathology department, based on morphology and IHC (STAT6 and/or CD34 positive) from August 2020 to September 2022 were reviewed and clinico-pathological correlation was done. The risk factors included age, tumor size, necrosis and mitosis. Results: The total number of cases reported were 14 which comprised of 8 males and 6 females. The most common location was lung, and the median age was 48 years. The youngest patient was 27 years old. Tumor size varied from 1 cm to 15 cm (average was 8 cm). Mitotic activity was more than 4/10 high power fields in most of the cases and only one case showed tumor necrosis. Conclusion: In this study, 4 point metastatic risk stratification was evaluated. Out of 14 cases, 7 cases were categorized as intermediate risk with resection being the modality of treatment. None of these cases fell under high-risk group. A follow up of these cases showed no local recurrence/metastasis.

Keywords: Metastatic risk stratification, solitary fibrous tumor

   AOP15 Top

   Bone/Joints/Soft Tissue Pathology Top

A gamut of chondroid tumors of humerus: Clinico-radiological dilemma and pathological consensus

Subarna Sharma, Geetika Khanna, Dharmendra Kumar Singh, Anju Sasidharan

Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India

Introduction: Chondroid tumors are a heterogenous group of benign (enchondroma), locally aggressive (atypical enchondroma, low grade chondrosarcoma) and malignant (high grade chondrosarcoma) entities, with clear cell chondrosarcoma being the commonest in humerus. The clinical presentation as nonspecific non-inflammatory lumps and the inconclusive radiological findings pose diagnostic difficulties. Histopathology is the gold standard that demonstrate chondrocytes embedded in a cartilaginous matrix and is the only reliable evidence in chondroid tumors to differentiate benign versus locally aggressive versus malignant lesions, irrespective of radiological findings. Objectives: 1. To illustrate the clinico-radiological spectrum of chondroid lesions in humerus. 2. To discuss the diagnostic dilemma of chondroid tumors. 3. To demonstrate the cases in which histopathology outperforms radiology in determining aggressiveness in these tumors. Materials and Methods: We collected radiological details of all patients presenting with nonspecific pain and lump in arm in the Orthopedics OPD, Safdarjung Hospital, New Delhi over last one year and identified those suspected to have chondroid tumors clinic-radiologically (n=10) and correlated with histopathology. All the cases were discussed in multidisciplinary team (MDT) meeting. Results: Out of the 12 cases, radiologically 3 were diagnosed as enchondroma, while 9 as chondrosarcoma. Histopathology was consistent with radiology findings in 2 cases of enchondroma (66.66%) and 7 cases of chondrosaroma (77.77%). The radiopathological discordance were discussed in MDT to identify potential pitfalls. Conclusion: Histopathology is superior over radiology in determining the aggressive nature in intra-osseous chondroid tumors.

Keywords: Chondroid tumor, chondrosarcoma, enchondroma

   AOP16 Top

   Bone/Joints/Soft Tissue Pathology Top

Correlation of bone marrow aspiration and trephine biopsy in various diseases: A 2 years study

S Shanmathi, H M Sudha

M S Ramaiah Medical College, Bengaluru, Karnataka, India

Introduction: Bone marrow examination is an important tool to evaluate various disorders such as neoplastic, non-neoplastic hematological diseases and other non-hematological diseases. Bone marrow aspirate and trephine biopsy are complementary to each other and when both are obtained, they provide a comprehensive evaluation of bone marrow. Objectives: The study was aimed to correlate the findings of bone marrow aspirate and biopsy in various hematological and non-hematological diseases. Materials and Methods: A retrospective study was done for 2 years in the Department of Pathology, M.S. Ramaiah medical college and hospital, Bangalore. 86 consecutive patients in which bone marrow biopsy was done, were correlated with bone marrow aspirate. Results: Commonly encountered diseases were nutritional anemia (31.3%) and acute leukemia (11.6%). Out of 86 cases, 77 cases were diagnosed on bone marrow aspiration with diagnostic accuracy of 90%. And 82 cases were diagnosed on trephine biopsy of bone marrow, with diagnostic accuracy of 95%. The study showed 93.9% positive correlation between bone marrow aspirate and biopsy. Conclusion: Bone marrow biopsy was superior to bone marrow aspirates in diagnosis of myelofibrosis, metastatic solid tumor, aplastic anemia and morphological remission of marrow in leukemia, where bone marrow aspirates yielded a dry tap or diluted marrow. Though cellular morphology is better appreciated in BMA, this study emphasizes the usefulness of BMB for the detection of fibrosis, pattern of distribution of the cells and metastatic deposits.

Keywords: Bone marrow aspiration, correlation, trephine biopsy

   AOP17 Top

   Bone/Joints/Soft Tissue Pathology Top

Evaluation of fine needle aspiration cytology and core needle biopsy in space occupying lesion of bone

Priyadarshini Swain, Kalpalata Tripathy, Aparajita Mishra, Doki Sunil

Department of Pathology, SCB Medical College and Hospital, Cuttack, Odisha, India

Introduction: Fine-needle aspiration cytology (FNAC) is a simple, cost-effective procedure which is widely used to diagnose lesions in various sites but has a relatively less utility in bone lesions. when compared with core biopsy, FNAC is painless and cost-effective, has a lower risk of tumor seeding and gives an early diagnosis with minimal disruption of lesion. The present study attempts to find out the efficacy and reliability of fine-needle aspiration cytology (FNAC) in preoperative diagnosis of bone lesions in correlation with radiological and histopathological findings and to determine the spectrum and morphological features of various bone lesions on FNAC. Objectives: 1. To evaluate the efficacy of FNAC in the pre operative diagnosis of bone lesions. 2. Statistical evaluation of sensitivity, specificity, positive predictive value & negative predictive value. Materials and Methods: FNAC has been done in patients who had diagnosed to have bone lesions clinically and radiologically in dept of pathology, scb medical college cuttack. A total of 74 cases of bone lesions were studied by FNAC over a period of 2 years. 48 procedures were performed by pathologists, and 26 procedures were guided. Patients were sent for core needle biopsy in dept of orthopaedics. Cytology findings were correlated with that of histology on subsequent surgical biopsies. According to IBM SPSS statiscitcs V21.0 method statistical analysis was done. Results: Out of 74 cases 12 were tumor like lesions, 25 were benign tumors and 37 were malignant. Corelation with histology was available in 67 cases. Conclusion: We conclude that FNAC is a simple and accurate preoperative diagnostic technique for assessment of bone tumors.

Keywords: Bone lesions, core needle biopsy, Fine-needle aspiration cytology

   AOP18 Top

   Bone/Joints/Soft Tissue Pathology Top

Exploring solitary fibrous tumors at a tertiary cancer center: Clinicopathological and immunomorphologic profile

Soumya Mishrikotkar, Pritika Nehra, Zachariah Chowdhury, Shashikant Patne, Ipsita Dhal

Tata Memorial Hospital, Varanasi, Uttar Pradesh, India

Introduction: Solitary fibrous tumor (SFT) is a rare subtype of soft tissue neoplasm, with incidence of 0.1/100,000 person years, predominantly occurring in the thoracic cavity and less commonly at other sites. Objectives: To analyse the clinicopathologic and immunomorphologic features of SFTs along with risk stratification. Materials and Methods: A retrospective analysis of SFTs was conducted at a tertiary cancer centre of North India over a period of 3 years. Results: 14 cases of SFT were identified, affecting middle aged men and women equally. 5 cases were found at lung/pleural sites while 9 at non-pleural sites, including gastrointestinal tract, kidney and penis. 10 cases were localized, 1 showed locoregional spread and 3 revealed distant spread, out of which 2 displayed lymph node metastasis. Features of malignancy were evident in 2 cases. STAT6 and CD34 were positive by immunohistochemistry (IHC) in all cases. BCOR and p53 status evaluated by IHC, revealed high BCOR expression (>5+) in 1 case and high p53 expression (≥2+) in 5 cases. Surgical resection was performed in 9 patients while chemotherapy and radiation therapy were administered in 3 and 1 patients respectively. On a reasonable median follow-up of 14.5 months in 8 patients, 7 patients manifested no evidence of disease while a single patient died of other causes. Conclusion: SFT must be considered in the differential diagnosis of soft tissue neoplasms seen at various sites other than the thoracic cavity. Meticulous histopathological evaluation along with positive immunoreactivity of the tumor for STAT6 are the harbingers for the precise diagnosis.

Keywords: Histopathology, solitary fibrous tumor, STAT6

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   Bone/Joints/Soft Tissue Pathology Top

Histopathological evaluation of retroperitoneal soft tissue tumors – Pearls and pitfalls

N Sivaranjini, Akkamahadevi Patil, G Champaka, M N Suma

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Retroperitoneal tumors are rare cancers representing 0.2% of all cancers in adults. Though majority of retroperitoneal soft tissue masses are sarcomas, the differential diagnosis remains broad. The wide differential diagnosis and the need to tailor treatment to tumour histology, dictate the need for needle biopsy before resection of retroperitoneal lesions. Objectives: To evaluate the histopathological spectrum of retroperitoneal soft tissue lesions and to emphasise the role of biopsies in these lesions. Materials and Methods: A retrospective analysis of retroperitoneal soft tissue lesions diagnosed between January 2020 to July 2022 was done. Demographic, histopathological and immunohistochemical parameters were assessed. Results: During the study period, 96 cases were diagnosed as soft tissue neoplasms of which liposarcoma (20 cases, 28%) was the most common diagnosis, followed by leiomyosarcoma (12 cases, 12%). 49 cases were diagnosed on biopsy alone in conjunction with immunohistochemistry and treatment was initiated based on the report. Of these 3 cases required a repeat biopsy. In 17 cases surgical resection was performed following the biopsy. Conclusion: A needle biopsy in conjunction with immunohistochemistry will suffice to arrive at a diagnosis in majority of cases and guide further management. However, the inherent heterogeneity of retroperitoneal sarcomas remains a diagnostic pitfall. Hence complemented by adequate, biopsy evaluation must be clinico radiological correlation.

Keywords: Biopsy, retroperitoneum, sarcoma

   AOP20 Top

   Bone/Joints/Soft Tissue Pathology Top

Case series of sarcomas in oral cavity: A 2 year study in a tertiary cancer care centre

Jasleenkaur Oberoi, Anjali Sharma

Bhagwan Mahaveer Cancer Hospital and Research Centre, Jaipur, Rajasthan, India

Introduction: In addition to squamous cell carcinoma, several other rare tumors can arise in the oral cavity. Detection of these other tumors requires a high index of suspicion and careful pathologic review. Oral cavity sarcomas form less than 1% of oral cavity malignancies. Objectives: To evaluate the demographics and prevalence of sarcomas in the oral cavity. Materials and Methods: The study comprised of retrospective review of 6 cases of oral sarcomas in a single centre. All histologically proven sarcomas in a tertiary cancer care centre were evaluated and analyzed on the basis of clinical, histopathologic, and immunohistochemical features. Results: We herein present a series of 6 cases of oral sarcomas, thus contributing regional data to the few series reported in the literature. A total of 6 oral sarcomas were identified. The most prevalent was pleomorphic sarcoma (3 cases) followed by one case each of chondrosarcoma, Ewing sarcoma and carcinosarcoma. Age range was 1 year to 57 years. Males were more commonly affected than females. The most commonly affected sites were mandible (4 cases, 66.66%), maxilla (1 case, 16.66%) and buccal mucosa (1 case, 16.66%). Conclusion: Soft tissue sarcomas occur less frequently in oral and maxillofacial soft tissue than in other tissues. Prior radiation is one known risk factor for oral cavity sarcoma. IHC plays an important role in diagnosis of spindle cell lesions of oral cavity.

Keywords: Immunohistochemistry, oral cavity, sarcomas

   AOP21 Top

   Bone/Joints/Soft Tissue Pathology Top

Histopathological and ultrasonographic correlation in characterising benign and malignant soft tissue lesions

Sandhyarani Kanna, Aakanksha Koul, B Girish, Archana Shetty

Dr Chandramma Dayanand Sagar Institute of Medical Sciences and Research, Bengaluru, Karnataka, India

Introduction: Ultrasound examination of soft tissue lesions is a common preliminary examination conducted for diagnosis. Role of high-resolution grayscale and color Doppler ultrasound was evaluated for histopathological correlation in characterising benign and malignant soft-tissue lesions. Objectives: 1. To study the ultrasound characteristics of soft tissue lesions 2. To correlate these characteristics with the nature of soft tissue lesions. Materials and Methods: Retro-prospective study done by department of Pathology and radiology during the period of 2 years. The ultrasound images were analysed for different characteristics - size, shape, margins, sound transmission, composition, echogenicity, calcification and vascularity. These were compared with the final histopathology diagnosis of soft tissue lesions - Inflammatory/reactive/benign/malignant. Positive and negative predictive rate was calculated. Results: There were a total of 75 benign and 4 malignant masses. 40 cases ultrasonographic evaluation reports were available for correlation. There were significant differences between the benign and malignant soft-tissue tumors in terms of parameters including tumor margin, shape and size while echogenicity didn't show significant correlation with the type of lesions. Conclusion: The utility of Ultrasound in characterisation of soft tissue lesions is significant with shape, size, margins having high predictive value and echogenicity with high negative predictive value in differentiation of reactive, benign and malignant soft tissue lesions.

Keywords: Histopathological, soft tissue lesions, ultrasound

   AOP22 Top

   Bone/Joints/Soft Tissue Pathology Top

Immunohistochemical expression of H3.3G34W in 67 giant cell tumors of bone and its diagnostic mimics: A single institutional study at a tertiary cancer referral centre, India

Bharat Rekhi, Vinayak Dave, Sonali Khetle, Ajay Puri

Department of Pathology, Tata Memorial Centre, Mumbai, Maharashtra, India

Introduction: Despite its classic histopathological features, sometimes there is a challenge in differentiating giant cell tumor of bone (GCTB) from its various mimics. Lately, Histone 3.3G34W has been identified as a useful immunohistochemical marker. Objectives: To evaluate H3.3G34W immunohistochemical staining in 67 GCTBs. Materials and Methods: Immunohistochemical staining for H3.3G34W (monoclonal, RM263, 1:100 dilution) was graded in terms of staining intensity (1+to 3+) and the percentage of tumor cells showing crisp nuclear staining. Results: Seventy-one (65.7 %) GCTBs occurred in patients 15-66 years old (average=32, median=9), in femur (26, 36.6%), proximal tibia (11, 15.5%), distal radius (9, 12.6%), pelvis, including sacrum (8, 11.2%) and other bones (17, 23.9%), including a single multicentric case. Out of 67 GCTBs, wherein H3F3G34W immunostaining worked, 55 (82.1%) cases showed positive staining in the mononuclear cells, including tumors with fibrous histiocytoma-like areas, sparing the osteoclast-like giant cells. The average percentage of tumor cells showing positive immunostaining was 69%, with 3+ staining intensity in 42/55 (76.4%) cases and 2+ in 13 (23.6%) GCTBs. All 4/4 (100%) malignant GCTBs showed positive staining, including the mononuclear and pleomorphic/sarcomatous cells. Three (4.3%) cases developed metastasis (axillary nodes, mediastinum and lung). All 3/3 (100%) metastatic GCTBs showed positive immunostaining in the metastatic lesions. Out of seven post-denosumab treated GCTBs, four showed no residual giant cells and lacked H3.3G34W immunostaining. None of the other 37 “giant cell-rich” lesions displayed H3.3G34W immunostaining. Conclusion: The diagnostic sensitivity of H3.3G34W for GCTB was 82.1% and specificity was 100%. The present study, constituting one of the first reports from our country, further validates the value of H3.3G34W in differentiating GCTB, including metastatic and malignant type from its diagnostic mimics. Its utility in identifying residual tumor cells in post-denosumab treated GCTBs is worth exploring.

Keywords: Giant cell tumor of bone, H3.3G34W

   AOP23 Top

   Breast Pathology Top

Study of non-neoplastic breast lesions with cytohistomorphological correlation

Saloni Patra, Shweta Kochar, Surekha Bhalekar, Rajiv Rao

D.Y. Patil School of Medicine, Nerul, Navi Mumbai, Maharashtra, India

Introduction: Palpable breast lump is a common presenting complaint. Fine needle aspiration cytology (FNAC) is one of the first line investigations in diagnosis of palpable breast lesions although histopathological examination is more valuable for confirmation. FNAC is inexpensive, offers advantages like minimal invasiveness, cost-effectiveness, rapid results and accurate means of diagnosing breast lesions. It is essential to categorize non-neoplastic breast lesions by correlating FNAC diagnosis with histopathological findings as non-neoplastic breast lesions form a significant percentage of breast lesions. Objectives: Study is aimed to: 1) Determine spectrum of non-neoplastic breast lesions. 2) Assess the diagnostic accuracy of the FNAC by comparing with histopathological diagnosis. 3) Study the exact etiology of granulomatous mastitis. Materials and Methods: FNAC and histopathological techniques in a prospective study over a period of 24 months with sample size of 100 cases. Results: FNAC had a sensitivity of 76.3% & specificity of 100% in our study. 25-35 years of age was most common age group showing majority of non-neoplastic breast diseases. Most prevalent lesion in our study is Mastitis. All granulomatous mastitis cases are not tuberculous. Cystic neutrophilic granulomatous mastitis is an entity difficult to distinguish from tuberculous mastitis. Conclusion: Most common condition non-neoplastic lesion being mastitis. Knowledge of etiologic agent is essential as giving unnecessary anti-tuberculosis drugs in cases of idiopathic granulomatous mastitis causes side effects and treating tuberculosis with steroids would aggravate the infection. FNAC is benefial in differentiating neoplastic from non neoplastic lesions.

Keywords: Breast, mastitis, non-neoplastic

   AOP24 Top

   Breast Pathology Top

Immunohistochemical expression of basal cytokeratins (CK5, CK6) in ER, PR, Her2Neu breast cancer and its clinicopathologic correlation – A prospective study

Ankita Kolhe, Anjali Bode, Balwant Kowe

Indira Gandhi Government Medical College, Nagpur, Maharashtra, India

Introduction: Breast cancer is leading cause of cancer death in developing nations. Breast cancer in India accounted for 10.6% of all deaths in the year 2020. Histological grading is an inexpensive and simple method to assess prognosis and tumour behaviour, thereby identifying patients who may require neoadjuvant therapy. Gene expression profiling has revolutionised our understanding of breast cancer by identifying four molecular subtypes: (i) luminal A (ii) luminal B (iii) HER2 enriched (iv) basal like subtypes. CK 5 and CK6 are basal/ myoepithelial markers and they imply a basal like molecular phenotype associated with an aggressive behavior and hence poor prognosis. Objectives: To analyze the expression of CK 5/6 in ER, PR, HER2neu breast cancers and its clinicopathological correlation. Materials and Methods: This was a observational study conducted on 74 patients over a period of 2 years. All female patients who were diagnosed on Modified radical mastectomy for invasive breast carcinoma were included. Results: To be presented at the time of presentation Conclusion: The study conducted shows majority of the patients in 4th and 5th decades with grade 2 invasive breast carcinoma. All the cases of grade 2 invasive breast carcinoma were positive for CK5/6. CK 5/6 positivity shows significant statistical correlation with the size of the tumor.

Keywords: Breast, CK5/6, malignancy

   AOP25 Top

   Breast Pathology Top

Molecular spectrum of PIK3CA mutation in Indian patients with breast carcinoma

Vyomika Teckchandani, Meetu Agrawal, Kaushal Kalra, Sheetal Arora, Mukul Singh, Sunil Ranga

VMMC and Safdarjung Hospital, New Delhi, India

Introduction: The availability of alpelisib (PIK3CA inhibitor) as a promising new therapy to improve progression free survival for hormone receptor positive breast cancer patients mandates the testing of PIK3CA mutations in patients of breast cancer. The phosphoinositol 3 kinase pathway is the most frequently altered pathway in HR positive breast carcinoma, and these cases are more prone to develop metastatic disease when compared to negative counterparts. With this background, this study was undertaken to understand the heterogeneity of the mutational landscape of PIK3CA. Objectives: To study the prevalence and spectrum of PIK3CA mutations in Indian patients with hormone receptor positive breast cancer. To correlate the mutational status with age, size and tumor grade. Materials and Methods: DNA material was extracted from paraffin blocks of 31 newly diagnosed patients with ER+/PR+/HER2neu- invasive breast cancer. PIK3CA mutation status was tested using ARMS-PCR based analysis for 6 mutations namely C420R on exon 7, E545x, E542x, E545A and Q546x on exon 9 along with H1047x on exon 20. Results were analyzed by calculating the Cycle threshold value using ABI 7500 Fast Dx software. Correlation was done with patient demographic data. Results: Six out of 31 cases (18.3%) showed PIK3Ca positivity with H1047x substitution on exon 20 being the hotspot mutation. Conclusion: The phosphatidyl inositol pathway (PIK3CA) is altered in a significant subset of HR positive breast cancer patients. Testing for the presence of these mutations is now important for identification and therapeutic guidance in these patients.

Keywords: Breast cancer, HR positive, PIK3CA

   AOP26 Top

   Breast Pathology Top

Analysis of epidermal growth factor receptor gene (EGFR) mutation in triple negative breast carcinoma: Implications for personlised medicine

Aiswarya Unnithan, Subashish Das, P N Sreeramulu, R Kalyani

SDUAHER, Kolar, Karnataka, India

Introduction: Triple-negative breast cancer (TNBC) is a subtype of breast cancers, has poor prognosis and discovered molecular targets The panel of markers used for molecular classification include estrogen receptors (ER), progesterone receptors (PR), human epidermal growth factor receptor (HER)2/neu, p53, Bcl2, epidermal growth factor receptor (EGFR). Objectives: 1. To evaluate the overexpression of EGFR in TNBC in our population. 2. To identify the number and level of mutations in the breast issue associated with EGFR expression in TNBC. 3. To assess the prognostic implications of EGFR overexpression in TNBC. Materials and Methods: Fifty samples of triple negative breast cancer (TNBC) were selected for EGFR mutation analysis by immunostaining for evaluation of EGFR protein expression. B. Representative unstained, formalin fixed & paraffin embedded (FFPE) sections are selected for genomic DNA extraction using QIAamp DNA extraction kit. PCR is performed from extracted DNA to amplify exon regions 18-21 of EGFR gene. Results: Out of 50 TNBC cases, there was positive correlation between EGFR protein overexpression and EGFR gene copy number in 30 cases. Over expression of EGFR is observed in two cases without gene amplification. Mutations observed were independent of IHC EGFR expression. Conclusion: This study reveals the presence and estimate the prevalence of EGFR mutations in triple negative breast cancers. EGFR is an important marker to categorise patients with breast cancer, for evolving clinical trials. EGFR directed therapy may be of utmost significance in future to benefit the patient population.

Keywords: Epidermal growth factor receptor, mutation, triple-negative breast cancer

   AOP27 Top

   Breast Pathology Top

Comparative analysis of cytological grading using Robinson grading system of breast carcinoma with histological grading by Nottingham modification of Scarff Bloom Richardson (SBR)'s method

Ashwarya Gupta, Savita Agarwal, Pinki Pandey, Vineet Chaturvedi, Kapil Trivedi

Uttar Pradesh University of Medical Sciences, Saifai, Etawah, Uttar Pradesh, India

Introduction: Cytological examination is useful for the diagnosis of breast carcinoma. Many cytological grading systems have been developed, still none of them is accepted as gold standard leading to lack of standardization of grading breast carcinoma on cytopathology necessitating histological examination for prediction of prognosis and planning treatment. Objectives: This study will evaluate the correlation between cytological grading done by Robinson grading system and histological grading by Nottingham modification of Scarff Bloom Richardson (SBR)'s grade. Materials and Methods: Total 60 cases of IDC with availability of both FNAC and histopathology from the period 2016-2021 were retrived. Cytological grading obtained by Robinson grading system was compared with Histological grading using Nottingham modification of Scarff Bloom Richardson (SBR)'s method by testing concordance, association and correlation. Significance of the different cytological parameters was assesed using bivariate correlation studies and multiple linear regressions to predict final cytological grade. Results: Robinson's system demonstrated good correlation (ρ =0.548; P =0.000 and τ =0.523; P = 0.000), and a fair kappa value of agreement (κ =0.319) with the SBR's grading system. In multiple regression analysis, all of the cytological parameters of Robinson's system except cell uniformity and chromatin had significance in predicting the final cytological grade. Conclusion: Robinson grading system can be used for grading Breast carcinoma as it is simple and done on FNAC smears which can be obtained without going into a tedious process of histopathological examination of biopsy.

Keywords: Bloom Richardson, breast carcinoma, Robinson grading

   AOP28 Top

   Breast Pathology Top

Rare breast tumors – A case series from a Northern Karnataka Tertiary Care Centre

S Sumitha, Purushotham Reddy, S M Chowkimath

Department of Pathology, Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Breast cancer is a heterogeneous disease in its presentation, pathological classification and clinical course. With increase in its incidence, a large number of uncommon entities are diagnosed, whose descriptions are mostly from case reports and small series. So, there are high chances of misdiagnosis in such cases. Objectives: This case series aims to discuss the histopathological description of rare breast neoplasms encountered in the pathology department of KIMS, Hubballi and we attempted to summarize the current available knowledge about these. Materials and Methods: Patients treated for breast lesions for one year from August 2021 to July 2022 either with excision or mastectomy were screened and cases that were reported as unusual breast neoplasms with rare incidence were identified. The gross, microscopy and relevant IHC of these specimens were reviewed and described in detail. Literature search of electronic databases via PubMed and the search engines Google/Google Scholar was performed. Emphasis on keywords based on the histology type was used to limit search. Results: Overall six cases were reported as unusual breast neoplasms. Among these two cases were benign that includes Pseudoangiomatous Stromal Hyperplasia and benign adenomyoepithelioma. The remaining four cases were malignant breast neoplasms consisting of primary angiosarcoma, malignant adenomyoepithelioma, IDC with apocrine differentiation and squamous cell carcinoma deposits in breast. Conclusion: This series aims to focus on the histopathological features of these rare breast neoplasms and hopes to add to the existing understanding. It will be helpful for pathologists for diagnosing these cases accurately, which is crucial as it affects the overall treatment and prognosis.

Keywords: Adenomyoepithelioma, PASH, rare breast malignancies

   AOP29 Top

   Breast Pathology Top

Malignant adenomyoepithelioma: A tertiary care cancer center experience of a rare malignant entity

Musheera Aziz, Ayushi Sahay, Asawari Patil, Tanuja Shet, Sangeeta Desai

Tata Memorial Hospital, Mumbai, Maharashtra, India

Introduction: Biphasic breast tumors with myoepithelial cell predominance constitute heterogeneous group, ranging from benign adenomyopeithelioma (AME) to malignant AME (M-AME). However, definition of M-AME not clear in literature due to rarity. Recently, Rakha et al proposed novel classification of M-AME: M-AME in-situ (AME+DCIS), M-AME invasive (malignant epithelial and/or myeoepithelial component), and AME with invasive breast carcinoma (IBC). Objectives: To review clinicopathological features of M-AME diagnosed at our tertiary cancer care institute. Materials and Methods: All M-AME diagnosed between 2012-2022 retrieved from departmental archives and reviewed. Results: Out of 82 AME, 18 were M-AME (22%). Median age 48 years (range 34-90). Size ranged from 0.7-22 cm. Sixteen were unilateral, one bilateral, while one unilateral M-AME with contralateral benign AME. As per Rakha classification, 3 were M-AME in-situ, 6 M-AME invasive, while 9 AME with IBC. Of latter, 6 were hormone receptor (HR)+Her2-, 2 HR-Her2+ and 1 triple negative. Core biopsy available in 8/18 showed M-AME (1), AME (1), intraductal papilloma (1), sclerosing adenosis (1) and IBC (4). In all these, subsequent excision revealed M-AME. All M-AME showed bilayered epithelial-myoepithelial cells, confirmed on immunohistochemistry. Increased mitoses (3-25/10 HPF), moderate-marked atypia, prominent nucleoli, necrosis and infiltration was noted. Three cases showed nodal metastasis. Eight patients received adjuvant therapy. On follow up, 2/18 showed lung metastasis and 3/18 local recurrence. Conclusion: M-AME may be misdiagnosed on core biopsy, and carries potential for metastasis and recurrence. Complete excision with clear margins, and adjuvant therapy may be indicated. Recognition of biphasic elements, confirmation with immunohistochemistry, and identification of aggressive histology is essential for accurate diagnosis.

Keywords: Adenomyoepithelioma, biphasic tumor, malignant adenomyoepithelioma

   AOP30 Top

   Breast Pathology Top

Digital versus manual assessment of Ki67 in breast cancer patients

Shahanaz Saiyed, Bhagyashri R Hungund, Ashwini Ratnakar

KLE University's Jawaharlal Nehru Medical College, Belagavi, Karnataka, India

Introduction: The Ki67 protein is substantially used as prognostic and predictive marker for cancer diagnosis, aggressiveness and treatment. WHO 2019 documented “Recommendations on digital interventions for health system strengthening”. Digital pathology provides reliable and reproducible results, thereby reducing inter- and intrapathologist variability and accredit automation to amplify routine practice. Objectives: To compare the Digital Vs Manual Assessment of Ki67 in histologically diagnosed cases of breast carcinoma. Materials and Methods: Fifty cases of histologically diagnosed breast carcinoma cases were taken in the year 2021 in Dr. Prabhakar kore Hospital. Formalin fixed paraffin embedded tissue blocks were taken H& E and Ki67 IHC staining done using Ki67 SP6 (vitro) antibody. Manual counting of Ki67 positive nuclear stained cells were done from the captured images. Hotspot areas were marked and Ki67 assessed in 500 cells from the same printed images by three observers. Blinding was done to prevent bias between the observers. Digital image analysis done by uploading images in Qupath open software. Results: Digital image analysis with Qupath software Ki67 measurement shows excellent correlation (r >0.93 pearson c.c) with manual Ki67 measurement. Manual assessment of Ki67 by three observers using same hotspot visual images was done and it shows excellent agreement between interobservers (r > 0.94). Intraclass correlation coefficient- average (r =0.977), single measurement (r=0.916). Conclusion: Digital Ki67 assessment exhibit same variability and reproducibility as manual Ki67 assessment. Given the non inferiority and substancial time saving, the present study supports more widespread adoption of automated Digital image analysis of Ki67 in routine clinical practice.

Keywords: Breast Ki67, digital, manual

   AOP31 Top

   Breast Pathology Top

A comparative study between conventional method and Robinsons grading system for reporting breast cytopathology and their histopathological correlation

Davinder Kaur, Jaspreet Singh, Vijay Mehra, Permeet K Bagga

Government Medical College, Amritsar, Punjab, India

Introduction: Breast cancer is one of the leading causes of death in women. FNAC is an important part of triple assessment of the palpable breast lump. cytological grading of breast carcinoma, which is simple, feasible and reproducible can be used for selection of neoadjuvant therapy. Objectives: The present study was conducted to compare conventional cytological examination of breast lesions with Robinsons grading system of reporting and to study the correlation with histopathology. Materials and Methods: The present study was conducted on 50 patients of swelling or lump in breast Fine needle aspiration cytology procedure was done and graded according to conventional method and by Robinsons grading and results were correlated with histopathology. Results: A total 50 female patients were included in the study, cytologically C4 lesions are found in maximum number of patients, with Robinsons Cytological grading, most common tumour was grade I, followed by grade II and Histologically, most common tumour was grade I. Conclusion: In the era of neo-adjuvant chemotherapy, grading of breast cancer should be incorporated in FNAC reports for prognostication. Robinsons grading correlates more with histopathology.

Keywords: Breast carcinoma, cytological grading, fine needle aspiration cytology

   AOP32 Top

   Breast Pathology Top

Retraction clefts in invasive carcinoma breast – Correlation with histopathological factors

Shachi Shrivastava, Anuradha C K Rao

Yenepoya Medical College, Deralakatte, Karnataka, India

Introduction: Retraction clefts (RCs) in breast neoplasms have recently piqued pathologists' interest because they can be easily identified and classified in hematoxylin-eosin (H&E) stained sections. It can be a predictor of nodal metastasis in breast carcinomas. It is the cavity in tumour sections with no endothelial cell lining around tumour glands or nests. Some of the studies proposed that the RC can be a marker of disease progression and nodal metastasis. We studied the amount of RCs in 50 invasive breast cancer specimens, as well as their relationships with histopathological factors. Objectives: 1. To assess the relationship of RC with histopathological factors 2. To determine the relationship of RC with hormonal status. Materials and Methods: 50 formalin-fixed paraffin-embedded samples of Invasive breast carcinoma no specific types were included. Conventionally stained H & E sections of 4 μm and 2 μm thick were studied for RC, and compared with HPE factors and hormonal status. Results: Out of 50 cases, 42 cases showed presence of RC. It showed significant correlation with the size of tumour (p-0.001), LVI (p-0.004), PNI (P-0.010), stage (p-0.001) and HER2 positive status (p-0.03). No statistically significant association was identified between the RCs and patients' age (p – 0.549), tumour grade (p-0.667), lymph node status (p - 0.281), ER (p-0.318), PR status (p - 0.18), DCIS (p-0.195), Necrosis (p-0.19). Conclusion: Retraction clefts in invasive breast carcinoma can be an important histopathological marker for detecting cases with poorer outcome based factors.

Keywords: Breast carcinoma, histopathological factors, retraction cleft

   AOP33 Top

   Breast Pathology Top

The prognostic indicator of tumor infiltrating lymphocytes in breast carcinoma

Sindhu Kumar, B N Navya, Sathyavathi R Alva, Anjali Rao

Department of Pathology, KVG Medical College and Hospital, Sullia, Karnataka, India

Introduction: Breast cancer is one of the common malignant tumors in women. Use of biomarkers will ensure breast cancer patients to receive optimal treatment. Tumor-infiltrating lymphocytes (TIL) are emerging as one of the key prognostic markers of breast cancer. Objectives: 1. To evaluate percentage of TILs in tumor tissue. 2. To study prognosis of breast carcinoma in relation to concentration of TILs. 3. To compare TILs with ER & PR status. Materials and Methods: All cases reported as breast carcinoma in histopathology laboratory between 2019 and 2021 were included in the study. TILs were analyzed by three pathologists separately based on recommendations by an International TIL Working Group 2014 to prevent inter-observer bias. The results were correlated with ER & PR status. Results: Among 30 study subjects, majority had TILs of intermediate grade (53.3%) followed by higher & lower grade in, 40% and 6.7% cases respectively. TILs expression was statistically significant for both ER and PR in positive & negative groups (p<0.05). Increase grades of TILs with negative molecular sub-type (ER & PR) statistically proved good prognosis in breast cancer (p<0.05). Conclusion: Our study concluded that higher grades of TILs correlated significantly with ER, PR negative status and showed favorable survival with good response to treatment.

Keywords: Breast cancer, prognostic factors, tumor-infiltrating lymphocytes

   AOP34 Top

   Breast Pathology Top

Expression of BRCA1 and BRCA2 in triple negative breast carcinomas

Zoya Hasan, Shramana Mandal, Nita Khurana, Sushanto Neogi

Maulana Azad Medical College, New Delhi, India

Introduction: Breast Cancer is the most common Cancer among women worldwide and a leading cause of cancer death among women. Triple Negative Breast Carcinomas (TNBC) accounts for 10-20% of all Breast cancer cases. BRCA1 and BRCA2 are Tumor suppressor genes and its mutation is frequently associated with progression to Breast Carcinomas. These tumors are poorly differentiated and commonly exhibit Triple Negative phenotype. Objectives: The aim of the study was to assess BRCA1 and BRCA2 expression in Triple Negative Breast Carcinoma cases and its correlation with Clinico-pathological parameters. Materials and Methods: This study included 30 TNBC cases with no prior therapy. Results: Patients age ranged from 25 to 73 years, with a mean age of 47.97 years. A total of 09 TNBC cases (30%) showed BRCA1 positivity and 12 cases (40%) showed BRCA2 positivity; Out of which, Five (16.67%) cases showed both BRCA1 and BRCA2 positivity. One case of Metaplastic carcinoma was observed which expressed both BRCA1 and BRCA2. Thirteen (43.33%) cases were Basal type TNBC, whereas 17 (56.67%) cases were of non-Basal type based on the expression of CK5/6. Statistically significant association was found between BRCA1 expression and Lymph node involvement (p=0.032), BRCA1 and Necrosis (p=0.049), BRCA2 & Lymph node involvement (p=0.034) and BRCA2 and Histological grade (p=0.05). Conclusion: These findings suggested that TNBC associated with poor prognostic factors are more likely to express BRCA1 and BRCA2. Immuno-expression of BRCA1 and BRCA2 in TNBC maybe an inexpensive and a valuable preliminary diagnostic modality.

Keywords: BRCA1, BRCA2, triple negative breast carcinomas

   AOP35 Top

   Breast Pathology Top

Characterisation and prognostic significance of tumor infiltrating lymphocytes (TILs) with special emphasis on the role of B-cells in breast carcinomas

Sarika Gupta, Kachnar Varma, Vatsala Misra, Neha Sharma

Moti Lal Nehru Medical College, Prayagraj, Uttar Pradesh, India

Introduction: There is increasing evidence of intricate interactions between tumor cells and the intra and peritumoral stromal infiltrating B lymphocytes helping in production of immunoglobulins that specifically recognize tumor antigens, eliciting anti tumor humoral immunity. TIL-Bs are also associated with major prognostic effect on the patient's clinical outcomes and disease free survival. Objectives: To evaluate the density and localisation of TIL-Bs alongwith their correlation to clinico pathological factors and prognosis. Materials and Methods: This prospective and retrospective study was conducted on over 100 samples including radical mastectomies and core biopsies. HE stained sections used for analysing histopathology. Tumor infiltrating lymphocytes counted in the centre of tumor (CM) and at the junctional area between the tumor and stroma (IM). CD20 lymphocytes analysed by dividing the tumor into- 1) The intratumoral, 2) within the distant stroma, and, 3) within the adjacent stroma. The total number of CD20 lymphocytes determined by combining the three counts. IHC panel applied- ER/ PR/ Her-2 neu for molecular classification of tumor, Ki-67 for degree of mitotic activity, CD20 for B lymphocytes. Results: The mean age of cases was 45 years. TILs increase in invasive breast cancer. IHC stained sections confirm that tumors of higher histological grade show higher count of tumor infiltrating B-lymphocytes and CD20 expression. Their proliferation was also higher in triple negative and HER-2 neu positive tumors. No significant association with lymph node involvement noted. Conclusion: In summary, this study analyses the role of TIL-Bs on the prognosis, correlation with clinico pathological variables and overall survival in cases of breast carcinoma, found to be significantly higher in triple negative and HER-2 positive tumors.

Keywords: Breast carcinoma, CD20, tumor infiltrating lymphocytes

   AOP36 Top

   Breast Pathology Top

Categorization of fine needle aspirates of palpable breast lumps using the International Academy of Cytology Yokohama System along with assessment of risk of malignancy and diagnostic accuracy in a tertiary care centre

Tanya Jain, Vandana Agrawal, Parul Gupta

L. N. Medical College, Bhopal, Madhya Pradesh, India

Introduction: The International Academy of Cytology Yokohama System has developed a standardized system of reporting breast cytology by classifying them into five categories (C1-C5) - Insufficient, benign, atypical, suspicious, and malignant. Objectives: The main objectives of our study were to classify breast fine needle aspirates as per the IAC Yokohama system and assess the risk of malignancy, sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy. Materials and Methods: This study was done over a period of 1.5 years (March 2021 to September 2022). A Total of 109 FNAC specimens were studied and reclassified as per the newly proposed IAC Yokohama system of reporting. Using a histopathological diagnosis as the gold standard, the Sensitivity, Specificity, PPV, NPV & Diagnostic accuracy was calculated accordingly & the ROM was also assessed for each category. Results: Out of the 109 breast FNAs, 54 correlates with histopathology. The risk of malignancy for benign, atypical, suspicious, and malignant categories were 3.4%, 0%, 66.6% & 95% respectively. Maximum sensitivity (96.4%) was noted when suspicious & malignant cases were considered as positive test results. The highest specificity (97.4%) was observed when benign and atypical cases were considered as true negative test results and maximum diagnostic accuracy noted was 97.1%. Conclusion: The IAC Yokohama system is an exceptional system for accurately diagnosing breast FNAs with great reproducibility of reports and better communication between the pathologist and clinician.

Keywords: Fine needle aspiration cytology, IAC, Yokohama system

   AOP37 Top

   Breast Pathology Top

The paired perils of breast cancer and diabetes

Pravitha Peter, Mayura Phulpagar

Topiwala National Medical College, and B.Y.L Nair Charitable Hospital, Mumbai, Maharashtra, India

Introduction: Diabetes is among the most important chronic conditions, and breast cancer is one of the most prevalent cancers in women worldwide. Measures related with early detection of breast cancer and its association with underlying lifestyle diseases are therefore of emerging significance in the current scenario. Objectives: This study analyses the association of diabetes mellitus with breast cancer in women. Materials and Methods: This retrospective observational study analyses total of 100 cases of breast cancer in women aged between 28-85 documented in the pathology logs. The data analyzed using SPSS software and P value calculated. Results: Among the 100 cases of breast carcinoma studied, 10 showed association with underlying diabetes mellitus whereas control group (100 non CA) 8 people had diabetes mellitus. The data showed a p value>0.05 rendering it statistically insignificant. Conclusion: Our study shows no significant relation between the two which is in contrast with published literatures which states that type 2 diabetes mellitus could be related with excess risk for breast cancer and could have deleterious effect on the natural history, diagnosis, and treatment of the condition. Since sample size was small we recommend similar study on larger number of cases. The study helped to raise the importance of creating awareness related to carcinoma breast and the need for its early detection and prevention. This study was done a few months away from celebrating Global Breast Cancer Awareness Month (October).

Keywords: Breast cancer, diabetes mellitus

   AOP38 Top

   Breast Pathology Top

Screening of breast lesions by triple assessment in a tertiary care hospital

M N Sushmitha, B R Rajalakshmi

JSS Academy of Higher Education and Research, Mysuru, Karnataka, India

Introduction: Accurate preoperative diagnosis of a breast lesion is essential to confirm or exclude cancer for optimal treatment planning. The diagnostic accuracy can be increased by employing multimodality tests as in Triple Assessment, which includes – Clinical breast examination (CBE), Imaging (Ultrasonogram (USG) and mammography) and Fine needle aspiration cytology (FNAC)/core biopsy. Objectives: To assess the performance characteristics of Triple test (Clinical breast examination, Imaging and Fine needle aspiration cytology) in the diagnosis of breast lesions by correlating with final histopathologic diagnosis. Materials and Methods: All women who underwent screening for breast lesions by triple assessment were included in the study. Pregnant women and inflammatory lesions were excluded from the study. Results: 110 cases were screened by clinical breast examination, where 33.6% were diagnosed as malignant, 46.4% as benign, and 20% were indeterminate. In FNAC, 39.1% cases were diagnosed as malignant, 47.3% cases as benign & 13.6% were indeterminate. By imaging, malignancy was diagnosed in 46.4% cases, while 42.7% cases were benign & 10.9% (n=12) were indeterminate. Radiological examination has better accuracy in diagnosing the breast lesions (81.5%) than FNAC (72.2%) and clinical examination (63%), while combined triple assessment has an improved diagnostic accuracy of 93%. Triple assessment diagnosis was concordant with final HPE diagnosis in 103 /110 cases (53 malignant & 50 benign lesions) cases, while 7 cases showed discordance with final HPE diagnosis. PPV of triple assessment was 90.19%, and the NPV was 90.38%, with a p-value of 0.001, which is highly significant. The sensitivity and specificity of triple test was 90.2% & 90.4%. Conclusion: Triple assessment of breast lesions is a valid and reliable method for accurate preoperative diagnosis.

Keywords: Breast, fine needle aspiration cytology, mammography, screening, triple assessment

   AOP39 Top

   Breast Pathology Top

Cytohistological indicators of neoplastic breast lesions with emphasis on tumor budding using estrogen receptor marker

Chandana Ullas, N M Nandini

JSS AHER, Mysuru, Karnataka, India

Introduction: Breast cancer is the most common cancer in urban Indian women with age adjusted rate of 25.8 and mortality of 12.7 per 1,00,000 women. Prognosis is greatly determined by clinicopathological and molecular characteristics of the tumor. Significance of tumor budding as prognostic factor is now being evaluated. Hormone receptor status evaluation has become a critical determinant for classification and management of breast cancer. Estrogen receptor (ER) status has emerged as an important criteria in determining the prognosis and treatment as it plays a major role EMT and thus in tumor metastasis. Objectives: 1) To correlate tumor budding with histopathological features and hormone status. 2) To assess ER status of tumor buds. Materials and Methods: Study design: Analytical study-Prospective study. Study duration: Eighteen months (October 2020- March 2022). Sampling technique: Purposive sampling. Core biopsy and resected breast specimens were considered for tumor budding counting in 20X objective. Tumor buds were also counted in corresponding ER stained slides. Tumor bud count was correlated with histopathological features and ER status. Results: Tumor budding is high in grade III and ER positive tumors and low in grade I and ER negative tumors. Conclusion: This study demonstrated that tumour budding is an adverse prognostic indicator of breast lesions. Significant correlation with estrogen receptor demonstrated that the tumour budding may be hormone-driven process.

Keywords: Breast cancer, estrogen receptors, tumor budding

   AOP40 Top

   Breast Pathology Top

Evaluation of accuracy of fine needle aspiration biopsy for breast lesions using the “International Academy of Cytology Yokohama System” for reporting breast cytopathology

SillaAlisha Patro, Lity Mohanty, Bhuban Mohan Das, Madhumita Debata

Department of Pathology, SCB Medical College and Hospital, Cuttack, Odisha, India

Introduction: Breast cancer represents the leading malignancy in women with a rising incidence worldwide. Fine Needle Aspiration Biopsy (FNAB) has been shown to be an accurate method of diagnosing breast lesions. However, due to the increasing popularity of core needle biopsy (CNB), the usage of FNAB has been decreasing. FNAB has other advantages, of time, economy, and lesser discomfort to the patient. The IAC Yokohama system of breast cytology was proposed recently to standardize breast cytology reporting. Objectives: 1. To classify and interpret the FNA smears of breast lesions using a new reporting system-”The international Academy of Cytology Yokahama system” for reporting Breast Fine needle aspiration biopsy cytopathology. 2. To calculate the risk of malignancy (ROM) of each category. 3. To calculate the sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy for all categories. Materials and Methods: FNAC is done in patients with clinically detected breast lumps. Cytological diagnosis is reported as per THE YOKAHAMA REPORTING SYSTEM (Cat I-Insufficient, Cat II-Benign, Cat III-Atypical, Cat IV-Suspicious of malignancy, Cat V-Malignant). Then the patient is referred to the department of surgery for core needle biopsy/lumpectomy/mastectomy followed by histological examination for the confirmation and to know the accuracy of breast Yokohama cytology grading system. Results: Out of 210 breast lesions reported on FNAB, the category wise distribution was-Category I, II, III, IV & V accounts for 4 (1.9%), 90 (42.8%), 20 (9.5%), 6 (2.8%), 82 (39.04%) respectively. Conclusion: To conclude, FNAB remains an accurate method for assessment of breast lesions by using IAC Yokohama reporting system.

Keywords: Core needle biopsy, fine needle aspiration biopsy, Yokahama reporting system

   AOP41 Top

   Breast Pathology Top

Giant benign mammary phyllodes

Shivani Bansal, Ipsita Panda, Nandita Kakker Arunanshu Behera, Divij Jayanta

PGIMER, Chandigarh, India

Introduction: Phyllodes tumor is uncommon fibroepithelial neoplasm accounting for less than 1% of all female breast tumors. The size of the tumor is variable and only handful of giant phyllodes of benign nature have been reported. Here we report a case of a massive phyllode (30 cm) with a benign histological nature. Objectives: To study histopatholoical characteristics of rare giant mammary phyllodes. Materials and Methods: Grossly, the mass measured 37×34x32 cm in size with clear resection margins. On cut-section, homogenous grey-white areas seen with no areas of necrosis. Forty sections examined microscopically showed a benign fibroepithelial neoplasm with extensive areas of hyalinization and myxoid degeneration. A low rate of mitosis was noted (3-4/10 HPF). Hence was reported as a 'Giant Benign Phyllodes Tumor'. Results: Hence was reported as a 'Giant Benign Phyllodes Tumor'. Conclusion: A giant size of phyllodes tumor does not translate into malignancy always and warrants extensive sampling of the tumor before establishing its benign nature.

Keywords: Breast, giant phyllodes

   AOP42 Top

   Breast Pathology Top

An investigation on stromal CD10 expression in breast carcinoma and its relation with ER, PR and Her2Neu using immunohistochemistry

Sayan Mukhopadhyay, Rajib Kumar Mondal, Anup Roy

Nil Ratan Sircar Medical College and Hospital, Kolkata, West Bengal, India

Introduction: CD10 is a zinc-dependent matrix metalloproteinase, that degrades many bioactive peptides. CD10 expression in tumour stroma is associated with biological aggressiveness of many epithelial malignancies. Objectives: The aim of this study is to evaluate stromal CD10 expression in breast carcinoma and to examine its correlation with ER, PR and HER2-neu. Materials and Methods: CD10 expression in modified radical mastectomy samples of one hundred and twenty patients was assessed by immunohistochemistry at Nil Ratan Sircar Medical College and Hospital, Kolkata, and scored as negative, weak and strong. ER, PR and HER2-neu expressions were also assessed in the present study. Results: CD10 was found to be positive in stroma of 79/120 (65.8%) cases. Stromal CD10 showed positive correlation with tumour grade (p<0.05), HER2-neu (p<0.01), and negative correlation with ER (p<0.01) and insignificant negative correlation with PR (p=0.256). Conclusion: CD10 expression correlated strongly with well-established negative prognostic markers, that is, HER2-neu positivity, ER/PR negativity, and higher tumour grade indicating the fact that CD10 can be used as an independent marker of poor prognosis and also can be used as target for the development of novel therapy.

Keywords: CD10, ER, HER2-neu, PR

   AOP43 Top

   Breast Pathology Top

Role of PDL1 and CD8 in predicting the pathological response to neoadjuvant chemotherapy

Kapil Trivedi, Pinki Pandey, Roopak Agarwal, Vineet Chaturvedi, Savita Agarwal, Asttha Kapoor, Ashwarya Gupta

Uttar Pradesh University of Medical Sciences, Saifai, Etawah, Uttar Pradesh, India

Introduction: Neo Adjuvant Chemotherapy (NACT) was first introduced in 1970 for inoperable tumors and now become the standard treatment approach in locally advanced breast cancer (LABC). Pathological complete response (pCR) following NACT is an important indicator for disease-free survival in all subtypes of breast cancer. An increased number of CD8+T cells in the tumor center is thought to indicate an effective antitumor immune response. Expression of PD-L1 in tumor micro-environment has shown a correlation with the presence of tumor infiltration lymphocytes. Objectives: This study will correlate the expression of PDL-1 and CD8 in pre-NACT core biopsies with the pathological response and assess their predictive role. Materials and Methods: All cases of IDC with available pre-chemotherapy core needle biopsy and post-chemotherapy mastectomy specimens in the period from 2016 to 2022 were retrieved. Evaluation of PD-L1 & CD8 expression was done on pre-NACT biopsies. Pathological response of NACT on post-NACT specimens was assessed and categorized into responders and non-responders. Results: In 47 cases of post-NACT Mastectomies with available pre-NACT biopsy, the pathological response was assessed. 11 (23.4%) & 36 (76.6%) patients were responders and non-responder respectively. PD-L1 & CD8 expression was found to be positively associated with pathologic response (p < .001, p=.002). PD-L1 expression in tumors also correlated with the presence of CD8+TILs (p =.042). Conclusion: The expression of PD-L1 and CD8+TILs correlates with response to chemotherapy. High PD-L1 and CD8+TILs may predict better response to neoadjuvant chemotherapy.

Keywords: Breast carcinoma, neoadjuvant chemotherapy, PD-L1

   AOP44 Top

   Breast Pathology Top

Concordance between cytological (Robinson's Grade) and histopathological grading (Nottingham Modification of Bloom Richardson Grade) in invasive breast carcinoma – An observational cross sectional study

Mrinal Chandrasen, Naveen Verma, R K Chandrakar

Shri Shankaracharya Institute of Medical Sciences, Bhilai, Chattisgarh, India

Introduction: *Breast cancer is the most common female cancer worldwide representing nearly a quarter (25%) of all cancers. *fine needle aspirations cytology is considered to be one of the simplest, cost effective and highly accurate technique for evaluation of breast lumps. Objectives: (A) To study the concordance Between Cytological and Histopathological Grading in Invasive Breast Carcinoma. (B) To find out the proportion of regional lymph nodes metastasis amongst various Cytological grades of breast carcinoma. Materials and Methods: *A one and half year study was conducted 2021-2022. *It included all the 80 cases of Invasive Breast Carcinoma. *PAP and HE stained smears were evaluated. *A score of 1-3 was given to each of these parameters. *Mitotic figures were counted. Results: Statistically Significant. r value 0.804. P value less than 0.01. Conclusion: *In the present study, a high degree of concordance is seen between Cytological and HG system.

Keywords: Fine needle aspirations cytology, prognostication grading

   AOP45 Top

   Breast Pathology Top

Expression of androgen receptor in breast carcinoma and its correlation with estrogen, progesterone and human epidermal growth factor receptor-2 status

Mayank Nangru, Ruchi Agarwal, Swaran Kaur Saluja, Parveen Rana Kundu, Monika B Gathwal, Sunaina Hooda

BPS GMC(W), Khanpur Kalan, Sonepat, Haryana, India

Introduction: Breast cancer is the second most common cancer overall, and by far the most common cancer in women. Immunohistochemistry (IHC) has been of immense value in the diagnosis and prognosis of breast carcinoma. Despite this, it is still difficult to find targeted therapies for triple negative breast cancer. Objectives: To study the expression of androgen receptor (AR) in cases of breast carcinoma and its correlation with estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor-2 (Her-2 neu). Materials and Methods: Cross sectional study was done for duration of one year in which 45 cases of breast carcinoma with known ER, PR and Her-2 neu status were included. All cases were studied for androgen receptor expression and androgen receptor status was compared with ER, PR and Her-2 neu status. Results: Out of total 45 cases, 28 were positive for androgen receptor. In cases with MBR grade I, all cases are positive, in cases with MBR grade II, 60% were positive whereas with MBR grade III 50% were positive for androgen receptor. In ER positive cases, 72%; in PR positive cases, 77%; in Her-2 neu positive cases, 73% were positive for androgen receptor. In triple-negative breast carcinoma (TNBC) cases, 48% were positive for AR. Conclusion: AR is expressed in a significant number of breast cancers and is associated with ER, PR and Her-2 neu positivity. It can be used as an additional marker in cases of breast carcinoma. In cases TNBC, AR can help in finding targeted therapies.

Keywords: Androgen receptor, breast cancer, immunohistochemistry

   AOP46 Top

   Breast Pathology Top

Robinson's cytological grading on aspirates of breast carcinoma and its correlation with Bloom Richardson's histological grading

Digvijay Deore, Nitin Sagare, Rachana Binayke, Sanjay Bijwe

Grant Government Medical College, Mumbai, Maharashtra, India

Introduction: The importance of histological typing and grading of breast carcinomas is well established and has gained a strong foothold. As neoadjuvant therapy is becoming increasingly common for treatment of early breast cancer, it's desirable to grade the tumor preoperatively on FNAC so that most appropriate medical regimen can be selected. Objectives: 1. To evaluate Robinson's cytological grading of aspirates of breast carcinoma and Bloom Richardson's histological grading. 2. To compare Robinson's cytological grading of aspirates of breast carcinoma with Bloom Richardson's histopathological grading. Materials and Methods: Present study of 30 cases of malignant breast lesions was carried out in cytopathology and histopathology department of pathology in a tertiary care hospital. Female patients referred for FNAC of palpable breast lump to cytology section and their histopathological specimen will be included in study. In each patient, clinical and investigational findings are obtained from medical records. Pathological findings are obtained by examination of hematoxylin and eosin-stained slides. Results: Majority patients have invasive ductal carcinoma 19 (63.3%), followed by invasive lobular carcinoma 6 (20%), ductal carcinoma insitu 2 (6.7%), papillary carcinoma 2 (6.7%) and medullary carcinoma 1 (3.3%). According to Robinson's staging, majority belong to stage1 i.e. 14 (46.7%), followed by stage 2 i.e. 11 (36.7%) and least is stage3 seen among 5 (16.7%). According to Bloom Richardson's score, majority belong to stage1 15 (50%), followed by stage2 14 (46.7%) and least is of stage3 among 1 (3.3%). Conclusion: In present study, there was high level of agreement between Bloom-Richardson's histological grading system and Robinson's cytological grading system. Breast cancer cytological grading is easy, practical and gives useful prognostic information.

Keywords: Bloom Richardson Grading, breast, Robinson

   AOP47 Top

   Breast Pathology Top

Role of E-cadherin and Vimentin as markers of epithelial mesenchymal transition of breast carcinoma and their effect on aggressiveness of tumour

Sumiah Syed, Anuradha C K Rao

Karpaga Vinayaga Institute of Medical Sciences, Maduranthakam, Chengalpattu, Tamil Nadu, India

Introduction: Breast cancer remains one of the most prevalent forms of cancer, globally. Epithelial- mesenchymal transition (EMT) is a physiological phenotypic shift in which epithelial cells break down and then migrate to other locations in the body. E-cadherin and vimentin are regarded as major conventional canonical markers of epithelial mesenchymal transition. Irrespective of available modern therapies, there is an increased morbidity and mortality due to breast cancer. Prognosis of breast cancer may be improved by focusing on EMT and metastasis, which lead to advanced stage of breast cancer. Objectives: 1) To study prognostic importance with the expression of E-Cadherin and Vimentin in the breast carcinoma using IHC. 2) To correlate the expression of E-Cadherin and vimentin with the histopathological parameters and hormonal status of tumor. Materials and Methods: A total of 56 cases of breast cancer were included in the study. E- cadherin and Vimentin protein expression was evaluated by immunohistochemistry and was correlated with clinical stage, pathological type, grade of the tumor, and lymph nodes status. Expression was scored for each antibody separately and semi quantitatively by assessing the stain localization, intensity, and the percentage of stained cells in the tumors. Results: In our study, there is statistical significant increased expression of vimentin in triple-negative cases. E-C expression decreases as the Grade and Stage of the tumor increases whereas it is vice-versa for Vimentin. In addition, Our study revealed that E-cadherin and vimentin protein is colocalized within the same tumor cells in invasive breast carcinoma. This study also reveals an inverse correlation between E-cadherin and Vimentin (r =-637) with a significant correlation (p. value = 0.001). Conclusion: E-cadherin and Vimentin could be useful as markers for EMT and tumor aggressiveness in breast cancers and may serve as predictive markers to aid in early treatment and prolonged survival rate among the patients.

Keywords: E-cadherin, Epithelial-mesenchymal transition, invasive breast carcinoma, Vimentin

   AOP48 Top

   Breast Pathology Top

Prognostic value of IMP3 and its role as an epithelial-mesenchymal transition promoter in breast

Tanvi Jha, Preeti Diwaker, Somnath Mahapatra, Sonal Sharma, Vinod Kumar Arora

University College of Medical Sciences, New Delhi, India

Introduction: Tumor metastasis is the most common cause of death in breast cancer. In view of the high mortality in breast cancer patients, there is a need to identify new prognostic biomarkers. Objectives: IMP3 has a role in induction of epithelial-mesenchymal transition (EMT) and SLUG is an EMT marker. They have both been implicated in cancer metastasis and hence, might prove to be potential therapeutic targets in breast carcinoma. Materials and Methods: This retrospective study was conducted on 60 breast carcinoma cases in a tertiary care center using tissue microarrays. Demographic and clinicopathological details were recorded. Immunohistochemistry (IHC) for IMP3 and SLUG was performed and evaluated for percentage cell-positivity and staining intensity. Data was recorded on a pre-designed proforma and analyzed. Results: IMP3 positivity was found in 87% and SLUG positivity was observed in 90% breast carcinoma cases. IMP3 positivity was significantly associated with tumor size (p = 0.03) and TNM stage (p = 0.024). SLUG immunoexpression was significantly associated with TNM stage (p = 0.006). A highly significant association (p <0.001) was found between IMP3 and SLUG positivity and their staining intensities (p <0.001). Significant correlation was also found between IMP3 and SLUG percentage cell positivities (p <0.001). Conclusion: Significant association of immunoexpression of IMP3 and SLUG with TNM stage proves them to be poor prognostic markers. Also, significant positive correlation between immunoexpression of IMP3 and SLUG highlights their proposed role in tumor invasiveness via EMT in breast carcinomas. Hence, IMP3 and SLUG based targeted therapies might prove to be useful in treatment of breast carcinoma.

Keywords: Breast carcinoma, IMP3, SLUG

   AOP49 Top

   Breast Pathology Top

Prognostic significance of neutrophil lymphocyte ratio (NLR) and platelet lymphocyte ratio (PLR) and their association with clinicopathological profile in patients with carcinoma breast

Abhiraami Muthuraman, J Sree Rekha, Biswajit Dubashi, Kadambari Dharanipragada

JIPMER, Puducherry, India

Introduction: The role of inflammation and immune response in cancer development, progression and treatment has gained enormous interest in the past decade. Peripheral blood Neutrophil - Lymphocyte Ratio (NLR) and Platelet - Lymphocyte Ratio (PLR) reflects the balance between pro-tumorigenic systemic inflammation and anti-tumor immune response of the patient. Objectives: To identify the association of peripheral blood NLR and PLR with clinicopathological characteristics, treatment response and survival in breast cancer patients. Materials and Methods: The clinicopathological, haematological and treatment data of 355 newly diagnosed breast cancer patients treated at our institute from July 2017 to June 2019 was collected retrospectively from the case records and hospital information system. These patients were followed-up till December 2021 for any disease recurrence, progression and mortality. Results: Elevated pre-treatment NLR>2.17 and PLR>118.89 could significantly predict a poor disease outcome such as disease recurrence or disease progression or death of the patient with a sensitivity of 78.7% (p<0.001) and 62.3% (p=0.001) respectively. Elevated pre-treatment NLR>2.17 (p=0.008) and lack of pathologic complete response (pCR) to neoadjuvant chemotherapy (p=0.04) proved to be independent and significant predictors of poor disease free survival after multivariate analysis. An increase in NLR at one month post neoadjuvant chemotherapy by >1.01 times the baseline value could significantly predict a poor pathological response to NACT with a high specificity of 82.2% (p<0.001). Conclusion: Pre-treatment NLR and PLR can be used as prognostic biomarkers in patients with carcinoma breast. Monitoring the trend in NLR at one month after the end of NACT can predict pathological response to NACT.

Keywords: Breast cancer, neutrophil/platelet lymphocyte ratio

   AOP50 Top

   Breast Pathology Top

Utility of dual-color dual in-situ hybridization (D-DISH) for Her2/Neu testing in breast cancer

Aditi Rathi, Tanuja Shet, Asawari Patil, Ayushi Sahay, Sangeeta Desai

Department of Pathology, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, Maharashtra, India

Introduction: HER2 gene amplification, a poor prognostic factor in invasive breast cancer (IBC), has shown substantial utility as predictive marker with significantly improved survival following anti-HER2 therapies like Trastuzumab. Dual-color dual in-situ hybridization (D-DISH) is recently introduced fully automated assay for HER2/neu evaluation on light microscopy with several advantages over traditional Fluorescence in-situ hybridization (FISH). Objectives: To standardize and validate D-DISH assay using FISH as gold standard and assess inter-observer variability. Materials and Methods: D-DISH was performed using the latest HER2 Dual ISH DNA Probe Cocktail assay by Ventana Medical Systems (2020) (Tucson, USA), in 148 cases of IBC, which were previously subjected to immunohistochemistry by Ventana PATHWAY anti-HER2/neu (4B5) antibody (Tucson, USA) and FISH assay by ZytoLight SPEC ERBB2/CEN17 Dual-Color Probe (Bremerhaven, Germany). D-DISH was interpreted by four pathologists blinded to FISH results. Cohen's kappa statistic and Intra-class correlation coefficient (ICC) were used to assess concordance between FISH and D-DISH and inter-observer variability between four pathologists. Results: Concordance of 98.65% and Cohen's kappa-value of 0.97 was observed between FISH and D-DISH. ICC (0.93-0.97) and kappa-values (0.98-1.0) for inter-observer variability showed almost perfect agreement by D-DISH. Interobserver variability evaluated for polysomy (kappa-values 0.40-0.64), genomic heterogeneity (kappa-values 0.42-0.74), and HER2 group categorization (kappa-value 0.89-0.93) showed moderate, substantial, and perfect agreement respectively. Conclusion: We validated the latest version of D-DISH as a substitute for FISH in ascertaining the HER2 gene status with significant interobserver reproducibility. D-DISH assay may be introduced as a reflex test for detecting HER2 gene status in routine diagnostic workflow.

Keywords: Dual-color dual in-situ hybridization, fluorescence in-situ hybridization, HER2/neu

   AOP51 Top

   Breast Pathology Top

Association of tumor infiltrating lymphocytes with clinicopathological variables and molecular subtypes of invasive breast cancer

Ritika Johri, Garima Goel, Swagata Brahmachari, Deepti Joshi

AIIMS, Bhopal, Madhya Pradesh, India

Introduction: Breast cancer is considered to be immunogenic and lymphocytic infiltration observed in breast cancer has been linked to its biological behavior. Routine tumor infiltrating lymphocytes (TIL) evaluation is not being used in clinical practice due to lack of reproducibility. Objectives: To quantify and subtype stromal TIL in invasive breast cancer and explore their relationship with clinicopathological variables of breast cancer. Materials and Methods: This observational study was conducted at AIIMS Bhopal. All the consecutive cases of invasive breast carcinoma received from January 2018 to June 2020 fulfilling the inclusion criteria were studied. Sample size was 75. TILs were quantified on Hematoxylin and Eosin stained sections. Immunohistochemistry was performed with ER, PR, HER2 and Ki67 for molecular classification and with CD4, CD8 and FOXP3 to subtype stromal TILs. The data was analyzed using appropriate statistical tests. Results: The highest TIL grade was associated with triple negative breast cancer. Amongst the TIL subtypes, CD4+ TILs were more frequently present in Her2neu positive cancers and triple negative breast cancers, CD8+ TILs in Luminal A type of breast cancer and triple negative breast cancers. FOXP3 was found to be expressed more in higher grades of tumor associated with high NPI. FOXP3+ infiltrate was found to have a positive correlation with NPI (p value=0.019). Conclusion: There is a differential expression of various TIL subsets across different histologic grades and molecular subtypes of breast cancer. TIL evaluation in histologic section can serve as a surrogate marker of tumor immunity and its subtypes may serve as forthcoming prognostic biomarker.

Keywords: Breast cancer, immunohistochemistry, tumor infiltrating lymphocytes

   AOP52 Top

   Breast Pathology Top

Prognostic value of marginal adipose tissue invasion in invasive breast carcinoma

Prajwala Nagaraju, K Amita, Geetanjali A Burbure, Ramya Suresh

Adichunchanagiri Institute of Medical Sciences, B.G Nagara, Karnataka, India

Introduction: Breast cancer with mortality of 12.7/100,000 women and age adjusted rate of 25.8 /100,000 women is ranked as number one cancer among Indian females. Functional lymphatics at peritumoral site is mainly responsible for lymphovascular invasion (LVI). Marginal adipose tissue invasion (ATI) of cancer cells leads to larger area of contact between peritumoral lymphatics and increased chances of LVI, ultimately leading to Lymph node metastasis (LNM). Objectives: To investigate whether ATI of cancer cells at tumor margin influenced lymph node status and prognosis in patients with invasive breast carcinoma (IBC). Materials and Methods: In this retrospective study, 43 cases of IBC were studied over a period of 3 years. Data for 33 cases with marginal ATI were clinicopathologically compared with data for 10 cases without ATI. The utility of combination of ATI and peritumoral LVI was also examined. Results: The frequency of LNM was 63.63% in patients with ATI (21/33) and 0% in patients without ATI (0/10) (p value < 0.001). The frequency of peritumoral LVI was 88.88% in patients with ATI (24/27) and 11.1% in patients without ATI (3/27) (p value < 0.01). ATI was an independent factor influencing LNM. In addition, patients without ATI or LVI had no LNM (n = 7). Conclusion: ATI positive cases were associated with adverse outcomes. ATI should be incorporated in surgical pathology report of IBC as ATI and peritumoral LVI will be useful in the framing therapeutic strategies and in assessment of prognosis.

Keywords: Lymph node metastasis, lymphovascular invasion, marginal adipose tissue invasion

   AOP53 Top

   Breast Pathology Top

Skin adnexal tumours of breast: A diagnostic dilemma

J S Reshmi, Tanuja Shet, Asawari Patil, Ayushi Sahay, Sangeeta Desai

Department of Pathology, Tata Memorial Centre, Mumbai, Maharashtra, India

Introduction: Skin adnexal tumors (SATs) comprise a wide spectrum of benign and malignant tumors that exhibit morphological differentiation towards one or more adnexal structures found in normal skin. In breast, these tumors may be clinically and histopathologically confused with Invasive breast carcinoma (IBC). Objectives: To review clinicopathological features of skin adnexal tumors of breast diagnosed at our institute. Materials and Methods: We retrieved and reviewed all cases of skin adnexal tumors of breast diagnosed from 2012 to 2022 (10 years). Results: We diagnosed 13 cases of SATs of breast in above duration. Patient age ranged from 31-73 years (median age 60) and 61.5 % were females. Out of the 13 cases, 2 cases were benign nodular hidradenomas, 8 cases were carcinomas with eccrine differentiation, and there was one case each of carcinoma with follicular differentiation, apocrine adenocarcinoma and sebaceous carcinoma. All the tumors had a dermal and subcutaneous location in the absence of a breast mass by imaging. On immunohistochemistry, GATA 3, Estrogen receptor and Androgen receptor were positive in 30 %, 23 % and 23 % of cases, respectively. All the cases underwent wide local excision. 2/13 cases showed nodal metastasis and received adjuvant radiation and chemotherapy. The rate of local recurrence was 30 % in malignant tumors. There was no evidence of distant metastasis. Conclusion: Awareness of spectrum of adnexal tumors in breast is essential as it is a great clinical mimicker of Invasive breast carcinomas, and the management is entirely different.

Keywords: Adnexal tumors, breast, invasive breast carcinoma

   AOP54 Top

   Breast Pathology Top

Diagnostic utility of catenins (p120 and β-Catenin) in invasive breast carcinoma

Sheenal Bhatia, Ayushi Sahay, Tanuja Shet, Asawari Patil, Sangeeta Desai

Tata Memorial Hospital, Mumbai, Maharashtra, India

Introduction: E-Cadherin (EC) immunohistochemistry (IHC) is considered diagnostic for differentiating invasive breast carcinoma (IBC) from invasive lobular carcinoma (ILC). However, aberrant retained EC expression reported in ILC (0-23.5%), or reduced/absent EC in IBC (18.7-43.5%) in tumours diagnosed based on morphology alone. Other cadherin-catenin complex proteins (p120, β-catenin) may be useful in such cases. ILC shows cytoplasmic relocalization of p120, while IBC shows membranous p120. β-catenin shows loss of membranous staining in ILC, similar to EC. Objectives: To assess diagnostic utility of catenins (p120, and β-catenin) versus EC in IBC. Materials and Methods: A total of 92 IBC cases where EC IHC had been performed previously were retrieved. p120 and β-catenin IHC was performed and compared with EC (gold standard). Results: p120 and β-catenin were interpretable in 71 (36 IBC, 35 ILC) and 72 cases (37 IBC, 35 ILC) respectively. p120 showed cytoplasmic positivity in 35/35 (100%) ILC and membranous positivity in 36/36 (100%) IBC. β-catenin showed retained membranous staining in 36/37 (97.2%) IBC and loss in 34/35 (97.2%) ILC. One IBC (EC retained) showed β-catenin loss, and one ILC (EC lost) showed retained β-catenin. Additionally, β-catenin stained blood vessels and connective tissues. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of p120 was 100% compared to EC; β-catenin showed 97.3% sensitivity, 97.14% specificity, 97.3% PPV, 97.14% NPV and 97.22% accuracy compared to EC. Conclusion: p120 and β-catenin may serve as useful adjuncts to EC for distinguishing ILC versus IBC. p120, being a positive marker, is particularly useful. β-catenin, though sensitive and specific, may have limited utility due to significant background staining.

Keywords: Catenins, E-cadherin, p120, β-catenin

   AOP55 Top

   Breast Pathology Top

A histopathological study of T regulatory cells in breast carcinoma using CD4 and FOXP3 immunohistochemistry markers

Nisarga Rani, Nirupama

Dr Chandramma Dayanand Sagar Institute of Medical Education and Research, Hubli, Karnataka, India

Introduction: T regulatory cells (Tregs) play an important role in regulation of tumour immunity and immune response. Cytotoxic T cells are equipped to control tumour growth and metastasis, tumour corrupted immune regulatory T cells impair effective immunity and promote tumour progression. There is dearth in histopathological studies, especially in their expression in human breast carcinomas, prognosis and their density in relapsed cases. In the present study we aim at morphological study of T-regs in various molecular subtypes of breast carcinomas, morphological associations and prognosis wherever possible. Objectives: 1. To study morphology of breast carcinomas and subtype the tumor based on molecular classification. 2. To Identify tumor infiltrating CD4 lymphocytes and estimating T cells /HPF at tumor margins and intra tumoral regions. 3. To identify Tregs expressing immunohistochemistry marker FOXP3 in breast carcinoma using tissue sections. Materials and Methods: Resected specimens of breast carcinoma received at the department of Pathology of our hospital were included in the study. Inclusion criteria: All primary cases and histologically proven cases of breast carcinoma are included. Exclusion criteria: Improperly fixed/ processed/ preserved paraffin blocks and blocks without adequate tissue. Data was analysed using analytical statistics chi-square method. Results: A total of 16 cases were studied, predominant cases were Invasive carcinoma with no special type and one mucinous carcinoma. Majority of them were Luminal subtype A. A statistical significance of FOXP3 noted with grade of tumour, hormonal study (ER, PR, and Her2/neu) 0.02, 0.04, 0.04, 0.018 respectively. However no association seen in size of tumour and lymph node metastasis. Intratumoural CD4 lymphocytes shown statistical significance with grade of tumour and molecular subtypes of tumour, 0.03 & 0.04 respectively. Peritumoural lymphocytes show no significance with grade, size of tumour and molecular subtypes. Conclusion: Breast carcinoma predominantly shown Luminal A subtype. Treg cells, FOXP3 and intratumoural CD4 lymphocytes expression increased with higher grade of tumour. However FOXP3, intra and peritumoural CD4 lymphocytes association with size of tumour and nodal status were not seen. Further study with larger number required to observe the statistical significance.

Keywords: Breast carcinoma, CD4, FOXP3

   AOP56 Top

   Breast Pathology Top

Expression of proteinase-activated receptor 2 (PAR2) as a correlate of concern in triple-negative breast cancer (TNBC)

Gargi Kapatia, Subhpreet Kaur, Sandeep Kumar, Ishita Laroiya, Gurpreet Singh, Maryada Sharma, Amanjit Bal, Manni Luthra-Guptasarma

Department of Pathology, AIIMS, Bathinda, Punjab, India

Introduction: Triple-negative breast cancer (TNBC), a highly aggressive cancer with poor outcome and lacking specific diagnostic, prognostic, or targeted therapeutic strategies, constitutes roughly 20% of all breast cancer cases. TNBC cells lack receptors for estrogen, progesterone, and human epidermal growth factor. Objectives: To find if PAR2 is suitable correlate that could serve as a TNBC biomarker. Materials and Methods: A retrospective study was performed with 88 TNBC and 74 non-TNBC patients who had undergone mastectomy/lumpectomy with axillary clearance for carcinoma breast. Immunohistochemical staining was carried out for levels of proteinase-activated receptor 2 (PAR2), encoded by F2RL1 gene, and staining scores were calculated, based on intensity and percentage positivity. Results: PAR2 levels were markedly upregulated in TNBC patients, compared with patients with other breast cancer subtypes. Amongst different non-TNBC subtypes, higher expression was noted in luminal B (88.8%) and HER2+ (100%), compared with luminal A (52.5%). PAR2 levels were significantly high in TNBC patients with age more than 40 years than corresponding patients of non-TNBC group (P= 0.0017). Furthermore, there was a statistically significant increase in levels of PAR2 expression in lymph node negative (P=0.0096) and early stage (P=0.005) of TNBC versus non-TNBC patients. PAR2 staining of ductal carcinoma in situ and invasive ductal carcinoma revealed lower expression in invasive component. Conclusion: Our data suggest that PAR2 levels constitute a correlate of concern for TNBC, tying in with a recent report that higher levels of F2RL1 gene expression correlate with poorer disease-free, as well as overall survival in TNBCs. Keywords: Immunohistochemistry, PAR2, triple-negative breast cancer

   AOP57 Top

   Breast Pathology Top

Is tumor budding, a potential histopathological marker to assess clinicopathological prognosis in breast carcinoma: A cross-sectional study

C Sindhu, R Kalyani, P N Sreeramulu

Department of Pathology, Sri Devaraj Urs Medical College, Kolar, Karnataka, India

Introduction: Breast cancer is the most common type of cancer, primary cause of mortality and the second leading cause of cancer induced death among women. Many prognostic factors are implicated in breast carcinoma including clinical, histomorphology and molecular parameters. Tumor budding is one such histomorphological parameter and evolving concept in several cancers. Objectives: To evaluate tumor budding in invasive breast carcinoma and its association with clinicopathological parameters. Materials and Methods: A retrospective study conducted in Department of Pathology, RL Jalappa Hospital, Kolar between January 2017 to December 2021. The demographic and clinical details with histopathology slides of invasive breast carcinoma slides were retrieved from the Archives of the Department, evaluated for tumor budding and associated with clinicopathological parameters like histological type, grade, necrosis, age, tumor size, lymph node status, lymphovascular invasion and hormonal receptor status. Results: 110 cases of Invasive breast carcinoma were analyzed. The majority of the patients were postmenopausal (67.2%), tumor size <5 cm (61.8%), Invasive Ductal Carcinoma NOS (94.5%) common histological type, had positive lymph nodes (54.5%), grades 1 and 2 (85.4%), NPI score of less than 5.4 (78.1%) and presented in stage 1 and 2 (57.2%). Association of tumor budding with lymph nodes status and lymphovascular invasion was found to be statistically significant. Conclusion: Tumor budding assessed on routine histologic sections was associated with lymph node metastasis and lymphovascular invasion. Tumor budding can be a novel histopathological marker that can be incorporated as a prognostic factor in evaluating cases of breast cancers.

Keywords: Breast carcinoma, peritumoral budding, tumor budding

   AOP58 Top

   Breast Pathology Top

Pathological factors responsible for recurrence in early breast cancer patients in a tertiary cancer centre – 3 year experience

Kandathil Philip, Feba Philo Jolly, Geetha Muttath, Maya Padmanaban, Adarsh Dharmarajan, Nandini Devi

Division of Oncopathology, Malabar Cancer Centre, Thalassery, Kannur, Kerala, India

Introduction: Breast cancer is the most common and leading cause for cancer among women globally accounting for 24.5% of the total number of new cases diagnosed in 2020. Objectives: To determine different pathological factors responsible for recurrence among early breast cancer patients. Materials and Methods: This is a retrospective study which included all invasive early breast cancer patients diagnosed from 1st January, 2015 to 31st December, 2017 who was under follow up with pathological stage upto stage IIB. Results: There were 399 early invasive breast cancer patients who underwent surgical treatment. Recurrence occurred in 34 patients (8.5%), majority of which were distant recurrence (83.3%) followed by regional recurrence (11.1%) and local recurrence (5.6%). The most common sites of distant recurrence was bone (21%). Among the recurrence patients, majority had tumour size of T2 (82.4%), higher tumour grade (grade 2/3) (88.2%) and almost half the cases (47%) had worst molecular subtypes like TNBC and Her 2 enriched. The overall survival of the recurrent early breast cancer patients was 49.6% as against 88.4% in the non-recurrent group which was statistically significant. Conclusion: In our study, recurrence among early breast cancer patients was found in 8.5%, predominantly distant recurrence affecting most commonly bone. Few pathological factors like higher grade (G2/G3), larger tumour size (T2) and worse molecular subtypes like TNBC and Her 2 enriched increased the risk of developing recurrence among early breast cancer patients although there was no statistical significance.

Keywords: Early breast cancer, molecular subtypes, recurrence

   AOP59 Top

   Breast Pathology Top

Correlation of cytological findings and BI-RADS system for ultrasonography with histopathology in breast masses

Monika Gathwal, Chiranjeev Gathwal, Ruchi Agarwal, Swaran Kaur Saluja, Deepti Agarwal, Parveen Rana, Nitika Chawla

BPS GMC (W), Khanpur Kalan, Sonipat, Haryana, India

Introduction: Breast ultrasound and FNAC play an important role in characterization of a breast lump as benign and malignant. There is a need to study radiological and cytological correlation of breast lesions with histopathology & to evaluate the diagnostic accuracy of BIRADS and FNAC. Objectives: To find correlation of cytological findings and BI-RADS classification in breast masses with histopathology. Materials and Methods: This prospective study included 120 patients with breast lump who underwent breast ultrasound, FNAC and histo-pathological examination from 2016 to 2018. Results: On sonography, 120 patients evaluated; 44 were classified as benign breast disease (BI-RADS II, III) and 76 were classified as suspicious of carcinoma or carcinoma of breast (BI-RADS IV, V). On histopathology 95.45% and 81.81% cases of BI-RADS III & IVa were classified as benign and 73.68% cases of BI-RADS IVb, 100% cases of BI-RADS IVc & V were classified as malignant. BIRADS show sensitivity (95%), specificity (75%), PPV (82%), NPV (93%) and diagnostic accuracy (86%) while FNAC breast show sensitivity (89%), specificity (98%), PPV (98%) and NPV (93%) and diagnostic accuracy (93%). On association analysis using Chi square test, the BIRADS and FNAC both show significant association with p value <0.001. Kappa value of FNAC was 0.867 and of BIRADS was 0.710 with significant p value of <0.001. Conclusion: Sub-categorization of BI-RADS 4 play an important role in management of breast masses and cases with BI-RADS 4b and above should be investigated thoroughly with biopsy to confirm or rule out malignancy.

Keywords: BIRADS, breast carcinoma, radio-cytological correlation

   AOP60 Top

   Breast Pathology Top

Diagnostic significance of micro lymph node metastasis and its clinicopathological correlation among the women harboring ductal carcinoma of the breast

Seema, Dayal

UPUMS, Etawah, Uttar Pradesh, India

Introduction: Lymph node metastasis is a significant prognostic marker in the breast cancer treatment and is associated with poor prognosis. Objectives: This study was undertaken to determine the lymph node metastasis status including its parameters and correlation with clinical and microscopic variables. Materials and Methods: This study was conducted in the Department of Pathology, UPUMS, Saifai, Etawah (U.P), India. The female patients diagnosed with ductal carcinoma breast with lymph node metastasis from 2008 to 2022 were included. The metastatic lymph node parameters and its other associated pathologies were evaluated and compared. Immunohistochemistry for ER, PR, Her2neu, vimentin, P53 was also correlated. Results: The maximum number of patients were < 50 years of age (58.6 %). The common tumor size was 5 cm and below 64 (69.5 %). The frequent histological grade 67 (72.8%), favorable prognosis was moderate 55 (59.7%). The largest lymph nodes retrieved was 3x2 cm and maximum no. of lymph nodes involved 17/19. The other lymph nodes pathologies were granulomatous lesion 2 (2.17%), microfilaria 2 (2.17%), calcification 4 (4.3 %). The immunohistochemistry positivity were ER (16.30%), PR (11.95%), Her2neu (21.73%), vimentin (16.30%) and P53 (6.52%). Conclusion: The lymph node status not only emulate chronological age of tumor but also tumor biology. It was also found associated with clinical parameters, tumor histology and immunohistochemistry. These variables may be used for better management of breast cancer patients.

Keywords: Ductal carcinoma, histopathology, lymph node

   AOP61 Top

   Breast Pathology Top

Role of cancer stem cell marker (CSC) CD44 in triple negative breast cancer (TNBC) and its association with clinicopathological parameters and angiogenesis

Reena Tomar, Garima Rakheja, Nidhi Verma, Shruti Thakur, Nita Khurana, Deepak Ghuliani

Maulana Azad Medical College, New Delhi, India

Introduction: Triple-negative breast cancer (TNBC) (ER–/PR–/Her2–), which accounts for 15–20% of all breast cancers. Triple negative breast cancers (TNBC) are aggressive tumors with limited options for targeted therapy. Objectives: This study aims to find out the expression of (CSC) marker CD44 in (TNBC) verses non triple negative breast cancers (NTNBC). Materials and Methods: CSC marker CD44 was put in total 58 patients and TNBC was further categorized into CSC+ / CSC-. Angiogenesis by CD34 was observed and microvessel density (MVD) was calculated for all cases. Association of (CSC) Cancer Stem cell marker with histological tumor grade (Nottingham's grade) was observed and statistical analysis was done using appropriate statistical methods. Results: Out of total 58 cases, 28 cases were TNBC and 30 cases were NTNBC. Out of 28 TNBC, 22 were CSC+ and 6 were CSC- . Out of 30 NTNBC, 16 were CSC+ and 14 were CSC- (p value 0.043). Averge microvessel density (MVD) was 9.73/mm2 in TNBC and was 11.5/mm2 in NTNBC. High histological grade was associated more with TNBC. Conclusion: CSC marker CD44 was expressed more in triple negative receptor type of breast cancers (TNBC). Their positivity correlate with high histological grade. This study will give novel tissue-based risk biomarkers for triple negative breast cancers and will lay the foundation for planning the advanced strategies for the treatment and follow up in triple negative breast cancer patients.

Keywords: Cancer stem cell, CD44, triple-negative breast cancer

   AOP62 Top

   Cytopathology Top

Cytomorphological correlational study of thyroid disorders with imaging and biochemical serum markers

Sumaiyya Firdous, Zeenath Begum

Faculty of Medical Sciences, KBN University, Kalaburagi, Karnataka, India

Introduction: Fine-needle aspiration cytology (FNAC) based on Bethesda system is a valuable test for diagnosing diseases of thyroid gland. Thyroid lesions can be categorized as euthyroid, hypothyroid or hyperthyroid based on levels of triiodothyronine, thyroxin and thyroid-stimulating hormone. Ultrasonography (USG), based on Thyroid Imaging Reporting and Data Systems (TIRADS) is the modality of choice to determine the risk of malignancy in thyroid nodules. Objectives: 1. To study the cytomorphological features of thyroid lesions based on Bethesda System-2014. 2. To correlate the cytomorphological features with radiological imaging based on TIRADS classification-2017. 3. To correlate the cytomorphological features with biochemical markers and histopathological findings whenever available. Materials and Methods: Retrospective correlational analysis of 70 cases over a period of one year from March-2021 to April-2022 was done. Each patient's clinical, biochemical, FNAC and USG findings were recorded from the reports. Histopathology findings noted wherever available. Results: Out of 70 cases, 57 were from Bethesda category-II, 4 cases were from category-III, 3 cases were from category-IV, 1 from category-V, 5 cases were from category-VI. Out of 70 cases, 54 cases were from TIRADS-2, 7 cases from TIRADS-3, 3 cases from TIRADS-4a, 2 cases from TIRADS-4b and 4 from TIRADS-5. Out of 70 cases, 12 cases were of hypothyroidism (Hashimoto's thyroiditis), 5 cases were of hyperthyroidism (2 colloid nodule, 1 follicular neoplasm, 2 papillary carcinoma) and 53 were euthyroid. 10 cases were available for histopathological correlation. 2 cases were of multinodular goiter, 1-case was Hashimoto's thyroiditis, 1-Hurthle cell adenoma, 1-Parathyroid adenoma, 1-Follicular adenoma, 4-Papillary thyroid carcinoma. Conclusion: A combined approach of clinical, cytological, biochemical, radiological and histopathological correlation of thyroid diseases enhances the diagnostic accuracy and improves management.

Keywords: Bethesda system, thyroid disorders, Thyroid Imaging Reporting and Data Systems classification

   AOP63 Top

   Cytopathology Top

Cytomorphological evaluation of lymphadenopathy and histopathological correlation

SyedaHafsa Fatima, Zeenath Begum, Pratima Sambrani, Uzma Alvi

Faculty of Medical Sciences, Khaja Bandanawaz University, Kalaburagi, Karnataka, India

Introduction: Lymphnodes are affected by a wide spectrum of diseases varying from infections to malignancy, which commonly manifests as lymphadenopathy. Various modalities available for diagnosis of lymphadenopathy are clinical evaluation, FNAC and excisional biopsy. Biospy is gold standard, but it is complicated and time consuming procedure. A recent trend in medical practice is to adopt a diagnostic technique which is both cost effective and minimally invasive, hence, FNAC is often used as a first line of investigation for lymphadenopathy, as it is simple, rapid, inexpensive and less invasive. Objectives: To study the role of FNAC in the diagnosis of lymphnode lesions. To evaluate the accuracy of FNAC, and correlate it with histopathology. Materials and Methods: This is a 2 year retrospective study including 235 cases of lymphnode FNAC presenting with lymphadenopathy to our department, out of which 50 cases were available for histopathological correlation. Results: Out of 235 cases, benign lesions were common (173 cases, 73.6%), followed by metastatic malignancies (35 cases, 14.9%) and primary malignancies (27 cases, 11.5%). Among 173 benign lesions, non-specific reactive lymphadenitis (80 cases, 46.2%) was most common and we also found 1 case (0.57%) each of Rosai-Dorfman disease, Kikuchi lymphadenitis and Sinus histiocytosis. On correlation with histopathology, out of 24 cases diagnosed as benign on cytology, one turned out to malignant and out of 26 cases diagnosed as malignant, one turned to benign. Overall sensitivity, specificity and accuracy of FNAC in diagnosis of lymphadenopathy was 95.8%, 96.1% and 96% respectively. Conclusion: FNAC was proven to be an excellent first line investigation for diagnosis of lymphnode lesions, as it is inexpensive and an easy alternative to open biospy.

Keywords: Fine-needle aspiration cytology, histopathology, lymphadenopathy

   AOP64 Top

   Cytopathology Top

A study of benign breast lesions – Cytology at tertiary care centre

Majji Sarayu, M Neeraja, V Sivasankara Naik, E Divya, P Deepthi

Government Medical College, Anantapuramu, Andhra Pradesh, India

Introduction: Breast FNAC has a significant value as a first line of investigation. This is probably due to its simplicity, minimal invasiveness, cost efficiency and fast turn around time. Therefore, patients presenting with breast lump at a tertiary care center can initially be evaluated by FNAC. Objectives: To study the various cytomorphological features of benign breast lesions. Materials and Methods: This study is a cross-sectional retrospective study conducted in the Department of Pathology from 2021 to 2019. Clinical details and cytology features were collected from the Department records. Results: A total of 200 cases were collected during the study period. Age groups ranged from 16-50 years. All the cases were females. The following were the cytological diagnosis: Fibroadenoma-110 (55%), Fibrocystic disease-28 (14%), Benign phyllodes -27 (13.5%), Fibroadenosis-10 (5%), Acute suppurative mastitis-9 (4.5%), Chronic granulomatous mastitis- 8 (4%), Lactating adenoma-5 (2.5%), Fat necrosis – 3 (1.5%). Conclusion: FNAC helps in rapid diagnosis and early management of lesions. It also helps in preventing unnecessary invasive surgeries in nonneoplastic and benign breast diseases making it a useful diagnostic tool.

Keywords: Benign breast lesions, cytology, fine-needle aspiration cytology

   AOP65 Top

   Cytopathology Top

Application of 2014 Bethesda system for reporting cervical cytology – An institutional study

Vallamsetty Nagasudharani, M Neeraja, V Siva Sankara Naik, E Divya, J Bhagyalakshmi

Government Medical College, Anantapur, Andhra Pradesh, India

Introduction: Cervical cancer is one of the leading malignancies among women in India. Pap smear (conventional method) is widely used cervical cancer screening test. In developing countries like India, there is a great need for mass screening program for detection of cervical lesion. Objectives: To evaluate the abnormal cytological entities detected by pap smear and to classify the cytological findings according to 2014 Bethesda system. Materials and Methods: This is a hospital based study of conventional cervical smears received between May 2020 to June 2022. For evaluating the Pap smears, the Bethesda system 2014 for reporting cervical cytology was used. Results: In the study period, 1418 cases were valuated. Age of women ranged from 19 to 92 years. Most common complaint was irregular bleeding per vagina and leucorrhea. 82.08% belonged to NILM category, 9.3% were unsatisfactory for evaluation. ASCUS accounted for 4.87%, ASC-H 0.56%, LSIL1.83%, HSIL 0.77%, AGC 0.28% and SCC 0.28% cases of cases. Conclusion: It was found that negative for intraepithelial malignancy cases were common in our set up.

Keywords: Bethesda system, cervical cytology, papanicolaou smear

   AOP66 Top

   Cytopathology Top

Spectrum of cytological abnormalities in lymphnodes at tertiary care hospital

Manthri Kavya, M Neeraja, V Sivasankar Naik, M Sreenivasulu, Y Sreedhar, B Aneela

Government Medical College, Anantapuramu, Andhra Pradesh, India

Introduction: Lymph node enlargement is common presentation in all age groups ranging from infections to malignancy. Therefore, management of cases depends on lymph node pathology, which can be studied by collecting material through fine needle aspiration method. Objectives: To study the cytomorphological features associated with lymphadenopathies. Materials and Methods: The present prospective study was carried out in the Department of Pathology at Government medical college, Anantapur, a Tertiary Care Centre. A total of 200 patients of all age groups underwent FNAC of enlarged lympnodes during study period. Results: FNAC diagnosis was found to be as follows: Tubercular lymphadenitis in 70 cases (35%), Reactive hyperplasia in 60 cases (30%), Metastatic deposits in 40 cases (20%), Lymphomas in 20 cases (10%), Salivary gland tumors were 10 cases (5%). Conclusion: FNAC of lymph nodes is an excellent first-line investigation to determine the nature of the lesion. Tubercular lymphadenitis is the most common lesion followed by reactive hyperplasia.

Keywords: Lymphadenopathy, malignant neoplasms, tuberculous lymphadenitis

   AOP67 Top

   Cytopathology Top

Cytological grading of salivary gland lesions according to Milan system in a tertiary care hospital

Salma Sultana, M Neeraja, V Siva Sankara Naik, P Sravani

Government Medical College, Anantapuramu, Andhra Pradesh, India

Introduction: Fine Needle Aspiration Cytology (FNAC) used routinely with clinical and radiological findings for evaluation of salivary gland lesions as it is diagnostically challenging due to significant cytological diversity and heterogenecity. Introduction of Milan System for Reporting Salivary Gland Cytopathology (MSRSGC) which is internationally accepted, helps in uniform reporting, improve overall patient care by early diagnosis and proper preoperative surgical planning. Objectives: To grade the salivary gland lesions according to MILAN System and to calculate risk of malignancy (ROM). Materials and Methods: A 4 yrs study was conducted on 100 cases, at government Medical college, hospital, Anantapuramu from july 2018 to july 2022. All the relevant data, records and slides were assessed and correlated with histopathology for available specimens. The cytological diagnosis was re-evaluated as follows: Category 1: Non diagnostic (ND), Category 2: Non neoplastic (NN), Category 3: Atypia of undetermined significance (AUS), Category 4a: Neoplasm: Benign (NB), Category 4b: Neoplasm: Salivary gland neoplasm of uncertain malignant potential (SUMP), Category 5: Suspicious of malignancy (SM) and Category 6: Malignant (M). Results: Total 100 cases were studied. Case distribution in various categories were ND (8%), NN (42%), AUS (0%), NB (35%), SUMP (2%), SM (9%) and M (4%). Overall ROM reported were ND (0%), NN (0%), AUS (0%), NB (10%), SUMP (100%), SM (50%) and M (100%). The sensitivity, specificity, positive predictive value and negative predictive value of the study was 33.33%, 96.77%, 50%, 93.75% respectively. Conclusion: The Milan system is an universal reporting system that helps with risk assessment and guides for appropriate management.

Keywords: Fine needle aspiration cytology, Milan system, risk of malignancy, SGL

   AOP68 Top

   Cytopathology Top

Significance of nuclear morphometry in salivary gland neoplasms – A cross sectional study

Jahnavi Reddy Yeduguri, Y Jahnavi Reddy, R Kalyani, S M Azeem Mohiyuddin

Sri Devaraj Urs Medical College, Kolar, Karnataka, India

Introduction: Fine needle aspiration is one of the commonest and efficient investigation done for salivary gland swellings, which helps in diagnosing various lesions of salivary glands. The morphology of nucleus mainly determines the behaviour of the cell/tissue. Morphometry analysis is done by using various softwares. Morphometry study of the cell consists of measurement of cell parameters such as area of the nucleus, perimeter of the nucleus, feret diameter, skewness etc. In this study we planned to consider only nuclear morphometry in FNAC samples of salivary gland neoplasms which may help in differentiating benign from malignant neoplasms. Objectives: 1. To study FNAC smears of all salivary gland neoplasms. 2. To study nuclear parameters of FNAC smears. 3. To correlate the nuclear morphometry finding with cytological findings. Materials and Methods: (a) Design of study – Retrospective Laboratory observational cross sectional study. (b) Total number of study subjects - All the cases in the span of 4 years (2018-2021). (c) Mode of selection of subjects - All the cases with salivary gland swellings. Procedure – All the smears of salivary gland neoplasms were taken, analysed using Image J software for nuclear morphometric parameters such as area of the nucleus, perimeter of the nucleus, feret diameter, minimum feret, skewness. These findings are correlated with histopathological diagnosis of all the available cases. The values were noted and analysed for cytomorphological diagnosis. Results: The average age of presentation in benign salivary gland neoplasm was 38.7 years and for malignant salivary gland neoplasms was 49.4 years. Among benign salivary gland neoplasms, pleomorphic adenoma was the commonest and among malignant squamous cell carcinoma deposits were the commonest lesions. The nuclear morphometry analysis showed that the mean values of area, perimeter, ferret diameter, minimum ferret and skewness of malignant lesions were comparatively higher than benign. Conclusion: This article concludes that nuclear morphometry along with regular cytopathological evaluation will improve the accuracy of diagnosis in all neoplastic lesions of salivary gland. This further aids in better treatment and predict prognosis for the patient. Digital morphological analysis also helps in getting quantitative values from qualitative data.

Keywords: Fine needle aspiration cytology, nuclear morphometry, salivary gland

   AOP69 Top

   Cytopathology Top

Spectrum of thyroid lesion on fine needle aspiration in tertiary care hospital

Ravina Purohit, Arpita Nishal, R N Hathila, Prashant Patel, Archana Patel

Goverment Medical College, Surat, Gujarat, India

Introduction: Thyroid swelling is a very common complain in routine practice. Fine needle aspiration cytology (FNAC) is the most common preoperative investigation for diagnosis of thyroid lesions and a helpful tool for intial diagnosis and workup of thyroid nodule. Objectives: To study the spectrum and frequency of thyroid lesions and categorise it according to Bethesda system for reporting thyroid cytology. Materials and Methods: This is a retrospective study conducted at department of pathology in tertiary care center from January 2021 to December 2021 that were referred to our cytopathology laboratory in this period. The data of 88 cases with enlarged thyroid gland were retrieved from past records. Thyroid lesions were classified according to Bethesda system for Reporting Thyroid Cytopathology (2017). Results: Out of 88 cases, Bethesda category I included 8 cases (9%), category II consisted of 71 cases (81%), category III (AUS/FLUS) consisted of 4 cases (5%), category IV (Follicular Neoplasm) included 2 cases (2%), category V (Suspicious of malignancy) included 0 case (0%), category VI (malignant) had 2 cases (2%). One case of thyroglossal cyst was also found but it is not included in Bethesda category. Conclusion: FNAC is safe, quick, cost effective and reliable method as first line evaluation in thyroid gland nodules and does not involves major complications. FNAC derives much of its significance from its ability to reliably identify benign thyroid nodules, thus sparing many patients with unnecessary surgery.

Keywords: Bethesda system, cytology, thyroid pathology

   AOP70 Top

   Cytopathology Top

Utility of the application of International Academy of Cytology Yokohama system of reporting breast fine needle aspiration cytology in patients with breast lump in comparison with radiological findings and the gold standard

M T Megha, Aswathy Chandramohan, Limi Mohandas

Department of Pathology, Sree Gokulam Medical College, Venjarammoodu, Kerala, India

Introduction: Breast cancer is the most common malignancy in women worldwide. There is a need to correlate the radiological and cytological findings of breast lesions with histopathological diagnosis to evaluate the diagnostic accuracy of ultrasound and breast FNAC. Objectives: 1) To assess the utility of breast FNAC in patients with breast lump according to Yokohama system and compare with radiological findings and with the gold standard. 2) To calculate the risk of malignancy (ROM) of each category. Materials and Methods: FNAC is done using a 22 G needle, the wet smears are stained with Papanicolaou stain and dry smears with Giemsa stain. The smears are studied under light microscopy (10x, 40x). A total of 63 breast FNAC samples were classified according to IAC Yokohama Reporting system. The cytology report is compared with radiological findings and histopathology findings. The risk of malignancy is calculated. Results: We correlate this IAC Yokohama category with histopathology diagnosis and radiological diagnosis. The ROM for each category was 44.4% for category 1, 8.8% for category 2, 16.6% for category 3 and 100% for both category 4 and 5. The sensitivity, specificity, positive predictive value and negative predictive value of FNAC were respectively 63.6%, 100%, 110% and 83.6% respectively. The sensitivity, specificity, positive predictive value and negative predictive value of USG were respectively 86.3%, 90.2%, 82.6% and 92.5% respectively. Conclusion: IAC Yokohama system of reporting breast pathology helps in standardizing and increasing reproducibility of breast FNAC.

Keywords: Histopathology, IAC Yokohama, risk of malignancy

   AOP71 Top

   Cytopathology Top

IAC Yokohama cytomorphology of breast lesions with histo-radio correlation and utility of P63 immunostaining in grey zone lesions

Nikita Jaju, R Shubha Sangeetha, Y A Manjunatha

DR B R Ambedkar Medical College and Hospital, Bengaluru, Karnataka, India

Introduction: In 2016, International Academy of Cytology (IAC) established a “Breast Group” which included pathologists, radiologists, surgeons, and oncologists and has proposed five-category classification, “The IAC Yokohama System for Reporting Breast Cytopathology (C1-C5)”. Aim is to bring the uniformity to reporting system and address cytomorphologic grey zone (C3/C4) uncertainties. True grey zone lesions pose diagnostic difficulty due to presence of atypical/ suspicious features. p63 positive immunocytostain is characterized as reliable marker of myoepithelial cells of breast and is recognized as a prominent feature of benign lesions. Objectives: 1) To correlate the cytomorphological spectrum of breast lesions according to IAC Yokohama reporting system with radiological reporting (BIRADS). 2) To implicate p63 immunocytostain in grey zone lesions and confirm with histopathology. Materials and Methods: Over a period of 18 months, 100 FNACs were performed with radiological pre-requisite and 48 cases had followed up by histopathological examination were included in study with implication of p63 in grey zone lesions. Results: Among 100 cases studied, Female:Male ratio was 99:1. Incidence ranged between 30-49 years (48%) of age. Lesions were then categorized according to IAC Yokohama reporting system. Most commonly encountered benign lesion was fibroadenoma (C2)(40 %) and malignant lesion was IDC (C5)(20%). BIRADS showed 78.12% specificity in correlation with FNAC findings. Grey zone lesions (C3/C4) accounted for about 20% of cases (fibroadenoma, fibrocystic disease, papilloma, papillary lesions, proliferative lesions, radial scar, sclerosing adenosis, breast abscess, phyllodes tumor, etc). Then, P63 immunocytostain on these lesions helped to efficiently categorize suspicious lesions as benign/malignant. Conclusion: The IAC Yokohama cytopathology reporting system offers a standardized approach by uniformly categorizing breast lesions and p63 immunocytostain removes the ambiguity in grey zone lesions with direct link to management recommendations to the clinicians.

Keywords: Grey zone lesions, IAC Yokohama, p63

   AOP72 Top

   Cytopathology Top

A comparitive study of granulomatous lymphadenitis in FNAC concentrates with acid fast bacilli staining and cartridge based nucleic acid amplification test in the diagnosis of tubercular lymphadenopathy

S Bhavya, C N Anushree, Y A Manjunatha

DR B R Ambedkar Medical College, Bengaluru, Karnataka, India

Introduction: India has the highest burden of tuberculosis (TB) with incidence of 2,590,000 in 2021. Extrapulmonary tuberculosis (EPTB) is an important clinical condition constituting 15- 20% of all TB cases. Its most common presentation is lymphadenitis. Diagnosis of EPTB is challenging due to its paucibacillary nature. Objectives: To compare Fine Needle Aspiration Cytology (FNAC) findings with Acid fast bacilli (AFB) staining and CB-NAAT in the diagnosis of tubercular lymphadenopathy. Materials and Methods: A prospective study of 11 months conducted at of Dr.B.R. Ambedkar medical college and hospital, Bangalore. It excluded known cases of TB and patients aged below 18 years. Smears were made with FNAC aspirates and stained with Papanicolaou and Ziehl Neelsen stain. Remaining aspirate sent for CB- NAAT test. This study included 60 cases showing granulomatous lymphadenitis. Results: Among 60 cases most affected age group is 18-30 yrs (67.3%). Majority cases manifested in cervical lymph node (69.1%) and showed purulent aspirates (43.4%). AFB and CB-NAAT showed positive in 38.3% cases and 81.6% cases respectively. 4 AFB positive cases showed CBNAAT negative. FNAC showed 36 cases of epithelioid granuloma with necrosis (47.2% AFB, 67.34%CB-NAAT - positive) and 24 cases of epithelioid granuloma without necrosis (25% AFB, 32.65% CB-NAAT - positive). Compared to CBNAAT ZN stain had 38.8% Sensitivity, 63.6% Specificity, 82.6% PPV and 81.1% NPV. Conclusion: CB-NAAT can be used as an initial and rapid diagnostic method for tuberculous lymphadenopathy. It is less sensitive to blood tinged samples.

Keywords: Cartridge based nucleic acid amplification test, granulomatous lymphadenitis

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   Cytopathology Top

Utility of conventional ZN staining and modified bleach techniques in detection of AFB in aspirates from suspected cases of TB lymphadenitis

Anjali Shandilya, K Indrani

MVJMC and Research Hospital, Bengaluru, Karnataka, India

Introduction: In 2016, the WHO estimated that the disease affected 10.4 million people and 1.3 million deaths. India has the highest TB burden for onefifth of global incidence (21%). It can involve any organ system and has a wide spectrum of clinical presentation. With the advent of (HIV), there is a global upsurge of mycobacterial infection and Tb has become a major cause of morbidity and mortality. Conventional ZN can be used in smears to detect AFB and sensitivity ranging from 20% to 43%. Modified bleach technique is a simple technique which requires no expertise and is inexpensive. Objectives: (1) To evaluate the efficacy of Modified bleach technique for AFB stain in diagnosing TB lymphadenopathy. (2) To assess the role of Modified bleach technique with conventional ZN stain in diagnosing suspected cases of tuberculous lymphadenopathy. Materials and Methods: FNAC was done using 22 gauge needle, under strict aseptic precautions. In each case, part of the aspirate was used for preparing 3 smears at least, one each for H & E stain, Giemsa stain and ZN stain. Remaining aspirate material in suspected cases of tuberculosis was flushed out for liquefaction with 5% sodium hypochlorite solution in a test tube at room temperature for 30 min. The aspirated material along with the bleach solution was then centrifuged for 15 min at 3000 rpm. The supernatant was discarded and smears were made from the sediment on clean glass slides, followed by ZN staining and half of the residual aspirated material sent for CBNAAT. Results: Total of 100 cases were studied. The age group range was from 1-90 yrs Site wise distribution of cases - Cervical were (91%), Axillary were (6%) and Inguinal were (3%). Majority of the cases Modified bleach technique shows AFB positivity where ZN stain shows AFB positivity. In addition Modified bleach technique method shows AFB positivity where ZN stain shows AFB negative cases. Conclusion: FNAC is a reliable and economical investigating modality in lymphadenopathy. FNAC coupled with Z.N staining should be the 1st line of investigation in cases with lymphadenopathy. The modified bleach method was more sensitive and safer than routine ZN staining. As the background was clear, the bacilli were easily visible and the time taken for screening and reporting was shorter. The present study highlights the utility of CBNAAT from FNAC material as an adjuvant in the diagnosis of Tuberculosis lymphadenopathy.

Keywords: Acid fast bacilli, Ziehl-Neelsen

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   Cytopathology Top

Cytological spectrum of body fluids in tertiary care hospital

Aishwarya Wankhade, Rakesh T Shedge, Vikas Kavishwar, Mayura Phulpagar, Sweety Shinde, Vrushali Patil

Topiwala National Medical College and BYL Nair Hospital, Mumbai, Maharashtra, India

Introduction: Cytological evaluation of body fluids is proved as a proficient technique in present age owing to its rapid, affordable and accurate results. It proves to be of great assistance in grading fluids into non-malignant and malignant for expediating clinical diagnostic and therapeutic interventions. Objectives: Our study aims at categorizing fluids into non-malignant and malignant based on their cytology. Materials and Methods: The study includes 211 body fluids (ascitic fluid, pleural fluid, CSF) received in cytology section of Pathology Department in Nair Hospital in year 2021. The samples were centrifuged and sediment was transferred on two glass slides. One stained with May-Grunwald Geimsa stain, other with Papanicolaou stain. The stained smears were studied under microscope and evaluated. In case of clear fluids cytocentrifuge was used to prepare smears. Results: Our study shows a slight female predominance (1.1:1) with maximum cases in age range of 41-60 yrs. Pleural fluids comprised majority of cases (52.6%), followed by ascitic fluids (41.2%) and CSF (0.6%). Majority cases had non-malignant etiology (79%). Lymphocytic (72.6%, 47.3%), neutrophilic (19.0%, 40.3%), eosinophilic predominance (1.1%, 1.7%) and mixed cell infiltrate (7.1%, 10.5%) were seen in pleural and ascitic fluid respectively.20 cases each of pleural and ascitic fluids contributed to the 21% of fluids with malignant etiology. Malignant fluids were reported as atypical cells seen (42.5%), positive for adenocarcinoma (25%), suspicious for malignancy (15%), signet ring cells seen (10%), neoplastic squamous cells seen (5%) and malignant round cells seen (2.5%). Conclusion: Cytological analysis of body fluids is acknowledged as a prompt method to simplify further clinical management of patients.

Keywords: Body fluids, malignant, non-malignant

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   Cytopathology Top

Comparitive study of expanded Masood's cytologic index and other cytologic scoring systems with modified bloom-Richardson's histopathological grading of breast carcinoma and its validity

S A Arpitha, Rahesh H Chandan, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: The incidence of breast carcinoma is increasing in developing countries due to adoption of western life-style. Fine-needle aspiration cytology is the initial method to evaluate the palpable breast lesions. The neoadjuvant therapy is helpful in treating high grade rather than low grade breast carcinomas. Masood cytologic index delineates all the breast lesions into four groups. Objectives: To compare various cytological scoring systems with gold standard histopathological grading system and its validity and use in prognosis. And to see which of the cytological grading systems have better correlation with histopathological grading. Materials and Methods: This is a prospective study for 2 years (Jan 2021- Jan 2023) of all patients referred to cytology section of department of Pathology, KIMS, Hubballi. Results: A total of 180 cases were examined at the present study period. The patients age ranges from 24- 86 years old with a mean age of 52.52 years. The most common histopathological grading seen was bloom Richardson grade 2, then being grade 1 and grade 3. Expanded Masood's cytological index had a concordance level od 82% with histopathological grading ay the present study period while Robinson's cytological index had a concordance of 86%. Conclusion: It is recommended that cytological grading be included in all FNAC reports of ductal carcinoma of breast, so appropriate treatment can be started timely and over treatment of low grade tumors be avoided.

Keywords: BR grading, expanded Masood's index, IDC

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   Cytopathology Top

Fine needle aspiration cytology of head and neck lesions

Ekta Gupta, S A Bolde, Shwetha, S S Dantakale

Dr. V.M. Government Medical College, Solapur, Maharashtra, India

Introduction: Fine Needle Aspiration Cytology (FNAC) is a relatively safe, easy and inexpensive technique to accurately diagnose head and neck lesions on an outpatient basis. Lesions of head and neck are easily accessible and include inflammatory lesions, benign and malignant neoplasms. Objectives: To study the spectrum of various head and neck lesions on FNAC. To correlate the cytological findings with other investigations. Materials and Methods: This is an observational study of 100 cases of FNAC of head and neck swellings performed as an OPD procedure from April to September 2022 at a tertiary care centre. Cytomorphological diagnosis was given. Data was compiled and analysed. Results: The most common organs involved were cervical lymph nodes (44%), followed by thyroid (26%), salivary glands (5%), head and scalp (6%), post auricular nodule (2%) and other tissues (17%). The most common lesions found are Reactive lymphadenitis (22%), TB lymphadenitis (17%), colloid goitre (19%), benign cystic lesions (5%). Rare malignant tumors like Non Hodgkin lymphoma, Thyroid undifferentiated carcinoma with osteoclastic giant cells and uncommon benign lesions like benign fibrous histiocytoma and Kimura disease were diagnosed on FNAC and confirmed on histopathology. Conclusion: FNAC provides important diagnostic information which helps in providing medical and surgical treatment to patients.

Keywords: Fine needle aspiration cytology, head and neck lesions, lymphadenitis

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   Cytopathology Top

Evaluation of immunohistochemical expression of PD-L1 on cell blocks in patients of non small cell lung carcinoma (NSCLC)

J Roshny, Ujjawal Khurana, Ashwani Tandon, Tanya Sharma, Abhishek Goyal, Alkesh Kumar Khurana

All India Institute of Medical Sciences, Bhopal, Madhya Pradesh, India

Introduction: Molecular testing (EGFR, ALK, ROS and PD-L1 etc.) in lung cancer patients has a pivotal role in targeted therapy and patient management. Majority of lung carcinoma patients present in advanced stages where the feasible mode of diagnoses are cytology and small biopsies. The expression of PD-L1 has been studied and standardized on biopsies, however its utility on cell blocks has not been evaluated considerably. Objectives: To analyse the utility of cell blocks in testing PD-L1 expression. Materials and Methods: This is a prospective study encompassing cases from September 2021 to August 2022. Cell blocks were prepared from aspirates of tumor mass/ metastatic lymph node, pleural fluid and bronchoalveolar lavage using plasma thromboplastin method. PD-L1 expression was quantified in cell blocks and small biopsies using the commercially available, IHC-based, SP263. The Tumor Proportion Score (TPS) was calculated using the recommended scoring algorithm. Results: A total of 77 lung cancer patients including 69 cases of NSCLC (90.4%), 4 small cell carcinoma (5.2%) and 4 other malignancies (4.4%) were included. Thirty eight cell blocks were made comprising of 30 cases of NSCLC (79%) including 28 adenocarcinoma and 2 squamous cell carcinoma, 4 small cell carcinoma (10.5%) and 4 other malignancies (10.5%). Out of these 38 patients, small biopsies were available for 28 patients. The results of immunohistochemistry with PD-L1 antibody on cell blocks and biopsies will be presented. Conclusion: It is pertinent to standardize PD-L1 expression on cell block specimens in lung cancer patients as many a times they may be the only diagnostic tissue available.

Keywords: Adenocarcinoma, cell blocks, lung carcinoma

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   Cytopathology Top

A comparison between conventional cytology, liquid based cytology and cell block in the analysis of peritoneal fluid in gynecological malignancy

Mousumi Biswas, Pranati Pradhan, Asaranti Kar, Bhagyalaxmi Nayak, Lity Mohanty

Department of Pathology, SCB Medical College and Hospital, Cuttack, Odisha, India

Introduction: Despite being the standard procedure for the analysis of peritoneal fluid in gynecological malignancies, the conventional cytology has many pitfalls. In this study we intend to compare the diagnostic efficacies of Conventional cytology, LBC and Cell block in the evaluation of peritoneal fluid in gynaecological malignancies. Objectives: 1. Preparation of conventional cytosmear, LBC & cell block from the peritoneal fluid collected from the patients with clinical or radiological suspicion of gynecological malignancy. 2. To compare different cytological parameters among conventional cytosmear, LBC & cell block preparation. Materials and Methods: We conducted a prospective cross-sectional study on the patients having ascites due to gynecological malignancy. We assessed each sample from the patients by conventional cytology, LBC and cell block. For each diagnostic tool adequacy, background, cellularity, cell cluster, cytoplasmic details, nuclear details and presence of malignant cells were studied and compared. Results: A total of 42 patients (14 endometrial cancer, 26 ovarian cancer, 2 benign cases) were included in the study. Mean age was 54.4±8.34 years. No significant difference of the cytological criteria was observed between conventional cytosmear and LBC except the background which was clearer in LBC. The nuclear detail and the cellular architecture were better seen in cell block compared to other two methods. No significant difference was seen between the three methods in detecting malignant cells. Conclusion: LBC had no significant advantage over cytosmear and cell block in the evaluation of peritoneal fluid, but their combined usage can yield more diagnostic precision to the peritoneal fluid samples in gynaecological malignancies.

Keywords: Endometrial neoplasm, ovarian neoplasm, peritoneal fluid

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   Cytopathology Top

Utility of extracellular matrix and background material in diagnosis of salivary gland lesions in the era of Milan

Geetanjali Burbure, K Amita, M Sanjay, T Rajini

Adichunchanagiri Institute of Medical Sciences, B.G Nagara, Karnataka, India

Introduction: Salivary gland (SG) lesions are challenging at cytology. We previously introduced pattern based analysis for diagnosing salivary gland lesions with high efficacy. The primary aim was to determine the utility of background elements, which includes different types of extracellular matrix and background material, in distinguishing various salivary gland lesions. Objectives: To determine the utility of background elements, which includes different types of extracellular matrix and background material, in distinguishing various salivary gland lesions. Materials and Methods: Retrospective diagnostic analytical study involving SG lesions, aspirated over a duration of five years. All the cases were reviewed and classified based on the predominant background extracellular matrix and background material. The cellular details were then taken into consideration and a final cytological diagnosis was rendered. All the cases were classified into six groups as follows group I- absence of extracellular matrix (clean background), group II- chondromyxoid fibrillary stromal matrix, group III-mucin, group IV-hyaline basement membrane material, group V- proteinaceous material and group VI-necrotic/dirty background. Results: Total of 233 FNAC cases were included. Histopathology correlation was obtained in 118 cases. False positives were two and false negatives were seven. Concordance was noted in 102 cases and discordant diagnosis in 16 cases. Sensitivity, specificity and diagnostic accuracy was 79.4%, 97.6% and 92.3% respectively. Conclusion: The high efficacy of FNAC, obtained by classifying lesions based on background elements like different types of extracellular matrix material and other background material, establishes the utility of this method in reporting SG lesions.

Keywords: Background, extracellular matrix, salivary gland lesions

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   Cytopathology Top

A prospective evaluation of thyroid lesions by fine needle aspiration cytology according to the Bethesda system for reporting thyroid cytopathology and its histopathological correlation

Himachal Mishra, Manika Alexander

Gadag Institute of Medical Sciences, Gadag, Karnataka, India

Introduction: Fine needle aspiration cytology (FNAC) is regarded as a gold standard diagnostic procedure in the assessment of thyroid lesions. Objectives: This study was carried out to evaluate thyroid lesions by FNAC based on the Bethesda system for reporting thyroid cytopathology and to correlate the cytological findings with histopathology. Materials and Methods: A prospective study was done on 203 patients with thyroid lesions based on the Bethesda system for reporting thyroid cytopathology between January 2021 to June 2022 and results were compared with histopathology wherever possible. Sample size was calculated and purposive sampling done. Data analysis was done in SPSS software. Results: Based on the Bethesda system for reporting thyroid cytopathology, thyroid lesions were classified. Out of 203 samples evaluated, 11 were categorised as category I, 171 cases were categorised as category II, 4 cases were categorised under Category III. 7 cases were categorised under category IV, 2 cases categorised as category V and 8 cases were categorised under VI. Out of 203 patients evaluated, only 32 patients underwent surgery. Out of 32 patients, 26 (81.25%) were confirmed as benign and 6 (18.75%) were found to be malignant on histopathology. Sensitivity, specificity and diagnostic accuracy are 83.33%, 96.15% and 96.78% respectively. Negative predictive value and positive predictive value were found to be 96.29% and 100% respectively. Conclusion: The study shows that thyroid lesions can be effectively categorised and reported by the Bethesda system for reporting thyroid cytopathology and hence it represents a reliable system.

Keywords: Bethesda, cytology, thyroid

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   Cytopathology Top

A comparison study of sonographic features and fine needle aspiration cytology (FNAC) with histopathology findings in the diagnosis of solitary thyroid nodule

Rajesh Meda, S Harsha Vardhan, B H Poorna Chandra Sekhar, G Baleswari, A Venkatalakshmi

Department of Pathology, Sri Venkateswara Medical College, Tirupathi, Andhra Pradesh, India

Introduction: Thyroid nodules can be a manifestation of a diversity of diseases affecting the thyroid gland. Ultrasonography (USG) is the first line of investigation in evaluation of euthyroid nodules. Thyroid nodules classified based on Thyroid imaging reporting and data system (TIRADS) - USG based risk stratification system, Subjects with high risk category of TIRADS undergo fine needle aspiration cytology (FNAC). Thyroid Bethesda system is used to report FNAC samples and risk of malignancy is determined. Objectives: To compare USG features according to TIRADS and FNAC (Bethesda classification) with Histopathology of excised samples of thyroid nodules. Materials and Methods: It is observational study carried out in tertiary care hospital, SVRRGGH Tirupathi. 60 patients from January – 2022 to June-2022 with thyroid nodules taken in this study. Subjects underwent - TIRADS, FNAC and proceeded to definitive surgery and histopathology report obtained. Comparison is done between TIRADS, FNAC with histopathology. Surgical biopsy report was used as gold standard for diagnosis. Results: Study population was predominantly female, age ranging from 17-70 years sensitivity, specificity, positive predictive value, negative predictive value, for FNAC is 82%, 90%, 86%, 86% respectively and for TIRADS is 82%, 49%, 52.%, 78% respectively. FNAC classification was equally sensitive and more specific than TIRADS. Among USG parameters micro calcifications was most sensitive and specific, followed by irregular margin of taller from wider shape. Four patients with benign cytology and suspicious USG features had malignancy in Histopathology report. Conclusion: USG & FNAC are equally sensitive in distinguishing malignant thyroid nodule but FNAC is more specific. FNAC is minimal invasive procedure it is used to distinguish benign from malignant with a high degree of accuracy. In patients with high risk features on USG, benign cytology, repeat FNAC and confirmation should be done by histopathology of surgically restricted specimen.

Keywords: Bethesda, fine needle aspiration cytology, thyroid nodules, thyroid imaging reporting and data system, ultrasonography

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   Cytopathology Top

Evaluation of The International System (TIS) of reporting serous fluid cytopathology using cell block and immunohistochemistry as gold standard in a tertiary health care center

Ayan Sarkar, Soumit Dey, Pranamita Ray, Mir Hassan, Ishita Ghatak, Sunita Ghosh

Ramakrishna Mission Seva Pratishthan Vivekananda Institute of Medical Sciences, Kolkata, West Bengal, India

Introduction: Serous effusion is collection of fluid found in the serous cavities of the body. It has the potential to detect the underline causes that may be neoplastic or non-neoplastic. In 2019 The International Academy of Cytology (IAC) and American Society of Cytopathology (ASC) sponsored a workforce of experts to create a standardized system for serous effusion cytology reporting named as “International System for Reporting Serous Fluid Cytology” (TIS). Objectives: 1. To evaluate use of TIS in serous fluid cytology. 2. Compare TIS with conventional smears, taking cell block as “Gold Standard”. 3. Immunohistochemistry staining was done using the markers in case of ambiguity. Materials and Methods: Effusion fluid sample was received, divided into two equal parts, one for conventional smear (CS) and other for cell block (CB). CS stained with MG-G and PAP, CB was stained with H&E and reporting as per TIS. Immunohistochemistry staining was done. Results: Out of 180 specimens 166 were adequate, 133 were Negative for malignancy, 11 were with atypical cells, 12 were suspicious for malignancy and 10 were malignant. IHC was done on 60 cell blocks and 54 were corroborative with cell blocks diagnosis and 6 were non-corroborative. Sensitivity of TIS was found 93.1% and specificity was 97.08%. PPV was 87.09% and NPV was 98.51%. Conclusion: After extensive searching the current literature, 18 similar studies were found, 6 studies corroborated with this study.12 studies were different due to smaller sample size. Larger sample size requires to arrive at convincing conclusion.

Keywords: Cell block study, serous effusion, The International System

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   Cytopathology Top

Cytopathological spectrum of pancreatic lesions

Parul Verma, Aminder Singh, Pavneet Kaur, Harpreet Kaur

Dayanand Medical College and Hospital, Ludhiana, Punjab, India

Introduction: Pancreas is a retroperitoneal structure and imaging techniques are often required to guide the biopsy needle to sample the target area. Although clinical presentation, radiological findings and location of pancreatic mass is helpful but fine needle aspiration and tissue diagnosis remains the gold standard for final diagnosis and to plan the management. Objectives: To categories the lesions according to Papanicolaou Society of Cytopathology system for reporting Pancreaticobiliary Cytology. Materials and Methods: This was both a retrospective and prospective study of two and half years in which 70 cases from pancreas were evaluated. Results: A total of 2265 cases of guided FNAC were received in the Department of Pathology of Dayanand Medical College and Hospital, Ludhiana over a period of 2 years and 6 months, out of which 70 were from pancreas. The majority of the patients were grouped under Positive or Malignant Category comprising 57.14% [n=40], followed by Suspicious (for malignancy) which were 14.28% [n=10]. Only 2 cases [2.85%] were reported as Non diagnostic. The commonest age group who underwent fine needle aspiration cytology of pancreatic lesions was 51-60 years [n=28, 40%] followed by age group of 61 to 70 years [n=17, 24.28%]. Conclusion: In our study the youngest patient was 10 years old while the oldest patient was 88 years old with mean age was 60.14 years. Most common location of lesion was head of pancreas and majority cases were diagnosed as Positive or Malignancy.

Keywords: Pancreatic cytopathology

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   Cytopathology Top

Utility and efficacy of rapid on-site evaluation of image-guided fine needle aspiration cytology from hepatobiliary tract lesions

Rohan Das, Evarisalin Marbaniang, Vandana Raphael, Donboklang Lynser

NEIGRIHMS, Shillong, Meghalaya, India

Introduction: Fine needle aspiration cytology (FNAC) is broadly accepted method for diagnostic evaluation of pathological lesions. Studies have shown with application of Rapid onsite evaluation (ROSE) of smears, there was statistically significant increase in diagnostic yield of image-guided FNACs. There are limited studies on ROSE of image-guided FNACs from biliary tract lesions in our country. Objectives: To determine the utility and efficacy of ROSE on FNAC from neoplastic and non-neoplastic lesions of Hepatobiliary tract. Materials and Methods: A prospective study was conducted wherein ROSE was performed for FNACs from 50 patients with hepatobiliary tract lesions. Samples were stained with toluidene blue for checking adequacy with a proposed adequacy criteria. Cytological slides were later analysed after staining with May grunwald giemsa and Papanicolaou stains. Results were compared with those obtained from FNAC from previous years without onsite evaluation. Results: During 2006-2017, of the 330 image-guided FNACs (without ROSE) from hepatobiliary lesions, 47 were non-diagnostic whereas during our study (2021-2022), of the 50 image-guided FNACs (with ROSE) from hepatobiliary lesions, 2 were non-diagnostic. An increase in diagnostic yield of FNAC from 85.76% to 96 % was noted (p= 0.044). Conclusion: ROSE with toluidine blue staining significantly increases diagnostic yield of hepatobiliary FNACs and abolishes need of repeat procedure.

Keywords: Fine needle aspiration cytology, hepatobiliary, rapid onsite evaluation

   AOP85 Top

   Cytopathology Top

An assessment of ovarian lesions by intraoperative pathological consultation

Nikhil Choudhary, Jaya Mishra, Bifica Sofia Lyngdoh, Caleb Harris, Wansalan Karu Shullai

NEIGRIHMS, Shillong, Meghalaya, India

Introduction: Intraoperative frozen section diagnosis of ovarian tumors is widely used in making diagnostic distinction and to decide course of surgery. In centers where frozen section facilities are also not available a simple and cost effective intraoperative FNAC or squash cytological diagnosis would help surgeon to decide whether to go for radical surgery and on further management. Objectives: To compare diagnostic accuracy of intraoperative consultation in ovarian lesions. Materials and Methods: A prospective study was conducted wherein FNAC, Imprint and Squash cytology was performed form 31 ovarian lesions. The samples were stained with rapid H&E and Giemsa. Cytology diagnosis was compared with HPE diagnosis. Results: During our study (2021-2022) from 31 ovarian samples, Diagnostic accuracy from FNAC, Imprint and Squash are 77.4%, 77.4% and 83.8% respectively, and sensitivity, specificity, PPV and NPV from FNAC, Imprint and Squash of malignant tumors were found to be more significant as compared to benign and borderline tumors. Conclusion: Intraoperative cytology is a relatively good method to diagnose nature of ovarian lesion if frozen section is not available.

Keywords: Accuracy, intraoperative, ovary

   AOP86 Top

   Cytopathology Top

Application of the proposed Sydney system for reporting lymphnode cytology with histopathological correlation

Devidutta Rout, Subhasish Mishra, Asaranti Kar, Lity Mohanty

Department of Pathology, SCB Medical College, Cuttack, Odisha, India

Introduction: Fine needle aspiration cytology is regarded as the initial and most commonly indicated procedure for lymphadenopathy of unknown etiology. There exists a lack of uniform reporting system. The Sydney system provides categorization of lymphnode aspirate diagnoses into 5 categories resulting in better communication between the cytopathologist and the clinician. Objectives: To categorize lymph node aspirate diagnoses according to The Proposed Sydney system into 5 categories and calculate the risk of malignancy (ROM) of each category. Materials and Methods: FNAC was done in patients with clinically detected lympadenopathies. One airdried smear and two fixed smears with ethanol were prepared. Airdried smear was stained with diff quik and fixed smears with H&E, Papanicoulau Stain. The cytological diagnosis was correlated with histopathological diagnosis wherever required. Results: 112 cases were studied, out of which 6 (5.35%) were categorized as inadequate (L1), 49 (43.75%) as benign (L2), 5 (4.4%) as atypical cells of undetermined significance (L3), 5 (4.4%) as suspicious for malignancy (L4) and 47 (41.9 %) as malignant. Conclusion: Lymph node Cytology as reported by Sydney system has high diagnostic accuracy and will help in achieving uniformity and ultimately better patient management.

Keywords: Lymphnode, malignancy, metastatic

   AOP87 Top

   Cytopathology Top

Cytological diagnosis of secondary malignancies involving thyroid – A retrospective study

V Chaitra, S Balu, D Priya, Soumya

Department of Oncopathology, Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Involvement of thyroid gland from either metastasis or by direct extension from adjacent organ malignancies are rare and can pose a diagnostic challenge. Objectives: To study the utility of thyroid FNA in diagnosing secondary thyroid malignancies. Materials and Methods: All thyroid FNAC diagnosed as metastasis/ infiltration from adjacent organ malignancies during the period of January 2019 to August 2022 were analyzed retrospectively. Results: During the study period a total of 990 thyroid FNACs were performed. 648 (65%) were non neoplastic, 311 (31%) - primary thyroid neoplasm. This study included 31 cases of secondary thyroid neoplasms consisting of 16 cases of metastasis (1.6%), 9 infiltrations by adjacent structures (upper aerodigestive tracts, 0.9%) and 6 malignant lymphomas (0.6%). The age ranged from 40-80 years. Male: female-1.07:1.0. Among metastasis, 7 cases were Squamous cell carcinoma from head and neck region, esophagus and one case from unknown primary. 6 were adenocarcinoma (1 case each from GE junction, stomach, endometrium, ovary, lung, gastrointestinal tract). Sonographic characteristics were diverse. Cytological diagnosis of secondary malignancy was offered in correlation with clinical and radiological findings. All the patients received chemotherapy and adjuvant radiotherapy. Most patients were lost for follow-up and one succumbed to death within 2 months of diagnosis. Conclusion: Although secondary thyroid malignancies are infrequent, needs to be kept in differential diagnosis in cases with unusual morphology. Correlation with clinical and imaging findings are essential to arrive at final diagnosis and to differentiate from primary as management protocols differs for both.

Keywords: Metastasis, secondary infiltration, thyroid cytology

   AOP88 Top

   Cytopathology Top

Morphological markers of chromosomal instability: As a predictor of malignancy in pleural effusions

Phiza Aggarwal, Uma Handa, Ranjeev Bhagat, Deepak Aggarwal

Department of Pathology, Government Medical College and Hospital, Sector 32, Chandigarh, India

Introduction: Malignancy in pleural effusions is the indication of poor prognosis and is of paramount importance for the patient and the attending physician. The distinction between malignant cells and reactive mesothelial cells in effusion cytology is difficult and requires ancillary techniques. Chromosome instability (CI) plays a major role in carcinogenesis. The presence of micronuclei, chromatin bridging, nuclear budding and multipolar mitosis on morphology are the indicators of CI that may help to distinguish benign from malignant effusions. Objectives: To evaluate the frequency of 4 morphological markers of CI in pleural effusions and to correlate these with the cytological diagnosis. Materials and Methods: This was a prospective observational study done in a tertiary care hospital. A total of 55 pleural fluid samples were received and analysed in patients with confirmed underlying malignancy. Four morphological markers of CI were counted in the MGG stained smears and the score was correlated with the conventional cyto-morphological diagnosis. Results: Out of 55 cases, there were 42 malignant and 13 benign effusions on cytological examination. The mean number of micronuclei, nuclear budding, chromatin bridging and multipolar mitosis in malignant effusions were 7.26 ±3.500, 9.55 ±5.531, 1.83±1.167 and 2.21 ±1.616 respectively that was significantly higher than the benign effusions (1±0.71, 1.1±0.86, 0.38±0.50 and 0.15±0.37 respectively) (p<0.05). Conclusion: The evaluation of CI markers is a simple, inexpensive method that can be done on routine MGG smears in suspected cases of metastatic effusions.

Keywords: Chromosome instability, effusions, pleural

   AOP89 Top

   Cytopathology Top

Epidemiological profile of thyroid nodules by FNAC in the Pathankot cantonment population of Punjab

Kunal Bagul, V Dahake, N Khandelwal, L Prasad, V Tewari, V Srivastava, K Rajmohan

Indian Army, 167 Military Hospital, Pathankot, Punjab, India

Introduction: Thyroid is the most commonly affected endocrine gland for endocrine disorders in India. Here we present study of thyroid nodule through fine aspiration cytopathology (FNAC) and its profile according to the Bethesda system for reporting of thyroid cytopathology (BSRTC). Objectives: This study is done to generate epidemiological data regarding prevalence of thyroid disorders in this military hospital which cover, health care of cantonment area of Pathankot, Punjab of India from December 2020 to August 2022. Materials and Methods: This study comprised of 51 cases of thyroid swelling irrespective of their age and sex. FNAC was done using 23 gauge needles. Smears were prepared and stained with May Grunwald-Giemsa (MGG) Stain as we as Papanicolaou (PAP) stain. Then cases were classified according to various categories as per BSRTC. Results: The commonest affected age group in this study was 31-40 years. Out of total cases seven were males and 44 were females. The youngest patient was 13 year old and oldest one was 80 years old. Benign category constitutes most of the cases total 41 out of which benign follicular nodule constitute 35 cases. Lymphocytic thyroiditis is the second most common thyroid disorder among benign category and it constitutes 11 cases. Suspicious for malignancy and malignancy categories include two cases each. Conclusion: In this study there is high prevalence of benign follicular nodule among all thyroid disorders. Other thyroid disorders like lymphocytic thyroiditis should be given equal importance because of its increasing prevalence.

Keywords: Cytopathology, Goiter, thyroid

   AOP90 Top

   Cytopathology Top

How useful are cervical smear in detecting endometrial carcinoma? A tertiary care centre experience

BrijMohan Singh, Varun Kumar Singh

Kasturba Medical College, Manipal, Karnataka, India

Introduction: The conventional Pap smear is a screening test for cervical cancer with a proven efficacy. The reporting of benign/malignant endometrial cells on Pap smear takes a back seat in this cervical cancer focused strategy. Objectives: To review Pap smears of biopsy proven endometrial adenocarcinoma for the detection of normal/atypical endometrial cells. Materials and Methods: A 2.5-year retrospective study was carried out from the archives of histopathology for all diagnosed endometrial carcinomas who underwent cervical Pap smear test within 6 months before histological diagnosis. The demographic details, Pap smear report and other histological parameters were noted. Results: A total of 24 cases were included. The age ranged from 44-75 years (median-59 years). Cervical cytology review was normal in 11 cases. The review picked up atypical cells in eight (08) cases and at least one endometrial cell cluster in 10 cases, none of which were reported in the initial smear impression. 13 cases had either atypical cell or endometrial cells, which were missed in the initial impression. Atypical cells were seen in 4/10 (40%) of FIGO grade 1, 2/7 (28%) of grade 2 and 2/5 (40%) of grade 3 tumours. Endometrial cells were reported in 3/10 (30%) of FIGO grade 1, 5/7 (71%) of grade 2 and 1/5 (20%) of grade 3 tumours. The major features limiting diagnostic accuracy of Pap smear were reactive changes associated with atrophy (n=5), inflammation (n=5), obscuring haemorrhage (n=2) and scant cellularity of atypical cells (n=4). Conclusion: All women with atypical/benign endometrial cells on Pap smear need endometrial sampling irrespective of age/menstrual status.

Keywords: Endometrial cancer, nuclear grade, Pap smear

   AOP91 Top

   Dermatopathology Top

Association of human leukocyte antigen class I and class II genotyping in autoimmune vesiculobullous lesion of skin

Shalini Rawat, Amit K Yadav

VMMC, New Delhi, India

Introduction: Autoimmune vesiculobullous diseases of the skin are characterized by autoantibodies against different intra-/extracellular structures within the epidermis and at the basement membrane zone. The human leukocyte antigen (HLA) also known as major histocompatibility complex, are involved in antigen processing and presentation, and usually show highly significant association with autoimmune diseases. Objectives: To study the association of HLA class I and class II alleles with cases of Autoimmune vesiculobullous lesion of the skin. Materials and Methods: A case control observational study comprising of 37 clinically diagnosed cases of AIBD was done. the diagnosis was confirmed by punch biopsy followed by histopathological and direct immunofluorescence examination. Furthermore, HLA genotyping study was done on these cases by PCR-SSOP method. Results: The patients ranged from 3-80 years of age with female preponderance (67.6%). Among 37 cases, pemphigus vulgaris (48.6%) was the most common followed by bullous pemphigoid (18.9%), pemphigus foliaceus (13.5%) and dermatitis herpetiformis (10.8%). The HLA genotyping study showed that among HLA class I, HLA-A*03, A*29 and A*31; and B*52 and among HLA class II, HLA-DR*03, DR*11, DR*14 and DR*15; and DRB5* showed stronger association with various AIBD cases. Conclusion: The present study showed association between specific HLA class I and class II alleles and AIBDs in a Noth Indian population.

Keywords: AIBDs, human leukocyte antigen genotyping, North Indian population

   AOP92 Top

   Dermatopathology Top

Comparative expression of Melan A and S100 in active and stable vitiligo lesions and correlation with CD4 and CD8 T cells

Sai Jyoti, Minakshi Bhardwaj, Kabir Sardana

ABVIMS and Dr RML Hospital, New Delhi, India

Introduction: Vitiligo is a chronic pigmentary disorder which is characterized by the onset of depigmented spots on various body areas. It is important to assess the disease status (active/stable) to guide therapeutic decisions. Objectives: Comparison of expression of Melan A and S100 in active and stable vitiligo lesions and correlation with CD4 and CD8 T cells. Materials and Methods: Skin biopsies from 45 clinically diagnosed vitiligo patients (25 active and 20 stable) were stained and studied using hematoxylin and eosin, Melan A, S 100, CD4 and CD8. All the sections were evaluated for histopathological findings and expression of the immunohistochemical antibodies. Results: Basal cell layer vacuolization and dermal melanophages were more common in active vitiligo (p =0.003). S100 and CD8 expression was increased in active cases while CD4 and CD4/CD8 ratio was more in stable cases but with no statistically significant p value. A moderate positive correlation between MELAN A with CD8 (p = 0.019) and S100 with CD4 (p = 0.013) in active vitiligo was noted. Conclusion: The present study shows that center and margins of active lesions of vitiligo show increased disease activity which is characterised by decreased expression of MELAN A, CD4 and an increased expression of S100, CD8 with concomitant increased basal cell layer vacuolization and dermal melanophages. However, the role of CD4 in pathogenesis of vitiligo needs further studies on larger cohorts.

Keywords: Active, stable, vitiligo

   AOP93 Top

   Dermatopathology Top

Mast cell profile in leprosy skin lesions

H G Sowjanya, Vijaya Mysorekar

M S Ramaiah Medical College, Bengaluru, Karnataka, India

Introduction: The disease pattern in leprosy is determined by the level of host response to Mycobacterium leprae. The role of mast cells in development of delayed hypersensitivity reaction has been studied in the last few years which has led to linking their role in development of skin lesions in leprosy. Objectives: To study mast cell profile (density, morphology and distribution) in indeterminate type and spectrum of leprosy skin lesions. Materials and Methods: A study was conducted on 80 untreated leprosy cases and 10 healthy individuals taken as control. In each case 4 mm punch biopsy from the skin lesion was taken, fixed in 10% buffered formalin, processed using routine technique, 4 microns sections cut and stained with H&E, Fite-Faraco stain for acid fast bacilli, and 1% aqueous toluidine blue for mast cells. Correlation of mast cell density of each type of leprosy skin lesion with the control was done. Similarly, correlation of mast cell profile between tuberculoid and lepromatous type was studied. Results: Mast cell density in indeterminate leprosy was significantly higher (p<0.01). Mast cell density in tuberculoid type was significantly lower than the lepromatous type (p<0.001). The lepromatous group showed altered morphology and increased mast cell degranulation. However, the distribution of mast cells in tuberculoid and lepromatous types were similar. Conclusion: Indeterminate leprosy cases show increased mast cell density, indicating mast cell response in the early stage of evolution of leprosy. Thus, in indeterminate leprosy lesions, mast cell density may be of diagnostic value.

Keywords: Mast cell density, toluidine blue

   AOP94 Top

   Dermatopathology Top

Prevalence of peripheral blood eosinophilia in bullous pemphigoid and it's correlation with age

Monika Gamit, Mahomed Hafeezulla Shariff

Yenepoya Medical College and Hospital, Deralakatte, Karnataka, India

Introduction: Bullous pemphigoid is most common autoimmune vesico-bullous disorder characterized by subepidermal blistering of the skin. It mainly affects the elderly age group. Peripheral blood eosinophilia is the common finding in bullous pemphigoid. Objectives: (a) To study the prevalence of peripheral blood eosinophilia in patients with bullous pemphigoid. (b) To demonstrate the statistical correlation between peripheral blood eosinophilia and age of the patient. Materials and Methods: This retrospective study was carried out in department of pathology at Yenepoya medical college and hospital, Mangalore over a study period of January-2016 to July-2022. Histopathology proven cases of Bullous pemphigoid were included. Clinical details and relevant data for the study was collected from the hospital's electronic data system. Results: In total, 46 cases of bullous pemphigoid were analysed. Mean± SD age at presentation was 56.5±17.5 years, 29 (63%) were females & 17 (37%) were males. Overall, 34 (74%) patients had pathological eosinophilia with Mean±SD (13.3±8.7) circulating eosinophils. A significant correlation found between peripheral blood eosinophilia and age of the patient. Patients with eosinophilia were significantly older at presentation with mean ±SD age of 66.95±13 (P-value= <0.001) compare to patient with normal eosinophil count with Mean± SD age of 46.2±2 (P-value =<0.001). Conclusion: Patients with peripheral blood eosinophilia were significantly older at presentation. Patients with a normal eosinophil count were younger at presentation.

Keywords: Bullous pemphigoid, eosinophilia, older age

   AOP95 Top

   Dermatopathology Top

Comparative study of histopathologic features of vulvar lichen sclerosus versus lichen planus

Sueallen DSouza, Gayatri Ravikumar, Rajalakshmi Tirumalae, Meryl Antony

ST Johns Medical College, Bengaluru, Karnataka, India

Introduction: Vulvar lichenoid dermatoses pose diagnostic challenges due to overlapping clinico-pathological features. Existing criteria for diagnosis and distinction between Lichen sclerosus (LS) and Lichen planus (LP) needs validation to understand similarities or differences that can confound diagnosis. Objectives: To study the differences in histopathological features between vulvar LS and LP. Materials and Methods: A 10-year retrospective study of biopsy proven vulvar LS and LP with relevant clinical and detailed histopathological analysis. Results: There were 28 cases of LP and 71 cases of LS. Frequently, LS presented as hypopigmented patches, while LP presented as ulcers/erosion. Epidermal findings like wedge-shaped hypergranulosis, Civatte bodies, basal vacuolation were significantly higher in LP whereas LS showed atrophic epidermis with hypogranulosis. The distinguishing features of: (i) LS with lichenoid inflammation (LS-L) versus LP with lichenoid inflammation (LP-L), (ii) Conventional LS (LS-C) versus LS with unconventional features (LS- UC) (iii) LS-UC versus LP were analyzed. The significant histological features differentiating LS-L (n=15) from LP-L (n=24) were hypogranulosis, melanin loss and papillary dermal homogenisation while melanophage clustering and dense inflammation were exclusive in LP-L. LS-UC (n=31) showed parakeratosis, thickned granular layer with normal or acanthotic epidermis in contrast to LS-C which had basal vacuolation and flattened rete. LS-UC (n=31) had significant basement membrane thickening, interstitial inflammation and dermal homogenisation, while LP (n=28) showed extensive basal vacuolar alternation, necrotic keratinocytes, saw-tooth rete pegs and lacked dermal homogenisation. Conclusion: LS is the most common vulvar lichenoid dermatosis. The unconventional variants may mimic LP and subtle histological clues help to arrive at the correct diagnosis.

Keywords: Vulvar lichen planus, Vulvar lichen sclerosus

   AOP96 Top

   Dermatopathology Top

A twelve years study on histoid leprosy in post elimination era

C Prabhashree, Bhavikatti Bharati Mohan

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: India achieved the goal of elimination of leprosy as a public health problem by 2005. Histoid leprosy (HL) is a rare variant of lepromatous leprosy, presenting clinically as localized crops of shiny nodules of different sizes, mimicking many other dermatoses and can be missed clinically. Objectives: 1) To study various clinical and histopathological parameters of HL. 2) To study and correlate between clinical and histopathological diagnosis of HL. Materials and Methods: This is a twelve year (2010-2022) study on HL, undertaken in the Department of Pathology, Karnataka Institute of Medical Sciences, Hubballi. Results: In this study we had 18 cases of HL (2.1% of leprosy cases) with 8:1 as male to female ratio. Maximum number of cases (66.6%) were in age group of 21–30 year. Nodules (100%) were the commonest skin lesion observed with earlobe infiltration in 3 cases and lepra reaction in 1 case. Histopathology showed epidermal atrophy, grenz zone and well-circumscribed dermal areas packed with spindle-shaped to polygonal histiocytes in all 18 cases, with mean bacterial index as 5.52. 17 of 18 cases (94.44%) were de novo cases of HL. 12 (66.6%) cases showed correlation between clinical and histopathological diagnosis for HL, remaining had sarcoidosis, dermatofibroma and other variants of leprosy as clinical diagnosis. Conclusion: HL is a rare entity, well-circumscribed collection of cells may show storiform pattern mimicking dermatofibroma making differentiation important. As such cases are highly bacilliferous and probably harbouring drug-resistant mutant strains, they pose a serious threat to national elimination programs.

Keywords: Dermatofibroma, histoid leprosy, skin nodules

   AOP97 Top

   Dermatopathology Top

Role of C4D immunohistochemical marker in the diagnosis of bullous pemphigoid

Raaga Sneha, B Vijaya

Department of Pathology, JSSAHER, Mysuru, Karnataka, India

Introduction: Bullous pemphigoid (BP) is diagnosed by clinical findings, histopathological study and Direct Immunoflourescence (DIF). Not all centers will have the facilities to conduct DIF for accurate diagnosis as it is expensive. This study is being done to assess the diagnostic ability of C4d Immunohistochemical (IHC) marker that highlights the deposition of complement along the dermoepidermal junction in BP. Objectives: Primary: To determine the diagnostic ability of C4d marker in cases of BP. Secondary: To correlate the expression of C4d marker with DIF. Materials and Methods: The present study has been undertaken in the Department of Pathology, JSSH, Mysore. A sample size of 35 cases was calculated. Fifty-nine cases of BP that were subjected to DIF were retrieved from the archives of Dept of pathology for a period of 8 yrs (from 2015-2022). The formalin fixed paraffin embedded blocks of perilesional skin in these cases were subjected to C4d marker and findings were correlated with DIF to calculate sensitivity, specificity and predictive values. Results: Out of 59 selected cases, 57 cases were DIF positive and 2 cases DIF negative. All cases were subjected to C4d staining, of which 47 stained positive. C4d deposits were seen as linear brown stain along the dermoepidermal junction (13), around the basal keratinocytes (8), combined linear staining and around basal keratinocytes (22), discontinuous linear staining (3) and focally around basal keratinocytes (1). These findings were correlated with DIF and sensitivity, specificity, positive predictive value and negative predictive values of 82.45%, 100%, 100% and 16.66% respectively were calculated. Conclusion: C4d is a reliable diagnostic IHC marker for bullous pemphigoid in the absence of DIF.

Keywords: Bullous pemphigoid, C4d immunohistochemical (IHC) marker

   AOP98 Top

   Dermatopathology Top

Granulomatous lesions – A diagnostic challenge to dermatopathologists

M K Lavanya, Sunita Vernekar, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Cutaneous granulomatous lesions are very commonly encountered by pathologist and dermatologists. Granulomatous dermatoses can be seen in infectious as well as non-infectious conditions leading to chronic inflammation. Granulomatous skin lesions often present as a diagnostic challenge to dermatopathologists as a single identical histopathological pattern can be produced by several etiologies and contrariwise, a single etiology may produce diverse histopathological patterns. Objectives: To determine the pattern of different granulomatous skin lesions and the clinico-histopathological correlation of the lesions to arrive at correct etiologic diagnosis. Materials and Methods: This is a 6 year retrospective study on 480 cases of clinically provisionally diagnosed granulomatous skin lesion biopsies. Results: Majority of cases were due to infectious etiology 458 cases (95.41%). Leprosy was commonest (90.2%) followed by cutaneous Tuberculosis (3.54%). Rest of the cases included various fungal and other lesions. There was male predominance. Conclusion: Histopathology is still a gold standard tool in classification of granulomatous lesions and with support of special stains we can arrive at exact etiology. A definitive diagnosis made by demonstration of the etiological agent is essential, which will bear an impact on patient management and outcome.

Keywords: Cutaneous tuberculosis, granulomatous dermatoses, leprosy

   AOP99 Top

   Dermatopathology Top

A clinico-histopathological study of non-neoplastic nodules involving the extremities

S Sreelakshmi, Savithri Ravindra

Kempegowda Institute of Medical Sciences, Bengaluru, Karnataka, India

Introduction: Skin is a complex organ which is composed of epidermis, dermis and subcutis. In a tropical and developing country like India inflammatory dermatoses are more prevalent than tumours, many of them present as nodules. Objectives: To study the histopathological spectrum of non-neoplastic nodular lesions in the extremities. To correlate with the clinical presentation and demographic data. Materials and Methods: This is a prospective study conducted in the department of pathology, in a tertiary care hospital, from January 2021 to June 2022. Included all skin biopsy specimens from nodular lesions involving upper and lower limb, excluded tumours and cases without proper clinical details. All H and E-stained slides were evaluated and correlated with clinical details. Results: This study consists of 55 cases. Male to female ratio was 1.12:1, with age range from 16-75. The lesions were predominantly on the lower limb (n=36; 65.5%) compared to upper limb (n=19;34.5%). Prurigo nodularis was the commonest lesion (n=31; 56.4%) followed by hypertrophic lichen planus (n=13; 23.6%). On correlating with the clinical diagnosis, concordance was observed in the 46 cases (83.6%) and 9 cases (16.4%) showed discordance. 5 cases clinically thought as prurigo nodularis and 4 were hypertrophic lichen planus on histopathology. Further work up and follow up were suggested for all the cases showing discordance. Conclusion: Non neoplastic nodular lesions have many overlapping features hence a good clinical and histological correlation is needed for the definitive diagnosis.

Keywords: Extremities, non-neoplastic nodules, skin nodules

   AOP100 Top

   Dermatopathology Top

Immunohistochemical study of melanocyte stem cells in vitiligo

Adil, Khan, Amit K Yadav, Niti Khunger

Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India

Introduction: Vitiligo is an acquired pigmentary disorder of unknown etiology characterized by marked reduction of the pigment producing cells. Re-pigmentation in vitiligo depends on the viable melanocyte stem cells (MelSCs) in lesional and perilesional skin. Very few studies have been conducted earlier on human vitiliginous skin to evaluate the presence and distribution of MelSCs. This study uses immunohistochemical markers to assess the MelSCs in vitiliginous skin and the findings can have the potential to aid in designing newer strategies for vitiligo therapy. Objectives: (1) To evaluate the expression of CD34 and c-kit in melanocyte stem cells and compare the expression between lesional and perilesional skin in patients of vitiligo. (2) To correlate the expression of these markers with vitiligo histological score. Materials and Methods: 33 Punch biopsies of lesions, clinically diagnosed with vitiligo, were sent for histopathological examination. It was formalin fixed, paraffin processed. Sections of 3-4 micron were taken and stained with H and E stain for vitiligo histological scoring and immunohistochemistry was performed for MelSCs evaluation. Results: Vitiligo histological score was significantly higher in lesional skin as compared to perilesional skin (p < .0001) suggesting an increased disease activity in lesional skin. However, we were unable to observe MelSCs due to decreased expression of CD34 and c-kit in lesional and perilesional skin. Conclusion: Vitiligo histological score was significantly higher in lesional skin as compared to perilesional skin (p < .0001) suggesting an increased disease activity in lesional skin. However, we were unable to observe MelSCs due to decreased expression of CD34 and c-kit in lesional and perilesional skin.

Keywords: Melanocyte stem cells

   AOP101 Top

   Dermatopathology Top

Proliferating pillar tumor with malignant transformation

Mohsin Patel, R Thiripurasundari

VVMMC and H, Karaikal, Puducherry, India

Introduction: PTT benign tumor originating from the outer root sheath of a hair follicle.[1] considered biologically benign, PTT may be locally aggressive. In rare instances, malignant transformation has been reported, evidenced by regional or distant metastases. Malignant transformation in case of PTT is very rare and unusual finding. It is usually confused with squamous cell carcinoma both sharing many common features. So the identification of malignant PTT is very essential. It is usually a solitary lesion and most commonly occurs in elderly women. First described by Wilson-Jones. It occurs most commonly on the scalp in women older than 42 years.[9]. Objectives: 42 year female presented with scalp- occipital region cyst (?dermoid) type Painless Swelling (cyst) since 1 year with recently rapid enlargement swelling is firm in consistency; measuring 2x2 cm No other h/o of neck node lymphadenopathy. Materials and Methods: Gross examination: 2X2 cm grey brown scalp biopsy on cut section: Grey brown firm in consistency. Microscopic examination: Tissue fragments with neoplasm composed of squamous epithelium with trichilemmal type keratinisation. Tissue showed cellular tumor comprised of lobules of squamous cells. Center of lobules were filled with keratinous material derived from abrupt keratinisation of large polygonal cells with abundant pale eosinophilic cytoplasm without any intervening granular cell layer (trichilemmal type). Cells shows moderate to marked pleomorphism areas of severe atypia seen severe mitosis some FOCI appears invasive. Results: Proliferating pilar tumor with malignant transformation. Site of biopsy: Scalp (scalp dermoid cyst). Conclusion: In summary, malignant proliferating trichilemmal cysts are rare and pose a diagnostic dilemma for the pathologist. It has also been stated that the tumor has tendency to recur and metastasize more frequently than squamous cell carcinoma. Wide surgical excision should be considered as the primary modality of treatment while alternative therapies require further evaluation.

Keywords: Proliferating pillar tumor malignant transformation

   AOP102 Top

   Dermatopathology Top

Diagnostic utility of direct immunofluorescence on paraffin-embedded skin biopsy samples for the diagnosis of autoimmune vesiculobullous lesions

B K Praveen, Hemlata Panwar, Deepti Joshi, Dinesh P Asati, Jai Kumar Chaurasia, Neelkamal Kapoor

All India Institute of Medical Sciences, Bhopal, Madhya Pradesh, India

Introduction: Autoimmune Vesiculobullous diseases (AIBDs) refers to a group of diseases characterized by blisters of skin/mucosa due to the presence of circulating autoantibodies against antigens in epidermis or dermo-epidermal junction. Direct immunofluorescence (DIF) for IgG, IgC3, and IgA on fresh-frozen tissue is the gold standard diagnostic test for AIBDs. However, DIF in the absence of frozen tissue technically gives challenges in the diagnosis of AIBDs. Objectives: Practical utility of DIF done on paraffin-embedded skin biopsy rather than fresh frozen tissue for the diagnosis of AIBDs. Materials and Methods: This Cross-sectional comparative study included 30 cases of AIBDs, DIF for IgG and IgA was performed on the paraffin-embedded tissue (PE-DIF) after proteinase digestion on histopathological confirmed 15 Pemphigus Vulgaris (PV), 3 Pemphigus Foliaceous (PF), 4 Bullous Pemphigoid (BP), 3 Dermatitis Herpetiformis (DH) and 3 Sub-corneal pustular dermatosis (SCPD), one case each of Linear IgA disease and Pemphigoid Gestationis (PG). PE-DIF staining pattern was compared with the DIF on fresh frozen tissue (FF-DIF). Results: All cases of PV and PF showed an intercellular chicken wire staining pattern similar to FF- DIF. However, background staining was more intense in PV cases while less intense in PF cases. 3 BP cases showed linear staining in PE-DIF. DH, SCPD, Linear IgA Disease, and PG cases didn't show positivity. It was noticed that the immunofluorescence in PE-DIF was rapidly deteriorating than FF-DIF. Conclusion: DIF done on Paraffin-Embedded tissue can be used as supplement and salvage technique with histopathology for the diagnosis of AIBDs, particularly when cryostat facility for frozen tissue is not available and patient is unable to undergo a second biopsy procedure.

Keywords: Autoimmune bullous, paraffin-embedded tissue, pemphigus

   AOP103 Top

   Dermatopathology Top

Histomorphological findings of skin lesions in leprosy, a comparative study between pretreatment and after fixed duration of treatment

Chandraprava Mishra, Priyadarshini Biswal, Dilleswari Pradhan, Asaranti Kar

Department of Pathology, S.C.B. Medical College and Hospital, Cuttack, Odisha, India

Introduction: Multi drug treatment has achieved elimination of leprosy as a public health problem and now the focus has shifted to handling the challenges after treatment. A better delineation of the histological features of treated leprosy lesions may enable us to understand more about the complex immunological changes in leprosy. So we considered worthwhile to study the histological findings at the end of FDT for paucibacillary, smear negative and smear positive MB leprosy and compare with the pretreatment status. Objectives: To study and compare the histopathological features of skin lesions of leprosy before and after fixed duration of treatment. Materials and Methods: Study design: Prospective study. skin biopsies obtained from patients clinically diagnosed as leprosy who attended skin OPD (Pre treatment cases). Again skin biopsy was taken after 3 months and 6 months (Post treatment cases). Histomorpholgical changes noted in H&E stain. ZN stain also done. Results: Total number of cases included in the study is sixty. Majority of patients had patches (36.7%) and nodules (33.3%). About 13.3% had macules and patches, 10% plaques and rest had macules and nodules. Extent of lymphohistiocytic infiltrate in the dermis was recorded. During follow up, significant reduction of lymphohistiocytic infiltration was noted, especially after 2nd follow up. Finally the histopathological changes between pretreatment and post treatment follow up were compared. Conclusion: Awareness of histomorphological changes that occur in skin lesions of leprosy after completion of treatment can aid the pathologists to determine whether the lesions are active or inactive histologically and assist the clinician to convince the patient that his disease is inactive and does not need further management.

Keywords: FDT, histomorphological changes, leprosy

   AOP104 Top

   Digital Pathology Top

Significance of nuclear morphometry in carcinoma breast – A cross-sectional study

Ankita Girdhar, R Kalyani, P N Sreeramulu

Sri Devaraj Urs Medical College, Kolar, Karnataka, India

Introduction: Fine needle aspiration is one of the reliable method in diagnosing Breast cancers. Morphometric studies are done in various organs benign and malignant neoplasms by using software which measures cellular, cytoplasmic and nuclear parameters. Objectives: 1. To evaluate FNAC smears of breast cancer. 2. To study nuclear morphometry parameters in aspirated smears. 3. To assess the smears cytological findings with nuclear morphometry parameters. Materials and Methods: 60 cases which were diagnosed with Carcinoma Breast were collected from July 2020 to June2022 from cytology section, Department of Pathology, Sri Devaraj Urs medical college, Tamaka, Kolar. The smears were analysed and were subjected to nuclear morphometry study. Nuclear parameters were captured in the Zen software and Image J software. The nuclear morphometric findings were correlated with cytological findings and histopathological diagnosis in the available cases and descriptive statistical analysis was done. Results: Nuclear morphometry was used to differentiate between benign and malignant aspirates with gradually increasing nuclear size parameters from benign breast cases to malignant breast cancers. Nuclear area, nuclear perimeter, nuclear feret diameter, minimum feret and shape factor for Benign Breast lesions were 25.16 ± 3.2 μm2, 21.58 ± 1.89 μm, 6.5 ± 0.94 μm, 4.87 ± 0.50 μm, 0.92 ± 0.02 and for malignant breast cases were 46.57± 12.24 μm2, 27.53 ± 3.26 μm, 10.08 ± 1.18 μm, 6.49 ± 0.88 μm, 0.93 ± 0.01 respectively. Conclusion: Nuclear morphometric study in breast cancer is a new concept which supplement FNAC in differentiating benign from malignant lesions. It helps in diagnosis and assess the prognosis in breast cancer.

Keywords: Breast cancer, fine needle aspiration cytology breast, nuclear morphometry

   AOP105 Top

   Endocrine Pathology Top

Diagnostic utility of immunohistochemical markers CK-19, CD-56, Ki-67 in solitary thyroid nodules

Arundhati Shit, Moumita Maiti

NRS Medical College and Hospital, Kolkata, West Bengal, India

Introduction: Both benign and malignant thyroid neoplasms and few non-neoplastic conditions may present with solitary thyroid neoplasm (STN). Intra and interobserver variation due to some overlapping histomorphologic features pose diagnostic dilemma in distinguishing benign and malignant nodule. Objectives: To analyse expression of CK-19, CD-56 and Ki-67 in different thyroid lesions presented as STN and their role in differentiating papillary thyroid carcinoma (PTC) from follicular neoplasm (FN). Materials and Methods: An observational, cross-sectional, study in Pathology Department, NRSMCH, Kolkata, from February 2021 to July 2022. Tissue sections, after processing and staining in H&E, CK-19, CD-56 and Ki-67 expression by immunohistochemistry was studied. Evaluation and correlation of expression of CK-19, CD-56, Ki 67 to distinguish between benign and malignant STN and determining their utility in reaching definitive diagnosis was tried. Statistical analysis software EPI INFO was used and p-value <0.05 considered significant. Results: Out of 60 surgically resected STN specimens, non-neoplastic-23, benign-10 and malignant-27 cases [PTC-21 cases, follicular carcinoma, anaplastic carcinoma, poorly differentiated carcinoma-2 cases each]. CK-19 was found to be a sensitive (85.71%) and highly specific (100%) positive marker for diagnosing PTC. Difference in expression of CD-56 (p=0.000023) and CK19 (p =0.00001) was highly statistically significant in differentiating PTC and follicular neoplasms (FN). CD-56 was found to be a highly sensitive (100%) and specific (71.42%) positive marker for diagnosing FN. Difference in Ki-67 Labeling Index was highly significant between benign versus malignant tumors (p=0.00035) and also well-differentiated versus poorly differentiated thyroid neoplasm (p=0.00001). Conclusion: The panel of three IHC markers CK-19, CD-56 and Ki-67 is helpful for definitive diagnosis of different solitary thyroid nodules in cases with overlapping & doubtful histomorphologic features.

Keywords: Solitary thyroid nodule

   AOP106 Top

   Endocrine Pathology Top

Study of utility of immunohistochemical expression of BRAF (VE1 clone) in a cohort of thyroid neoplasms

Galeti Revathi, Vanditha Singh, Rupa Mehta, Nithin M Nagarkar

AIIMS, Raipur, Chhattisgarh, India

Introduction: BRAF V600E mutations are seen in 40 - 50% of papillary thyroid carcinomas, ~15% of poorly differentiated thyroid carcinomas, 20 - 60% of anaplastic thyroid carcinomas. These mutations portend a poor prognosis with higher rates of recurrence, extrathyroidal extension, lymph nodal metastases, resistance to Radioactive Iodine RAI treatment and areresponsive to targeted treatment. BRAF V600E mutations can be evaluated by immunohistochemistry (VE1 clone), molecular testing (PCR, Sanger sequencing, NGS). Objectives: Evaluating BRAF V600E mutation by using IHC as surrogate on thyroid tumour biopsies. Materials and Methods: Thyroid tumour biopsies received between 2014 to 2021 were classified using WHO Classification of tumours of the thyroid 2017. BRAF VE1 IHC performed on representative section was scored as described by Zhang et al into intensity (0 to 3+) and distribution (homogenous/ heterogenous) and compared between the histopatholgical categories. Results: 76 thyroid tumours out of which 18 were benign (17 FA and 1 HA), 56 were malignant (43 PTC, 2 FC, 1 PDC, 4 ATC, 5 MTC, 1 lymphoma) and 2 were borderline (1 AFA and 1 FT-UMP). Of 43 PTCs, 27 were classic, 10 were FVPTCs (5 encapsulated, 5 infiltrative sub-types), 2 columnar, 1 tall cell, 1 oncocytic, 1 solid and 1 diffuse sclerosing variants. BRAF VE1 IHC staining was seen in 41 out of 43 PTCs, 1 poorly differentiated carcinoma and 2 out of 4 anaplastic carcinomas, and NOT observed in any other neoplasm. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of BRAF VE1 IHC in diagnosis of Papillary thyroid neoplasms was 83%, 100%, 100%, 95.4%, 96.2%. Conclusion: BRAF VE1 antibody IHC can be used as diagnostic and prognostic marker for papillary thyroid carcinoma.

Keywords: BRAF IHC, papillary carcinoma, VE1 antibody

   AOP107 Top

   Endocrine Pathology Top

Risk of malignancy in thyroid aspirates with special emphasis on atypia of undetermined significance/follicular lesion of undetermined significance and its correlation with ancillary techinques

Beant Kaur, Aparajita Goel

S.N.M.C, Agra, Uttar Pradesh, India

Introduction: Fine needle aspiration cytology of the thyroid has been shown to be able to categorise many benign and malignant lesions & is also especially useful in clinically equivocal cases and cases where biochemical and immunological parameters are not helpful. Objectives: To determine the concordance of diagnosis of thyroid lesions on fine needle aspirates with help of ancillary techniques and establish the risk of malignancy. Materials and Methods: The study was conducted at Pathology department of SNMC, Agra in patients with thyroid swellings. FNAC was done and cytologic diagnosis was given as per Bethesda which was further worked-up on histopathology and IHC. Results: Out of total 66 cases, 2 cases were non-diagnostic, 31 cases were diagnosed as benign (Bethesda 2) and 12 were Bethesda 3 (, AUS/, FLUS) while 13 cases were diagnosed as follicular neoplasm or suspicious of follicular neoplasm (Bethesda 4). 09 cases were categorized as either malignant or suspicious for malignancy (Bethesda 6 and 5). Histopathologic and IHC correlation was done in most cases. For Bethesda 1 category, 1 was benign and 1 was malignant on histopathology. For Bethesda 2 category, 2/31 cases were found to be malignant on final histopathology. For Bethesda 3, 6/12 Cases were malignant. Bethesda 4 revealed to have 2/13 cases to be malignant. For Bethesda 5 and 6 categories, 100% concordance was found. Conclusion: Risk of malignancy was found to be significantly high in Bethesda 3 category which warrants the use of such strict cytologic criteria providing adequate information for it to fit into benign or malignant categories.

Keywords: Bethesda system, fine needle aspiration cytology, thyroid nodule

   AOP108 Top

   Endocrine Pathology Top

Expression of CK 19, CD 56 and CD 57 in follicular adenomas-thyroid

M Bharanidharan, Meeta Singh

MAMC, New Delhi, India

Introduction: The follicular variant is the most common variant of PTC and the encapsulated forms have been proposed to be termed as NonInvasive Follicular Thyroid neoplasm with Papillary like nuclear features (NIFTP). Both Follicular adenoma (FA) and NIFTP are the two entities which shows encapsulation and previously reported cases of FA needs to be revisited due to overlapping morphology to find out the prevalance of NIFTPs. Objectives: This study aimed to evaluate the incidence of NIFTP in cases diagnosed as FA and to correlate morphological criterias of NIFTP and IHC findings, to find out the incidence of NIFTP in the cases reported as FA. Materials and Methods: In this retrospective study, 40 tumors were investigated, which where histologically diagnosed as FA and CK 19, CD 56 and 57 immunostaining were performed on all cases. The criterias were encapsulation, follicular growth pattern, nuclear features of PTC, defined by a nuclear score of 2 or 3. Exclusion findings were psammoma bodies, more than 30%solid pattern, no lymphovascular or capsular invasion, no tumor necrosis and no high mitotic activity (≥3/hpf). Results: Out of 40 cases of FA, five cases (12.5%) were reclassified as NIFTP. Conclusion: After application of all IHC and morphological criterias, some of the FA cases can be reclassified as NIFTP.

Keywords: Criterias, follicular adenoma, NIFTP

   AOP109 Top

   Endocrine Pathology Top

Cytogenetic abnormalities and correlation with hormonal profile in non-obstructive azoospermia: A case control study

Aatish Saraswat, Lt Col Paresh Singhal

Department of Pathology, Armed Forces Medical College, Pune, Maharashtra, India

Introduction: WHO defines primary infertility as a disease of the reproductive system wherein there is a failure to achieve a clinical pregnancy after 12 months or more of regular unprotected sexual intercourse. Affects 3-15% of couples. Male factor contributes to 50% of cases. The frequency of chromosomal abnormalities in the general population is 0.6-1.2%. High prevalence in primary infertile males: 2-14%. Objectives: To assess the prevalence, type of chromosomal abnormalities and correlation with hormonal profiles in primary infertile males with non-obstructive azoospermia (NOA) or Oligospermia when compared to normal age matched controls. Materials and Methods: A case-control study which included 100 cases and 50 healthy controls. Detailed medical history was taken, physical examination with investigations including semen analysis, hormonal profile and scrotal sonography was done for both the groups. Infertile males with obstructive/surgical causes were excluded. Chromosomal analysis was performed on peripheral blood T-lymphocytes cultured for 69-72 hours. 20 metaphases were analysed for each patient, especially when mosaicism was suspected. Special assays like C-banding and FISH were performed wherever required. FSH, LH, and Testosterone measured from early morning serum samples by electrochemiluminescence using VIDAS ®2016 serum chemistry analyser. Major chromosomal abnormalities were further subdivided into structural and numerical. Results: Total 81 patients out of 100 had NOA of which 63 (77.77%) had normal karyotype, 7 (8.64%) had numerical chromosomal abnormalities whereas 5 (6.17%) had structural abnormalities. 13 out of 100 patients had Oligospermia of which 2 (15.38%) had numerical and1 (7.69%) had structural abnormalities. Control population had no abnormality. The hormonal assay had a significant correlation with chromosomal karyotyping when compared between normal and abnormal karyotyping in infertile patients (p value < 0.05). Sperm count was also significantly reduced in infertile patients with abnormal karyotype (p value < 0.05). Conclusion: Azoospermia is the most common presentation. A significant association between chromosomal aberrations in infertile males and non-obstructive semen abnormalities. Serum gonadotrophin levels are higher and Serum Testosterone levels are lower in males with primary infertility compared to controls. This study recommends the cytogenetic workup to be considered in non-obstructive azoospermia patients with deranged hormonal profiles as screening of underlying genetic defects.

Keywords: Azoospermia, chromosomal aberration, hormonal profile

   AOP110 Top

   Endocrine Pathology Top

BRAFV600E mutation in papillary carcinoma thyroid

M S Bhuvitha, G Krishna

Government Medical College, Thiruvananthapuram, Kerala, India

Introduction: Thyroid malignancies comprises 1% of total human malignancies, Papillary thyroid carcinoma (PTC) being the most common malignant tumour of thyroid cancers. It is estimated that 80% of thyroid malignancies are comprised of PTC. The most important etiology of PTC is imposed on activating mutation of RET/RAS/BRAF/MAPK pathways in PTC. Single hotspot mutation at nucleotide 1799 of the BRAF gene has been identified as the most common genetic event in PTC, with a prevalence of 29-83%. The use of molecular markers for thyroid cancer diagnosis, prognosis, targeted therapy and surveillance has been exciting area of the study and change. Objectives: A) Primary objective: To know the proportion of BRAFV600E mutation positivity in Papillary carcinoma thyroid specimens in Department of Pathology, Government Medical College, Trivandrum. B) Secondary objective: To study association between the mutation status of BRAFV600E and tumor aggressiveness in papillary carcinoma thyroid. Materials and Methods: Study design: cross sectional study. Study setting: Government Medical College Thiruvananthapuram. Study period: May 2017 to May 2020. Study Population: histopathologically proven cases of papillary carcinoma thyroid from the thyroidectomy specimens in department of Pathology. Method and collection of data: record paste. Sample size: 56 cases. Sampling method: first 56 consecutive specimens meeting the inclusion and exclusion criteria during the period of analysis where included in the study. Study Procedure: Thyroidectomy specimens received in the department of Pathology where diagnose test PTC during the study period was formalin fixed, paraffin embedded and thin sections made. Slides were stained using H and E stain. The blocks of those slides with a tumor were collected. Unsteen sections are made from the same and IFC for BRAFV600E where done on those slides and their expression was studied. Results: Among the PTC cases, the population with PTC in our study should equal proportion of patients <45 and 45 years of age. There was a female propondence. Size of the primary tumor was more than 1 cm for majority of cases. 44% of the total cases studied showed lymph node metastasis. On evaluating BRAFV600E expression in PTC, 80% showed positive expression, and mutation association with elderly age and limford metastasis was evident in present study. Conclusion: The study concludes, BRAFV600E mutation is common among primary PTC patients. It is strongly correlated with elderly age and lymph node metastasis. BRAF IHC in daily practice can be considered as an alternative for essential to for the BRAFV600E mutation analysis. It was observed that BRAFV600E mutation was associated with age tumor stage and prognosis.

Keywords: BRAFV600E, lymphnode metastasis, papillary thyroid carcinoma

   AOP111 Top

   Endocrine Pathology Top

Expression of hTERT, CD56 and HBME-1 in thyroid neoplasms

Anjali Mittal, Meeta Singh, Shyam Lata Jain, Ishwar Singh, Rajdeep Singh

Maulana Azad Medical College, New Delhi, India

Introduction: Thyroid cancer is the most common malignancy of endocrine glands. Due to overlap in the morphological features of different thyroid neoplasms, neoplasms with atypical morphology, unknown metastasis and for small biopsy/FNAC samples there is a need for a set of markers that may aid in differentiating various thyroid neoplasms. We studied the expression of three markers hTERT, CD56 and HBME-1 and their utility in differentiating the thyroid neoplasms. Objectives: 1) To study the expression of hTERT, CD 56 and HBME-1 in thyroid neoplasms. 2) To correlate these markers with histological subtyping. Materials and Methods: In this retrospective study in a tertiary care hospital with 20 cases of thyroid neoplasms and the clinical data was collected and IHC was done on the tumour sections followed by interpretation of the results. Results: CD56 was expressed in 3 (100%) cases of FA, 1 (8.3%) cases of PTC, 3 (100%) cases of FC and 1 (50%) cases of FVPTC. HBME-1 was expressed in only one (33.3%) case of FA, 12 (50%) cases of PTC, 2 (66.7%) cases of FC and one (50%) cases of FVPTC. hTERT expression was seen only in 10% of all the cases. with only two cases (16.7%) cases of PTC showing hTERT positivity. Conclusion: In this study we found significant correlation of lymph node metastasis with CD56 expression and HBME-1 expression. A significant correlation was observed between hTERT expression and extrathyroidal extension of thyroid neoplasms. A significant correlation was seen between CD56 and HBME-1 expression.

Keywords: CD56, HBME-1, hTERT, thyroid cancer

   AOP112 Top

   Endocrine Pathology Top

Borderline thyroid tumours: Nuclear score inclusion – To clear diagnostic confusion

Tejas Vidwans, Varsha Dhume, Vikas Kavishwar

Topiwala Nair Medical College, Mumbai, Maharashtra, India

Introduction: Follicular patterned thyroid lesions include various entities ranging from Adenomatous goitre to follicular variant of papillary carcinoma. In 2022 WHO, Follicular Neoplasms are classified into:- Benign, Low Risk and Malignant. Low risk group includes the borderline tumours from 2017 WHO classification. It introduced these categories with the purpose to prevent the potential overtreatment and to reduce the burden of a “CANCER “diagnosis. Objectives: Classify the follicular patterned neoplasms in borderline category based on histomorphological features and to study their nuclear parameters according to the proposed nuclear scoring system. Materials and Methods: 52 borderline neoplasms (NIFTP, WDT-UMP, FT-UMP) showing predominantly follicular architecture and intact /questionable capsular/vascular invasion were studied by 3-point scoring scheme (Nuclear size and shape, nuclear membrane irregularities, chromatin: each assigned a score of 0 or 1) with range of total scores 0 to 3. Nuclear and cytoplasmic characteristics were statistically compared with follicular adenoma. Association between variables was assessed by Chi-square test. Results: NIFTP (41) and WDT- UMP (7) had nuclear score 2. In FT-UMP (4) nuclear score was 1. Nuclear score was lower in follicular adenoma and follicular carcinoma whereas it was higher in FVPTC. As compared to follicular adenoma, in borderline tumors, statistically significant differences were observed. Conclusion: High nuclear score in NIFTP differentiates it from other follicular patterned neoplasms except FVPTC. Separation of borderline category from follicular adenoma on morphology is justified as per statistical analysis.

Keywords: FT-UMP, NIFTP, Nuclear scoring system, WDT-UMP

   AOP113 Top

   Endocrine Pathology Top

Comparative study of two successive American Thyroid Association Risk Stratification Systems in patients with differentiated thyroid cancers in India

Malavika Satheesh, Malini Eapen, Usha Menon, Vasantha Nair, Niveditha Kartha

Amrita Institute of Medical Sciences, Kochi, Kerala, India

Introduction: The risk of recurrence following treatment in differentiated thyroid cancer (DTC) patients follows American Thyroid Association Risk Stratification System (ATA RSS) guidelines - 2009 followed by revision in 2015. Studies comparing the two systems are negligible in India. Objectives: To assess the risk stratification change between ATA-RSS 2009 and 2015 in a cohort of DTC patients. Further, compared the performance of the two systems with outcome. Materials and Methods: The study group comprised 221 adults with DTC who had total thyroidectomy with subsequent radioactive iodine ablation. Clinico-pathologic parameters were analysed following initial therapy to assign ATA RSS strata. At 1 year, response to treatment (ATA RTR-2015) and clinical end point at last follow-up were analysed. Results: The intermediate group of ATA RSS-2009 was re-categorised as low risk (3.1%) and high risk (6.3%) under ATA RSS-2015. ATA RSS-2015 had better PPV (90.79%), NPV (41.38%), sensitivity (44.81%), specificity (89.55%) and accuracy (58.37%). However, ATA RTR-2015 had a higher PPV (94.02%), NPV (57.69%), sensitivity (71.43%), specificity (89.55%) and accuracy (76.92%). Extranodal extension had OR = 4.45 (p = 0.001) and microscopic extrathyroidal extension (ETE) had OR = 2.93 (p = 0.013) while predicting persistent disease status. Conclusion: A substantial shift was seen in our study from ATA RSS-2009 to ATA RSS-2015. Even though ATA RSS-2015 was found to be better than ATA RSS-2009 in predicting outcome, ATA RTR-2015 was seen to be far superior. We also observed that microscopic ETE and ENE significantly predicted persistent disease status at the clinical end point.

Keywords: American Thyroid Association Risk Stratification System, recurrence, thyroid cancer

   AOP114 Top

   GastroIntestinal Pathology Top

Histopathological panorama of neoplastic and non-neoplastic lesions of gastrointestinal tract and hepatopancreatobiliary system at a tertiary care hospital

Priyanka Sharma, Vijay Suri

Adesh Institute of Medical Sciences and Research, Bathinda, Punjab, India

Introduction: Diseases of the Gastrointestinal & Hepato-Pancreato-Biliary tract are one of the most commonly encountered problems in everyday clinical practice. They are responsible for a great degree of morbidity and mortality. These lesions maybe neoplastic or non-neoplastic. Diagnostic biopsies of upper GI tract & specimens of gallbladder followed by colon account for a major number of histopathology reporting. Neoplastic lesions occur commonly in the colon but can be present anywhere in the gastrointestinal tract. Some lesions have an increased risk for malignancy so it is important to determine the histologic type and characteristic for treatment. Objectives: To study the histopathological panorama of all lesions of the gastrointestinal tract, hepatobiliary system and pancreas. To analyze the association between demographic patterns of individuals and their lesions. Materials and Methods: This is a prospective observational study. 350 cases of tissue specimens and biopsies from gastrointestinal tract, hepatobiliary system and pancreas were studied. All specimens were routinely processed. Histopathological findings are reported as percentages or frequencies. The categorical data such as histopathological findings and demographic pattern is compared using Chi-square test & correlation statistics are applied. Results: A total of 350 cases were included in the present study. Out of these, 156 (44.6%) were males and 194 (55.4%) were females with Male: Female ratio of 0.8:1. Out of 350 cases, 145 (41.4%) were from Gastrointestinal tract and 205 (58.6%) were from hepatopancreatobiliary system. Gall bladder 191 (54.6%) was the most common site, followed by Large intestine 45 (12.9%), Appendix 42 (12%), Small intestine 24 (6.9%), Tongue and buccal mucosa 11 (3%), Pancreas 09 (2.6%), Esophagus 8 (2.3%) and Liver (1.7%). Neoplastic lesions accounted for 12.9% cases and non neoplastic lesions for 87.1 Benign neoplasia accounted for 90.3% of the total neoplastic lesions and malignant lesions constituted 9.7%, Adenocarcinoma being the most common malignant lesion (60%). Conclusion: Due to their wide histological presentation and aggressive malignant potential, histopathological typing of various lesions is mandatory to confirm the diagnosis.

Keywords: Gastrointestinal, hepatopancreatobiliary, histopathology

   AOP115 Top

   GastroIntestinal Pathology Top

Histopathological spectrum of lesions in upper gastrointestinal endoscopic biopsies

GouriPriya Rajeev, Sharvani Setty, C Narayana Murthy, V L Ramesh

Basaveshwara Medical College and Hospital, Chitradurga, Karnataka, India

Introduction: Upper gastrointestinal tract (GIT) disorders are one of the most commonly encountered problems in clinical practice with significant morbidity and mortality and endoscopic biopsy is common procedure performed for diagnosis of a variety of benign and malignant lesions. Endoscopic biopsy is the current gold standard for the assessment of patients with upper GIT symptoms. Objectives: To determine the spectrum of histopathological lesions of upper GIT and to evaluate the use of endoscopic biopsies as an effective tool in the diagnosis and management of various upper GIT lesions. Materials and Methods: A retrospective study was conducted on the upper GIT endoscopic biopsies and their histopathological assessment done at the department of pathology, Basaveshwara Medical College and Hospital, Chitradurga from January 2021 to August 2022. Results: Of 107 endoscopic biopsies studied, 65 were from males and 42 were from females. Age range was 19-90 years. There were 38 (35.5%) biopsies from esophagus, 10 (9.34%) from GE junction, 36 (33.6%) from stomach, 20 (17.7%) from duodenum and 7 (6.54%) biopsies from other sites. 62 (57.9%) biopsies were non-neoplastic, 8 (7.47%) were benign neoplasms while 36 (33.6%) were malignant neoplasms. Most frequently diagnosed inflammatory lesion was chronic gastritis and neoplastic lesion was squamous cell carcinoma of esophagus. Conclusion: Endoscopic findings along with histopathological examination of endoscopic biopsy helps in proper diagnosis and better management of upper GI lesions. Knowledge of spectrum of lesions will be crucial in diagnosing the lesions on histopathology.

Keywords: Endoscopic biopsy, histopathology, upper GI

   AOP116 Top

   GastroIntestinal Pathology Top

Expression of stem cell biomarker CD44 in oral squamous cell carcinoma and it's association with lymph node metastasis and TNM staging

Satadruti Chakraborty, T N Suresh, S M Azeem Mohiyuddin

Sri Devaraj Urs Medical College, Kolar, Karnataka, India

Introduction: Oral squamous cell carcinoma (OSCC) is one of the most prevalent cancers in the world. OSCC is a highly invasive lesion associated with high morbidity and significant mortality, attributed to resistance to therapy, metastasis and recurrence driven by specific populations of cancer stem cells (CSC). The evidence of association of expression of CD44, a stem cell biomarker and metastatic potential of the tumor is inconclusive in OSCC and hence needs further evaluation. Objectives: To determine immunohistochemical expression of CD44 in oral squamous cell carcinoma (OSCC) & to find its association with lymph node metastasis & TNM staging. Materials and Methods: 105 histologically proven cases of OSCC were studied. Histopathological parameters like depth of invasion, presence of lymph node metastasis, grading and TNM staging was done according to the new AJCC staging criteria. Both intensity and proportion of CD44 expression were recorded. Results: 49 cases (46.6%) showed a depth of invasion more than 10 mm. 52 out of 105 cases (49%) had nodal involvement. TNM staging was 5.7%, 7.6%, 44.7% and 42% for stages I, II, III and IV respectively. Majority of the cases (87.5%) showed CD44 expression in tumor. There was a significant association between the CD44 expression and lymph node metastases (p<0.001). Higher CD44 expression was seen in Stages III & IV (p<0.001). Conclusion: CD44, a stem cell biomarker is significantly associated with higher TNM stage and lymph node metastases. This can help predicting the tumor behavior in small biopsy.

Keywords: CD44, lymph node metastases, oral cancer

   AOP117 Top

   GastroIntestinal Pathology Top

Insulinoma-associated protein 1 expression in primary neuroendocrine neoplasms of the gastrointestinal and pancreaticobiliary tracts: A study from a tertiary care hospital of Coastal Karnataka

Emerine Edwin, Pooja K Suresh, Sneha Rao, Hema Kini, Jyoti Kini, Saraswathy Sreeram

Kasturba Medical College, Mangalore, Karnataka, India

Introduction: Neuroendocrine neoplasms (NENs) are uncommon tumours that can arise from the gastroenteropancreatic (GEP) system. The 2019 WHO GEP-NEN classification classifies them into grades of neuroendocrine tumours (NETs), neuroendocrine carcinomas (NECs) and mixed neuroendocrine-non-neuroendocrine neoplasms (MiNENs). There is no single best immunohistochemical marker for their diagnosis. Insulinoma-Associated Protein 1 (INSM1) is a novel marker found in developing neuroendocrine tissues and tumours. Objectives: To evaluate the expression of INSM1 in primary GEP-NENs. Materials and Methods: Retrospective review (2016-2021) was used to assess clinical features and histomorphology of 70 cases of GEP-NENs. Paraffin-embedded tissue was available in 49 GEP-NENs which along with 10 other non-neuroendocrine gastrointestinal neoplasm controls were subsequently stained with INSM1, Syn, CgA and Ki-67. Results: The patients ranged from 13-87 years of age with majority of them over 50 years (68.5%) and mostly men (62.8%). Most common site for GEP-NENs was duodenum (34.3%), followed by rectum (20%). INSM1, CgA and Syn expression was noted in 71.4%, 57.1% and 87.8% of GEP-NENs respectively. INSM1 was more sensitive than CgA but less sensitive than Syn and as specific as CgA and SYN (Sensitivity – 71.4%, 57.1% and 87.8% and specificity – 90%, 90% and 90% of INSM1, CgA and Syn respectively). Conclusion: INSM1 is a promising novel marker for GEP-NENs. Compared to traditional neuroendocrine markers, INSM1 is more sensitive and as specific. In addition, we found that INSM1 showed positivity in G3 NETs as compared to CgA and Syn. Further studies on INSM1 in NENs of the entire body is recommended to validate INSM1 as a single neuroendocrine diagnostic marker.

Keywords: Gastrointestinal neoplasms, INSM1, neuroendocrine tumour

   AOP118 Top

   GastroIntestinal Pathology Top

Correlation of scrape cytology and histopathology in diagnosis of gastrointestinal tumors

Patil Shashirekha, Nagarekha Kulkarni, C Bharath

Vijayanagar Institute of Medical Sciences, Ballari, Karnataka, India

Introduction: Gastrointestinal tumors, mainly esophagogastric and colorectal carcinomas constitute one of the major causes of morbidity and mortality worldwide. The early detection and treatment of gastrointestinal tumors has been shown to improve patient's survival significantly. The current study is carried out to correlate scrape cytology and histopathology in diagnosis of gastrointestinal tract tumors. Objectives: To study the cytohistopathologic correlation and its utility in diagnosis of gastrointestinal tract tumors. Materials and Methods: Thirty surgically removed gastrointestinal tract tumor specimens were studied. Scrapings were taken from representative areas of each specimen and spread on slides, these slides were examined immediately after staining with H/E and then reported. Later the specimen was fixed in 10% formalin, then grossing was done and routine histopathological processing was carried out, followed by staining with H/E, then was examined and reported. The diagnosis obtained by scrape cytology and histopathology was correlated and analysed. Results: A total number of 30 specimens were received and majority of them were of colon cancer (33.3%), followed by stomach and rectal tumors with male: female ratio of 3:2. Overall the scrape cytology diagnosis showed total concordance (100%) with histopathology diagnosis. Conclusion: Scrape cytology is simple, rapid and economically feasible technique and can be useful as an adjunct for frozen section in intraoperative consultation. The material and knowledge obtained from this study can be used as a teaching material and provide a basis for implementation of this technique in day to day practice.

Keywords: Gastrointestinal tumors, histopathology, scrape cytology

   AOP119 Top

   GastroIntestinal Pathology Top

Expression of CD34 in tumoral and peritumoral areas of colorectal carcinoma and its association with TNM staging

Ritu Raghav, Sachin Kolte, Chintamani, Sunil Ranga

VMMC and Safdarjung Hospital, New Delhi, India

Introduction: CD34 is an endothelial antigen which is used to highlight the microvessel density (MVD) as a marker of angiogenesis. There is a significant increase in microvessel density in colorectal carcinogenesis. Objectives: 1. To quantify microvessel density within tumoral areas and peritumoral areas. 2. To correlate microvessel density with TNM staging. Materials and Methods: A total 40 cases of colorectal carcinoma were included in this cross-sectional study, done at Department of Pathology, VMMC and Safdarjung Hospital, New Delhi. To assess the MVD within tumoral and peritumoral area, anti CD34 antibody was used. Results: The number of microvessels were counted in all colorectal carcinoma specimens after identifying the “hot spot”. In this study mean (±SD) of MVD in tumoral area was 4.64 (±2.52) and mean (±SD) of MVD in peritumoral area was 3.33 (±2.58). There was significant difference between CD34 stained blood vessels in the tumoral and peritumoral area (p=<0.001). Association of MVD with TNM staging and also to the various parameters like age, gender, site, size, locality and differentiation of tumor was assessed. Conclusion: CD 34 labelling have higher specificity and accuracy than regular HE staining and can be used as molecular biological indicator for stratifying these patients for planning appropriate treatment including antiangiogenic and anti tumor antibody directed therapy.

Keywords: CD34, microvessel density, TNM staging

   AOP120 Top

   GastroIntestinal Pathology Top

Does the buds foretell pollination in lymph nodes?

Anjali Balu, S M Choukimath, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Tumor budding is defined as single cells or clusters of <5 cells at the invasive margin of colorectal cancer. Convincing evidence suggests that tumor budding may be the visible correlate of an Epithelial Mesenchymal Transition (EMT)-related process in the tumor microenvironment. The current study was an attempt to find out the relationship between tumor budding in colorectal carcinoma and their respective lymph node status. Objectives: 1. To determine tumor budding and its score in colon and rectal resection specimens. 2. To correlate tumor budding with lymph node metastasis in resection specimens. Materials and Methods: A 2-year single-centre study on 33 colorectal resection specimens were analysed. Formalin-fixed paraffin-embedded tissues were stained with routine H&E and studied under a 20X objective. Evaluation of tumor budding was done using the International Tumor Budding Consensus Conference guidelines. Results: Out of 33 colorectal-carcinoma cases, 13/33 (40%) showed low-score of tumor budding, 4/33 (12%) with intermediate, 4/33 (12%) with high-score and 12/33 (36%) with zero-score. Metastatic lymph node deposits were observed in 9/13 (69%) cases of low-score, 4/4 (100%) of intermediate, 4/4 (100%) of high-score and 2/12 (17%) of zero-score. There was a statistically significant association between the scoring of tumor budding and lymph node metastasis (p < 0.001%). Conclusion: Assessment of tumor budding in resection specimens will predict the possibility of lymph node metastasis, especially when fewer are dissected. Also, nodal positivity is most certain with intermediate and high score tumor budding. Therefore, tumor budding can be considered as a significant prognostic factor in colorectal-cancer.

Keywords: Budding, colorectal carcinoma, metastasis

   AOP121 Top

   GastroIntestinal Pathology Top

Morphological and immunohistochemical analysis of gastrointestinal tract lymphomas

N Sandhya, M N Suma, S Balu

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Gastrointestinal tract is the most common site for occurrence of extranodal lymphomas, accounting for 30-40% of all extranodal lymphomas. The most commonly involved site is stomach, followed by small intestine and large intestine. DLBCL is the most common type of lymphoma occurring in the gastrointestinal tract. Objectives: Histomorphological and immunohistochemical evaluation of gastrointestinal lymphomas. Materials and Methods: A descriptive retrospective study is conducted over a period of 4 years (2019 – 2022). All cases with morphological diagnosis of gastrointestinal lymphomas, confirmed on immunohistochemical studies are included in the study. Results: A total of 31 cases comprising of 22 biopsies and 9 resection specimens are included in the study. Male: female ratio is 1.6:1 with peak incidence during 4th to 6th decade. The most common lymphoma diagnosed is Diffuse large B cell lymphoma (45%) followed by High grade B cell lymphoma NOS (16.1%), Mantle cell lymphoma (12%), Marginal zone lymphoma (9.6%), Burkitt lymphoma ((9.6%) and intestinal T cell lymphoma (6.4%). The most frequent site is stomach (42%), followed by small intestine (35%) and large intestine (23%). Conclusion: DLBCL is commonly encountered gastrointestinal lymphoma, involving the stomach most frequently. Differentiating low grade lymphomas from reactive conditions and high grade lymphomas from other malignant neoplasms based on morphology alone is challenging. Hence Immunohistochemical evaluation is good adjuvant to morphology and necessary to arrive at final diagnosis.

Keywords: DLBCL, gastrointestinal lymphomas, stomach

   AOP122 Top

   GastroIntestinal Pathology Top

Role of tumor-stroma ratio and tumor budding in colorectal adenocarcinoma – A three year institutional experience

Shaivy Malik, Adil Aziz Khan, Sufian Zaheer

Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India

Introduction: Colorectal cancer (CRC) is the second most common cancer worldwide and third in causing cancer-related deaths. The pTNM classification is based on anatomical evaluation and lacks predictive accuracy. Tumor-stroma ratio (TSR) and Tumor budding (TB) is validated by many international study groups as a prognostic marker in various epithelial cancers. However, there are limited studies in Indian cohort and its role in prognostication requires more validation. Objectives: Here we aimed to correlate the prognostic role of TSR and TB with pTNM staging, histopathological grading and tumor size in patients of colorectal adenocarcinoma. Materials and Methods: The study is conducted in Department of Pathology at a tertiary care hospital. A total of 40 patients who underwent curative surgery for CRC between, September 2019 to September 2022 were retrospectively included in this study. H & E stained sections of tumor were examined and TSR was calculated using Java based open access image processing software (Image-J) and TB was calculated in hot spot region and their values were correlated with pTNM and pathological grading of adenocarcinoma. Results: We observed that low TSR (< 0.5) and high TB were significantly more associated with poorly differentiated CRC (p < 0.0001 and p = 0.0001 respectively). Also, low TSR (< 0.5) was significantly higher in cases having high T and N stage. Overall, TSR showed higher sensitivity and specificity than TB and better correlated with pTNM and pathological grading. The findings will be discussed further in detail. Conclusion: Our study shows, TSR and TB can be better prognostic markers in CRC.

Keywords: Colorectal cancer, tumor budding, tumor-stroma ratio

   AOP123 Top

   GastroIntestinal Pathology Top

Histopathological spectrum of lesions in upper gastrointestinal endoscopic biopsies

Jagadeswari Kukkala, M Bharathi, Anjali Rao, Sathyavathi R Alva

KVG Medical College and Hospital, Sullia, Karnataka, India

Introduction: The upper gastrointestinal (GI) endoscopy is a simple, safe and well tolerated procedure for the visualisation of various lesions. When combined with biopsy it aids in the early detection of the pathologic process and institution of appropriate therapy. Our study aims at finding out the spectrum of lesions in endoscopic biopsies from upper GI tract. Objectives: 1. To enlist the spectrum of histopathological lesions in upper GI endoscopy. 2. To correlate the clinical diagnosis with histopatholoical diagnosis. Materials and Methods: A two year retrospective study was carried out at the department of pathology of our tertiary care institute using the data received between the years 2020 and 2022. The histopathological data of upper GI tract endoscopic biopsies was assessed and results were obtained. The statistical data analysis was performed using SPSS V 16. Results: In our study the mean age of presentation was 61 yrs. 34 (67%) were males and 17 (33%) were females. Among 51 cases, 30 (59%) were malignant and 21 (41%) were benign. The most common malignancy in esophagus was squamous cell carcinoma (20%). Among gastric malignancies, classic adenocarcinoma (18%) was more compared to signet ring cell carcinoma (12%). Most common benign lesion reported in our study was gastric ulcer (18%). Around 4% cases were reported to be severe dysplasias. Conclusion: Histopatholological examination in adjunct with upper GI endoscopy should be considered as much more valuable diagnostic tool rather than endoscopy alone and helps in giving an accurate diagnosis.

Keywords: Biopsy, endoscopy, histopathology

   AOP124 Top

   GastroIntestinal Pathology Top

Association between the immunohistochemistry expression of E-cadherin, beta-catenin and CD-44 in colorectal adenocarcinoma

Amrutha Tunuguntla, T N Suresh, P N Sreeramulu

Department of Pathology, Sri Devaraj Urs Medical College, Kolar, Karnataka, India

Introduction: Colorectal cancer ranks 9th among cancers in Indian population. Epithelial-mesenchymal transition (EMT) is an important process during cancer metastasis. In EMT there is downregulation of Ecadherin which is intracellular adhesion molecule. Mutations in Betacatenin genes and Wnt signalling pathway causes accumulation of the Betacatenin protein leading to classic adenoma-carcinoma sequences. The stem cell differentiation is by the expression of CD 44, which is strongly associated with EMT and is useful in targeted therapy. Objectives: To determine the ImmunoHistoChemistry (IHC) expression of E cadherin, betacatenin, CD44 in Colorectal Adenocarcinoma and to correlate the IHC expression of E cadherin, betacatenin and CD44 with Histopathological stage, lymphnode metastasis and grade of Colorectal adenocarcinoma. Materials and Methods: 50 histologically proven cases of colorectal adenocarcinoma from 2016 -2021 were included in this study. Clinical, demographic data, TNM staging and lymphnode metastasis was obtained. Immunohistochemical staining for E cadherin, Betacatenin and CD 44 was be done and analysed. Results: Cases belonging to stage I, II, III and IV are 23%, 41.1%, 33.4%and 2% respectively. Lymphnode metastasis was present in 37.2% of the cases. There is statistically significant association between tumor staging with Betacatenin (p<0.01) and CD44 (p<0.01) and mild association with Ecadherin (p=0.66). In Lymphnode metastasis significant association was found between Ecad (p<0.01) and CD44 (p<0.04). In grading mild association was found between all the markers. Conclusion: There is a significant increase in the expression of CD44 in lymphnode metastasis and in advancing stages and a significant loss of Ecadherin as the stage advances and significant increase in Betacatenin expression in higher stages and high grade carcinoma.

Keywords: Betacatenin, CD44, colorectal adenocarcinoma, E cadherin

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Clinico-histomorphological study of lesions of ampullary and periampullary area

Karishma Dhingra, Deepika Rawat, Vatsala Misra

MLN Medical College, Prayagraj, Uttar Pradesh, India

Introduction: Ampullary tumors are rare, accounting for only 0.2% of gastrointestinal cancers and approximately 7% of all periampullary tumors. Males affected more than females with median age of 60. They present with wide clinical spectrum, most common being jaundice and abdominal pain. Different histological types have variable prognosis. Objectives: To study in detail the clinical and histomorphological features of patients presenting with ampullary and periampullary lesions. Materials and Methods: The study was a prospective and retrospective study of two years from 2020-2022 conducted in the Department of Pathology. Study included 56 cases presenting clinically and endoscopically as periampullary lesions. Endoscopic biopsies were taken, processed and-3 μm thick sections were stained with H and E. IHC was done for CK7, CK17, MUC1 and CDX2 in all cases to differentiate between intestinal and pancreatobiliary type. Results: Males are five times more affected than females with mean age of 54 yrs. CBD is commonly involved followed by periampullary and ampullary lesions. Histological diagnosis was intestinal type (48 %), pancreatobiliary (25 %), mixed (8 %) and other types (19 %). IHC was done to further confirm the type as intestinal and pancreaticobiliary type. Conclusion: The histological features showed intestinal type being more common than pancreatobiliary type that was further confirmed by IHC. Former had better prognosis and survival.

Keywords: Ampulla, CBD, duodenum

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Correlation of colorectal mucosal histology with development of post infection irritable bowel syndrome: A pilot study

Yogamaya Pattanayak, Pallavi Bhuyan, Prasanta Parida, S P Singh, Lity Mohanty

Department of Pathology, SCB Medical College, Cuttack, Odisha, India

Introduction: Irritable bowel syndrome (IBS) is one of the most common gastrointestinal disorder. One third cases of IBS occur following acute gastroenteritis (AGE), known as Post infectious IBS (PI-IBS). The aim of the study was to find out the relation of colorectal mucosal histology with development of PI-IBS and associated risk factors. Objectives: 1. To correlate the colorectal mucosal histology of patients of acute gastroenteritis during the episode with that of after 6 months of follow up. 2. To compare the colorectal mucosal histology of the patients who are developing PI-IBS after follow up with that of the patients who are not developing PI-IBS. 3. To study the histological factors influencing the development of PI-IBS. Materials and Methods: This prospective cohort study was performed between Oct-2020 to Aug-2022 in 74 patients of acute gastroenteritis, attending the Infectious and diarrhea and gastroenterology ward, SCB MCH, Cuttack by performing detail history taking, routine examination and colonoscopic biopsies with follow up evaluation after 6 months. Results: One fourth patients (25%) suffered from IBS after 6 months of AGE. The colorectal biopsies revealed defect in epithelial integrity, persistent mucosal microscopic inflammation, elevated mucosal lymphocytes and mast cells, cryptitis and crypt abscess. Other contributory factors were younger age, long duration of gastroenteritis, cramping abdominal pain, watery and bloody stool, family history of IBS. Conclusion: From our study we found that the risk of development of IBS was increased following AGE. Histological investigations should be performed in patients with persistence of symptoms beyond 3 months. This is important to avoid unnecessary, excessive and long term drug use as well as to reduce morbidity.

Keywords: Acute gastroenteritis, irritable bowel syndrome, post infectious irritable bowel syndrome

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Role of HER2 overexpression in diagnosis of gastric adenocarcinoma by IHC

Amit Jaiswal, Ritu Mehta

Armed Forces Medical College, Pune, Maharashtra, India

Introduction: Gastric cancer remains the second most common cause of cancer mortality in the world. HER2 over expression in gastric adenocarcinoma has proven beneficial for targeted treatment with trastuzumab along with chemotherapy in patients. Objectives: To ascertain the role of HER2 over expression in diagnosis and its prognostic use in Gastric adenocarcinoma patient. Comparison of HER 2 expression by immunohistochemistry. Materials and Methods: H&E stained slides of gastric adenocarcinoma were reviewed for HER2 immunohistochemistry. Results were assessed with the help of Fisher test, Student 't' test and Chi-Square test. P value < 0.05 was considered statistically significant. Results: A total of 50 cases were included out of which10 cases showed HER2 positivity of 3+ and 10 cases show HER2 positivity of 2+ on immunohistochemistry. Peak age of incidence was between 40-60 yrs with mean age of 50 years. There was no statistical correlation between age, gender, tumor location, lymphovascular invasion, perineural invasion and HER2 positivity with respect to tumor subtype, differentiation and stage. Conclusion: With introduction of HER2 molecular targeted therapy for patients with gastric adenocarcinoma, HER 2 testing is crucial to find patients who can be benefitted by this treatment. HER2 testing should be done in all patients with gastric adenocarcinoma.

Keywords: Gastric cancer, Her2, targeted therapy, trastuzumab

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The utility of a prognostic scoring system based on histomorphological parameters in low grade colorectal cancer

Akhil Chandran, Aneesha Asok, M Feroze

MES Medical College, Perinthalmanna, Malappuram, Kerala, India

Introduction: Grading of colorectal cancer (CRC) is traditionally based on the percentage of gland formation. High-grade CRC is subjected to more precise molecular grading strategies. However, low-grade cases have inhomogeneous outcome due to insufficient categorization. Tumour budding and tumour-infiltrating lymphocytes (TIL) have developed as additive prognostic factors in CRC. Categorization of the inhomogeneous low grade CRC is being attempted in recent times. Objectives: (a) To assess tumour budding and tumour infiltrating lymphocytes in low-grade CRC and its association with pT, pN, and lymphovascular invasion (LVI). (b) To categorize low grade CRC into low, intermediate and high risk based on a prognostic scoring system. Materials and Methods: 105 WHO low-grade CRC cases were analysed in terms of gland formation, budding, and TIL. For each parameter 1 to 2 points were given resulting in a sum score. A scoring system was developed to categorize low grade CRC cases into a low, intermediate, and high-risk group. Results: Higher tumour budding is associated with high pT stage (p= 0.022), high pN stage (0.042) and more chance for LVI (p=0.038). Low TIL is associated with high pN stage (p=0.042). By applying the scoring system, high score corresponds to high pT stage (p=0.027), high pN stage (0.035) and high chance for LVI (p=0.015). Conclusion: Even in the age of molecular pathology, it is important to pay special attention to tumour morphology for additional information on tumour behaviour and prognosis. Combining morphological parameters of tumour and tumour environment can help to subdivide low grade CRC into new prognostic groups.

Keywords: Low grade colorectal cancer, prognostic scoring system, tumour budding, tumour-infiltrating lymphocytes

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SMAD4 expression and its correlation with E-cadherin and N-cadherin AS markers for epithelial mesenchymal transition in colorectal carcinoma

Murugaiyan Elamurugan, Jyoti R Kini, K B Vatsala

Kasturba Medical College, Manipal, Karnataka, India

Introduction: Epithelial-Mesenchymal Transition (EMT) is one of the driving mechanisms in the pathogenesis of invasion and metastasis of colorectal carcinoma (CRC). The process of EMT is regulated by one of the signal transducers TGF-β/SMAD4 pathway. During EMT there will be loss of cellular adhesions associated with loss of membranous E-cadherin and gain of mesenchymal markers (N-cadherin) along with loss of SMAD4 which culminates into invasion and metastasis. Objectives: 1) To assess the immunohistochemical expression of E-cadherin, N-cadherin, SMAD4. 2) To correlate their expression with each other and clinicopathologiacal features. Materials and Methods: This was a retrospective study of two years duration conducted in a tertiary care center. We evaluated the immunohistochemical expression of SMAD4, E-cadherin and N-cadherin in resected specimens of CRC. Results: We evaluated 70 cases of CRCs. Majority of cases were T3 and T4 (40%, and 35% respectively). Most cases had nodal status N0 (46%), N1 and N2 stage was seen in 36% and 21% cases respectively. Accordingly, AJCC stage III cases were the maximum in our series (56%). E- cadherin showed statistically significant stronger expression in cases with stage N0 and N1 (p-0.027). A weaker expression of membranous n-cadherin was seen in greater proportion of cases with stage1 and was found statistically significant (p-0.006). Strong expression of N-cadherin was significantly associated with female gender (p-0.014). Smad 4 expression was positively associated with larger tumor size (p-0.023). Conclusion: Loss of E cadherin and gain in N cadherin expression is associated with higher Stage, serving as markers for tumor aggressiveness and highlighting the role of EMT in tumor progression.

Keywords: Colorectal carcinoma, epithelial mesenchymal transition, immunohistochemistry

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Comprehensive clinicopathological study of Barrett's oesophagus

Pawan Khandre, B R Vani, N Anvita, B Deepak Kumar, V Srinivasa Murthy

ESIC MC and PGIMSR, Rajajinagar, Bengaluru, Karnataka, India

Introduction: Barrett's oesophagus (BE) is a premalignant esophageal lesion occurring in 10% of patients with gastro-oesophageal reflux disease (GERD) and in 2% of general population. Endoscopic appearance of salmon-coloured mucosa ≥1 cm proximal to the gastroesophageal junction and presence of intestinal metaplasia (IM) on biopsy is diagnostic. Various histological features occurs in BE. IM with goblet cells have higher risk for oesophageal cancer. Objectives: 1) To study the various histomorphological features in Barrett's oesophagus 2) To correlate with clinico-endoscopic features. Materials and Methods: The present study was a cross-sectional study conducted in the histopathology department over a period of 6 months. All cases diagnosed as BE were taken. Clinical details were collected from departmental case files and medical record archives. H&E, AB-PAS stained slides were retrieved and studied for morphology. Giemsa stained slides were studied for presence of H.pylori. Results: A total of 20 cases of BE were studied. Majority of patients are (40%) in age group of 50-60 years with male to female ratio of 5.6:1. Most common endoscopic finding noted was reddish mucosa at the lower end of esophagus (90%). Microscopic features of esophagitis, type of metaplasia, dysplasia, carcinoma and H.pylori presence were studied and tabulated. Conclusion: BE runs an indolent course and is a potent forerunner of malignancy. Hence endoscopic examination coupled with histopathological examination of altered esophageal mucosa is mandatory for a definitive diagnosis and follow up.

Keywords: Barrett esophagus, dysplasia, histomorphology

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To study the expression of mismatch repair (MMR) protein in colonic carcinomas (CRC) using immunohistochemistry

Amrutraj Patil, Rachana Lakhe, Preeti Doshi, Milind Kesarkhane, R C Nimbargi

Bharati Vidyapeeth Medical College, Pune, Maharashtra, India

Introduction: CRC is the 3rd most common cancer in both men & women. In 10-15% of sporadic CRC, Microsatellite Instability (MSI) has been reported due to defective MMR genes. In sporadic CRC, MSI is linked to methylation of MMR genes & BRAF mutations. Microsatellites are short, tandemly repeated sequences found throughout the genome that are used as markers for dMMR deficiency. Objectives: To classify CRC using MSI status into MSI-H and MSI-L & clinicopathological correlation with MMR gene expression. Materials and Methods: We studied 72 cases of CRC (Biopsies and resections). H&E & IHC was performed by using MLH1, MSH2, MSH6 and PMS2 antibody. Results: 40 CRC (55.55%) had loss of any 1 or more of the MMR gene proteins & 32 CRC (44.44%) had intact expressions. These includes: MLH1 only (sporadic)– 24 cases (33.33%), MLH1 and PMS2 (Type 1)–05 (6.94%), PMS2 only (Type 2)– 02 (2.77%), MSH2 & MSH6 only (Type 3)– 05 (6.94%), Loss of MSH6 (Type 4)-04 (5.55%), pMMR- All antibodies are Intact 32 (44.44%) Maximum cases were found in age group of 61-70 years with loss of MLH1. Our study confirms loss of MLH1 (33.33%) which is highly suggestive of Lynch syndrome. Conclusion: CRC with abnormal MMR gene expression were associated with MSI-H. About 33.33% demonstrated absence of MLH1 only. All CRC patients should be tested for MMR gene defects using IHC and/or PCR. This study shows MMR IHC expression in CRC suggesting the utility of IHC and distinct expression of MMR protein.

Keywords: Colorectal carcinoma, lynch syndrome, mismatch repair

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Human papilloma virus, programmed death ligand-1 and tumor microenvironment: Implications for patients with head and neck squamous cell carcinoma

Asttha Kapoor, Pinki Pandey, Savita Agarwal, Vineet Chaturvedi, Roopak Agarwal, Kapil Trivedi

U.P.U.M.S, Saifai, Etawah, Uttar Pradesh, India

Introduction: Oral squamous cell carcinoma is a major cause of death throughout the developed world. It is associated with smoking, paan and alcohol consumption. Human papillomavirus (HPV) has been considered as an etiopathogenic factor of head and neck squamous cell carcinoma (HNSCC). PD-L1 is expressed on tumor cells, and is able to bind to programmed cell death-1 (PD-1) on CD8+TILs thereby terminating their function. PD-L1 is considered to play a pivotal role in HNSCC by enabling tumor cells to evade immune system. Objectives: To study correlation among Immunohistochemical expression of PD-L1, CD8, HPV and various clinicopathological parameters. Materials and Methods: In this prospective study, total of 100 cases of HNSCC were taken. Tumor was graded according to World Health Organization (WHO) criteria. PD-L1, CD8 and HPV expression was determined by immunohistochemical staining. The obtained results were evaluated using Chi-square test. Results: Out of 100 cases, 73% cases were positive for PD-L1, HPV expression was seen in 90% cases and CD8 positivity was noted in 94% cases. A statistically significant association was observed between PD-L1 and alcohol abuse, abnormal sexual habits, multiple sexual partners, histological grade. In addendum to these CD8 was correlated with gender and smoking. Whereas, HPV was associated with abnormal sexual habits only. A significant correlation was found between expression of PD-L1 and CD8 (p=0.002). Conclusion: HNSCC that were highly infiltrated by CD8+TILs, and that had high PD-L1 expression may show better outcome, superior survival and cancer endurance.

Keywords: CD8, human papilloma virus, PD-L1

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Cluster of differentiation 44 expression in gastrointestinal malignancies. A study from South India

Ayesha Bhavikatti, G Guruprasad, C Panduranga

M S Ramaiah Medical College and Research Hospital, Bengaluru, Karnataka, India

Introduction: Cancer stem cell markers are now being tried in various cancers as prognostic markers including GI cancer but these kinds of studies are sparse in Indian population. Objectives: 1. To study the expression of CD44 among gastrointestinal cancer by immunohistochemistry. 2. To correlate the CD44 expression with various clinicopathological parameters. Materials and Methods: This study conducted over a period 50 months. Haematoxylin and eosin stained slide were screened for grading of the tumour, extent of invasion of tumour, confirmation of metastasis and staging was done. Immunohistochemical expression of CD44 was graded on the basis of percentage of tumor cells positive for staining. Statistical analysis was done and results were tabulated. Results: A total of 40 cases of GI cancer were studied. Ascending colon (37.5%) was the common site involved, 37 cases (92.5%) showed invasion beyond the muscularis external. Most of the tumours were poorly differentiated (37.5%). 50 % of lymphnodes showed tumour deposits. Majority of the cases were in stage II (40%). There was a significant correlation between histopathological type of differentiation with lymphnode metastasis and staging of tumour, lymphnode metastasis also had significant association with staging. Grade 2, CD 44 expression was most common followed by Grade 3. Significant association was observed between histopathological differentiations of tumour with CD44 expression. Tumours which are invading beyond muscularis externa and lymphnode positive cases showed moderate to high CD44 expression. Conclusion: CD44 expression was significantly noted in poorly differentiated tumors. Increased expression was also noted in cases of tumours invading beyond muscularis externa and lymphnode metastasis. Combination of CSC markers will increase the sensitivity and specificity and predict better overall survival in GI tumours.

Keywords: CD44, cancer stem cells, immunohistochemistry

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The histomorphological features of intestinal neuronal dysplasia – A rare diagnosis

Chiterlekha Kundal, Kim Vaiphei

PGIMER, Chandigarh, India

Introduction: Intestinal neuronal dysplasia (IND) is one the developmental enteropathies with clinical presentation of pseudo-obstruction. It is of 2 types: IND type A and IND type B. IND type A histology shows hyperplastic neural changes limited to myenteric plexus and normal submucosal plexus. The histological features of IND type B include hyperplasia of submucosal plexus, giant ganglia, ectopic ganglion cells at muscular and mucosa layers, and increased acetylcholinesterase activity in lamina propria indicating immaturity of the enteric nervous system (ENS). Here, we described histomorphological features in 6 such rare cases. Objectives: To study the histo morphological features of intestinal neuronal dysplasia. Materials and Methods: Full thickness biopsies and resected specimens from appendix, caecum, sigmoid and rectum were reviewed in six cases of IND retrieved over a period of 8 years from the departmental record. An autopsy case with coexistent adrenal hyperplastic syndrome and IND was also found. Clinical details were retrieved from the record. Slides were reviewed. Results: Age of diagnosis ranged from 5 months to 14 years. However, majority of patients were more than 1 year of age. All the cases were males. The most common presentation was constipation. In one of the cases, IND occurred in association with Hirschsprung's disease. In all cases, large ganglia with many ganglion cells as well as ectopic ganglion cells in lamina propria and muscular layer were present. Conclusion: IND is mostly diagnosed in colon. It can be associated with other anomalies of enteric nervous system ENS.

Keywords: Enteric nervous system, giant ganglia, intestinal neuronal dysplasia

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A retrospective analysis of appendiceal mucinous neoplasms: A study from tertiary oncology centre

Silpa Ray, Ashwini Nargund

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Appendiceal mucinous neoplasms account for 1% of appendectomy specimens. They include low grade appendiceal mucinous neoplasm (LAMN), high grade mucinous appendiceal mucinous neoplasm (HAMN) and mucinous adenocarcinoma (MAC). Recognition of these entities are important for proper management. Objectives: To study the spectrum of appendiceal mucinous neoplasms diagnosed in our center in the last 7 years. Materials and Methods: A retrospective analysis of appendiceal mucinous neoplasms diagnosed between January 2015-september 2022 was done. Demographic and clinical data, gross findings were retrieved from departmental archives. The tumours were reviewed and classified according to the 2019 WHO digestive tumours classification. Results: We analyzed 23 cases comprising of 4 males and 19 females with age ranging from 40-50 years. The study included 15 (65.3%) cases of LAMN, seven (30.4%) cases of MAC and one (4.3%) case of goblet cell adenocarcinoma. MAC was associated with MAC of ovary and colon in three and two cases respectively. Two of these appendiceal MAC showed signet ring cells. Three cases of LAMN had ovarian MAC as well. One LAMN was incidentally diagnosed in a case of intestinal sarcoma. HAMN was part of the spectrum in one MAC. Nine cases (34%) showed pseudomyxoma peritonei (PMP), out of which four cases had LAMN alone. Immnunohistochemical slides were reviewed in concurrent ovarian and colonic MAC cases to determine primary vs secondary. Conclusion: LAMN is the bulk of these neoplasms with a propensity for PMP. Many a time, it is difficult to ascertain the primary in extensive disease involving the ovary and colon.

Keywords: Adenocarcinoma, peritonei, pseudomyxoma

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Study of expression of cytokeratin 7 and 20 in gastrointestinal epithelial malignancies

N Chaitra, H K Sharath Kumar

Department of Pathology, Sri Siddhartha Medical College, Tumakuru, Karnataka, India

Introduction: Cytokeratins (CKs) are intermediate filaments found in cytoskeletal component of epithelial cells. The expression of individual keratins in gastrointestinal tract is specific to their location and is retained in malignancy. This property can be used to identify the origin of various tumors. Numerous studies are available on individual expression of CK7&20 and have shown variable expression ranging from 38-85%. Current study is designed to demonstrate usefulness of combined expression of CK7&20 in esophageo-gastrointestinal malignancies. Objectives: To study the association between CK7&20 expression with anatomical site, tumor grade and stage. Materials and Methods: 40 cases of biopsies and resected specimen of esophageo-gastrointestinal malignancies were stained for CK7&20 and assessed using three-tiered-system of scoring. Chi-square test was used to assess statistical significance. Results: CK7+/CK20- expression was seen in esophageal adenocarcinoma and CK7-/CK20- was expressed in esophageal squamous cell carcinomas. Gastric malignancies were negative for CK7 and 44% positive for CK20. CK7-/CK20+ phenotype is highly correlated with colorectal carcinomas. Majority of Grade-1&Grade-2 carcinomas showed negative and Grade-3 tumors showed variable expression of CK7. Majority of Grade-1 carcinomas showed score-3, while Grade-2&3 showed variable CK20 expression. Only three cases of Stage-2 and two cases of Stage-3 showed positive CK7. 50% of stage-1 showed weak CK20 expression. Majority of stage-2 and 3 malignancies showed moderate to strong positivity for CK20. Conclusion: The combined use of cytokeratin 7 and 20 has superior benefit in differentiating the site of origin, especially in metastatic tumors of unknown origin. No statistically significant correlation was found between CK7&20 expression and grade or stage of the tumor (p>0.05).

Keywords: Cytokeratin7 & 20, esophageo-gastrointestinal malignancies, immunohistochemistry

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Primary gastrointestinal lymphomas in adults: An institutional experience of 15 years from Northern India

Sumyra Qadri, Nissar Hussain Hamdani, Azra Shah, Khalil Mohammad Baba, Asifa Andleeb, Mohammad Hussain Mir

Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India

Introduction: Primary Gastrointestinal Lymphomas (PGIL) are uncommon and distinct Gastrointestinal Tract (GIT) malignancies which vary in their epidemiologic and clinicopathologic features worldwide. Since gastrointestinal malignancies are commonest malignancies in Kashmir valley, we intended to study PGIL in our hospital set up. Objectives: To study the clinicopathological and demographic profile of PGIL. Materials and Methods: This combined retrospective (eight and a half years) and prospective (six and a half years) descriptive observational study of 15 years was performed at Sher-i-Kashmir Institute of Medical Sciences, Srinagar from April 1998 to March 2013. Total 93 cases were included as per Dawson's criteria. All the cases were reviewed and reclassified according to the World Health Organisation (WHO)-Revised European American Classification of Lymphoid Neoplasms (REAL). Statistical analysis was done using GraphPad Prism 8. Results: Ninety-three patients (Male:Female ratio, 3:1) with a mean age of 47 years (age range=18-70 years) were studied. Patients mostly presented with non specific symptoms like abdominal pain (n=79) and anorexia (n=45); and small intestine (n=30), stomach (n=27) and large intestine (n=26) were the major sites affected. Histopathological and Immunohistochemical (IHC) studies revealed most of them to be Non Hodgkin's Lymphoma (NHL) of B-cell type (n=89) with Diffuse Large B-Cell Lymphoma (DLBCL), the commonest subtype (n=66) followed by Mucosa Associated Lymphoid Tissue (MALT) lymphoma (n=21). Stage distribution included 64 stage IE, 27 stage IIE and two stage IIIE patients. Surgical resection was performed in 53 patients along with chemotherapy (25), chemoradiotherapy (20) and anti-Helicobacter pylori (anti-H. pylori) treatment (2). Forty patients received chemotherapy without surgery, along with anti-H.pylori treatment (14) and radiotherapy (7). Mean survival of patients was 42 months. Conclusion: Primary Gastrointestinal Lymphomas (PGIL), although uncommon GIT malignancy, can clinically mimic any GIT disease and can involve any part of GIT. A high degree of suspicion can pick up these lymphomas early and a judicial combination of surgery, chemotherapy and radiotherapy can be offered for better survival. Further, IHC studies including genetic and molecular studies along with long term follow-up studies, are required to have a better understanding of these lymphomas in our region.

Keywords: Diffuse large B-cell lymphoma, Helicobacter pylori

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Histopathological study of premalignant and malignant lesions of intestinal tract

Haritha Bellapu

Sri Devaraj Urs Medical College, Kolar, Karnataka, India

Introduction: Colorectal cancer is a major cause of morbidity and mortality throughout the world. Prevention is based on early endoscopic detection of potentially curable cancers or precursor conditions such as dysplasia, which have a significant risk of progression to malignancy. Objectives: To analyse the spectrum of pre malignant and malignant lesions of intestinal tract by histopathological examination To determine distribution of these lesions in different sites of intestinal tract. Materials ands Methods: All the endoscopic biopsies & surgically resected specimens of intestinal tract received in the department of pathology, S.V.Medical College, Tirupathi Special stains and Immunohistochemistry are done wherever required Study Design & Duration- The present study is a Cross-sectional study for a period of 2 years from November 2016 to October 201. Results: Out of 113 endoscopic biopsies eight (8) were premalignant lesions and 25 were malignant. Out of 759 surgically resected specimens 37 specimens one (1) was premalignant and 36 were malignant. Conclusion: Malignant lesions are diagnosed more frequently than premalignant lesions. Most of the lesions were from large intestine followed by Anal canal. Premalignant and malignant lesions of small intestine are uncommon.

Keywords: Colorectal cancer, endoscopic biopsies, premalignant

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Evaluation of faecal calprotectin S100A8/S100A9 levels in patients suffering from irritable bowel syndrome with histopathologcal correlation

MD Mali, Ashfaq Ahmed, B Zeenath

Khaja Bandanawaz University's Faculty of Medical Science, Kalaburagi, Karnataka, India

Introduction: Irritable bowel syndrome (IBS) is a common functional bowel disorder which remains a clinical challenge with limited therapeutic options. The diagnosis of IBS is made by the Rome IV criteria. Objectives: To evaluate faecal calprotectin S100A8/ S100A9 levels (FCLs) in IBS patients and to compare those levels with Inflammatory bowel disease (IBD) patients sub-group and to determine the prevalence of Microscopic colitis in IBS patients with elevated FCLs. Materials and Methods: This prospective study was done from June 2021- July 2022. A total of 130 patients with chronic diarrhea were included. 90 patients belonged to IBS subgroup and 40 patients into Inflammatory bowel disease (IBD) for comparison. Diarrhea due to infections, drugs, pancreatic/bile acid deficiency and any organic gastrointestinal diseases were excluded. Patients were subjected to clinical details and FCLs were analysed by fluorescence immunoassay. Colonoscopic evaluation and segmental biopsies were taken. FCLs, colonoscopic and histopathological findings were evaluated in IBS patients and were compared with IBD patients. Results: The age ranged from 18 to 65 years with M:F ratio of 1:2.21. In 90 IBS patients, IBS-diarrhea predominant was the largest subgroup (72.23%) clinically.17 IBS patients showed elevated FCLs in which 10 patients showed features of Microscopic colitis[ lymphocytic colitis (07), collagenous colitis (02), indeterminate (01)] on histopathology respectively. Conclusion: This study shows FCLs can be elevated in subgroup of IBS Patients in which anti-inflammatory treatment can be a validated option. Also subgroup of IBS patients showing microscopic changes with elevated FCLs need further follow-up for considering these cases as probable future IBD patients.

Keywords: Faecal calprotectin, irritable bowel syndrome

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Screening for microsatellite instability (MSI) in sporadic colorectal adenocarcinoma: IHC versus PCR

Aditya Agnihotri, Anitha Javalgi, Vidisha Athanikar

SDMCMSH, Dharwad, Karnataka, India

Introduction: Microsatellite instability (MSI) is a key biomarker in colorectal cancer (CRC), with crucial diagnostic, prognostic, and predictive implications. Detection of microsatellite instability (MSI) is currently recommended by National Comprehensive Cancer Network (NCCN) for all patients with colorectal cancer (CRC). Current clinical reference methods are immunohistochemical staining of mismatch repair proteins and/or PCR analysis of frequently mutated short tandem repeat regions of DNA. Objectives: To compare the MSI Status performed via IHC and PCR techniques in CRC endoscopic biopsies. Materials and Methods: 42 colonoscopic endoscopic biopsies pathologically diagnosed as CRC were collected from the Department of Pathology, between June 2019 and June 2020. All samples satisfied the following criteria: (1) sporadic colon or rectal cancer confirmed by pathological diagnosis, (2) no preoperative therapy. We assessed two commonly used methods, IHC determination (Automated Roche Ventana) of loss of MMR gene products viz MLH1, MSH2, MSH6 and PMS2 against PCR amplification and subsequent fragment analysis of microsatellite markers, BAT25, BAT26, D2S123, D5S346 and D17S250 (Bethesda markers). Categorical variables would be tested by Chi-square, while continuous variables would be tested with Independent Samples T-test for parametric data and Mann-Whitney U test for non-parametric data respectively. Cohen's kappa (κ) coefficient statistic would be used for studying agreement. Results: Results under process. Conclusion: Both methods can be used in determining microsatellite instability in colorectal carcinomas. The preference of one method over the other would depend on the expertise and type of laboratory. Nonetheless, it is prudent to acknowledge and understand the advantages and shortcomings of each method.

Keywords: Colorectal cancer, microsatellite instability, PCR

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Tissue expression status of leptin and adiponectin in colorectal carcinoma – Development of a biomarker

P Priyanka, U S Dinesh

SDM Medical College and Hospital, Dharwad, Karnataka, India

Introduction: Colorectal carcinoma (CRC) is showing rising incidence of metastasis due to its advanced stage of presentation. Unfortunately, the existing biomarkers for prognosis have low sensitivity and hence there is a need for a definitive biomarker development. Leptinand Adiponectin are studied as potential biomarkers in CRC. Existing data on expression of Leptin and Adiponectin is ambiguous and limited. This study helps to understand the correlation of leptin and adiponection with other clinicopathological parameters. Objectives: To correlate the clinico-pathological parameters with tissue expression of leptin and adiponectin in CRC. Materials and Methods: It is a cross-sectional study design (March 2021 – February 2022). Paraffin embedded formalin fixed blocks for all diagnosed radical resection CRC cases were collected from the department of Pathology at tertiary care hospital to carry out IHC studies (leptin, adiponectin). Noncancerous colonic tissues were used as controls. Tumor characteristics and clinical data such as gender, age, occupation, tumor size, type, stage, grade, extension, lymph node involvement and metastasis were collected from the Hospital medical records. Suitable statistical tests were used. Results: Immunohistochemistry was performed on 41 cases of CRC and 10 cases of non-cancerous colonic tissues. Higher expression of leptin was observed in CRC when compared to non-cancerous tissues. Significant correlation was seen between leptin expression and histological type, advancing stage and metastasis (P<0.01). Significant negative correlation was found seen between adiponectin expression and advancing stage and grade of tumour. Conclusion: The study demonstrates the upregulation of leptin in CRC. The immunohistochemical staining of leptin and adiponectin could be a useful biomarker for prognostication of CRC.

Keywords: Adiponectin, colorectal carcinoma, leptin

   AOP142 Top

   GastroIntestinal Pathology Top

Histopathological predictors of microsatellite instability in colorectal cancer – A tertiary care center experience

Aminder Singh, Ankita Soni, Vikram Narang, Kunal Jain, Bhavna Garg, Harpreet Kaur

Department of Pathology, Dayanand Medical College and Hospital, Ludhiana, Punjab, India

Introduction: Microsatellite instability (MSI) has a therapeutic and prognostic implication in colorectal carcinomas (CRC). It can be detected either by immunohistochemistry (IHC) or molecular studies. Objectives: We aimed to identify the possible clinicopathological variables which can be used as predictors of microsatellite instability in such patients. Materials and Methods: CRC cases received for MSI detection by IHC (for one and half years) were included. A panel of four IHC markers (anti-MLH1, anti-PMS2, anti-MSH2, and anti-MSH6) was used. Confirmation by the molecular study was recommended in all the IHC-proven MSI cases. Various clinicopathological parameters were evaluated as predictors of MSI. Results: Microsatellite instability was detected in 40.6% (30/74) cases with MLH1 & PMS2 dual loss in 27% cases, MSH2 & MSH 6 dual loss in 6.8%, loss of all four MMR proteins in 2.7%, and isolated PMS2 loss in 4.1%. 36.5% cases showed MSI-H expression with only 4.1% cases showing MSI-L expression. ROC curve showed an area under the curve of 0.65 (95% CI, 0.515-0.776; p-value=0.03). On univariate analysis age <63 years, colon site, and absence of nodal metastasis were significantly higher in the MSI group. However, on multivariate analysis, only the age <63 years was found to be significantly higher in the MSI group. Conclusion: MSI detection can be performed either by IHC or molecular study. In this study, no histological parameter appeared to be the independentpredictor of MSI status. The age <63 years might predict the microsatellite instability, yet larger studies are needed for its validation. Thus, we recommend that IHC testing should be performed in all CRC cases.

Keywords: Colorectal carcinoma, microsatellite instability, microsatellite stable

   AOP143 Top

   GastroIntestinal Pathology Top

Molecular profiling of colorectal malignancies

Shalini Saurabh, Milind Chanekar, Shaikhali Barodawala, Kirti Chadha

Metropolis Diagnostics, Mumbai, Maharashtra, India

Introduction: Clinical applications of Genomic medicine, molecular diagnostics and Targeted therapies requiring specific molecular testing for selection of candidate patients are becoming a reality in clinical practice. Objectives: The incidence of colorectal cancer in India in men is 4.4 per 100000 and 3.9 in women. Recent advances in the treatment of colorectal cancers include frequent aberrant activation of EGFR signaling pathways (MAPK and PI3K). The most common mutations in the MAPKinase pathway are KRAS (40-50%), NRAS (2.5%), BRAF (5-10%) and PIK3CA. The RAS and RAF mutations are mutually exclusive. The Tyrosine kinase inhibitors targeting EGFR namely are Erlotinib and Geftinib. About 15% of all Colorectal cancers show underlying defects in DNA mismatch repair involving (MSH2, MLH1, hPMS2, MSH6). Materials and Methods: We conducted in our laboratory retrospective study based on comprehensive genome profiling of tumours from 220 patients with colorectal cancer. Results: Overall KRAS mutation was seen in 30.43% of patients of colorectal cancers, BRAF V600 mutation in 11.36% and PIK3CA gene mutation was seen in 18.18% of patients.79% of cases were MSI stable and MSI-H was four times more common than MSI-L. One case of metastatic adenocarcinoma was positive for KRAS codon 61 mutation (negative for codon 12/13 KRAS mutation) and exon 9 PIK3CA mutation. The KRAS mutation was most common in 51-60 years age group with preponderance in females whereas the BRAF V600 and PIK3CA mutation were more common in 20-30 years age group, being more common in males. Conclusion: The treatment of colorectal cancer was the first to be influenced by molecular profiling and has become a standard of care for predicting response to treatment and prognosis.

Keywords: BRAF V600, KRAS, PIK3CA

   AOP144 Top

   GastroIntestinal Pathology Top

Histopathological features of Meckel's diverticulum with clinical correlation

A N Shelashree, Suravi Mohanty, A M Shubha

St. John's Medical College, Bengaluru, Karnataka, India

Introduction: Meckel's diverticulum is a remnant of Vitello-intestinal duct, present in 2%–4% of the population. Clinical presentation varies from being an incidental finding at laparotomy to significant lower gastrointestinal bleed or lead point in intussusception. The clinical features may or may not mirror the histopathological changes in the diverticulum. The objective of the study is to evaluate the histopathological features of Meckel's diverticulum and correlate these with clinical findings. Objectives: 1. To evaluate histopathological features of Meckel's diverticulum. 2. To correlate histopathological features with clinical findings. Materials and Methods: Slides were retrieved from archives of Department of Pathology of all the cases of Meckel's diverticulum from 2012 to 2022. Clinical features were collected from the medical records of St Johns medical college Hospital. The slides were assessed for various histopathological features and correlation with clinical features was statistically analysed. Results: 44 cases were studied. Age of the patients ranged from 4 days to 27 years with male to female ratio of 5.2:1. Intestinal obstruction was present in 15 cases, gastrointestinal bleed in 17 and peritonitis in 14 cases. 23 cases had mucosal ulceration, 41 cases had inflammation, 26 cases had heterotopic mucosa. Significant statistical correlation was observed between the presence of heterotopic mucosa and gastro-intestinal bleed (p-value: <0.001) and perforation (p-value: 0.014). Presence of perforation and non-viability of margins was also statistically significant (p-value:0.034). Conclusion: Our study highlights the importance of careful histopathological assessment of resected specimens of Meckel's diverticulum for heterotopic mucosa. Sampling of the resected intestinal margins for viability dictates clinical severity in these patients.

Keywords: Gastrointestinal bleed, heterotopic mucosa, Meckel's diverticulum

   AOP145 Top

[TAG:2]Hematopathology [/TAG:2]

Expression of CD34 and CD31 in hypoplastic marrows with mast cell quantification

Soumya Gupta, G V Chaithra

Kasturba Medical College, Mangalore, Karnataka, India

Introduction: Bone marrow failure conditions characterized by hypoplastic marrow (HM) like Aplastic anemia (AA) and Hypocellular myelodysplastic neoplasm (MDS-h) are diagnostic dilemmas as they show significant clinico-morphological overlap. They have a common pathogenetic-pathway with the hematopoietic progenitor stem cells being the targets of autoimmune attack in AA and further neoplastic progression in MDS. An accurate diagnosis is critical as the prognosis and treatment differ. Objectives: 1. To analyse the expression of CD34 and CD31 in HMs using immunohistochemistry to distinguish AA from MDS-h. 2. To quantify mast cells in HMs for determining its diagnostic significance. Materials and Methods: A retrospective study of 65 cases diagnosed as HM from 2017-2021 were evaluated for the expression of CD34 and CD31 using immunohistochemistry. Mast cell quantification was done using Toluidine Blue special staining. Results: Total of 65 cases were evaluated, out of which 21 cases were of AA, 12 of MDS-h and 32 of other HMs. CD34 positive progenitor cells were absent/significantly decreased in 90.50% cases of AA, whereas it was relatively more in number (>0.5% of all the hematopoietic cells) in 43.80% cases of HM and 33.30% cases of MDS-h. Increased number of mast cells was seen in 76.19% cases of AA followed by 50% in HM and 33.33% in MDS-h. Conclusion: The expression of CD34 can be utilized to distinguish AA from MDS-h and other causes of HMs. Enumeration of CD34 positive stem cells can guide the management of the cases of AA and MDS-h. Mast cell quantification can be used as a supplementary diagnostic tool for HMs.

Keywords: Aplastic anemia, hypocellular-myelodysplastic neoplasm, mast cell

   AOP146 Top

   Hematopathology Top

Clinico-hematological study in leukemia

M Vidhyasini, N S Kamakeri, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Hematological Disorders Are Quite Frequent In Population. Leukemias Represent One Of The Most Important Problems In The Field Of Haematology. Early Diagnosis Of Various Subtypes Of Leukemia And Intervention Has Improved The Prognosis And Disease Free Survival. Objectives: (1) To Determine The Pattern Of Basic Hematological Parameters In Leukemia. (2) To Determine The Distribution Of Cases According To Age, Sex And Clinical Feature. Materials and Methods: All Patients Referred To Central Lab, Department Of Pathology, Kims Hospital, Hubballi As A Diagnosed Leukemia Taken Irrespective Of Age And Sex From September 2017 To September 2022. For All The Cases Peripheral Smear And Complete Hemogram Considered. Cytochemistry Was Done when required. Results: The Study Included 53 Cases Of Leukemia. In almost all age groups and types, 27/53 males predominated. The commonest type of leukemia observed in this study was 18/53 (34%) CML; 17/53 (32%) cases were AML; 10/53 (19%) Cases Were ALL. CLL was less frequent 7/53 Cases (13%). Conclusion: Leukemia is prevalent in this region of North Karnataka as in other parts of India. In almost all age groups and types of leukemias, males are predominated. The commonest type of leukemia observed in this study is18/53 (34%) CMLwhile, CLL was less frequent 7/53 Cases (13%). MPO and SBB done to differentiate AML from ALL. PAS is done to diagnose ALL. While peripheral smear examination remain key to the diagnosis of leukemia, conventional cytochemical techniques need to be used for typing and subtyping of leukemias.

Keywords: Cytochemistry, leukemia, peripheral smear

   AOP147 Top

   Hematopathology Top

Expression of CD34 blasts and mast cell tryptase in myeloproliferative neoplasms and its association with marrow fibrosis

Kesiya Thomas, Ranjitha Rao, Sharada Rai

Kasturba Medical College, Mangalore, Karnataka, India

Introduction: Myeloproliferative neoplasms (MPNs) are clonal hematopoietic stem cell disorders defined by the growth of cells from one or more of the myeloid lineages. The magnitude of blasts and fibrosis in bone marrow may indicate poor prognosis or progression towards acute myeloid leukaemia (AML). Recently, mast cells are gaining importance as literature suggested their role in progression to AML. Studies expressing the correlation of blasts, fibrosis and mast cell count in MPN cases are rarely reported. Objectives: To study the expression of CD34 blasts and mast cell tryptase (MCT) in MPNs and its association with bone marrow fibrosis. Materials and Methods: The present study was carried out in 66 patients over a period of 6 years, which comprised of 32 Chronic myeloid leukaemia (CML), 31 Primary myelofibrosis (PMF) and 3 Essential thrombocythemia (ET) cases. Routine staining along with immunohistochemistry (IHC) using CD34 and MCT were performed. Results: Expression of higher score of CD34+ blasts were seen in CML (n=19/32), when compared to PMF and ET where it was minimal or totally absent (p<0.001). Mast cells of grade 2+ was found in PMF (n=17/31), followed by CML and ET, which was statistically significant (p=0.006). CD34 (p=0.001) and MCT (p=0.001) counts were observed to be higher with greater degree of fibrosis in all MPNs. Conclusion: CML enumerated significant CD34 blasts in comparison to PMF and ET. Whereas, mast cells were expressed more in PMF compared to other subtypes. Enhanced expression of both markers was seen in association with increased marrow fibrosis.

Keywords: CD34, mast cell tryptase, myeloproliferative neoplasms

   AOP148 Top

   Hematopathology Top

Man versus machine in the diagnosis of anaemia – A tertiary care experience

Rose Thoams, S Sateesh Chavan, Purushotham Reddy

Karanataka Institute of Medical Sciences, Hubli, Karnataka, India

Introduction: The impact of early diagnosis and management of anaemia on patient outcome has been tremendous. Although conventional microscopic examination of blood using peripheral smear is unequivocal, the advent of automated haematology analyser has revolutionised the field of clinical haematology and its practice. This study highlights the efficiency and utility of specific parameters obtained by haematology analyser and its comparison with peripheral smear in diagnosis of anaemia. Objectives: 1. To determine the degree of correlation between automated haematology analyser and peripheral smear findings in the diagnosis and typing of anaemias. 2. To determine the sensitivity of the automated haematology analyser in detecting red blood cell morphology. Materials and Methods: 500 EDTA blood samples collected from the central laboratory, KIMS, Hubli during one and half years from February 2021 to August 2022, were run on the Sysmex 6 part automated haematology analyser and simultaneous peripheral smears prepared and stained using Leishman stain based on standard operating procedures. Results: 53.7% was normocytic normochromic anemia in which 82.9% showed normal curve, 28.9% was microcytic hypochromic anemia in which 92.4% showed shift to left, 6.2% was dimorphic anemia in which 61.3% showed normal curve contrary to the expected double peak and 4.8% was macrocytic anemia in which 83.3% showed shift to right. Conclusion: The Automated haematology Analyser provides highly valuable information with limited sample and is an essential tool in diagnosis, classification and management of anaemia.

Keywords: Anaemia, automated haematology analyser, peripheral smear

   AOP149 Top

   Hematopathology Top

A study on peripheral blood, bone marrow findings and cytogenetics of myeloproliferative neoplasms

Rani Aishwarya, Manisha Mohapatra, M Ruth Prasanna, Swarupa R Didla, V Shreni

GSL Medical College and Hospital, Rajamahendravaram, Andhra Pradesh, India

Introduction: Myeloproliferative neoplasms refers to heterogenous group of disorders resulting from abnormal proliferation of one or more terminal myeloid cell lines in the bone- marrow. It comprises of four classic types of myeloproliferative neoplasms i.e Chronic myeloid leukemia (CML), Polycythemia vera (PV), Essential thrombocythemia (ET), and primary myelofibrosis. The present study was undertaken to evaluate the peripheral blood, bone marrow findings and cytogenetics results in patients with myeloproliferative neoplasms diagnosed in our hospital. Objectives: To study the peripheral blood, bone marrow aspirations/biopsy findings and associated genetic mutations in patients with myeloproliferative neoplasm. Materials and Methods: The study was conducted over a period of 22 months (October 2020- August 2022). Out of a total 245 bone marrow studies, 54 patients were diagnosed as myeloproliferative neoplasm. Detailed clinical history along with complete blood picture, bone marrow aspiration/biopsy findings and cytogenetic results done in our hospital were analyzed. Results: These 54 patients, diagnosed as myeloproliferative neoplasm comprised of 30 (55.6%) cases of males and 24 (44.4%) were females. The age range varied from 15 to 79 years with a mean of 45 years. Majority, 44 (81.5%) cases were of chronic myeloid leukemia, 6 (11.1%) cases of polycythemia vera, 4 (7.4%) cases of essential thrombocytosis and 1 (1.8%) case of primary myelofibrosis. Of the 49 patients studied for cytogenetics, BCR- ABL Major gene was detected in 43 cases of CML and JAK-2 mutations in 6 patients. Conclusion: The complete blood picture, bone marrow and cytogenetics provides valuable information in the diagnosis and management of patients with myeloproliferative neoplasms.

Keywords: Bone-marrow study, cytogenetics, myeloproliferative

   AOP150 Top

   Hematopathology Top

Clinico-hematological study in pre-eclampsia

N P Nima, N S Kamakeri, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubli, Karnataka, India

Introduction: Pre-eclampsia is a multisystem disorder, unique to pregnant women after twenty weeks of gestation. There is a constant ongoing search for better predictors and prognostic factors to assess the progression and severity of the disease, hematological parameters being one among them. Objectives: (1) To know the changes in hematological parameters in pre-eclampsia. (2) Relationship between changes in different hematological parameters and progression to severity. (3) Predictability of complications of preeclampsia in terms of hematological parameters. (4) To find the major hematological parameters in relation with preeclampsia and its complications. Materials and Methods: A prospective study on 200 pregnant women diagnosed with preeclampsia whos 2 cc EDTA blood samples has been received in the central lab, Department of Pathology, KIMS Hubballi during the period of 18 months (January 2021 to June 2022). Relevant clinical history, general and per abdominal examination and urine examination findings were recorded. Complete hemogram was determined using Sysmex K-1000 automated blood cell counter. Smears were prepared and the values were crosschecked and additional microscopic findings were noted if any. Results: Total 200 blood samples studied. The age of the patient ranged from 18- 38 in which 44% of them came for routine obstretic examination. Out of 200, multiparous women are 36% and out of which 61% are showing Pre-eclampsia in previous pregnancy. 37%, 29% patients show relative neutrophilia and neutrophilic-leucocytosis respectively. Neutrophil/lymphocyte ratio and platelet/lymphocyte ratio are highly variable. Conclusion: This study concludes neutrophilia is the major haematological change in pre-eclampsia. No haematological parameter can predict the severity and complication of pre-eclampsia.

Keywords: Haematology, PIH, pre-eclampsia

   AOP151 Top

   Hematopathology Top

Correlation of haematological parameters and CD4 counts in HIV infected patients before and after initiation of highly active antiretroviral therapy (HAART)

T B Renuka, Sateesh S Chavan, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: HIV infection is a multisystem disease and haematological abnormalities are among the most common complications of HIV. These abnormalities reflect the underlying immune status and may be prevented or corrected by use of highly active antiretroviral therapy (HAART). Objectives: To correlate the haematological parameters and CD4 counts among HIV infected individuals before and after initiation of HAART. Materials and Methods: This is a prospective study on total 50 cases of newly diagnosed HIV patients over a period of 1.5 years. Complete haematological profile of each patient was evaluated at 0-, 6- and 12-months following initiation of HAART. Results: Out of 50 patients included in this study 4 died. Among 46, 56.5% (26) were females and 43.5 (20) were males. There was a significant improvement in haemoglobin, red cell count, haematocrit, platelet counts and in CD4 counts after 6 months and 12 months of therapy. Improvement in total count, absolute neutrophil count and absolute lymphocyte count was observed after 12 months of HAART. Conclusion: This study shows that there was improvement in all parameters in these patients and also in the stage of disease when they were followed for one year. So, regular follow up and compliance of patient is of utmost importance in checking the progress of disease as well as ensuring better quality of life. It can be concluded that people living with HIV/AIDS, HAART is useful for modifying the mortality and morbidity.

Keywords: CD4 counts, haematological parameters, HIV

   AOP152 Top

   Hematopathology Top

Clinico-hematological study of severe acute malnutrition in children

Anne Xavier, N S Kamakeri, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Severe acute malnutrition is a leading cause of childhood disease and death. According to NFHS-5 19% of children under five years are wasted. Children with SAM face a risk of morbidity and mortality eleven times greater than their healthy counterparts. Objectives: 1] To assess various clinical profiles associated with SAM. 2] To determine prevalence of anemia, its morphological patterns and its severity. 3] To study other changes in hematological parameters. 4] To determine the relationship between altered hematological parameters and progression to severity. Materials and Methods: All blood samples of children admitted with severe acute malnutrition received in the Department of Pathology were analysed in sysmex XN automated complete blood count machine while simultaneous peripheral smears were prepared and stained using Leishman stain over a period of 1.5 years. Results: Out of 80 cases 61.3% are males and 38.8% females. 9 cases are in the category of <-5 Z score (weight for height), 26 in -4.99 to -4 and 26 in -3.99 to -3 with maximum cases in 6-11 months (37.75%). Most common clinical presentation was fatigue constituting 19 cases (23.75%) followed by bronchopneumonia 16 cases (20%) and acute diarrheal disease 11 cases (13.75%). Mean hemoglobin level is 7.67. Prevalence of anemia is 93.75% out of which 32 cases (40%) were having severe anemia, 33 cases (41.2%) moderate and 10 cases (12.5%) had mild anemia with majority having microcytic hypochromic anemia. Conclusion: Almost all cases had anemia as comorbid condition along with significantly lower red cell indices and higher white cell count. So primary target should be to prevent malnourishment and secondary should be early diagnosis and treatment.

Keywords: Anemia, hematological changes, severe acute malnutrition

   AOP153 Top

   Hematopathology Top

Role of total leucocyte count and differential leucocyte count in patients with type 2 diabetes mellitus

T Sujithra, M Usha

M S Ramaiah Medical College, Bengaluru, Karnataka, India

Introduction: Diabetes Mellitus (DM) is a metabolic disorder leading to hyperglycemia. Although there is increased prevalence of both Type 1 and Type 2 diabetes, prevalence of Type 2 diabetes is increasing rapidly. The pathogenesis of Type 2 DM is characterized by low grade systemic inflammation. The adipocytes produce inflammatory markers like Interleukin-6 and C Reactive Protein (CRP) which can lead to insulin resistance in obese patients. Several studies have shown that Total Leucocyte Counts (TLC) are increased in Type 2 DM and can be used to predict development of micro and macro vesicular complications. Objectives: The objective of the study is to investigate the role of Total Leucocyte Count and Differential Leucocyte Count (DLC) in patients with Type 2 diabetes. Materials and Methods: The study was conducted on 153 Type 2 DM patients with increased Glycated hemoglobin levels (HbA1C). Patients with hypertension, coronary artery disease, pregnancy and other acute illness were excluded from the study. TLC and DLC were assessed from venous blood samples collected in EDTA vacutainer. Results: Results showed that the patients with Type 2 DM had increased Total Leucocyte Count and neutrophilia with relative lymphopenia. There was no variation in Monocyte, Basophil and Eosinophil count. Conclusion: TLC and neutrophils are increased in patients with Type 2 DM indicating that Diabetes Mellitus is a inflammatory state which can lead to insulin resistance.

Keywords: Diabetes mellitus, total leucocyte count

   AOP154 Top

   Hematopathology Top

Prevalence and causes of anemia in critically ill patients

Anju Rose, N S Mani

Bharati Vidyapeeth Deemed to be University, Pune, Maharashtra, India

Introduction: WHO defines anemia as hemoglobin level < 13 g/dl in men, < 12 g/dl in women. Cause of anemia in critical illness is complex and multifactorial. Primary mechanisms: loss of red blood cells due to phlebotomy, bleeding or trauma; decreased and increased production of RBCs, nutritional deficiency. Objectives: To evaluate course, causes of anemia and to analyze hematological, biochemical and clinical pathological parameters in critically ill patients. Materials and Methods: 300 cases were studied. Details like age, sex and clinical history was noted, hemogram and peripheral smears were studied. Other parameters were studied and correlated with the anemia. Progress of anemia, volume of phlebotomy and transfusion history was noted. Final cause of anemia was determined at completion of stay. Results: Of 300 patients, 63% (188) patients were males and 37% (112) were females. Total 265 patients were anemic and 35 were not anemic at the time of admission. During stay 35 patients also became anemic. During the stay anemia worsened in 99 cases and improved in 88 cases but continued to be anemic. The predominant reason for admission was surgery, chronic renal diseases and others. Most predominant morphological anemia was NCNC. 67 cases had shock on admission; most frequent was septic shock. Co-relation of anemia with clinical presentation, other hematological parameters will be co-related and presented. Conclusion: Anemia is a significant cause of morbidity in critically ill patients specially in background of sepsis, post operative period and chronic renal diseases.

Keywords: Anemia of critically illness, parameters, shock

   AOP155 Top

   Hematopathology Top

To evaluate the role of reticulin stain measured fibrosis in chronic myelogenous leukemia

Kamalpreet Kaur, Vijay Mehra, Rajiv Kumar Devgan, Permeet Kaur Bagga

Government Medical College, Amritsar, Punjab, India

Introduction: Chronic myelogenous leukaemia is a clonal, myeloproliferative disease that develops when a single, pluripotent, haemopoetic stem cell acquires the Philadelphia chromosome, is frequently associated with myelofibrosis. CML has a triphasic or biphasic clinical course: a chronic phase; an accelerated phase; and blast crisis. However, about 20% to 40% of patients die of complications of marrow failure associated with fibrosis. Objectives: To study the role of reticulin stain measured fibrosis in patients with chronic myelogenous leukemia and its prognostic significance. Materials and Methods: Bone marrow biopsies were performed on 50 cases of CML including both newly diagnosed cases as well as cases already on treatment and stained with reticulin stain to grade the fibrosis. Results: Reticulin fibrosis was graded on a scale of 1 to 4. Significant (Grade 3 or 4) fibrosis was noted in 23 patients (46%), out of which 12 were newly diagnosed usually in accelerated phase or blast crises and 11 were already on treatment with poor response. Grade 1 or 2 reticulin fibrosis seen in 27 patients (54%), 15 out of which were newly diagnosed mostly in chronic phase and 12 were on treatment with favourable response. Conclusion: This study concluded that reticulin stain measured fibrosis is the indicator for poor prognosis of patients of cml and their responsiveness to treatment.

Keywords: CML, myelofibrosis, reticulin stain

   AOP156 Top

   Hematopathology Top

Correlation of automated hematology analyzer obtained RBC indices and histogram with manual peripheral smear examination

Anjani Gupta, P Supriya, Anuradha Calicut Kini Rao

Yenepoya Medical College and Hospital, Deralakatte, Karnataka, India

Introduction: Anemia a common medical condition, is mostly diagnosed in developing nations. Peripheral smear examination is the gold standard for diagnosing most RBC disorders, which may fail to get diagnosed by automated cell counters. Few studies exist that compare automated analyzer based histogram, CBC parameters with peripheral smear examination. Present study was carried out to correlate these parameters. Objectives: The aim of the study is to correlate RBC indices and histograms obtained from automated analyzer with peripheral smear examination. Materials and Methods: Blood samples collected from 775 anaemic patients, over a 6 months period, in the Hematology laboratory, were typed for anaemia separately by haematology analyzer based parametes (RBC indices & histogram) and by peripheral smear examination. The diagnoses thus obtained were analysed to find out the significance of the mean difference in various parameters. Results: 775 cases examined on peripheral blood smear showed the highest number of caes to have NCNC (58.2%), followed by MCHC (34.1%),. Dimorphic (4%) and macrocytic anemia (3.1%) and haemolytic anemia (0.6%) had fewer cases. Analysis by erythrocyte indices and histogram showed similar frequency of various types of anemia, however the percentages varied in individual methods. The correlation between the diagnosis made by these methods was statistically significant, p<0.0001. Conclusion: Histogram based data provide early intimation of subtle changes in numerical parameters; subjective evaluation. RBC histogram and indices should be interpreted in the light of peripheral smear.

Keywords: Histogram, peripheral smear, RBC indices

   AOP157 Top

   Hematopathology Top

Renal disease and marrow fibrosis, a clinicopathological study at a tertiary care center

Ahely Ghosh, Meenakshi Balasubramanian, Jyothi Shetty

Topiwala National Medical College and BYL Nair Charitable Hospital, Mumbai, Maharashtra, India

Introduction: Refractory cytopenias are common in renal diseases and require bone marrow examination. There can be variable fibrosis in the marrow of renal disease patients. We attempted to study the clinicopathological spectrum of renal disease cases with variable degree of marrow fibrosis. Objectives: To understand the pathophysiology of bone marrow fibrosis in kidney disease patients by studying the marrows and recording fibrosis grade in the background of clinical picture, hemogram and other investigations. Materials and Methods: We studied the trephine biopsies of 14 patients of renal disease with cytopenias. We tabulated the clinical picture, clinical diagnosis, hemogram findings, RFT, other investigations, fibrosis grade and the impression on trephine biopsies. Results: 11 cases had low HB ranging 2.7 g -10 g. 5 had leucopenia with TWBC < 4000/cumm.7 had platelets counts <1.5 L/cu mm, lowest 10,000/cumm, 5 had pancytopenia. Serum creatinine were raised ranging 1.7 mg% - 8.6 mg%. Clinical symptoms were variable with weakness, edema, breathlessness and other symptoms. Marrow findings were variable. Fibrosis was grade 1 in 7, grade 2 in 4, grade 3 in 2 and grade 4 in 1 case. Conclusion: Marrow fibrosis is associated with refractory cytopenias. Many CKD patients have mineral and bone disorders with elevated PTH levels. Secondary hyperparathyroidism can cause secondary myelofibrosis and refractory anemia in CKD patients. Trephine biopsy and fibrosis grading can be important for diagnosis and management of refractory cytopenias.

Keywords: Cytopenias, marrow fibrosis, renal diseases

   AOP158 Top

   Hematopathology Top

Expression of CD30 in diffuse large B-cell lymphoma by immunohistochemistry

Rashmi Gautam, Sarika Singh, Sandeep Garg, Anubhav Vindal, Meeta Singh

Maulana Azad Medical College, New Delhi, India

Introduction: Expression of CD30 in Diffuse large B-cell lymphoma (DLBCL) has better performance status in many studies. As this subset has a promising Immune therapy, it becomes more relevant to study CD30+ve DLBCL. The present study included 25 retrospective cases of DLBCL. Objectives: 1. To detect CD-30 positivity in patients with DLBCL using Immunohistochemistry (IHC). 2. To determine its prognostic significance and co-relate with other biomarker(s). Materials and Methods: IHC was put on all 25 cases for expression of CD30 and 24 cases for CD10, CD20, BCL-6, MUM-1, FOXP-1, CD38, BCL-2, MYC and LMP-1. 2-years follow-up in term of Overall survival (OS) and Progression free survival (PFS) was recorded. Results: CD30 (<0% cut-off) and (≥20% cut-off) was observed in 40% and 12.5% cases, respectively. Advanced stage presentation and CD30 positivity had significant correlation (p=0.03). Double expression of MYC and BCL-2 was observed in 45% cases and showed statistically significant expression of CD30 <0% cut-off (p-value= 0.028). LMP-1 positivity was seen in 12.5% cases. Significant correlation was exhibited between CD30 ≥20% cut-off positivity and LMP-1 expression (p-value <0.001). CD30+ve and CD30-ve cases did not show remarkable difference in outcome. Significantly poorer OS was seen in patients of double expresser DLBCL with CD30-ve, when compared to CD30+ve (p= 0.031). Significantly worse outcome was observed with conjoint expression of LMP-1 and CD30 (p= 0.032). Conclusion: It is recommended to investigate the patient of DLBCL for CD30 immunoexpression along with status of EBV-infectivity and double expression of MYC and BCL-2. CD30 cannot be an independent biomarker for prognostication of DLBCL.

Keywords: CD30, double expresser diffuse large B-cell lymphoma, LMP-1

   AOP159 Top

   Hematopathology Top

Evaluation of acquired hemoglobinopathies in leukemias

Swati Tyagi, Anshul Singh, Vatsala Misra

Department of Pathology, Moti Lal Nehru Medical College, Prayagraj, Uttar Pradesh, India

Introduction: Leukemias constitute 3% of the global cancer burden and presents with myriad of symptoms including anemia. Although hemoglobinopathies are nearly always inherited, occasionally persons with previously normal hematologic function develop aberrant hemoglobin synthesis as an acquired abnormality, within the context of hematologic malignancies. Objectives: To study the association between Acquired Hemoglobinopathies and Leukemias. Materials and Methods: The subjects were newly/previously diagnosed cases of Leukemias. Patients who were diagnosed with congenital hemoglobinopathies or those already on treatment were excluded. Complete work3 up included CBC, supra vital staining and histochemical stains (Sudan Black B and Periodic Acid Schiff). All cases were subjected to HPLC (High Performance Liquid Chromatography) using BIO RAD 'VARIANT'. Patients presenting with Leukemoid reaction were taken as controls. Results: We included 8 age and sex matched controls and 34 cases. The mean age of cases was 33.8 years with a M:F of 1.2: 1. 15 cases were of ALL, 8 cases of AML, 9 cases of CML and 2 cases of CLL. Out of the 23 cases of Acute Leukemias, 13 cases showed evidence of beta thalassemia and 3 cases of alpha thalassemia whereas rest of the 7 had normal electrophoretic findings. None of the cases of chronic leukemias or controls had abnormal electrophoretic findings. The mean values of HbA2 and HbF were significantly different in acute leukemias compared to the controls. Conclusion: Acute haematological malignancies showed a significant predisposition towards development of acquired hemoglobinopathies especially beta thalassemia as compared to chronic ones. Hence, all such cases should also be screened for hemoglobinopathies.

Keywords: Haematological malignancy, haemoglobin variants, thalassemia

   AOP160 Top

   Hematopathology Top

Evaluation of pancytopenia through hematological parameters and bone marrow studies

Pratima R Aradhyamath, S Rukmini, Rajesh H Chandan

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Pancytopenia is a relatively common hematological entity. It is a striking feature of many serious and life threatening illnesses, ranging from simple drug-induced bone marrow hypoplasia, megaloblastic anemia to fatal bone marrow aplasia and leukemias. The severity of pancytopenia and the underlying pathology determine the management and prognosis. Objectives: 1. To study the bone marrow morphology in cases of pancytopenia. 2. To study the etiological factors of pancytopenia. Materials and Methods: Total 207 pancytopenic patients were evaluated clinically, along with hematological parameters and bone marrow aspiration in Hematology Unit, Department of Pathology, KIMS HUBLI, during the period of January 2020 to June 2022. Results: Among 207 cases studied, Most of the patients were in the age group of 21-30 years (22.2%) with male to female ratio 2.18:1. Most of the patients presented with generalized weakness (72.9%) and fever (39.6%). The commonest physical finding was pallor (97.2%) followed by edema (23.6%) and splenomegaly (20.28%). Megaloblastic anemia was the predominant blood picture. The commonest marrow finding was hypercellularity with megaloblastic erythropoiesis. The commonest cause for pancytopenia was Megaloblastic Anemia (40.57%), followed by Dimorphic Anemia (28.98%). Conclusion: Bone marrow study with good clinical correlation is very important to study the causes of pancytopenia and planning further investigations and also to determine the management and prognosis of the patients. However, in view of a wide array of etiologies, pancytopenia continues to be a challenge for hematologists.

Keywords: Bone marrow aspiration, megaloblastic anemia, pancytopenia

   AOP161 Top

   Hematopathology Top

Comparative study of hemostatic profile between conventional methods and 'Sonoclot coagulation and platelet functional analyzer' in patients with septicemia

Fairooz Musthafa, Vani, Akshay

JSS Academy of Higher Education and Research, Mysuru, Karnataka, India

Introduction: Comparative study of hemostatic profile between conventional methods and 'Sonoclot coagulation and platelet functional analyzer' in patients with septicemia. Objectives: Primary Objective: To compare hemostatic Status (Normal or Deranged) of patients with septicaemia, between Sonoclot coagulation and platelet functional analyser and conventional hemostatic parameters. Secondary Objective: To correlate the activated clotting time on Sonoclot with Prothrombin and Activated thromboplastin time. Materials and Methods: It is a prospective analytical study conducted on septic patients admitted to ICU. Conventional hemostatic tests were compared with the results of Sonoclot analysis. Results: 61 subjects were studied. 45.9% were females. Median (IQR) PT was 15.5 (13.3 – 19) secs, INR was 1.18 (1.0 – 1.5), APTT was 35.7 (30.2 – 42.4) secs. 45.9% had abnormal INR and 29.5% had prolonged APTT. Overall, 52.5% had prolonged coagulation. Median (IQR) platelet count was 2.06 (1.07 – 2.99) Lakh/cumm. 32.8% had thrombocytopenia. Median (IQR) Activated clot time on Sonoclot was 104 (80.2 – 129.5) secs. ACT was abnormal in 29.5% of our subjects. Median (IQR) clot rate was 53 (43.3 – 70). 91.8% showed hypercoagulability. Median (IQR) Platelet function was 3.9 (2.9 – 4.4). PF abnormality was seen in 90.2%. 62% cases showed graph abnormality with normal platelet count. There was good correlation between the hemostatic abnormality detected by Sonoclot and that of conventional tests (Cramer V (Phi) of 70.7%, P = 0.000). Conclusion: Sonoclot analysis will identify significantly higher proportion hemostatic abnormality in septic patients compared to conventional tests.

Keywords: Coagulation profile, sepsis, sonoclot

   AOP162 Top

   Hematopathology Top

Thalassemia – The dawning of a new era

Gauri Dadich, P Pallavi

JSS Academy of Higher Education and Research, Mysuru, Karnataka, India

Introduction: Thalassemia is an autosomal recessive disorder characterized by decreased synthesis of one or more hemoglobin polypeptide chains. In clinical practice, beta thalassemia has been categorized as either Transfusion Dependent Beta Thalassemia or Non Transfusion Dependent Beta Thalassemia. The major goals of transfusion therapy are: • Use of donor erythrocytes with an optimal recovery and half-life in the recipient. • Achievement of appropriate hemoglobin level. (To maintain pre-transfusion hemoglobin level above 9-10.5 g/dl.)• Avoidance of adverse reactions, including the transmission of infectious agents. Objectives: To evaluate the transfusion programme in our institute through evaluating: Pre – transfusion haemoglobin, Ferritin levels, Alloimmunization and Autoimmunization. Materials and Methods: An exploratory study was carried out among 70 patients over 18 months. Patients who met clinical and hematological criteria, were confirmed by gel electrophoresis, and enrolled in a regular blood transfusion program were considered cases. 3 ml of whole blood was sampled and analyzed using Mindray BC 3000. Serum ferritin was assayed using Two site Sandwich immunoassay. The detection of Auto-antibody was performed by DCT and Allo-antibody by ICT. The Data collected was analyzed using SPSS version 26.0. Association and relationship between the hemoglobin, Ferritin level and antibodies with transfusion was assessed by Chi Square and T-test. Results: The average hemoglobin value ranged from 8 gms/dl to 9 gms/dl. Ferritin levels were above 1100 micrograms/L in 90% of the patients and none showed auto-antibodies and allo-antibodies. Conclusion: The mean hemoglobin in the patients was below the recommended reference value. Ferritin was elevated in all patients with alloantibodies and autoantibodies being negative. These patients due to repetitive transfusions are more prone to iron overload, which leads to cardiac dysfunction.

Keywords: Ferritin, hemoglobin, thalassemia

   AOP163 Top

   Hematopathology Top

Clinicopathological evaluation of anaemia in pregnant women

Sumanlata Sahu, Deepa Rani

Institute of Medical Science, Banaras Hindu University, Varanasi, Uttar Pradesh, India

Introduction: Anaemia in pregnancy is a serious public health problem worldwide. The aim of this study was to study the complete clinicohaematological pattern of anaemia in pregnant women and assess its effect during pregnancy. Objectives: To study the complete haematological profile of pregnant women, to categorize the type of anaemia during the different trimester and assess its effect on pregnancy. Materials and Methods: A prospective observational study was done in Sunderlal Hospital, BHU. In this study we studied the clinical and haematological profile of 100 pregnant women. The laboratory tests include CBC, PBS, reticulocyte count, iron profile and HPLC along with their complete clinical and obstetric history. Results: In our study conducted the mean haemoglobin was of 8.1 gm/dl (range 2.5-11 gm/dl). 20% had mild anaemia, 61% had moderate anaemia, 19% had severe anaemia. Morphologically most common type of anaemia prevalent in all the trimester was Microcytic Hypochromic anaemia. In HPLC, we found 3 cases of thalassaemic trait. Conclusion: This study concludes that, good socio economical status, good antenatal care, good nutrition, birth spacing, complete clinical and haematological examination is required to improve the health during pregnancy as well as pregnancy outcome.

Keywords: Anaemia, haemoglobin, pregnancy

   AOP164 Top

   Hematopathology Top

Morphologic changes in circulating blood cells of COVID-19 patients

N Nilopher, Sridhar

Ramaiah Medical College, Bengaluru, Karnataka, India

Introduction: Since the beginning of the coronavirus disease 2019 (COVID-19) pandemic, many studies have described the quantitative peripheral blood findings seen in COVID-19 patients. However, morphologic changes have been described by only a few studies. We report morphologic changes in peripheral blood of COVID-19 patients. Objectives: The aim of the present study was to investigate the morphological changes affecting all peripheral blood cells of COVID-19 patients that could help in the early diagnosis and/or prognosis. Materials and Methods: The peripheral blood smears of all 200 patients were retrieved and morphological features of white blood cells, red blood cells, and platelets were reviewed and documented. Appropriate pictures were taken. Results: Of the 200 patients reviewed, 120 were males and 80 were females. The most common quantitative haematological abnormalities noted on complete blood count (CBC) were anaemia followed by neutrophilia, neutrophilic left shift, and lymphopenia. The most significant morphologic changes noted were neutrophils with clumped chromatin, multiple abnormal nuclear shapes, pseudo-Pelger-Huet deformity, and smudged neutrophils. Lymphocytes showed abundant blue cytoplasm and/or lymphoplasmacytoid morphology and dysmorphic monocytes. Platelet clumping was the most significant finding noted. The red blood cells were normocytic and normochromic with few nucleated red blood cells. Conclusion: Our study identifies and describes significant morphologic changes in the peripheral blood cells of COVID19 patients. An understanding of these morphologic changes along with established hematologic parameters can aid in the diagnosis of COVID-19. The prominent morphological changes can be predictive of undesirable outcomes.

Keywords: COVID-19, morphological changes, peripheral blood smear

   AOP165 Top

   Hematopathology Top

Reed Sternberg-like cells in non-Hodgkin lymphomas: A case series

Ruovinuo Sachu, Geeta Yadav, Anurag Singh, Mili Jain, Rashmi Kushwaha, S P Verma

King George's Medical University, Lucknow, Uttar Pradesh, India

Introduction: Reed Sternberg cells are characteristic of Classical Hodgkin Lymphoma. However, large cells with similar morphologic and immunophenotypic features are occasionally seen in T cell and B cell Non-Hodgkin Lymphomas (NHL), known as Reed Sternberg-like cells (RSLC). Objectives: To study the diagnostic difficulty in NHL having RSLC. Materials and Methods: Lymph node biopsy of four patients who presented with multiple lymphadenopathies, and underwent excisional biopsy were studied which showed the presence of RSLC in the lymph node biopsy. Detailed Immunohistochemistry was done in all four cases. Results: Lymph node biopsy of all cases showed the presence of large pleomorphic mononucleate and multinucleate cells with prominent nucleoli which on IHC were positive for CD30 (strong) and PAX5 (weak). The background population was composed of medium to large-sized atypical lymphoid cells with round nuclei, vesicular chromatin, 1-2 prominent nucleoli, and scant to moderate amount of cytoplasm which was positive for CD20 and PAX-5, unlike Hodgkin lymphoma. Of four cases, one was diagnosed as small lymphocytic lymphoma and the rest three were diagnosed as diffuse large B cell lymphoma with RSLC. Conclusion: RSLC with similar morphologic and immunophenotypic profiles can also be seen in Non-Hodgkin Lymphomas necessitating the meticulous morphological and immunohistochemical approaches to these lymphomas in order to differentiate them from Hodgkin lymphoma.

Keywords: Lymphoma, Reed Sternberg like cells

   AOP166 Top

   Hematopathology Top

Acquired coagulation abnormalities in patients of plasma cell myeloma

Safaa Tasneem, Dilleswari Pradhan, Lity Mohanty, Manmohan Biswal

S C B Medical College and Hospital, Cuttack, Odisha, India

Introduction: Coagulation abnormalities are the virtue of the pathophysiology of the Plasma Cell Myeloma or due to the treatment modalities for plasma cell myeloma. Acquired coagulation abnormalities have been reported in few scattered studies. This study is an attempt to find the proportion of various acquired coagulation abnormalities in the patients with Plasma Cell Myeloma. Objectives: 1. To investigate the abnormalities of coagulation factors in patients of Plasma Cell Myeloma. 2. To compare the observed acquired coagulation abnormalities in patients of Plasma Cell Myeloma with healthy controls. 3. To evaluate the risk of thrombosis and bleeding and give a message to the treating physician for the prognosis of the disease. Materials and Methods: An observational study was conducted on 50 patients with Multiple Myeloma attending the out-patient department of a tertiary care hospital in Orissa. We collected data on clinical profile, coagulation profile and routine investigations of the patients of Multiple Myeloma and compared with 50 healthy controls. Coagulation profile included factor V assay, factor VIII assay, serum Fibrinogen levels, D-dimer levels, partial thromboplastin time (aPTT) and prothrombin time (PT). Results: The proportion of acquired coagulation abnormalities were 24% with anaemia 24% had raised aPTT, 20% had raised PT, about 10% had factor V deficiency, 14% had factor VIII deficiency and 10% had hyperfibrinogenaemia. Conclusion: Our study indicates that various coagulation abnormalities occur in patients with multiple myeloma, which may contribute to the increased the risk of venous thromboembolism and bleeding which is observed in these patients.

Keywords: Acquired, coagulation abnormalities, plasma cell myeloma

   AOP167 Top

   Hematopathology Top

To evaluate hematological indices in chronic liver disease in adults

Kiranpreet Kaur, Navjot Kaur, Vijay Mehra, Permeet Kaur Bagga

Government Medical College, Amritsar, Punjab, India

Introduction: Chronic liver diseases is characterized by decreased hepatic function as a result of chronic inflammation or insult to the liver for more than 6 months. It is one of the major causes of death the western countries is now the fifth leading cause of early loss of life. Abnormalities in hematological parameters are common in patients with cirrhosis. Two important markers for liver fibrosis are AST-to-platelet ratio index (APRI) and Fibrosis-4 (FIB4). Objectives: To evaluate the haematological indices and type of anemia in chronic liver disease patient. To evaluate liver function test in cld patient and to calculate marker of liver fibrosis and to correlate it with fibroscan. Materials and Methods: This prospective study was carried out in the department of pathology, Government Medical College, Amritsar, from April 2021 to June 2022. The written informed consent was taken from the patient who presented with sign and symptoms of chronic liver disease. They had first undergone complete blood count, liver function test and fibroscan if needed. Results: Most common anemia in cirrhosis is normocytic normochromic. Macrocytic anemia and thrombocytopenia is common with alcoholics. FIB 4 and APRI index was calculated using haematological indices and was correlated with fibroscan. Patient with increased score value had increased liver stiffness measurements on fibroscan above 12.5 kPa. Conclusion: Early evaluation and management of these easily available parameters and non- invasive scoring system may be more appropriate in prevention and development of complications among chronic liver disease patient.

Keywords: Chronic liver disease, fibrosis, hematological indices

   AOP168 Top

   Hematopathology Top

A comparative study of INR values between automatic (ACL elite pro) and semiautomatic (ERBA) coagulometers

Ruchir Uttam, Prakash Patil

Shri B M Patil Medical College, Vijayapura, Karnataka, India

Introduction: INR is the preferred test of choice for patients pre-operatively and also to assess the risk of bleeding or coagulation status of patients. The INR is derived from prothrombin time (PT) which is calculated by ratio of patients PT to control PT standardized for the potency of the thromboplastin reagent. Objectives: To compare INR values between automatic (ACL Elite Pro) and semiautomatic (ERBA) coagulometers. Materials and Methods: This prospective hospital based study of 250 samples is conducted in central laboratory of Shri B. M. Patil Medical College, Vijayapura for testing of PT, aPTT and INR. Blood sample is collected and is processed in respective coagulometers for the value of PT, aPTT and INR. Results: The baseline PT, INR and APTT value in (Automatic) coagulometer was found to be 14.51 ± 5.29 seconds, 1.22 ± 0.47 seconds and 29.47 ± 8.30 respectively whereas, semiautomatic instrument was 13.5 ± 4.60 seconds, 1.15 ± 0.46 seconds and 27 ± 6.96 seconds respectively. Conclusion: Study showed good agreement between automated and semi-automated instruments based on mechanical endpoint detection method and photo-optical endpoint detection method for assessing coagulation test including PT, aPTT and INR values. Efficiency wise, automatic coagulometer (ACL COAGULOMETER) provides slight edge over semi-automated coagulometer (ERBA).

Keywords: Coagulometer, INR value, semiautomatic

   AOP169 Top

   Hematopathology Top

Pattern of thalassemia and other hemoglobinopathies – A hospital based correlative study of HPLC retention time and red blood cell parameters

Prafulkumar Koradhanyamath, B P Bommanahalli

Gadag Institute of Medical Sciences, Gadag, Karnataka, India

Introduction: Hemoglobinopathies are most common monogenic hereditary disorders. β-thalassemias and their interaction with hemoglobin E (Hb E) and hemoglobin S (Hb S) are a considerable health problem in India. The data on prevalence of thalassemias and hemoglobinopathies in different caste/ethnic groups in the region of north Karnataka is scarce or nonexistent. Objectives: To evaluate the spectrum and propotion of thalassemia and other hemoglobinopathies. To assess predictive value of red cell indices and discriminant indices in identification of thalassemia and hemoglobinopathies. Materials and Methods: A Hospital based cross sectional study from December 2020 to august 2022 in a tertiary care center. Universal sampling of all anemia patients as per who criteria, with a sample size of 1068. All the samples will be screened for hemoglobin disorders by HPLC (High Performance Liquid Chromatography). Discriminant Indices like MENTZER INDEX, SIRDAH INDEX, ESHNAI INDEX, and ENGLAND AND FRASER INDEX etc will be calculated using the red cell indices. Results: Proportion of thalassemia and other hemoglobinopathies was found to be 4.45% among anemia patients that is 48 cases among 1068 anemia patients. Cases of β-thalassemia trait (BTT) – 26, sickle cell trait – 11, Hb D heterozygous – 5, HbE Heterozygous – 2, Delta BTT / Heriditary persistence of Foetal hemoglobin (HPFH) Trait – 3, Sickle cell anemia – 2, Double heterozygus for HbS and BTT – 1, others – 1. Shine and lal index had highest sensitivity of 96.1%. England and fraser had highest specificity 95.0%. Conclusion: The discriminant indices can serve as effective screening too for β-thalassemia trait in resource limited areas.

Keywords: Beta thalassemia, discriminant indices, hemoglobinopathies

   AOP170 Top

   Hematopathology Top

Predictive value of neutrophil CD64 in patients of sepsis from adult ICU

Rohan Waykole, Amit Nisal, Anu Christopher, Ravindra Nimbargi

Bharati Vidyapeeth (Deemed to be University) Medical College Hospital and Research Centre, Pune, Maharashtra, India

Introduction: Sepsis is a common cause of morbidity and mortality in both paediatric as well as adult patients admitted in intensive care units (ICU). Neutrophil CD64 (nCD64) expression corresponds to inflammatory responses during infection or tissue injury. Expression of CD64 in neutrophils is up-regulated under the influence of inflammatory-related cytokines such as interleukin 12, interferon gamma, and granulocyte colony-stimulating factor. Neutrophil CD64 is a highly sensitive and specific marker for systemic infection and sepsis. In this study we intend to monitor the expression of CD64 in neutrophils as a prognostic marker for sepsis. Objectives: To know the predictive value of nCD64 in patients of sepsis from adult ICU and its correlation with clinical and other biochemical parameters traditionally used for diagnosis of sepsis. Materials and Methods: rospective analytical study was conducted in Bharati Hospital, Pune for a period of two years. A total of ninety-one sepsis cases from the adult intensive care unit (ICU) were included. Routine laboratory parameters like complete blood count, PT, APTT, Fibrinogen, CRP and Procalcitonin were included and presented in a tabulated format. Expression of neutrophil CD64 was analysed on Beckman Coulter Navios flow cytometer. MFI was calculated for these cases. Appropriate statistical analysis was done. Results: MFI (Median fluorescence intensity) scores were higher in diagnosed cases of sepsis based on clinical and other biochemical parameters. The increase and decrease in MFI scores on day 4 correlated with the other clinical and biochemical parameters. Conclusion: nCD64 result is an independent prognostic factor and can be used as an alternative for other biochemical parameters in predicting the outcome of sepsis patients in ICU.

Keywords: Median fluorescence intensity, neutrophil CD64, sepsis

   AOP171 Top

   Hematopathology Top

Modified Leishman stain – A new insight in the peripheral smear staining

B E Chaitra, Mohit Kumar, K P Athira

HIMS, Dehradun, Uttarakhand, India

Introduction: Leishman stain (LS) modified Romanowski stain, routinely used in hematology fails to give a rapid diagnosis in medical emergencies. In this Era of rapid diagnostic methods, there is a need for a stain which reduces the turnaround time without compromising the quality of the peripheral smears. Modified leishman stain (MLS), a modification in LS where PHENOL an accentuating agent is added to reduce the staining process to 4 mins. Field's stain (FS) is a rapid version of Romanowski stain where smears are stained with in 1 min. This study aimed to asses and compares the staining quality of MLS, LS and FS. Objectives: This study was performed to compare the staining quality of MLS, LS and FS on peripheral smears. Materials and Methods: 3 set of peripheral smears were made from each blood sample, one for MLS, LS and FS respectively A Total of 300 smears were studied and scored based on five parameters that is overall staining, cytoplasmic and nuclear staining of leucocytes, RBC staining and platelet staining. The results were recorded, statistical analysis (ANOVA) was done using software SPSS20. Results: It is observed that MLS showed a significant high average score compared to LS and FS (P value - 0.0001). However there was no statistical significance in the staining quality between FS and LS (P value - 0.2). Conclusion: MLS showed excellent staining results in 4 mins. MLS stand out as a great alternative to LS in a medical emergencies as a rapid diagnostic methods.

Keywords: Field's stain, modified leishman stain, phenol

   AOP172 Top

   Hematopathology Top

High CXCR4 expression predicts extramedullary disease in B-ALL

Tina James

Kodagu Institute of Medical Sciences, Madikeri, Karnataka, India

Introduction: Acute lymphoblastic leukemia (ALL) can infiltrate the liver, spleen, lymph nodes and brain. One of the mechanisms proposed for extramedullary infiltration in leukemia is the interaction between chemokine receptor CXCR4 and its ligand CXCL12. Overexpression of CXCR4 causes trafficking of leukemia cells to extramedullary organs, where CXCL12 is constitutively produced by stromal cells. Objectives: To correlate CXCR4 expression on blasts with extramedullary organ infiltration, and with prognosis in B-ALL. Materials and Methods: 106 adult and pediatric patients diagnosed with B-ALL by bone marrow aspirate examination and flowcytometry were included in this prospective study. After diagnosis, patients received chemotherapy and were followed up for up to two years for hematological remission status and clinical outcome. CXCR4 expression was analysed by flowcytometry and compared with different clinical presentations and with outcome. Results: CXCR4 expression of varying median fluorescence intensity (MFI) and percentage positivity was noted in all 106 cases. Both these parameters significantly correlated with the degree of splenomegaly. Among the various clinicopathological parameters related to outcome, age at diagnosis, degree of hepatomegaly & splenomegaly, and CD 10 & CD 5 expression were all significantly related to event free survival and overall survival. Conclusion: This study has examined the clinicopathological, hematological, biochemical, immunophenotypic & cytogenetic characteristics of B-ALL and has assessed their impact on prognosis. CXCR4 expression on blasts was conclusively shown to correlate with extramedullary organ infiltration in B-ALL, which is known to affect prognosis. CXCR4 is therefore a potential new prognostic marker in B-ALL.

Keywords: B-ALL, CXCR4, prognosis

   AOP173 Top

   Hematopathology Top

Cytogenetic evaluation of Burkitt type translocations: A 5 years study at regional cancer center

R Subhan Ali, B L Kavitha, S Shanthala

Department of Pathology, Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Burkitt lymphoma (BL) is a high grade B-cell non-Hodgkin lymphoma (NHL) occurring in children and adults. The t (8;14)(q24.1;q32) and its variants – the t (2;8)(p12;q24.1) and t (8;22)(q24.1;q11.2) are associated with B-cell neoplasia and result in MYC/Immunoglobulin (IG) gene rearrangement which drive the oncogenesis. Objectives: 1. To study the frequency and clinic-pathological features of Burkitt type translocations found in hematolymphoid malignancies. 2. To study the additional cytogenetic findings in these cases. Materials and Methods: All hematolymphoid cases showing Burkitt type translocations diagnosed from 2016 and 2021 were included in this study. Clinical details were collected from archived case files. Peripheral blood, bone marrow smears, histopathology and IHC slides were reviewed. Bone marrow flowcytometry and cytogenetic analysis was done. Results: This study included 55 cases showing Burkitt type translocations, comprising of 43 males and 12 females. We had 33 cases of Burkitt lymphoma (60%), 18 cases of Pre B – ALL (32.7%), three cases of Multiple Myeloma (5.4%) and one case of CLL (1.8%). Majority of the cases presented with fever (43.6%) followed by abdominal pain and swelling (41.8%). Conventional karyotyping revealed t (8;14)(q24;q32) in 52 (94.5%) cases, t (8;22)(q24;q11) in 2 (3.6%) cases and t (2;8)(p12;q24) in one case. Complex karyotype and additional abnormalities were seen in 12.7% and 47.2% cases respectively. Conclusion: Burkitt lymphoma is an aggressive disease, but it has high curable rates. The frequency and distribution of Burkitt type translocations in our series and association with 1q and 6q abnormalities is similar to the literature. Patient with 8q24 abnormalities and complex karyotype carry poor prognosis.

Keywords: 8q24, Burkitt lymphoma/leukemia, cytogenetic

   AOP174 Top

   Hematopathology Top

Evaluation of a spectrum of platelet function disorders diagnosed on a coagulation analyser platform

Eliz Thomas, Prasanna N Kumar

Department of Pathology, PSG Institute of Medical Sciences and Research, Coimbatore, Tamil Nadu, India

Introduction: Platelet disorders comprise a heterogeneous group of rare diseases caused by acquired factors or molecular anomalies in genes that are relevant in platelet formation and/or function. The clinical complications are highly variable, ranging from almost negligible to life-threatening. Early and accurate diagnosis and close medical follow-up is long known to be of great importance. The most widely used test to assess platelet function is by using the principle of light transmission aggregometry (LTA). However, using a dedicated aggregometer is time consuming and labor intensive. This study utilized automated LTA method using a coagulation analyzer. Objectives: To study the spectrum of platelet function abnormalities assessed in our laboratory over a period of 3 years. Materials and Methods: Platelet rich plasma was prepared from venous blood collected in 3.2% citrate tube and the platelet function tests (PFT) were carried out in Sysmex coagulation analyzer CS-2400. Results: A total of 33 cases with abnormal PFT were analysed in our set up. All the patients presented with complaints ranging from ecchymoses, epistaxis, easy bruising, gum bleeding, epistaxis, hemarthrosis and vaginal haematoma. The most common acquired condition was drug-induced thrombocytopathia. Rare inherited disorders like Bernard-Soulier syndrome and Glanzmann thrombasthenia were also diagnosed. Conclusion: It is recommended that PFT be added onto the investigative profile for patients who present with abnormal bleeding and/or unexpected thrombocytopenia. The CS-2400 analyser is a cost- efficient standardized analyser to assess platelet function even when the platelet count is low. It requires a shorter turnaround time for samples and also has the advantage of being a walk-away technology.

Keywords: Platelet function disorders, platelet tests, thrombocytopathias

   AOP175 Top

   Hematopathology Top

Assessment of haemostatic abnormalities in patients of chronic kidney disease

Anshu Palta, Aprajita, Anita Tahlan, Pooja Sharma, Sanjay D'Cruz

GMCH-32, Chandigarh, India

Introduction: Chronic Kidney Disease (CKD) is a global health problem with a prevalence of approximately 8-16% affecting more than 300 million people worldwide. CKD has also been found to be associated with various haemostatic abnormalities due to derangement in all three aspects of haemostasis. Objectives: (1) To evaluate haemostatic abnormalities in patients of CKD. (2) To evaluate and correlate haemostatic parameters with severity of CKD. Materials and Methods: Blood samples obtained from 125 diagnosed CKD patients and 25 healthy controls were analysed on automated haematology analyser (Beckman LH-780) and automated coagulation analyser (ACL TOP TM 500 CTS). Results: The mean age of the study population was 52.98 ± 15.10 years. Anaemia was noted in 103 (82.4%) patients. Thrombocytopenia and thrombocytosis were detected in 37 (29.6%) and 6 (4.8%) patients respectively. vWF:Ag and vWF:RCo levels were statistically significantly increased in CKD patients. No significant difference was found for vWF:Ag/vWF:RCo with worsening of the disease. PT and APTT were increased in 23 (18.4%) and 15 (12%) patients of CKD respectively (p- value=0.02 & <0.01). Fibrinogen levels were significantly increased in stage V of CKD patients as compared to stage I (p-value=0.04). D-Dimer was significantly increased in 106 (84.8%) as compared to control group (p-value <0.01). Factor V, VII and VIII were increased in 17 (13.6%), 41 (32.8%) and 41 (32.8%) CKD patients respectively. Levels of antithrombin, protein C and protein S were found to be decreased in 30 (24%), 15 (12%) and 45 (36%) patients respectively. Conclusion: The above data suggests that CKD is hypercoagulable state and coagulopathy is multifactorial.

Keywords: Chronic kidney disease, haemostasis

   AOP176 Top

   Hematopathology Top

Acute myeloid leukemia: Comparing French-American-British classification, immunophenotype and cytogenetics

Mekhala Rao, Girish Kamat, Deepak Goni, Girish Balikai

Introduction: Acute myeloid leukemia (AML) is a heterogeneous disease as it affects multiple lineages of hematopoietic cells. Objectives: (1) To correlate the subtypes of AML according to French–American–British (FAB) classification with immunophenotypic findings, (2) Comparing subtypes of AML according to FAB classification with findings of genetic studies, and (3) Correlate the immunophenotypic findings in AML patients with genetic studies. Materials and Methods: The study is a retrospective cross sectional study. 73 patients with a final diagnosis of AML, whose immunophenotyping and/or cytogenetic study results were available, were included in the study. Patients diagnosed between January 2017 and December 2020 were included in the present study. Results: 21 (31.81%) out of 66 patients with AML aberrantly expressed lymphoid antigens. The lymphoid antigens expressed were CD7, CD19, TdT, and CD5 which were found in 13 (19.69%), 9 (13.6%), 2 (3%), and 1 (1.51%) patient, respectively. Two out of three patients with t (8;21)(q22;q22) had CD19 aberrant expression. This association was found to be statistically significant with Fisher exact test, with a statistic value of 0.0277 (P < 0.05). Co expression of two lymphoid antigens such as CD7 and CD19 was associated with monosomy 7 and was found to be statistically significant with a Fisher exact test, with a statistic value of 0.0217 (P < 0.05). In our study, t (8;21) (q22;q22) was found in AML M2 and AML M1. Conclusion: This study highlights the importance of morphological, immunophenotypic, and cytogenetic evaluations in the diagnosis of AML.

Keywords: Acute myeloid leukemia, cytogenetics, immunophenotype

   AOP177 Top

   Hepatobiliary and Pancreatic Pathology Top

Prognostic significance of CD24, SOX2 and tumor budding in gall bladder cancer

Devanshi Dubey, Preeti Agarwal, Ajay Kumar Singh, Malti Maurya, Akshay Anand, Sameer Gupta

King George Medical University, Lucknow, Uttar Pradesh, India

Introduction: Risk stratification of gallbladder cancer (GBC) is based on grade, stage, tumor size etc. The current work was carried out to study GBC in our region and to explore CD24, SOX2 by immunohistochemistry and tumor budding morphologically. CD24 is an immunomodulator molecule involved in interference with mRNA stability and prevention of tumor cell death. SOX 2 is a stem cell marker which identifies the cases which may fare well with chemotherapy. Objectives: To study the immunohistochemical expression (intensity and percentage area) of CD24 and SOX 2 along with tumor budding in different grade and stages of gall bladder cancer and correlate its expression with clinico-pathological parameters. Materials and Methods: Detailed histomorphological and immunohistochemical study of 83 resection specimens of GBC. Results: Mean age of presentation 53 years ± SD with female predominance. Significantly poor outcome was seen in cancers with higher grade, larger tumor size, high total and direct bilirubin levels and nodal metastasis. Though not significant but patients with tumor budding of score 3 fared poorly when compared to score 1 and 2 (47% Vs 23% and 25%). CD24 expression was seen in 78.31 % (65/83) of the studied cases. Its expression was not significantly related to survival. SOX2 expression was seen in only one case. Conclusion: The above study supports addition of tumor budding to gall bladder cancer may provide with additional risk stratification information. CD24 is expressed in significant percentage of GBC and can be further explored in terms of immunotherapy target. SOX2 expression is insignificant in GBC.

Keywords: CD24, prognostic significance, tumor budding

   AOP178 Top

   Hepatobiliary and Pancreatic Pathology Top

Spectrum of drug induced liver injury – A histomorphological study in a tertiary care centre

P G Dhanya, Pushpa Mahadevan

VPS Lakeshore Hospital, Kochi, Kerala, India

Introduction: A wide range of drugs are attributed towards causing liver injury. Drug induced liver injury (DILI) account for 10% of acute hepatitis and is perhaps the most common cause of cholestatic hepatitis. Objectives: 1) Study histomorphological patterns of DILI 2) Correlation of morphological patterns with commonly implicated drugs. Materials and Methods: This is a four year retrospective descriptive study of patients with biopsy proven liver injury following drug intake. 48 patients with clinical history of drug usage were included. Causality evaluation was completed in 10 (20%) cases with final assessment of probable in 9 (90%) of these cases and highly probable in one (10%) case. Liver biopsy slides were retrieved, studied and categorized into different histological patterns. Results: There were 29 males and 19 females, with an age range of 7-75 years. Most common implicated agents was complementary and alternative medicines (CAM)(47.9%) followed by immunosuppressants (10.4%). Other drugs included anticonvulsants (4.1%), antibiotics (4.1%), NSAIDS (4.1%) and chemotherapeutic drugs (4.1%). Most common histological patterns were cholestatic hepatitis (35.4%) and acute hepatitis (27.1%) respectively. Chronic hepatitis, steatohepatitis, and vascular injury were other patterns encountered. 42% of patients on CAM presented with acute hepatitis. Conclusion: Cholestatic hepatitis and acute hepatitis accounted for most common patterns of injury. DILI can mimic any other liver disease histomorphologically. Hence, history of drug intake and other supportive laboratory parameters are essential to arrive at a conclusive diagnosis.

Keywords: Cholestatic hepatitis, drug induced liver injury

   AOP179 Top

   Hepatobiliary and Pancreatic Pathology Top

Analysis of Whipple's resection specimens: A histopathological study

Yash Saxena, Sudha Iyengar, Reema Bhushan, Rajesh Gaur

Gajra Raja Medical College, Gwalior, Madhya Pradesh, India

Introduction: Pancreaticoduodenectomy, often referred as Whipple procedure, is the most common surgery performed to treat pancreatic cancer's. Periampullary carcinomas are described as tumors arising within 2 cm of the major papilla in the duodenum. These tumors are of four different types: ampullary carcinoma, biliary carcinomas (intrapancreatic distal bile duct), pancreatic (head–uncinate process), and duodenal (mainly from the second part). Objectives: The aim of this study was to analyze the histopathological features of Whipple-resected specimen received at Gajra Raja Medical College, Gwalior. Materials and Methods: A retrospective study was done of 22 cases of Whipple resections received during 2020-22. All relevant histopathological details pertaining to gross, microscopic diagnosis, grading, staging were comprehensively studied. Reporting was done in accordance to CAP protocol. Results: Out of 22 patients 12 (54.54%) were males and 10 (45.45%) females. Ratio was 1.2:1. Mean age was 58.4 years. Most common site of tumour was periampullary, 14 [63.6%] followed by pancreatic head, 4[18.18%]; distal common bile duct, 3 [13.6%]; duodenum 1 [4.5%]. Most common histological type was adenocarcinoma, 21 [95.45%]. Lymph node was positive in 5 cases [22.72%]. Lymphovascular invasion was seen in 5 cases (22.72%) & Perineural invasion was seen in 6 [27.27%] cases. Conclusion: The current analysis shows that periampullary adenocarcinoma is the most common subtype.

Keywords: CAP protocol, periampullary adenocarcinoma, Whipples's procedure

   AOP180 Top

   Hepatobiliary and Pancreatic Pathology Top

Epithelial neoplasms of the gall bladder: A 10-year study with emphasis on the rarer subtypes

Suyog Ratnaparkhi, Rachana Chaturvedi, Manjusha Karegar

Seth GSMC and KEM Hospital, Mumbai, Maharashtra, India

Introduction: The incidence of gall bladder neoplasms is higher in India in comparison to western world, with wide geographic variation. Epithelial neoplasms are more common, of which benign include pyloric gland adenoma, biliary intraepithelial neoplasia (BilIN), intra-cholecystic papillary neoplasm (ICPN) and mucinous cystic neoplasm. Adenocarcinoma (pancreaticobiliary type) is the commonest malignant neoplasm. Rarer histological sub-types include, variants of adenocarcinoma, adenosquamous carcinoma, squamous cell carcinoma and other differentiation patterns. Objectives: To study clinicopathological spectrum of gall bladder epithelial neoplasms, with emphasis on tumors with rare histology. Materials and Methods: Gall bladder resection specimens showing primary epithelial neoplasms (excluding BilIN) on histology, in a 10-year period, were reviewed for their clinical presentation and pathological features, and categorized according to the WHO 2019 classification. Results: Total 6214 gallbladder specimens received of which, 117 (1.88 %) (28 males, 89 females) showed neoplastic pathology (age range 25-80 years), 26.53% detected incidentally. Most common symptom was pain in right hypochondrium. Mass lesion was seen in 78.57% and gall stones in 44.89 %. Benign tumors were intra-cholecystic papillary neoplasms (10.20%) and pyloric gland adenoma (1.02%). Malignant were pancreaticobiliary adenocarcinoma (74.48%), ICPN with invasion (3.06%); and 2.04 % each of intestinal type adenocarcinoma and adenosquamous carcinoma. Others were, 1.02 % each of neuroendocrine tumor, mucinous, clear cell, poorly cohesive, sarcomatoid, lymphoepithelioma-like and squamous cell carcinoma. Conclusion: Gall bladder epithelial neoplasms have wide histopathological spectrum. Recognition of rare types is important as they are often characterized by aggressive behaviour. Detection of incidental tumors may indicate the limitations of radiological imaging.

Keywords: Epithelial neoplasm, gall bladder, rare histology

   AOP181 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Correlation of inflammatory markers (IL-6, IFN-γ, TNF-α, ferritin, hs-CRP) with HbA1c in type 2 diabetes mellitus

Kavya Varshney, Mukul Singh, Sunil Ranga

VMMC and Safdarjung Hospital, New Delhi, India

Introduction: Type 2 diabetes mellitus is a chronic, multifactorial and multisystemic disease. Systemic inflammation plays a key role in the pathogenesis of insulin resistance and type 2 diabetes mellitus. Inflammatory biomarkers can be a valuable tool for the assessment of risk of complications among type 2 diabetes mellitus. Objectives: The objective was to compare serum levels of IL-6, IFN-γ, TNF-α, ferritin, hs-CRP, and HbA1c in type 2 diabetes mellitus and controls and to find the correlation of IL-6, IFN-γ, TNF-α, ferritin, hs-CRP with HbA1c in type 2 diabetes mellitus. Materials and Methods: A case control study was done consisting of 50 cases of type 2 diabetes mellitus and 50 age & sex matched healthy controls. The patients were selected based on ADA criteria and after applying inclusion and exclusion criteria. Serum levels of HbA1c, IL-6, IFN-γ, TNF-α, ferritin and hs-CRP were measured. Results: Results were calculated using Wilcoxon-Mann-Whitney Test and statistically significantly increased values were seen in the serum levels of IL-6, IFN-γ, TNF-α, ferritin and hs-CRP of type 2 diabetes patients as compared to normal healthy individuals (p<0.001). Serum ferritin levels were found positively correlated (p<0.001, Spearman Correlation test) while serum levels of IL-6, IFN-γ, TNF-α and hs-CRP showed no correlation with HbA1C levels in type 2 diabetes patients (p>0.05). Conclusion: This study establishes the role of inflammation in type 2 diabetes mellitus patients and also correlation of inflammatory markers with HbA1c will help to identify the inflammation at the subclinical stage.

Keywords: Diabetes mellitus, ferritin, HbA1c

   AOP182 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Role of ratios of neutrophil and monocyte to lymphocyte and of red cell distribution width to platelet in the detection of early onset neonatal sepsis in comparison with serum C-reactive protein

Tanvi Agarwal, H B Sridevi

Kasturba Medical College, Mangalore, Karnataka, India

Introduction: Early onset neonatal sepsis (EONS) is a preventable cause of neonatal mortality and morbidity. Blood culture, the current gold standard has a turn-around time of 48 to 72 hours and high false negatives. Thus there is a need for reliable biomarkers for its early detection. Objectives: The present study aims to find the utility of complete blood count ratios in detecting EONS. Materials and Methods: The laboratory investigations of neonates with and without clinical sepsis were recorded with purposive convenience sampling technique. The total leucocyte count, absolute neutrophil count, C-reactive protein (CRP) and ratios of leucocyte counts were compared by Mann Whitney test. Their performance was then analysed by receiver operating characteristic analysis. Results: A total of 196 neonates aged 0 to 3 days were considered for the study, out of which 98 cases were study group. Serum CRP and red cell distribution width to platelet (RPR) ratios were found to have statistical significance, albeit low sensitivity and specificity in detection of EONS. Positive correlation of RPR was found with CRP (p value = 0.001). The area under the curve for CRP and RPR was 0.661 and 0.628 respectively. However, there was no statistical difference in the neutrophil and monocyte to lymphocyte ratios between the study and comparative groups. Conclusion: In the background of low yield of blood culture positivity and its longer turnaround time, RPR can be used as reliable biomarker in diagnosis of EONS in conjunction with CRP.

Keywords: Biomarker, leucocyte ratios, neonatal sepsis

   AOP183 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Study of laboratory parameters in neonatal jaundice

Arya Prakash, B N Navya, Anjali Rao

Department of Pathology, KVG Medical College, Sullia, Dakshina Kannda, Karnataka, India

Introduction: Neonatal jaundice (NNJ) is one of the vexing concerns in neonates during first week of life. Majority of these cases are physiological due to high RBC turnover, other known pathological causes include ABO incompatibility, septicemia, Rh incompatibility, prematurity, G6PD deficiency, etc. Objectives: The study aims to analyze different causes of NNJ and various laboratory parameters in NNJ. Materials and Methods: In this retrospective study, 150 cases of term and preterm neonates having NNJ were enrolled. Laboratory profiles like Serum bilirubin levels, blood group of mother and neonate, Coomb's test, Complete blood count with peripheral smear examination, Reticulocyte count, and C-reactive protein levels of the neonates were obtained and various etiological causes of NNJ were analyzed. Results: The mean age of presentation of jaundice in newborns is around day 2 of life. 76 (51%) were males and 74 (49%) were females. Physiological jaundice constituted 58 % of cases followed by ABO incompatibility (22%), Septicaemia (12%), and Rh incompatibility (11%) respectively. Highest level of serum bilirubin was found in septicaemia and Rh incompatibility. Direct Coomb's test and Indirect Coomb's test were found to be positive in all cases (100%) with Rh incompatibility while they were positive only in 27 % of cases with ABO incompatibility. CRP was raised in all cases of septicaemia. Conclusion: Even though majority of the cases are due to physiologic jaundice, an array of hematologic investigations are usually necessary to accurately rule out other serious causes of neonatal jaundice.

Keywords: Neonatal jaundice, neonatal sepsis, physiological jaundice

   AOP184 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Study of nonspermatozoal round cells in semen analysis

Rashmi Budha, N Anvita, V Srinivasamurthy

ESIC MC and PGIMSR, Rajajinagar, Bengaluru, Karnataka, India

Introduction: Worldwide surveys have shown that almost one in every seven couples faces problems of infertility. Of the total infertility cases, 50% are due to the male factors. Semen analysis form the basic, cost-effective and non-invasive investigation for screening. In a semen sample, apart from spermatozoa there may be a variable number of non-sperm cells. In a routine report, they are mentioned as “round cells” without further differentiating them into leucocytes or immature germ cells. Objectives: 1. To study the morphology of sperms and nonspermatozoal cells in the semen. 2. To study the significance of these nonspermatozoal cells in evaluation of Semen Analysis. Materials and Methods: This is a prospective study from April 2022 with a sample size of 100. Samples received for routine analysis, after liquefaction, will be smeared on the slide by feathering technique and subjected to Papanicolaou staining. A total of 100 cells will be counted for each sample in oil immersion objective. The proportion of immature and mature germ cells will be counted. Results: Out of the 100 cases, majority were Normozoospermia (58%) followed by Asthenozoospermia (17%) and Oligozoospermia (10%). Nonspermatozoal cells were identified according to the size, shape and morphology. Immature Germ Cells were the predominant nonspermatozoal cells in Asthenozoospermia, whereas in Oligozoospermia and Azoospermia, they were lymphocytes. Conclusion: The differentiation of “round cells” into cells of spermatogenic and non-spermatogenic origins is important. For a better outcome on infertility, there is a need for initial screening and differentiation of round cells into immature germ cells and leucocytes with a simple and cost-effective method.

Keywords: Germ cells, non spermatozoal cells

   AOP185 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Study of haematological and biochemical profile with special reference to inflammatory markers and its correlation with clinical outcome in patients with COVID 19

Deepika Rawat, Vatsala Misra, Akanksha, Richa Singh, Chandrawati, Kshama Tiwari

Department of Pathology, Moti Lal Nehru Medical College, Prayagraj, Uttar Pradesh, India

Introduction: SARS CoV2 (COVID19) has emerged as pandemic viral infection in the world with the main and strong impact on respiratory airway & other systems. Objectives: To assess the prognostic values of haematological and inflammatory markers in COVID 19 related mortality by comparing these markers in survivors and non survivors in Indian population. Materials and Methods: An observational cross sectional study was conducted in COVID-19 referral hospital in MLN medical college, from July 2020 to February 2022, RT-PCR confirmed cases in consequently 3 waves. Patients were divided into two main groups:survival and expired group. Age, gender, ongoing medical conditions, hematological and inflammatory laboratory results such as the total leukocyte count, neutrophil to lymphocyte ratio, D-dimer, C-reactive protein, PT-INR, procalcitonin levels were compared between these groups. Results: 300 random patients on the basis of available clinical and laboratory records were taken. Male to female ratio in first, second and third wave was 1.32, 2.57 and 1.08. Survivors and non-survivors ratio in each wave was 4, 0.5 and 4. Mean±SD values among survivors and non-survivors in first, second and third waves for Total leukocyte count were (8381±473), (9017±418) & (14233±974) and non-survivors (16293±941), (11840 ±663) & (20095 ± 108), N/L ratio among survivors (5.10±5.2), (4.5±2.9) & (9.77±11.3) and non-survivors (12.9±8.8), (9.16±5.5) & (16.828±15.8), absolute lymphocyte count among survivors (1659±897), (1609±474) & (1708±120) and non-survivors (1745±174), (1456±103) & (1386±858), C-reactive protein among survivors (33.01±20.5), (28.0±37.9) & (82.4±53.7 and non-survivors (113.2±36.7), (110±46.5) & (127.45±38.30), d-dimer among survivors (0.53±0.5) (1.49±3.1) & (2.34±2.6) and non-survivors (5.36±2.9) (4.96±3.4) & (6.64±3.3) values were respectively. Conclusion: The difference in the values of parameters (TLC, N/L ratio, CRP, d-dimer) was found to be statistically significant among survivors and non-survivors.

Keywords: COVID-19, hematological parameters, inflammatory markers

   AOP186 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Correlation of RBC indices, peripheral blood smear and Iron profile in anemia of chronic disease

Viona Dcunha, M H Prabhu, S S Hiremath, Girija Patil

S. Nijalingappa Medical College and HSK Research Centre, Bagalkote, Karnataka, India

Introduction: Anemia of chronic disease is often observed in patients with infections, inflammatory and neoplastic diseases that persists for more than 1 or 2 months. There is hypoferremia in presence of ample reticuloendothelial iron stores. Objectives: 1. To correlate RBC indices and Peripheral Blood Smear (PBS) in patients with chronic disease. 2. To estimate Serum Iron profile in patients with chronic disease. Materials and Methods: 50 cases were evaluated in SNMC and HSK hospital. Inclusion criteria: Patients examined and diagnosed by the clinician with active disease state were included in the study. Exclusion criteria: Pregnant and lactating women were excluded in the study.


Peripheral blood smear in 74% of patients showed Normocytic and Normochromic blood picture. The presence of toxic granules, Burr cells and thrombocytosis were suggestive of cases of sepsis, chronic renal disease and in cases of post-infections respectively. Conclusion: The RBC indices are within normal limits, the elevated ferritin correlated with disease activity, with majority of the patients (74%) showing Normocytic Normochromic blood picture on peripheral blood smear. Serum Ferritin, a non-specific marker has a significant role in diagnosis of Anemia of chronic disease correlating with peripheral blood findings.

Keywords: Anemia, chronic inflammation, ferritin

   AOP187 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Study of inflammatory markers and clinical biochemistry parameters in COVID-19 patients

Vinayak Sharma, C R Gore

Dr. DY Patil Institute, Pune, Maharashtra, India

Introduction: Coronavirus disease (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. Objectives: The objective of the study is to estimate the significance of biochemical parameters and to correlate with disease severity, with various comorbid conditions and presence of lung ground glass opacities (GGO). Materials and Methods: This prospective observational study included 603 covid-19 patients confirmed by RT-PCR. All patients above 18 years were included, average age being 48 +/-14 years. Male predominance was seen (M:F was 2:1). Majority of the patients had GGOs in severe stage. Among patients with comorbidities, significantly more patients had diabetes 22% and hypertension (10%) and most of them were in severe category. They were subjected to various investigations which included renal and liver function markers and inflammatory markers (Sr. LDH, CRP and Ferritin). An attempt was made to compare these results with mild (45%), moderate (20%) and severe (25%) category cases. Results: Significantly more patients showed raised CRP levels (76.5%), followed by raised LDH levels (81% male and 73% female). Ferritin was significantly raised in 37% of female cases, and 3% of male cases. Among renal function markers, creatinine was observed to be higher in 16.3% of male and 21.6% female cases. Majority of the cases showed normal levels of serum urea (85%), conjugated, unconjugated and total bilirubin levels. Similar results were observed in serum AST, ALT and ALP levels for both the sexes. Conclusion: These Biochemical indices and CRP and LDH in particular can be used to assess severity in COVID 19 disease.

Keywords: Biochem, clinical, COVID-19

   AOP188 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Assessment of haematological and biochemical parameters in petrol pump workers in Amritsar, Punjab

Vivek Bharmota, Mandeep Randhawa, Permeet Kaur Bagga, Jaspreet Singh

Government Medical College, Amritsar, Punjab, India

Introduction: Petroleum products are used for various reasons by human beings at homes, in manufacturing and petrochemical industries with individuals most frequently exposed are those working in petrol pumps. Petrochemical workers are exposed to many noxious substances present at their work places. The most dangerous effects are due to benzene which is mostly found in petrochemical vapours with its exposure having significant deleterious health effects. Objectives: This study aimed to evaluate the impact of petroleum products on hematological and biochemical parameters in petrol pump workers. Materials and Methods: This study was done on 50 petrol pump workers from different areas of the city. The whole blood and serum samples were analyzed by fully automated hematology and biochemistry analyzers (TransAsia H 360 and EM 360) respectively. The parameters were Complete Blood Counts (Hb, TLC, DLC, RBC Count, PCV, MCV, MCH, MCHC, RDW, Platelet count), Liver Function Tests (S. Bilirubin, AST, ALT, ALP, γ-GGT, TSP, DSP), Renal Function Tests (B. Urea, S. Creatinine), S. Na+, S. K+. Results: Out of 50 petrol pump workers, 10 (20%) having working duration of <5 years showed no significant changes in parameters, 24 (48%) having working duration of 5-10 years showed mild changes in haematological parameters, 16 (32%) having working duration of >10 years showed significant changes in haematological parameters (Hb, PCV, MCV), liver enzyme levels and serum electrolytes. Conclusion: This study reflects that years of working in petrol pumps do have significant bearing on hematological, hepatic, renal parameters.

Keywords: Benzene, blood tests, petrol station workers

   AOP189 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Immature platelet fraction in patients infected with COVID-19

Neha Parashar, Rashmi Jain Gupta, Divya Gupta, Alka Siroha, Sandeep Garg, Suresh Kumar

Lok Nayak Hospital, New Delhi, India

Introduction: The recent pandemic of Coronavirus has been found to be associated with thromboembolic events. Patients infected with COVID-19 have been found to have thrombocytopenia. Decrease in platelet count may be caused by increased destruction and consumption of platelets or by decreased platelet production in the bone marrow. Assessment of the cause of thrombocytopenia can help in avoiding unnecessary transfusion of platelets. Thrombopoietic activity can be assessed by proportion of immature platelets. Immature platelet fraction is a new platelet parameter which is found to be raised in conditions associated with peripheral destruction and consumption of platelets. Objectives: To study the immature platelet fraction (IPF) of patients infected with COVID-19, admitted during the COVID wave in January- February 2022. Materials and Methods: The present study is a retrospective study where secondary data obtained from previous laboratory records of COVID positive patients admitted at Lok Nayak Hospital from 4 January to 4 February 2022 were analyzed. Sixty-eight COVID-positive patients were included. Platelet parameters included from automated hematology analyzer Mindray were platelet count, mean platelet volume (MPV), immature platelet fraction (IPF) and platelet large cell ratio (P-LCR). Data was analyzed using IBM SPSS Version 25. P value less than .05 was considered statistically significant. Results: IPF was found to have a statistically significant correlation with platelet count, MPV and P-LCR (p-value<0.05). There was an increase in IPF with decrease in platelet count. Increase in IPF was noted with increase in MPV and P-LCR. Conclusion: The present study suggests thrombocytopenia in COVID patients may be due to peripheral destruction of platelets.

Keywords: COVID-19, immature platelet fraction, thrombocytopenia

   AOP190 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

AMA-M2 positivity on immunoblot testing: An audit

Donna Philip, Renuka Malipatel

St. John's Medical College, Bengaluru, Karnataka, India

Introduction: Anti-mitochondrial antibodies (AMA) are closely linked to primary biliary cholangitis (PBC). The prevalence of AMA-M2 is low and AMA-M2 positivity may precede PBC. Up to 1% of healthy individuals can show positive AMA-M2. Objectives: To determine AMA-M2 positivity on immunoblot testing (Euroline ANA profile 3) in various hepatic and extrahepatic diseases. Materials and Methods: Immunoblot test results were retrospectively audited for AMA-M2 positivity during the period January 2018 to July 2022 (3.5 years). Patient details were obtained from the laboratory information system (LIS). Demographic details, indication for testing, presence of antibodies against other extractable nuclear antigens and liver function tests (LFT) noted. Results: A total of 5595 immunoblot tests were done over three and a half years. Among these 255 patients tested positive for AMA-M2; majority were females (n=207, 81%). Mean age was 37 years. Hepatic causes accounted for 1.5% of the cases (0.7% PBC, 0.7% non-PBC). The extrahepatic causes were rheumatic (139, 54.5%), hematologic (n=15, 5.8%), neurologic (n=10, 3.9%) and other systems (11, 4.3%). The cause was unknown in 76 cases (29.8%). Conclusion: AMA-M2 positivity was noted most commonly in patients with rheumatic diseases (54.5%). 29.8% of cases had normal baseline LFT and no known etiology. Further follow up is required in these patients to determine development of PBC in the future.

Keywords: AMA-M2, imunoblot, primary biliary cholangitis

   AOP191 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

ANA testing by indirect immunofluorescence versus immunoprofile – Is old still gold?

Shadma Shahin, Vijaya Mysorekar

MS Ramaiah Medical College, Bengaluru, Karnataka, India

Introduction: Autoimmune disease makes the immune system unable to recognize self-antigens and produces autoantibodies damaging several organs. Identification of these autoantibodies called antinuclear antibodies (ANA) helps in arriving at a diagnosis. Indirect immunofluorescence assay (IIF) has been the gold standard for detecting ANA. However, due to its easy availability ANA immunoprofile is now being increasingly utilized. Objectives: This study was conducted to compare ANA immunoprofile results with IIF patterns. Materials and Methods: We studied 98 patients clinically suspected to be having autoimmune disease who had undergone both Indirect Immunofluorescence as well as Immunoprofile by test strip during the period from September 2019 to August 2022. Immunofluorescence correlation with immunoprofile for ANA was analyzed. Results: Of the 98 samples processed 60 samples were negative for both ANA IIF and immunoprofile. 30 (79%) out of the 38 positive samples were from female patients. 22 cases (58%) were ANA-IIF positive whereas 34 (89%) were immunoprofile positive. 4 (18%) of the ANA-IIF positive samples showed negativity with immunoprofile. ANA-IIF negativity was observed in 76 of the total 98 samples under study. Of these, 16 (16.3%) exhibited positivity with immunoprofile. The homogenous pattern was the most common (59%) ANA pattern. Various combination and intensity of specific ANAs were detected and Ro52 was the most common antibody detected. The sensitivity, specificity, NPV, and PPV for IIF was 81%, 98%, 93%, and 95% respectively whereas the same for Immunoprofile was 88%, 85%, 95%, and 68% respectively. Conclusion: Indirect Immunofluorescence though requires expertise shows better results than immunoprofile in detecting ANA for diagnosing autoimmune diseases.

Keywords: Antinuclear antibodies immunoprofile, antinuclear antibodies indirect immunofluorescence assay, autoimmune diseases

   AOP192 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Prognostic markers in haematological and biochemical parameters among COVID-19 patients

Sunita Nyamagoudar, B H Ramesh

Raichur Institute of Medical Sciences, Raichur, Karnataka, India

Introduction: Coronavirus 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a global threat. Various biomarkers help us in identifying the severity of the patient. This study was formulated to assess the prognostic factors in these biomarkers conducted on the COVID-19 affected patients. A comparative study was done between the survivors COVID-19 and non-survivors of COVID-19. Objectives: To study the various haematological and biochemical markers in COVID-19 patients. Assessment of prognosis in the patients who survived COVID-19 compared to the non-survivors. Materials and Methods: Detailed investigations of COVID-19 positive pateients from January 2021 to December 2021 was collected. Among the 155 cases included in the study, 92 patients had survived COVID-19 and 63 patients had died due to COVID-19. Comparison of values of various biomarkers was done between the survivors and deaths. Chi-square test was used as a test of significance for qualitative data. A P value of <0.05 was considered as statistically significant. Results: Mean age of survivors was 50±14 and those of deaths was 51±13. Mean sPO2 was significantly higher in survivors compared to dead cases (p<0.05). Haematological profile showed significant difference (p<0.05) in absolute neutrophil count (ANC) and neutrophil to lymphocyte ratio (NLR). C reactive protein (CRP) showed significantly higher values in non-survivors compared to survivors (p<0.05). Significantly higher values were seen with serum ferritin in non-survivors compared to survivors (p<0.05). SPO2, ANC, NLR, CRP and ferritin are found to be poor prognostic factors in our study. Conclusion: Our study compared the haematological and biochemical parameters between the survivors and deaths in COVID-19. SpO2, ANC, NLR, CRP and serum ferritin were significantly higher in non-survivors than the survivors. Hence these parameters could be considered as poor prognostic markers and higher values of these markers can alarm clinician towards an intensive care management.

Keywords: Biomarkers, COVID-19, prognosis

   AOP193 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Measurement of oxidative burst activity in leucocyte subpopulations in whole blood

Shilpi Saxena, Maya Gupta, Ashita Gada, Ankita Parab, Priyanka Setia, Manisha Madkaikar

Command Hospital Western Command, Panchkula, Haryana, India

Introduction: The oxidative burst activity releasing Reactive oxygen species (ROS) constitute an important part of innate immune system. ROS have been extensively researched for its role in killing microorganisms via oxidative burst and diagnosis of chronic granulomatous disease (CGD). Its role have also been implicated in various conditions like ageing, autoimmune diseases, carcinogenesis, cardiovascular diseases and reperfusion injury. The ROS are produced by various leucocytes like neutrophil, eosinophils and monocytes subsets. Objectives: To measure oxidative burst activity in various types of leucocytes. Materials and Methods: Venous anticoagulated whole blood collected in EDTA from five healthy donors was used and the samples were processed within one hour of collection. Surface staining of CD45, CD16 and CD14 was done using stain-lyse-wash method and change in Mean channel fluorescence intensity (MFI) of Dihydrorhodamine 123 (DHR 123) after Phorbol Myristate Acetate (PMA) stimulation was measured using DxFlex flow cytometer (Beckmen-Coulter). Neutrophils were gated using CD45 dim SSC high and CD16 positive cells, while eosinophils were gated using CD45 moderate and SSC high and CD16 negative cells. Monocytes were gated using 45 bright SSC low and were further divided into classical (CD14+ CD16-), intermediate (CD14+ CD16+) and non-classical (CD14- CD16+). Results: The neutrophils revealed maximum SI followed by classical monocytes, eosinophils, intermediate and non-classical monocytes in that order. Conclusion: The oxidative burst activity is different in various subsets of leucocytes and this may be exploited to study pathogenesis of many diseases and explore tailored therapies to these in future.

Keywords: DHR, oxidative burst, phorbol myristate acetate

   AOP194 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

A comparative analysis of manual versus automated semen analysis: A prospective double blinded study at a newly established tertiary care centre in Central India

Hemlata Panwar, Deepti Joshi, Tanya Sharma, Ujjawal Khurana, Ankur Joshi, Neelkamal Kapoor

AIIMS, Bhopal, Madhya Pradesh, India

Introduction: Infertility affects 8–12% of couples worldwide, approximately 40–50% due to “male factor” infertility. According to the WHO, manual semen analysis is the gold standard diagnostic tool assessing male factor infertility. However manual semen analysis is time consuming and suffers from analytical variability and lack of standardization. The introduction of automated sperm analyser (SQAVision) has provided an alternative approach to manual semen analysis with quality results in less time and minimises the need of highly trained professionals. Objectives: The aim of this prospective double-blind study was to compare the automated semen analysis to the conventional manual method in terms of accuracy and precision. We also studied abnormal semen parameters and clinicopathological variables of male infertility. Materials and Methods: In this Hospital based study, a total of 135 semen samples were included. After obtaining informed consent, information regarding reproductive history and lifestyle factors noted and patients were instructed regarding collection of semen sample. All semen parameters such as total sperm concentrations, motility, vitality and morphology were assessed simultaneously and independently both manually as well as by using automated analyser. Results: Precision, accuracy and agreement between automated and manual semen analysis methods were assessed. Preliminary data analysis revealed good agreement between the sperm counts obtained by both the methods. Conclusion: Semen analysis is fundamental diagnostic tool to assess the male factor infertility. The automated semen analyser can be used interchangeably with manual semen analysis for sperm concentration and as a reliable high throughput system for semen analysis which will promote standardization of laboratory results.

Keywords: Automated analyser, male infertility, semen analysis

   AOP195 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Non-nuclear and rare nuclear ANA patterns in indirect immunoflourescence testing and their clinical associations

Gayatri Ravikumar, Anshika Gupta, Judah Pereira

Department of Pathology, St. John's Medical College, Bengaluru, Karnataka, India

Introduction: Antinuclear antibody (ANA) testing by indirect immunoflourescence (IIF) is routine screening test for suspected autoimmune diseases (AIDs). A standardized nomenclature is given by the International consensus on ANA pattern (ICAP). When the conventional nuclear patterns pose no diagnostic difficulty, the cytoplasmic, mitotic and rare nuclear patterns are infrequently reported. Objectives: The study was done to understand the clinical significance and frequency of these unconventional patterns in our population. Materials and Methods: Retrospective one year blinded study of ANA patterns in patients serum samples, tested at 1:100 dilution. Results: ANA samples received over one year was 4730. After excluding cases with repeat samples (n=162), there were 4568 cases, of which ANA positivity was seen in 673 cases (14.7%). Cytoplasmic patterns were found in 184 cases (27.3%) and mitotic pattern in 16 (2.4%) cases. Exclusive cytoplasmic patterns was seen in 100 cases (14.3%) and exclusive mitotic pattern in 14 cases (2.08%). Rare nuclear patterns was seen in 30 cases (4.5%). The most common exclusive cytoplasmic pattern was filamentous (n=39), whereas the common cytoplasmic pattern associated with nuclear positivity (mixed pattern) was cytoplasmic homogeneous (AC-19). The rare nuclear patterns included Topo-I (n=9), nuclear envelop (n=5), multiple (n=6) and few (n=8) nuclear dots. While some of the common cytoplasmic patterns like filamentous and homogeneous were more frequent in AIDs the uncommon patterns showed varied clinical associations. Conclusion: Non nuclear patterns on ANA IIF carry clinical significance. These patterns can occur independent of nuclear patterns or as mixed patterns. It is equally important to identify and report rare nuclear patterns as many of these have known autoimmune associations.

Keywords: Antinuclear antibody patterns, cytoplasmic, mitotic, rare nuclear

   AOP196 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Role of complete blood count (CBC) parameters as a prognostic assister in management of hospitalised COVID 19 patients: Study from a tertiary care center

Priyadarshini Biswal, Asaranti Kar, Lity Mohanty

Department of Pathology, SCB Medical College, Cuttack, Odisha, India

Introduction: Complete blood count (CBC) has always been an important and crucial tool in diagnosis and management of infectious diseases. It is also a rapid, cheap and simple procedure without much inconvenience to patients. CBC parameters of CoVid 19 patients recently has shown variations in comparison to other viral infections. Objectives: The aim of our study is to observe the various CBC parameters of CoVid 19 patients and assess them in the diagnosis and prognosis of disease. Materials and Methods: CBC reports of around 500 hospitalised CoVid 19 patients were examined which also included ICU patients during the 2nd wave from April 2021 to December 2021. Different parameters like TLC, DC, Platelet count, neutrophil to lymphocyte ratio (NLR) and platelet to lymphocytes ratio (PLR) were observed and correlated with severity. Results: Total 500 CBCs were examined and correlated with normal range in CBC. Most (92%) of the patients showed neutrophilia along with lymphopenia. Eosinopenia was observed in 91% of patients. NLR was also higher in 92% of Covid 19 patients. Platelet count was lower in ICU patients (88%) as compared to ward patients. PLR was also significantly high in ICU patients. Conclusion: Various CBC parameters can help in diagnosing CoVid 19 patients and also can be adopted as a prognostic marker during management. Hence, CBC parameters have a crucial role along with other inflammatory markers in assessing severity of CoVid 19 patients.

Keywords: Complete blood count, COVID 19, neutrophil to lymphocyte ratio

   AOP197 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Significance of differentiating round cells in human semen into leucocytes and immature germ cells

Sneha Jawalkar, Savitri M. Nerune

Department of Pathology, Shri B M Patil Medical College, Hospital and Research Centre, BLDE (Deemed to be University), Vijayapura, Karnataka, India

Introduction: Worldwide surveys have shown that almost one in every seven couples faces problems of infertility. In India, the infertility rate is 9% of the reproductive population and 50% are due to the male factors. Semen analysis forms the basic, cost-effective investigation for screening. In a semen sample there are cells apart from spermatozoa which in routine semen analysis reports are often labelled as 'round cells'. These cells could be either inflammatory cells or cells of spermatogenic origin (immature germ cells). These cells need to be differentiated as the significance of increase in each cell type is different. Objectives: The aim of our study was to differentiate between leukocytes and immature germ cells using peroxidase staining method. Materials and Methods: A total of 40 semen samples were studied. Complete semen analysis was done with special stains like peroxidase stain and pap stain were done. After peroxidase staining, round cells that were stained brown were counted as leucocytes and unstained round cells were further differentiated on Pap stain. Counting was done on Neubauers chamber. Results: Round cells were seen in all the semen samples but only a few percentages of cells were stained positive for peroxidase in each case. Conclusion: Not all round cells in semen sample are leucocytes but often are reported as leucocytes in semen analysis reports which misleads the clinician to treat patients with antibiotic therapy which might harm the normal flora of genital tract leading to other complications. Hence, round cells must be confirmed as leucocytes with special stains before reporting leucocytospermia.

Keywords: Human semen, leucocytes, round cells

   AOP198 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Seroprevalence and co-infection of Hepatitis B and Hepatitis C virus in a rural tertiary care hospital

Sumit Kumar, Surinder Kumar

BPS, GMC (W), Khanpur Kalan, Sonepat, Haryana, India

Introduction: Hepatitis B virus (HBV) and hepatitis C virus (HCV) have several important similarities including worldwide distribution, hepato-tropism, similar modes of transmission and the ability to induce chronic infections that may lead to liver cirrhosis and hepatocellular carcinoma. Co-infection with both HBV and HCV is linked with higher morbidity as well as mortality and impact healthcare resource utilisation. Objectives: This study aims to understand the occurrence of HBV, HCV and its co-infection in a rural tertiary care Hospital. Materials and Methods: Study was done from March 2022 to August 2022 in the department of microbiology of a rural tertiary care Hospital. Patients who visited outdoor patient departments or were admitted to the indoor patient departments were advised to undergo HBV and HCV test for screening before any invasive/surgical procedure were included in the study. ELISA and rapid immunochromatographic test were performed for HCV antibody and (Hepatitis B surface antigen) HBsAg. Results: Total 10765 samples were tested for hepatitis B and C virus. Out of them 483 (4.48%) samples were positive for HCV antibody. HBsAg was positive in 195 (1.81%) samples. Co-infection of both Hepatitis B and C was found in 10 (0.09%) cases. HCV antibody was detected in 257 (53.21%) males and HBsAg was detected in 127 (65.13%) male samples. Conclusion: Strict adherence to universal precautions, and proper disposal of used material should be implemented to decrease the risk of transmission of HBV and HCV. Immunization with HBV vaccine. Regular surveillance and precise estimate of the occurrence of Hepatitis B, Hepatitis C and HBV/HCV co-infection would be needed to formulate policy decisions and plan communal health. interventions.

Keywords: Co-infection, hepatitis B, hepatitis C, seroprevalence

   AOP199 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

ALIFAX-200: Is modified Westergren method for erythrocyte sedimentation rate a valid alternative to the gold standard?

Renuka Nagaraj, Radhika Shivanna

National Institute of Unani Medicine, Bengaluru, Karnataka, India

Introduction: Background: In order to assure comparability with the reference Westergren technique, the development of automated erythrocyte sedimentation rate (ESR) analyzers has necessitated thorough validation prior to their implementation in ordinary practise. Objectives: Our study's objective was to carry out the analytical validation of the Alifax 200 automated ESR analyser with gold standard method. Materials and Methods: Methods: The validation was carried out in accordance with the most recent recommendations of the International Council for Standardization in Hematology, and it involved determining intrarun and interrun precision, evaluating sample carryover, hemolysis interference, determining sensitivity to fibrinogen, comparing the method to the gold standard Westergren method, and performing a stability test. Results: The total number of samples after evaluation were divided into low (n = 69), intermediate (n = 85), high (n = 329), and very high range of ESR (≥100 mm/hr; n = 17). Mean difference between values of automated and Fabry method ESR for the low, intermediate, high, and very high ESR range was -2.97±3.93, -2.12±5.49, 1.22±5.94, 0.625±6.8 mm/hr, respectively.. Using the Bland–Altmann analysis the limit of agreement between the two methods for low-ESR range was (-9.60- to 2.00). For low ESR, (correlation coefficient, r= 0.472, P<0.001), while for high ESR, range was (correlation coefficient r= 0.521, P = 0.038) which indicates moderate positive correlation between the two methods. For normal ESR range correlation was (r= 0.412, P<0.01) and for intermediate ESR (r= 0.923, p<0.001). Overall, A strong positive correlation was seen between two methods (r= 0.952, p<0.001) which suggests that analytical is equally good than manual. Conclusion: The obtained results proved the that the results were highly comparable with the Analyser ALIFAX Roller 20 LC and the gold standard Westergren method, showing obvious improvements in standardization of ESR methods.

Keywords: Erythrocyte sedimentation rate, standardization, T, validation, westergren method

   AOP200 Top

   Laboratory Medicine (Biochemistry, Clinical Pathology, Immunology, Microbiology) Top

Cytomorphological study of cervical smears

Aditi Parkhe

Apollo Diagnostics, Bengaluru, Karnataka, India

Introduction: According to National Cancer Registry Program of India, cancers of uterine cervix are leading malignancies seen in the females. The burden of cervical cancer in India is enormous accounting for about 20% of all cancer related deaths in females. The study of cervical cytology assists in diagnosing various lesions of cervix. The most sensitive and widely used test for this is cervical smear (Pap smear). Regular screening and early diagnosis with a cervical smear are two of the most effective ways to prevent and control cervical cancer. Therefore, the study of cervical smears is included in this research plan. Objectives: A) To study the usefulness of cervical cytology in screening and diagnosis of diseases of cervix. B) To screen cervical smears for detection of precancerous lesions and carcinoma of uterine cervix. C) To ascertain reliability of cytological findings and its comparison / confirmation with histopathological examination whenever possible. D) To analyze the cases studied. Materials and Methods: A total of 1001 instances made up the sample size. The reporting was completed according to “THE BETHESDA SYSYTEM for reporting cervical cytology (2001)”. Smears were stained with Papanicolaou stain and were microscopically analyzed. Results: The study findings revealed 804 out of total 1001 smears to be satisfactory for evaluation. Among non neoplastic findings, 563 smears showed normal cytology with no specific pathology followed by inflammatory smears (149/1001). Most common complaint was per vaginal discharge followed by pain in abdomen. The mean age at which squamous cell carcinoma 9 SCC) cases presented was 49.11 years. Post coital bleeding was the main complaint for these patients. This cervical growth bleeds on touch in all cases of SCC on per speculum examination. Conclusion: Early detection predicts better prognosis. One of the most effective way of preventing and controlling carcinoma cervix is regular screening and early diagnosis with the help of cervical smear. Organised population based cervical smear screening linked to the treatment of detected neoplasia can lead to more than 70% reduction of disease related mortality.

Keywords: Cancer, cervical cytology, squamous cell carcinoma

   AOP201 Top

   Lymphoreticular Pathology Top

CD30 and P53 expression by immunohistochemistry in DLBCL And its association with cell of origin classification and other prognostic variables – A retrospective study

Aadhya Sharma, Ranjini Kudva

Kasturba Medical College, Manipal, Karnataka, India

Introduction: Diffuse Large B-cell Lymphoma (DLBCL) is the commonest type of adult Non-Hodgkin Lymphoma with a standard treatment regimen of R-CHOP. However, treatment outcomes are extremely variable due to molecular heterogeneity of DLBCL in conjunction with expression of various oncogenic and tumor suppressor proteins which impact therapeutic decisions. Non-GCB subtype has also shown inferior outcomes following standard immunochemotherapy. Our research aims to study the expression pattern of two IHC markers, CD30 and p53 in DLBCL cases classified according to cell of origin along with their association with other prognostic factors to identify patients who may benefit from therapies other than R-CHOP. Objectives: To study the expression pattern of p53 and CD30 in DLBCL cases subtyped as per Hans algorithm. To evaluate the association of CD30 and p53 expression with other prognostic variables in DLBCL (International Prognostic Index, Histologic Subtype, Ki67, c-MYC, c-MYC/Bcl-2 double expression). Materials and Methods: A 3 year retrospective study of 47 histopathologically diagnosed and subtyped cases of DLBCL excluding primary CNS lymphoma, primary mediastinal large B cell lymphoma, slide reviews and cases with prior chemotherapy. IHC staining with CD30 and p53 were done and analysed with other prognostic variables. Results: Overall association was found between p53 and cMYC IHC expression (p value<0.05). In Non-GCB cases, statistically significant associations found between p53 and Ann Arbor Stage and cMYC IHC expression (p<0.05). Conclusion: p53 expression seems to be associated with cMYC expression in Non-GCB subtype of DLBCL.

Keywords: CD30, diffuse large B-cell lymphoma, p53

   AOP202 Top

   Lymphoreticular Pathology Top

Orbital lymphoma: A series of cases from a tertiary cancer centre

L Pavan, Akkamahadevi Patil, Usha Amirtham, S Balu, Linu Jacob

Kidwai Memorial Institue of Oncology, Bengaluru, Karnataka, India

Introduction: Orbital lymphoma refers to, lymphoma occurring in the conjunctiva, eyelid, lacrimal gland, and ocular musculature. Majority of non-Hodgkin's lymphomas of the orbit are extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) type. Primary non-Hodgkin's lymphoma (NHL) of the orbit is a rare presentation, representing 8-10% of extranodal NHL and only 1% of all NHL. Objectives: 1. To study the clinicopathological spectrum of primary and secondary orbital lymphoma. 2. To further subtype these neoplasms using ancillary techniques. Materials and Methods: A descriptive retrospective study conducted over a period of 3 years (2020–2022). All cases with morphological diagnosis of orbital lymphomas, confirmed of immunohistochemical studies are included in this study. Results: A total of 16 cases were included in the study. Mean age of the patients was +SD. M:F ratio was 1:1.3. Orbital mass was the most common presentation (75%), followed by conjunctival (18 %) and eyelid mass (6%). Our series included 10 cases of primary and 6 cases of secondary lymphomas. All of the primary lymphomas were of B cell type, consists of 56% marginal zone lymphoma, 18% Diffuse large B cell lymphoma, 12% mantle cell lymphoma and 12% follicular lymphoma. The most common secondary lymphomas are marginal zone lymphoma. Conclusion: Extranodal marginal zone lymphoma constitute the bulk of orbital lymphomas. Accurate histopathologic evaluation is the most critical diagnostic step in the management of orbital lymphomas.

Keywords: Diffuse large B-cell lymphoma, marginal zone lymphoma, orbital lymphoma

   AOP203 Top

   Lymphoreticular Pathology Top

Bilateral lacrimal gland enlargement – Primary presentation of sarcoidosis

Kriti Singh, Deepti Gupta, Anjali Tewari

Regency Hospital, Kanpur, Uttar Pradesh, India

Introduction: Bilateral lacrimal gland enlargement is uncommon; however, its presence induces brainstorming process and intensive discussion between a clinician and an imaging specialist, leading to exploration of multiple systemic disease patterns such as lymphoproliferative disorders, sarcoidosis, Sjögren's syndrome, and tuberculosis. Thoughtful analysis and diagnostic work-up are required to confirm the diagnosis. Sarcoidosis is a rare systemic disease, with ocular involvement being still rarer. Objectives: To find the incidence of cases of sarcoidosis presenting as primary lacrimal gland swelling. Materials and Methods: Here, we present two cases of bilateral lacrimal gland enlargement which were diagnosed as sarcoidosis based on further work up. Results: Lacrimal gland swellings revealed non caseating granulomas on aspiration specimens/biopsies. Further work up showed mediastinal and hilar lymphadenopathy with pulmonary nodules which along with biochemical tests supporting the diagnosis of sarcoidosis. The cases highlight the ocular symptoms in sarcoidosis and clinico-radiological approach to bilateral lacrimal gland enlargement. Conclusion: Evaluation of a case of bilateral lacrimal gland enlargement is important as it may be tell-tale sign of an undiagnosed systemic disease. Every disease entity related to lacrimal gland enlargement should be given an equal consideration, and one should not be biased by the common condition which may lead to unnecessary management.

Keywords: Lacrimal gland enlargement, lymphoproliferative disorders, sarcoidosis

   AOP204 Top

   Lymphoreticular Pathology Top

Histiocytic/dendritic cell neoplasms and stroma-derived neoplasms of lymphoid tissues: A retrospective analysis from a tertiary cancer centre of North India

Kavya Kadali, Zachariah Chowdhury, Shashikant C U Patne, Ipsita Dhal

Mahamana Pandit Madan Mohan Malviya Cancer Centre and Homi Bhabha Cancer Hospital, Varanasi, Uttar Pradesh, India

Introduction: Histiocytic/dendritic cell neoplasms and stroma-derived neoplasms of lymphoid tissues (HDCN & SDNLT) are rare, accounting for <1% of all neoplasms occurring in lymph nodes and extramedullary locations. Objectives: To illustrate the clinicopathological characteristics of the complete gamut of HDCN & SDNLT. Materials and Methods: A retrospective analysis was undertaken over a period of 3 years at a tertiary cancer centre. Results: A total of 33 cases of HDCN & SDNLT were found during this period, amongst which majority were adult males. Langerhans cell histiocytosis [LCH] was the predominant neoplasm, followed by histiocytic sarcoma [HS], follicular dendritic cell sarcoma [FDCS], Langerhans cell sarcoma [LCS], Rosai-Dorfman disease, and Erdheim-Chester disease. Unisystem LCH predominantly involved the skeletal system whereas in multisystem LCH, lymph nodes and skeletal lesions were the commonest sites. The handy immunohistochemistry markers for diagnosing HDCN & SDNLT were CD68, CD163, CD4, CD1a, CD21 & CD23. Chemotherapy (CT) was the main modality of treatment (63.6%) due to multisystem involvement in the case of LCH or metastasis in other disease. Surgery was performed in 36.4% patients while radiotherapy was administered post-operatively in 9.1% patients. On a mean follow-up of 16.1 months, 5 patients were found to have no evidence of disease, 16 are alive with disease and 3 died of disease. Conclusion: HDCN & SDNLT are extremely rare and intriguing diseases that ought to be ascertained using a combination of meticulous morphological examination and an array of immunohistochemistry markers to rule out mimics such as carcinoma, lymphoma, and neuroendocrine tumours and for precise subclassification of these lesions.

Keywords: Dendritic, histiocytic, lymphoid stroma derived

   AOP205 Top

   Lymphoreticular Pathology Top

Testicular lymphoma – A study at tertiary care center

Teena Murthy, Ashwini Nargund, Usha Amirtham

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Malignant lymphoma of the testis is uncommon. It accounts for 1 to 8 percent of all testicular tumors, it is the most common type of testicular tumor in men over the age of 60 year. Objectives: In this study we present 12 cases of Malignant Lymphoma of the Testis. Materials and Methods: Present study is a retrospective study done at KIDWAI memorial institute of Oncology for a period of 9 years from 2014 to 2022. All cases of Malignant Lymphoma of Testis were included in the study. Results: In this study age group of patients were 10 yrs to 76 yrs. 10 excision biopsy specimens out of 12 were diagnosed as Malignant Lymphoma of Testis. The neoplastic cells expressed CD 10, CD20, MUM 1, BCL 6, BCL-2 and were negative for CD 3, Tdt, Cyclin D1 and ALK and were diagnosed as Diffuse large B cell Lymphoma GCB type/ Non GCB Type on immunohistochemistry, one case was T cell Lymphoma of Testis with IHC positive for T cell markers and one case was Acute Lymphoblastic lymphoma involving the testis with IHC positive for PAX 5 and Tdt. Bilateral involvement was seen in case of Acute lymphoblastic lymphoma involving the Testis. And Unilateral involvement of Testis was seen in other cases of Lymphoma of Testis. Conclusion: Lymphomas of testis are rare neoplasms of which the commonest was mostly seen in elderly and was Diffuse Large B cell Lymphoma- Non GCB type.

Keywords: Immunohistochemistry, lymphoma, testis

   AOP206 Top

   Lymphoreticular Pathology Top

Epstein-Barr virus – Positive diffuse large B-cell lymphoma in paediatric age group – A case series

Vishesha Adhvaryu, Uma Sakhadeo, Sridhar Epari, Sumeet Gujral, Tanuja Shet

Tata Memorial Hospital, Mumbai, Maharashtra, India

Introduction: Epstein - Barr virus (EBV) - positive diffuse large B-cell lymphoma (EBV+DLBCL), not otherwise specified (NOS) is a B cell neoplasm in immunocompetent individuals. It was previously designated as EBV+DLBCL of the elderly as it was first described above age of 60 years. Subsequently cases were described in younger individuals who showed better treatment outcomes and survival. Objectives: To study the clinico-pathological features of EBV+DLBCL in peadiatric population. Materials and Methods: We retrospectively reviewed clinico-pathological details of 11 cases of EBV+DLBCL, NOS diagnosed at our institute in the paediatric age group (<18 years) between 2012 and 2022. Results: The median age at diagnosis was 15 years. Male to female ratio was 1.7:1. Clinical and imaging details were available for 8 cases. There was no documented predisposing immunodeficiency. Six cases presented with nodal disease, and 2 cases were extranodal (bone and soft tissue). All the 8 cases presented in advanced stage (Stage III/IV). Serum EBV DNA result was available for a single case (positive). Histologically, all the cases showed variable number of mono/bi/multinucleated Hodgkin/Reed-Sternberg-like cells. Ten out of 11 cases were polymorphic resembling classical Hodgkin lymphoma (cHL) (microenvironment of small lymphocytes, plasma cells, and histiocytes) and one was monomorphic type. Immunohistochemistry revealed diffuse expression of pan-B cell markers (CD20/PAX5/CD79a) and EBV-LMP1 in the neoplastic cells in all cases. CD30 showed variable expression in 9 cases. CD15 was positive in a single case. Median survival was 34 months and one patient died of disease. Conclusion: Recognition of EBV+ DLBCL in paediatric population from cHL carries therapeutic and prognostic implications.

Keywords: Diffuse large B-cell lymphoma, Epstein - Barr virus, paediatric

   AOP207 Top

   Lymphoreticular Pathology Top

Subcutaneous panniculitis-like T-cell lymphoma – A review of pathological and clinical features of 15 cases from a tertiary care center

Priyadarsani Subramanian, Uma Sakhadeo, Epari Sridhar, Sumeet Gujral, Tanuja Shet

Tata Memorial Hospital, Mumbai, Maharashtra, India

Introduction: Subcutaneous panniculitis-like T cell lymphoma (SCPTCL) is a rare cutaneous cytotoxic T cell lymphoma mimicking immune-related panniculitis clinically and histologically. We report 15 cases of SCPTCL, the largest series from India till date. Objectives: To study the clinico-pathological features of SCPTCL diagnosed in the last 6 years at our institute. Materials and Methods: All cases diagnosed as SCPTCL from 2017 to 2022 were retrieved. The clinical details were obtained from institutional electronic medical records database. Histological and immunohistochemical features were reviewed. Results: 15 cases (20 biopsies) of SCPTCL were diagnosed in the study period. The mean age at diagnosis was 28.4 (age range 9-65 years). Fourteen patients presented with typical subcutaneous nodules; one patient presented with systemic hemophagocytosis and pancytopenia who subsequently developed skin lesions. Lupus work-up was negative in all cases. Histology showed lobulocentric lymphoid infiltration of subcutaneous fat by atypical lymphoid cells (15/15), fat necrosis (5/15), rimming of adipocytes by neoplastic T cells (8/15) and karyorrhectic debri (5/15). Hemophagocytosis within the lesion was noted in 4/15 cases. Additionally, two cases showed dermal involvement. The atypical cells were immunopositive for CD3, CD7, CD8 (15/15), TIA (2/2); with loss of CD5 (4/5); negative for CD56 (14/14) and CD30 (8/8). All nine cases for which bone marrow biopsy was available did not show lymphoma involvement. Three cases (3/9) relapsed after first line therapy, however responded well to second line chemotherapy. Conclusion: Careful clinical correlation with histological and immunohistochemical evaluation is essential for diagnosis and management of this rare entity.

Keywords: Cutaneous T cell lymphoma, hemophagocytosis

   AOP208 Top

   Miscellaneous Top

Efficacy of Xylene-free Haematoxylin and eosin staining as compared to conventional Haematoxylin and eosin staining

Khushbu Girdher, Roma Issacs, Saurabh Donald, Nalini Calton

Christian Medical College, Ludhiana, Punjab, India

Introduction: The routine histopathological diagnosis is based on Haematoxylin and Eosin (H&E) stained sections. Xylene is used as a deparaffinising agent in the haematoxylin and eosin staining procedure. Hazards associated with exposure to xylene are of concern, therefore substitutes like Liquid Dish Washing Soap (LDWS) and refined mineral oil have been evaluated in the past which can replace xylene. Objectives: To compare the efficacy of LDWS as a possible replacement for xylene in H&E staining procedure. Materials and Methods: The present study was conducted in the Department of Pathology, Christian Medical College, Ludhiana over 1 year and 6 months from 1st November 2019 to 30th April 2021. It was a single-blinded experimental study. Paired 225 paraffin sections from different tissues were taken for the study. From each tissue block, 2 sections were cut and stained with H&E stain: one section was dewaxed by a conventional method using xylene while the other by xylene free method using LDWS. Slides were scored using five parameters: nuclear staining, cytoplasmic staining, clarity, uniformity and crispness of staining. Results: The total time for staining by the conventional method was 41 minutes whereas it was 27 minutes for the LDWS method, which is 14 minutes less compared to the conventional method. Conclusion: The difference in staining by both methods was minimal and was not statistically significant, suggesting that LDWS is at par with the conventional xylene method in producing adequate staining.

Keywords: Dishwashing soap, Haematoxylin & Eosin, xylene

   AOP209 Top

   Miscellaneous Top

A case series of mucormycosis in post COVID patients

AiuseeNasreen Pooja, Adarsh Sanikop

Jawaharlal Nehru Medical College, Belagavi, Karnataka, India

Introduction: Coronavirus disease (Covid-19) infection has found its association with many types of bacterial and fungal co-infections. The exact incidence of mucormycosis is not known due to lack of population based studies. The estimated prevalence of mucormycosis is around 70 times higher in India than globally. Most identified patients suffering from mucormycosis also have pre or co-existing diseases weakening the immune system and hence raising the susceptibility for mucor infection. Objectives: To study the biopsy specimens of Post-Covid 19 patients for histopathological diagnosis of Mucormycosis using routine H&E and Special Stains. Materials and Methods: In this study, we have taken into consideration 100 biopsy specimens from rt-PCR positive confirmed cases of Covid-19 patients in our tertiary care hospital. The specimens were received at our Histopathology department, all were fixed in 10% in Neutral Buffered Formalin and were processed using LIECA Automated tissue processor. Paraffin blocks were made and thin 3.5 micron ribbon section were taken on the slide and projected to Hematoxylin& Eosin (H&E) staining and the findings were confirmed by Gomori Methenamine Silver (GMS) and Periodic Acid Schiff (PAS) staining. Results: Results obtained from the 100 biopsy specimens were as follows - 78 patients were male and 22 were female; Mean age was 52 years. Out of 100 suspected mucormycosis patients, 81 biopsy specimens turned out to be positive for mucormycosis on histopathology and 19 patients as Sinusitis. Among the 100 suspected mucormycosis patients, 72 patients were immuno-suppressed and 28 patients immunocompetent. Among immunosuppressed patients, majority having Diabetes Mellitus followed by hematological malignancies and post-pulmonary tuberculosis. Nasal cavity middle meatus followed by maxillary sinus remained the most common site involved. Among immunocompetent patients only unilateral paranasal sinus site was affected. Conclusion: In conclusion, Mucor infection is associated with Covid 19 pre infection as well as other immunosuppressing diseases which weakens the immune system and leads to an aggressive disease with poor outcome.

Keywords: Gomori methenamine silver, periodic acid schiff

   AOP210 Top

   Miscellaneous Top

Immunohistochemical expression of programmed death ligand 1 in oral squamous cell carcinoma

Neha Prasad, Jyoti R Kini, Hema Kini

Kasturba Medical College, Mangalore, Karnataka, India

Introduction: Programmed cell death ligand 1 (PD-L1) is an immune checkpoint molecule that weakens the host response against tumors. Blockade of PD-L1 enhances the host response in a variety of neoplasms hence has been incorporated in anti-cancer therapy. Objectives: The aim of this study was to evaluate the expression of PDL-1 in oral squamous cell carcinoma (OSCC) and its association with clinicopathological features. Materials and Methods: A retrospective study was conducted including all resected specimens of OSCC received in the Pathology department over a three year period. Clinical and morphological features were correlated with the immunohistochemical expression of PD-L1. Results: Of a total of 101 resected cases of OSCC, paraffin blocks with tumor were available for 94 cases. Increased expression of PD-L1 was found in 38.3% cases. The tumor proportion score (TPS) (p value= 0.019) and combined positive score (CPS) (p value=0.009) were significantly associated with PDL1 staining intensity. Nearly one third of the cases of well, moderately and poorly differentiated OSCC were positive for PDL-1. Marked lymphocytic response was noted in 53% of the cases with positive PDL1 staining. A highly significant association was seen between CPS, TPS and lymphovascular invasion (p value = 0.005) and TPS with perineural invasion (p value = 0.030). Conclusion: PDL-1 expression was seen in nearly two- fifths of the cases of OSCC. Perineural and lymphovascular invasion were seen to be significantly associated with PDL1 expression, indicating an aggressive nature in these tumors.

Keywords: Oral squamous cell carcinoma, perineural and lymphovascular invasion, programmed cell death ligand 1

   AOP211 Top

   Miscellaneous Top

Detection of human papilloma virus infection in cases of oral carcinoma using p16 immunohistochemistry

Thangjam Ruchika, Sunita Y Patil

JNMC, Belagavi, Karnataka, India

Introduction: Oral cancer is a global health problem with high prevalence and mortality rate across the world. It is the 6th most prevalent cancer worldwide. Oral squamous cell carcinoma accounts for 40-50% of all malignancies in India. The primary known risk factor for oral cancer includes tobacco and alcohol consumption. Recently infection with high risk Human papilloma virus (HPV) has become evident and is linked to the development of oral cancers etiologically. Objectives: To detect human papilloma virus infection in cases of oral carcinoma using p16 immunohistochemistry and to determine the association of human papilloma virus with the histological grading of oral carcinoma using p16 immunohistochemistry. Materials and Methods: A 1 year prospective study of 50 cases of oral squamous cell carcinoma (OSCC) were taken and histological grading was done as per modified Broder's grading system. All the OSCC cases were studied for p16 expression. Cases showing p16 positivity and its association with histological grading were studied. P value <0.05 was considered significant. Results: p16 was positive in 48% of cases with male preponderance. There was no statistically significant association of human papilloma virus infection with the histological grading of OSCC using p16 IHC. (p=0.304). Conclusion: p16 IHC is an important biomarker, particularly to high-risk HPV types infection, which makes it useful in evaluating HPV associated squamous carcinoma. Knowing the HPV status by IHC is a more feasible, easy, cost effective, reproducible test and could impact patient management and survival and the development of prophylactic vaccines based on their viral capsids.

Keywords: Human papilloma virus, immunohistochemistry, oral carcinoma

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Tumor associated tissue eosinophilia and CD117 positive mast cell density in lip and oral cavity squamous cell carcinoma

Jashmine Sanasam, Nadeem Tanveer, Sonal Sharma, Sandeep Kumar

University College of Medical Sciences and GTB Hospital, New Delhi, India

Introduction: Tumor associated tissue eosinophilia and mast cell density have been evaluated as prognostic marker in a variety of tumors, e.g. Breast, prostate, colorectal carcinoma. Recently, few studies have found the difference in prognostic significance of intratumoral and peritumoral mast cell density. Objectives: To study tumour associated tissue eosinophilia (TATE) and CD117 immunopositive intratumoral and peritumoral mast cell density in Lip and Oral cavity squamous cell carcinoma. Materials and Methods: This was a retrospective study done on archival blocks from oral and lip squamous cell carcinoma (50 cases each). Tumor associated tissue eosinophilia was evaluated on Hematoxylin and eosin slide and immunohistochemistry for CD117 was performed for mast cell density. All the cases were graded according to Anneroth's multifactorial grading system and Brandwein and Gensler scoring system of grading. The data was analyzed to find any correlation between these tumors, mast cell and eosinophil density. Results: In oral cavity squamous cell carcinoma, there was a statistically significant correlation between the tumor grade and the Anneroth grade and the intratumoral CD117 mast cell density. However, tumor associated tissue eosinophilia and peritumoral mast cell density did not correlate with the Anneroth grade or Brandwein Gensler score. In case of lip squamous cell carcinoma, only tumor associated tissue eosinophilia had a significant correlation with tumor grade. Conclusion: Intratumoral CD117 mast cell density can provide useful information regarding prognosis of oral cavity squamous cell carcinoma. This can have implication for anti-mast cell therapy in future.

Keywords: Mast cells, tumor associated tissue eosinophilia

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An analysis of quality indicators in a NABL accredited hematology laboratory

Shraddha Kankariya, Arpita Nishal, Sejal Gamit, Deepshikha Dave, Rahul Modi

Government Medical College, Surat, Gujarat, India

Introduction: As stated by the ISO 15189:2012, “The laboratory shall establish Quality Indicators to monitor and evaluate performance throughout critical aspects of pre-examination, examination and post examination processes”. This principal is an important aspect for NABL accreditation, to assure that the laboratory is upto the updated standards to prevent any kind of patient mismanagement. Objectives: 1) To monitor and access QI in various aspects of Lab. 2) To analyze and periodically review QI. Materials and Methods: Different QI at pre analytic, analytic and post analytic levels were evaluated and studied at our Hematology lab during the year of 2021 and non conformities that arose with respect to NABL criteria were resolved with due importance. Results: Pre analytic indicator- Maximum sample rejection rate of a year was 4.7% and minimum was 0.7%, which were within laboratory's acceptable limits. Analytic indicator- Majority of EQAS results for our instruments were within +/- 2 of Z score. Periodically Interlaboratory Quality Control done for various parameters was found to be satisfactory. Internal laboratory comparison – CV% charting is done daily for all the instruments. Interobserver quality check is also done periodically. Regular critical value reporting was between 0.7% to 4% monthly. Instrument breakdowns were noted and prompt action were taken. Post analytic indicators- Monthly TAT was maintained well within the lab limits. Conclusion: QIs are needed for monitoring satisfactory results and continuous improvement of the lab for better patient management.

Keywords: Hematology lab, NABL, quality indicators

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Evaluation of Cyclin D1 and Her2 Neu protein expression in head and neck squamous cell carcinoma

Snigdha Sinha, Subashish Das, S M Azeem Mohiyuddin

Department of Pathology, Sri Devaraj URS medical college, Kolar, Karnataka, India

Introduction: Head and neck cancer are one of the most common cancer worldwide and are highly aggressive. Various studies have documented the usefulness of molecular markers in carcinogenesis and proven helpful in diagnosis, treatment of head and neck cancers. Objectives: 1. To determine the proportion of expression of cyclin D1 and HER2 neu in head and neck squamous cell cancer and to analyze the association between its expression with histological grading and TNM staging. 2. To analyze the association between expression of HER2neu and Cyclin D1 with site and nodal status of the tumor and clinical outcome such as to locoregional control, depth of invasion and regional metastasis. Materials and Methods: Total of 69 histologically proven cases of head and neck squamous cell carcinoma were taken and immunohistochemical staining for cyclinD1 and HER2neu was done for these cases. For intensity scoring of HER2neu, 10 fields were chosen, based on which the intensity score was calculated and IHC was graded according to ASCO guidelines. For scoring analysis of cyclin D1, percentage of positive cells were calculated and averaged and the intensity score was graded. Results: 52 cases out of 69 (75%) showed strong and moderate positivity for cyclin D1 with a significant p value of 0.008. Only 5 out of 69 cases were positive for HER2neu and the results were not significant with a p value of 0.6643. Conclusion: The score for cyclinD1 expression increases with increase in tumor stage and metastases. The role of HER2neu should be studied more extensively to further establish it as a known marker.

Keywords: CyclinD1, Her2neu, squamous cell carcinoma

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Expression of mismatch repair (MMR) protein in endometrial carcinoma and endometrial hyperplasia

Belani Vaishali, Ranjitha Rao, Sharada Rai, Kausalya Kumari Sahu

Kasturba Medical college, MAHE, Mangalore, Karnataka, India

Introduction: Endometrial carcinoma (EC), in particular the most common endometrioid form (EEC), contributes significantly to the worldwide cancer burden, and its molecular categorization has lately gained significant importance. As theses molecular changes are associated with lynch syndrome (LS). Identification of these proteins also serve as screening in suspected cases for LS. Objectives: To study the expression of mismatch repair proteins (MLH1, PMS2, MSH6, MSH2) in EC and Endometrial hyperplasia (EH). Materials and Methods: The current study was carried out on 52 cases EC and 65 cases EH (7 cases of disordered proliferative, 12 cases of EH with atypia and 46 cases EH without atypia) retrieved from January 2016 to April 2022. H&E staining and immunohistochemical staining with MLH1, PMS2, MSH6, MSH2 were done. Results: Of 52 cases 78.84%were EEC. 42 cases showed negative expression of both MLH1/PMS2 (42/52, 80.77%). MLH1 negative expression was significant (0.005) compared to other markers. There was significant statistical correlation (0.004) between lower FIGO grade and negative expression of MLH1/PMS2. Only 6 cases had significant family history. All (12/12) the cases of EH with atypia were MMR deficient (MMR d) and 69.23% (32/46) were MMR d cases in EH without atypia. Conclusion: Of all the EC's, EEC are the most common cancers to be associated with MMR deficiency. MLH1 /PMS2 combination was commonest protein deficiency seen in our study and MLH1 loss was the commonest seen in EH with atypia. Significant correlation was seen with MMR deficiency and low FIGO grade.

Keywords: Endometrial carcinoma, endometrial hyperplasia, MMR

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Clinicopathological spectrum and utility of MMP-7 and Bmi-1 in non-small cell lung carcinoma

Farheen Chishti, Yookarin Khonglah, Biswajit Dey, Vijay Noel Nongpiur

Departments of Pathology and 1TB and Respiratory Diseases, North-Eastern Indira Gandhi Regional Institute of Health and Medical Sciences, Shillong, Meghalaya, India

Introduction: Non-small cell lung carcinoma constitutes majority of lung cancer cases. It is imperative to study biomarkers like MMP-7 and Bmi-1 in a high incidence region like North-East India to assess their relevance. Majority of the studies in the past were done using either immunohistochemistry on biopsy or by detection of mRNA by RTPCR which are either invasive or labor-intensive. Objectives: To assess the levels of MMP-7 and Bmi-1 in patients of NSCLC to determine a cut-off value compared to normal population and to correlate them with clinicopathological parameters and assess their relevance. Materials and Methods: Blood samples from 50 cases and 50 controls were collected and serum separated. Serum levels were measured by ELISA method for autoantibodies against MMP-7 and Bmi-1. Results: 62% of the cases were in stage IV and had metastatic disease. Levels of MMP-7 were significantly higher in cases of NSCLC as compared to healthy controls (Median 1.54 vs 1.12, p = 0.00016). Sensitivity and specificity for NSCLC detection were 80% and 28% respectively. Levels of Bmi-1 were significantly higher in cases as compared to controls (Median 135.4 vs 91.6, p=0.00056). Sensitivity and specificity for NSCLC detection were 68% and 64% respectively. Higher MMP7 levels correlated with metastases p value= 0.0182 (p<0.05). There was no significant difference between Bmi-1 level and clinicopathological parameters. Conclusion: The cut-off values of MMP-7 and Bmi-1 were1.54 ng/ml and 135.4 pg/ml respectively. They can be a potential screening tool for diagnosis of NSCLC since they are easily accessible in blood and have a longer half-life than other protein biomarkers which are currently used.

Keywords: Bmi-1, lung cancer, MMP-7

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A study of expression of cyclooxygenase-2 in cervical intra epithelial neoplasia and cervical cancer

S Nuzhath, Savitri Mallikarjun Nerune

Shri B.M Patil Medical College, Vijayapura, Karnataka, India

Introduction: Cervical cancer is the second most common cause of death in women worldwide, and in developing countries. Isoenzyme, cyclooxygenase (COX)-2 is induced by growth factors, tumour promoters, oncogenes, carcinogens. It promotes the proliferation, growth, spread of the tumour and overexpressed in transformed cells and malignant tissues. In this study, we evaluated the expression pattern of COX-2 in CIN, and in different types of cervical cancer and its correlation with tumour differentiation for therapeutic benefit. Objectives: To analyze the differential expression pattern of COX-2 in CIN and different types of cervical cancer and its correlation with tumour differentiation. Materials and Methods: 34 cases of histologically diagnosed in-situ and malignant lesions of the cervix were taken and paraffin blocks were retrieved from the Histopathology Section. Tissue was processed routinely. One section was stained with H & E for morphologic diagnosis and the other sections were subjected to COX-2 immunohistochemical staining. Colon carcinoma cases were taken as positive controls. Cytoplasmic and membrane staining of tumour cells were considered as positive and the grading was done. Results: COX-2 expression studied in 34 cervical cancer cases show overexpression in squamous cell carcinoma 20.6% (7 cases showing >50% positive tumor cells) when compared with the insitu and adenocarcinoma. High grade tumors have higher expression. Conclusion: COX-2 expression is directly proportional to the level of grading of tumor. Higher the grading, higher is the expression. Selective COX-2 inhibitors increase the efficacy of chemotherapy or radiotherapy.

Keywords: COX-2, high grade, overexpression

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Histopathological diagnosis of secretory carcinoma of salivary gland: Algorithmic approach; a single institution experience

P K Pravanya, K R Anila, Anitha Mathews, K Jayasree, Malu Rafi, Bipin T Varghese

Regional Cancer Centre, Thiruvananthapuram, Kerala, India

Introduction: Secretory carcinoma (SC) is a type of salivary gland carcinoma which was primarily segregated out from acinic cell carcinoma (ACC) based on morphological features and characteristic ETV6-NTRK3 gene fusion. Objectives: To study the histopathological features of secretory carcinoma cases diagnosed at our centre from January 2016 to December 2020. To assess the utility of S100, DOG-1 immunohistochemical panel in diagnosis of secretory carcinoma. Materials and Methods: Clinicopathologic data, haematoxyllin & eosin slides and IHC slides of cases of SC diagnosed from January 2016 to December 2020 were retrieved from our data base and reviewed. Results: 13 cases of SC were identified with male to female ratio of 12:1. Age ranged from 14-65 years with majority cases, eight involving parotid. Characteristic histopathological pattern included papillary cystic pattern with luminal secretions. The basic IHC panel, S100 and DOG-1 showed consistent S100 positivity and DOG-1 negativity in all cases. In addition mammaglobin immunostain done in one case showed positivity. Conclusion: Papillary cystic pattern with luminal secretion was the characteristic histomorphological pattern observed in all cases of SC.IHC panel, S100 and DOG-1 were contributory in confirming the diagnosis. All patients underwent surgery. Apart from two patients, other patients received postoperative radiotherapy due to indications like lymph node metastasis, perineural invasion, high grade transformation and positive margin. All patients are on follow up with follow up period ranging from two to six years. The follow up period has been uneventful. Tyrosine kinase inhibitors (TRK), larotrectinib and entrectinib, have recently emerged as new alternatives for treating NTRK fusion gene-driven diseases. Hence diagnosis of SC and segregating it from ACC is of therapeutic importance.

Keywords: Acinic cell carcinoma, salivary gland, secretory carcinoma

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Immunohistochemical spectrum of ovarian neoplasms

Mehar Ghuman, Harpreet Kaur, Aminder Singh

Dayanand Medical College and Hospital, Ludhiana, Punjab, India

Introduction: Ovarian cancer is the sixth most common cancer and the seventh leading cause of cancer deaths among women worldwide with worst prognosis among all gynecological malignancies. Ovarian tumors are heterogenous so an accurate histopathological diagnosis is required. World Health Organization classifies ovarian tumors as per their most probable cell of origin and histomorphological features. Immunohistochemistry is helpful where the morphology do not allow definite diagnosis of tumor. Objectives: To study immunohistochemical spectrum of ovarian neoplasms and correlation with histopathology. Materials and Methods: Study was conducted in DMCH, Ludhiana from 1st March 2020 to 28th February 2022, for two years. All primary and secondary ovarian neoplasms undergoing immunohistochemistry for definite categorization were included in the study. The ovarian neoplasms which did not underwent immunohistochemistry test and all other neoplasms were excluded from the study. A total of 20998 specimens were received during this study period and 88 cases were of ovarian neoplasms on which immunohistochemistry was done. Results: All the tumours were classified according to the WHO classification 2020 for ovarian neoplasms. Epithelial tumours formed the largest group with 56 cases (63.7%). Second most common type were sex cord stromal tumours comprising of 15 cases (17.0%) followed by metastatic tumours in 10 cases (11.3%), germ cell tumours 6 cases (6.8%). Least common were mesenchymal tumours comprising of 1 case (1.2%). Conclusion: The need to ascertain a correlation between the histomorphological features and IHC findings wherever possible is needed for a definitive diagnosis.

Keywords: Histomorphology, immunohistochemistry, ovarian tumors

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TP53 immunoexpression in mucinous tumors of the ovary

Joanna George, Mary Mathew

Kasturba Medical College, Manipal, Karnataka, India

Introduction: Ovarian mucinous tumors are a spectrum of tumors that demonstrates stepwise accumulation of genetic events during transformation from benign to malignant tumors. Mutations of TP53 have been studied in progression of mucinous benign and borderline tumors to mucinous ovarian carcinoma (MOC). Objectives: To study the pattern of expression of p53 on immunohistochemistry in ovarian mucinous tumors. Materials and Methods: The slides of 57 patients diagnosed with ovarian mucinous tumors were retrieved and reviewed. Relevant sections were taken and a tissue microarray (TMA), was prepared and were stained with p53 IHC. The pattern of expression was studied using the criteria as by Kang et al. into normal or abnormal (overexpression {5%, 50%, 70%, 100%}, complete absence or cytoplasmic). Results: In our study, mucinous cystadenomas, 26/31 (78%) showed a normal pattern of staining, while 3 cases showed complete absence and 2 showed cytoplasmic staining. 11 (57%) of 19 MBOT showed abnormal p53 staining and all 7 of the MOCs had abnormal TP53 staining of which 5 showed overexpression. Conclusion: P53 immunohistochemical pattern of expression in ovarian mucinous tumors show varied expression and is useful in studying the mutational patterns in the spectrum of benign, borderline, and malignant tumors, taking account of tumoral heterogeneity and terminal differentiation.

Keywords: Immunohistochemistry, mucinous ovarian tumors, TP53 expression

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Morphological spectrum of lupus nephritis

Ramneek Kaur, Pavneet Kaur Selhi, Sumit Grover

Dayanand Medical College and Hospital, Ludhiana, Punjab, India

Introduction: Renal disease manifests in up to 40-75% of SLE patients, most often occuring within five years of disease onset and hence is considered one of the strongest predictors of an unfavourable outcome. Renal biopsy at the time of initial presentation and subsequently during follow up may be needed to decide the therapeutic approach. Histopathological features play a significant role in patient management. Objectives: 1. To study in detail the histopathological features of kidney biopsies in patients of SLE 2. To correlate the histopathological features with the clinical profile. Materials and Methods: This study will be conducted in the Department of Pathology at Dayanand Medical College and Hospital, Ludhiana for a period of 2 years. All the biopsies were histologically analysed and classified into various classes based on light microscopy findings using routine, special stains and direct immunofluorescence. Results: A total of 415 renal biopsies were received, out of which 27 were diagnosed as lupus nephritis. Out of the 27 cases studied, 16 patients (59.3%) belonged to the age group of 21-40 years with a mean age of 38.2 years and Male: Female ratio being 1:4.4. The most common clinical complaint was synovitis in 16 (59.2%) followed by fever in 12 (44.4%), non-scarring alopecia in 11 (40.7%) and then malar rash in 10 patients (37%). Among the 27 patients, 12 patients (44.5%) had moderate degree of anaemia, 8 (29.6%) having mild degree of anaemia and 5 suffered from severe anaemia. 16 cases (59.3%) had Positive Anti dsDNA antibodies by ELISA. Most of our cases (n=13, 48.2%) were having Diffuse Proliferative Lupus Nephritis i.e Class IV LN. The second most common type was Focal Proliferative Lupus Nephritis i.e Class III LN with 5 cases (18.5%). Conclusion: Kidney biopsy is an essential tool to obtain good database for assessment and diagnosis of Lupus Nephritis based on the histomorphological findings.

Keywords: EULAR classification, kidney biopsy, lupus nephritis

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Changing trends in fungal rhinosinusitis – A study during pre-COVID, COVID and post-COVID era

L Nayana, Rajesh H Chandan, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: Fungal rhinosinusitis (FRS) comprises a spectrum of disease processes, which vary in clinical presentation, histologic appearances and biological significance. Although fungal infection of nose and paranasal sinuses is uncommon, its incidence has increased in the recent years. This change is mainly due to international traffic and opportunistic infections as a consequence of use of powerful cytotoxic drugs, steroids and antibiotic therapy. Objectives: (1) To study the morphological spectrum of fungi causing FRS & to categorise the types of FRS during pre-covid, covid & post-covid period. (2) To correlate clinically & with culture reports. Materials and Methods: This is a 7 yrs study (5 yrs retrospective & 2 yrs prospective) from 2015 to 2022 conducted in dept of Pathology, KIMS, Hubli. All biopsy specimens from nose & paranasal sinuses of the patients above 18 yrs of age were studied. For retrospective study, histopathology slides diagnosed as FRS were studied. Special stains like GMS, PAS were done. Results: A total of 415 cases of rhinosinusitis were studied out of which 235 cases were of FRS (51 precovid, 171 covid, 13 postcovid) with more than 1/3rd of cases being Acute Invasive FRS. FRS was more common in elderly age groups with involvement of maxillary sinus in most of the cases and Mucor being the commonest species. Other species identified were Aspergillus, Curvularia & Candida. Pathological examination had a higher sensitivity than microbiology examination in the diagnosis. Conclusion: COVID-19 was not the only factor contributing to FRS rather other factors such as DM, steroid use etc. were also contributory.

Keywords: Aspergillus, fungal rhinosinusitis, mucormycosis

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Correlation of intraoperative frozen section with histopathology

Devika Bansal, Harpreet Kaur, Bhavna Garg

Dayanand Medical College and Hospital, Ludhiana, Punjab, India

Introduction: Tissue specimen taken from the patient during surgery where surgeons need pathologic information urgently, they request for an intra-operative consultation on that tissue. Objectives: To study the indications, sensitivity, specificity, PPV, NPV and the diagnostic accuracy of frozen section. Materials and Methods: This study was done from 1st March 2021- 28th February 2022. Fresh tissue sent for frozen section, was examined grossly and representative sections were taken. Sections were cut at a thickness of 4-6 micron in the cryostat and were immediately stained with rapid hematoxylin and eosin and Toluidine blue stain. Frozen section diagnosis was made and immediately conveyed to the operating surgeon. For the remaining tissue, it was fixed in 10% formalin, grossed and representative sections were processed. The frozen section diagnosis was then compared to left over frozen section and permanent sections. Results: ighty five cases of frozen section were received for primary diagnosis [46.5%], 64 cases to look for metastasis [34.9%] and 34 cases to know the resected margin status [18.6%]. 156 cases were concordant with the final histopathological diagnosis [85.3%] and 26 cases were [14.2%] discordant. 1 case was deferred. The overall sensitivity, specificity, PPV and NPV of frozen section were 72.5%, 96%, 93.5% and 81.6%. Conclusion: Frozen section remains an invaluable tool for intraoperative diagnosis.

Keywords: Frozen section, sensitivity, specificity

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Experience with targeted NGS (Next Generation Sequencing) analysis of non small cell lung cancer: Detection of rearrangements and actionable mutations of EGFR, ALK, ROS1, MET, BRAF and its correlation with clinico – Pathological variables and clinical benefits

Ankita Pal, Rupanita Biswal, Soumya Surath Panda, Debahuti Mohapatra

IMS and SUM Hospital, SOA University, Bhubaneswar, Odisha, India

Introduction: Lung cancer is the leading cause of cancer morbidity and mortality in men and in women, it ranks third for incidence. Different types of Non small cell lung cancer (NSCLC) has certain gene mutations associated with its pathogenesis which can be detected by an innovative method known as 'Next Generation Sequencing '(NGS) owing to the evolution of targeted therapies and show consistent improvements in progression – free survival compared to conventional cytotoxic agents. Objectives: Application of NGS in Non small cell lung cancer (Adenocarcinoma) for the detection of mutations in the paraffin embedded tissue sections with reference to clinical, demographical, radiological & pathological parameters in each case. Materials and Methods: The prospective study of two years from July 2020 – July 2022 was done on the 46 samples. The cases were diagnosed as NSCLC after histopathological and immunohistochemical study then sent for NGS (Ion S5 system and Ion ampliseq technology). Results: 46 cases were referred for testing by NGS comprising of 38 biopsies and 8 cell block. EGFR mutations were most commonly detected in non smokers female, with papillary pattern. BRAF & ALK showed both cribiforn and micropapillary pattern. MET showed solid pattern. Some of the cases show co - mutation of EGFR-BRAF & EGFR-PIK3CA showing acinar & papillary pattern and adenosquamous type respectively. Conclusion: The advanced/metastatic stage of NSCLC are inoperable. Histopathological pattern can help in deciding the molecular panel & NGS can dictate those genetic aberrations that will individualize further treatment protocol for each patient and improves the quality of life.

Keywords: EGFR, mutation, next generation sequencing

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Bone marrow findings in geriatric patients

Nishav Garg, Harpreet Kaur, Bhavna Garg, Vikram Narang

DMCH, Ludhiana, Punjab, India

Introduction: Hemograms, Peripheral blood smears along with Bone marrow examination are important investigations carried out for the diagnosis and work-up of various hematological and non-hematologic disorders in patients across all age groups. With growing number of elderly people, increased incidence of leukaemias, lymphoma, myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPNs) and multiple myeloma is being observed. It seems likely that the indications and diagnoses made, as well as the impact of these diagnoses on therapy, would be different for this subpopulation. Objectives: To evaluate the hematological disorders in geriatric patients. Materials and Methods: The study was of 12 months (March 2021 to march 2022) prospective study and was conducted in the Department of Pathology, Dayanand Medical College and Hospital, Ludhiana. The study included all the patients 60 years and above who underwent bone marrow examination (aspiration/ biopsy or both) along with Hemogram, peripheral blood smears in the department. Special stains like Ziel-Neelsen, PAS and GMS, Perls prussion blue, Masson's trichrome, Reticulin stain was done wherever necessary. The hemograms and PBFs of all cases was also studied and analysed in detail and correlated with the bone marrow examination. Other ancillary tests like flow cytometry and immunohistochemistry was also included wherever available to reach a definite diagnosis. Results: Total 206 cases underwent bone marrow examination and the age range of the patients was 60 to 92 years, 128 were males and 78 were females. The most common indication being suspicion of multiple myeloma (23.0%) followed by pancytopenia (17.%). Multiple myeloma was most common pathological diagnosis in 18.9% cases followed by Myeloproliferative neoplasm in 12.1%, Lymphoproliferative neoplasm in 11.1%, Normocellular normoblastic marrow in 9.6%, Idiopathic thrombocytopenic purpura in 8.7%, acute leukemia in 7.7%, Myelodysplastic syndrome in 5.8%, Megaloblastic anemia in 5.3%, Aplastic anemia in 3.80%, Anemia of chronic disorder in 3.30%, Hemophagocytic lymphohistiocytosis in 2.90%, Leukomoid reaction 2.40%, Hemolytic anemia in 1.40%, Hypereosinophilic syndrome in 0.97%, Dual deficiency anemia in 0.90%, Iron deficiency anemia in 0.40%, Leishmaniasis in 0.40%, Granulomatous inflammation in 0.40%. Conclusion: In this study we found that Diagnosis of Multiple myeloma is the most common findings in bone marrow studies in elderly patients. Diagnosis of Multiple myeloma prompt for early institution of treatment resulting in reduced morbidity and mortality in such patients.

Keywords: Bone marrow, geriatrics, hematologic disorder

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Bcl-2 and Ki-67 in endometrial hyperplasia and disordered endometrium

Akalya Manimaran, Savita Dongapure

MVJ Medical College and Research Hospital, Hoskote, Karnataka, India

Introduction: Endometrial hyperplasia affects both pre & postmenopausal women which is a pathologic lesion that ranges from mild, reversible glandular proliferations to direct cancer precursors. Bcl-2, an anti-apoptotic gene involved in the regulation of apoptosis and Ki-67, the proliferation marker is the indicator for cell proliferation in endometrial carcinoma. Objectives: To study the expression of Bcl-2 and Ki-67 in endometrial hyperplasia and Disordered endometrium. Materials and Methods: The prospective study consists of total 33 patients out of which 15 cases are disordered proliferative endometrium and 18 cases are endometrial hyperplasias. Immunohistochemical staining was performed using Bcl-2 and Ki-67. The study was done at the Pathology department in MVJ Medical college and research hospital from September 2020 to July 2022. Quantitative variables are presented using Mean+/- SD. Results: Bcl-2 showed high positivity (>75%) in 43.75% of the Non-atypical hyperplasia cases and 53.33% of the Disordered proliferative cases show 50-75% positivity. Ki-67 showed high positivity (50- 75%) in 13.33% of the Disordered proliferative endometrium cases and 12.50% of the cases of Non-atypical hyperplasia and 60% of the disordered proliferative cases show <25%. 50% of the cases of Non atypical hyperplasia shows 25% positivity of Ki-67. Conclusion: Over expression of Bcl-2 expressions in Endometrial hyperplasia is considered to progress towards frank malignancy. Ki – 67, the proliferation marker is used to determine the progression of endometrial hyperplasia to carcinoma.

Keywords: Bcl-2, endometrial hyperplasia, Ki-67

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A sudden surge of tissue invasive fungal infection in post COVID 19 patients: A review of 236 cases

Heer Dabhi, Komal Patel, Mubin Patel, Pallavi Chaudhari, Suresh Padasala, Mandakini Patel

Goverment Medical Collage, Surat, Gujarat, India

Introduction: Health care sector across India reported significant increase in cases of covid linked mucormycosis and other fungal infections in covid 2nd wave. Mucormycosis is an opportunistic fulminant fungal infection in immunocompromised host. Concomitant diabetes and use of high dose steroid, further add to already compromised immunity in covid patients. It may manifest as rhino-orbital or cerebral mucormycosis. Fungus invades the arterioles and causes tissue necrosis, which is the hallmark of mucormycosis. Objectives: To study frequency and histomorphology of various tissue invasive fungal infections and their correlation with other parameters. Materials and Methods: This is retrospective observational study. Here we present histological and other findings in 236 cases of clinically suspected post covid fungal infection at our institute during 2-month periods. Results: Out of 236 specimens received, 211 showed presence of tissue invasive fungal infection with positivity rate of 89.4%, where male to female ratio was 2.2:1, mean age was 51 years. Most common organism detected was Mucor species in 72%, Aspergillus in 4.3% and 23.7% had Mixed infection. Angioinvasion was seen in 38.86%. The most common site of involvement was sinonasal (84.36%) followed by orbital (15.16%) and cerebral (0.47%). 68.18% patients were diabetics, 39% had history of moderate to severe covid requiring oxygen, 72.7% had received steroid. Conclusion: Mucor and Aspergillus were commonly observed tissue invasive and angioinvasive fungal species. Diabetes mellitus and rampant use of corticosteroid in a background of Covid-19 appears to increase opportunistic fungal infection.

Keywords: COVID-19, mucormycosis, tissue invasive fungal infection

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Study of diagnostic accuracy of immunohistochemical markers in endometrial carcinoma and endometrial hyperplastic disorders in a tertiary healthcare centre

Nishtha Danu, Smita Mulay, Anuja Patil, R C Nimbargi

Bharati Vidyapeeth Deemed to be University, Pune, Maharashtra, India

Introduction: Atypical endometrial hyperplasia is a precursor of endometrioid adenocarcinoma. The distinction between atypical hyperplasia and adenocarcinoma is difficult at morphological and cytochemical levels. WHO divides endometrial carcinoma into endometrioid carcinoma and others. Endometrioid carcinoma is low grade, others are high grade. IHC helps in determining diagnosis and prognosis in carcinogenic lesions. Objectives: To study the characterization of ER, PR, p53 and Ki67 in hyperplastic endometrial disorders and correlate the histopathological findings in endometrial hyperplasia and carcinoma, clinically. Materials and Methods: We studied 86 cases of endometrial hyperplasia and carcinoma (hysterectomy and biopsy specimens). Hematoxylin-eosin and immunohistochemistry staining for p53, ER, PR and Ki67 were performed. Results: Among 86 cases, 57 were typical hyperplasia (66.2%), 13 were atypical hyperplasia (15.1%) and 16 were endometrial carcinoma (18.6%). Of these 16 carcinoma cases, we got 12 endometrioid carcinoma and 4 clear cell carcinoma cases from 11 hysterectomy and 5 biopsy specimens. P53 positivity was seen in all the cases of clear cell carcinoma and 16.6% cases of endometrioid carcinoma. Absent ER, PR with high Ki67 expression was seen in clear cell carcinoma. All cases of hyperplasia were ER, PR positive and p53 negative with low Ki67 index. The results were correlated with age, grade and stage. Positive hormonal history was correlated in 50 cases. Conclusion: High ER, PR and low p53 expression helps in differentiating endometrioid carcinoma from clear cell carcinoma which shows high p53 and absent ER, PR expression. Ki67 expression was higher in clear cell carcinoma.

Keywords: Endometrial carcinoma, endometrial hyperplasia, immunohistochemistry

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Tip of the 'eyes'berg: A clinicopathological study of eyelid mass lesions

Saudamini Agarwal, Mithila Bisht, Anjana Arya, Nitesh Mohan

Rohilkhand Medical College and Hospital, Bareilly, Uttar Pradesh, India

Introduction: Geographical location, age, gender, and skin type are a few variables that affect the great array of lesions that can develop in the eyelid. While the final diagnosis is determined by histopathology, the provisional working diagnosis depends largely upon the clinical and demographic presentation of the patient. Objectives: To determine the clinicopathological characteristics of all patients with eyelid lesions diagnosed histopathologically in a tertiary case institute in western Uttar Pradesh restrospectively over a 12-month-period. Materials and Methods: Methods: Of the total 28 cases, all inflammatory, infectious, and cystic lesions were classed as non-neoplastic, whilst the neoplastic group was classified into benign and malignant. Demographics, clinical features, histopathological diagnosis, and special tests were documented and subjected to descriptive statistical analyses. Results: 16 lesions (57.2%) were non-neoplastic while 12 (42.8%) were neoplastic. The non-neoplastic category, led by epidermoid cyst, affected males more frequently, and was encountered most between the ages of 11 and 30 years. Benign tumours were observed in 5 cases (41.7%) and showed a 100% predilection towards the upper eyelid. Malignant neoplasms had a higher occurrence between the ages of 61 and 70 years. 5 of the 7 cases (71.4%) were sebaceous gland carcinoma. Conclusion: Our study demonstrates that eyelid mass lesions are largely non-neoplastic and present most in adolescence and young adulthood. Malignant tumours, the most common of which in Asian countries is sebaceous gland carcinoma, are more frequent in the elderly. It can, thus, be inferred that knowledge of clinical characteristics facilitates prompt clinical recognition, prevents misdiagnosis and also offers corroboration for histopathological confirmation.

Keywords: Epidermoid cyst, eyelid, histopathology

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Hansen's disease revisited – An immunohistochemical expression of CD74 in spectrum of leprosy

Neha Reddy, Sharada Rai, S Anusha

Kasturba Medical College, Mangalore, Karnataka, India

Introduction: Skin and peripheral nerves are both impacted by leprosy, a chronic disease brought on by the Mycobacterium leprae. Self-limiting tuberculoid leprosy and highly infectious lepromatous leprosy, which is characterized by a variety of symptoms, are the two distinct poles. The pro-inflammatory cytokine macrophage migration inhibitory factor (MIF) has a receptor called CD74, and when it interacts with the receptor, it increases the bactericidal activities of macrophages. Recently, migratory inhibitory factor and CD74 were discovered in skin biopsies of patients with lepromatous leprosy. These results led us to explore CD74 immunoreactivity in the entire spectrum of leprosy. Objectives: The objective of this study was to compare CD74 expression in entire spectrum of leprosy obtained from people undergoing dermatological consultation. Materials and Methods: Present study evaluated 60 cases of leprosy over a five-year period, comprising of 17 tuberculoid, 13 borderline tuberculoid, 11 lepromatous, 10 borderline lepromatous, 7 indeterminate and 2 borderline borderline leprosy. Routine examination and CD74 immunohistochemistry were analyzed according to different lineages of skin. Results: In the epidermis we found enhanced expression of CD74 in keratinocytes in the skin lesions of tuberculoid spectrum compared to lepromatous spectrum (p<0.005), whereas on the other hand CD74 expression in endothelial cells was significantly increased in lepromatous spectrum compared to tuberculoid spectrum (p<0.026). Conclusion: We conclude that CD74 has an important role in the localized inflammatory response in the leprosy; however, its participation is unclear. Further research is needed to understand how macrophage inhibitory factor, contributes to the immune microenvironment in leprosy.

Keywords: CD74 immunohistochemistry, leprosy, skin

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An institutional experience of neuroendocrinal neoplasm in par with recent classification – A retrospective study

Akriti Singh, D C Deepak, V Srinivasa Murthy

ESIC MC and PGIMSR, Rajajinagar, Bengaluru, Karnataka, India

Introduction: Neuroendocrine neoplasms (NENs) are tumors of neuroendocrine cells which are endocrine cells with neuron like features. The spectrum includes Pitutary, Parathyroids, Thyroid, Lungs, Gastrointestinal-tract, Thymus, breast, Paraganglia and other rare sites. The annual incidence varies between 1-5/1,00,000 worldwide. Majority of the NENs are neuroendocrine tumors (NETs) whereas 10–20% of NENs are neuroendocrine carcinomas (NECs). Objectives: To study the histomorphological spectrum of Neuroendocrine tumors with their Immunohistochemistry. To Classify the Neuroendocrine neoplasms based on the recent 2022 WHO classification. Materials and Methods: This is a five-year retrospective study from Jan 2017-Jan 2022. Demographic details like Age, Gender and relevant clinical history were documented. The tumors were categorized as per the anatomical site along with Immunohistochemistry study and Grade based on mitosis and Ki-67 index. The cases were classified according to the recent WHO Classification. Results: A total of 55 Neuroendocrine neoplasms were studied of which 34 were males and 21 were females of which GIT and pancreaticobiliary were 36, Lung and thymus were 4, Thyroid and parathyroid were 8, Pheochromocytoma were 3, metastatic were 2; and cervix and paraganglia were 1 case each. 17 cases were Neuroendocrine Carcinomas. NETs were classified into Grade1, Grade2 and Grade3 based on Mitotic counts and Ki-67 index. Conclusion: Neuroendocrine neoplasms are termed as zebras of oncology world because of their heterogenecity and confusing terminology. Hence, accurate classification and prognostication are of utmost importance to guide the clinicians towards best available treatment for patients with Neuroendocrine neoplasms.

Keywords: Ki67, neutroendocrine neoplasms, neuroendocrine tumors

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Histomorphological study of endometrium and immunohistochemical expression of estrogen and progesterone receptor in women with abnormal uterine bleeding

Anuja Mool, Kalpana Rathod, Leena Nakate

BJMC and Sasoon General Hospital, Pune, Maharashtra, India

Introduction: Abnormal Uterine Bleeding is defined as any uterine bleeding that is more than normal volume, of longer duration and varying in regularity or frequency. It is one of the commonest symptoms with which women present in gynaecology OPD. It causes impact on patient family, personal and day to day life. Objectives: 1) To determine various endometrial causes of abnormal uterine bleeding. 2) To evaluate estrogen and progesterone receptor expression by endometrium in cases of Abnormal uterine bleeding. Materials and Methods: Study design- Observation Cross Sectional; Study Setting-Tertiary health care center; Duration-March 2021 to January 2022; Sample size-55; Dilatation and Curettage Endometrial samples. The scoring of Estrogen and progesterone receptor was evaluated using Quick score. Results: 55 endometrial samples of patients diagnosed with abnormal uterine bleeding were studied. 49.1% females (27/55) fell in the age group 41-50 years. Histopathological diagnosis revealed 31.4% neoplastic lesion which involved 3 endometrial carcinoma, 1 adenocarcinoma, 3 cases each of complex and simple hyperplasia with atypia, 6 cases of simple cystic hyperplasia. The average QUICK score for glandular estrogen receptor was significantly higher than the stromal estrogen receptor whereas glandular progesterone receptor was significantly lower than stromal progesterone receptor. (p<0.05). Conclusion: The results of present study conclude that expression of estrogen and progesterone receptor is useful to plan a therapy and avoid any surgical intervention in cases of Abnormal Uterine Bleeding. It can also prognosticate in endometrial carcinoma.

Keywords: Abnormal uterine bleeding, estrogen receptor

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Comparison of PD-L1 and p16 in head and neck squamous cell carcinoma

Richa Kareer, Rashmi Arora, Neena Chaudhary, Sunil Ranga

VMMC and Safdarjung Hospital, New Delhi, India

Introduction: Understanding of pathology and molecular details of tumors, in the past two decades has become better. With the detection of cancer immunotherapy and better understanding of the T cell responses towards the targeted immune check point therapies, and also the successful clinical trials of drugs that blocks immunological checkpoints, identifying precise biomarkers of genomically targeted agents has become popular. Immunological check point inhibitors, which target T cell regulatory pathways to increase anti tumor efficiency, have bought great clinical advances and presents new methods for treatment. Objectives: To evaluate immunohistochemical (IHC) expression of PD-L1 and p16 in HNSCC. To compare the expression of PD-L1 with p16 (+) & p16 (-) HNSCC. To evaluate the association between PD-L1 and p16 with site and grade of tumor. Materials and Methods: 40 patients who were clinically and histologically diagnosed with head and neck squamous cell carcinoma were included. The immunohistochemical markers PD-L1 and p16 were used for this study. Immunohistochemical expression of PD-L1 in head and neck squamous cell carcinoma was analysed and association of PD-L1 with p16 was also assessed. Results: Expression of PD-L1 was compared with p16 positive and p16 negative cases and a positive correlation was seen in oropharyngeal location (p=0.035) and in oral cavity location (p=0.06). Conclusion: PD-L1 can be used as a powerful biomarker for immune mediated check point inhibition in head and neck squamous cell carcinomas.

Keywords: HNSCC, programmed death ligand-1, p16

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PD-L1 expression and Tumor infiltrating lymphocytes in a spectrum of carcinomas: A preliminary study

Viyati Vithlani, N S Mani

B.V.D.U Medical College, Pune, Maharashtra, India

Introduction: Many tumors infiltrated by the tumor-infiltrating lymphocytes (TILs), are associated with good clinical outcome. PD-L1 (programmed death ligand-1) on tumor cells binds to PD-1 inhibitory receptor on TILs and causes immune escape of tumor. Objectives: To characterize the expression of PD-L1 in carcinomas and associated presence of TILs. Materials and Methods: 40 cases of biopsies and resections for carcinomas were studied immunohistochemically for expression of PD-L1 in tumour cells and CD3 and CD8 in lymphocytes in tumour and stroma. Membranous/cytoplasmic staining of PD-L1 in >1% tumor cells was considered positive. TILs were graded as 0 (absent), low (<25/hpf) and high (>25/hpf). This was correlated with histopathology and other parameters. Results: Out of 40 cases, PD-L1 was expressed in 18 (45%) cases. Stromal CD3+ and CD8+ TILs were detected in 35 (87.5%) cases. Whereas intratumoral CD3+ and CD8+ were detected in 25 (61%) cases. CD3+ was the predominant subset of TILs. Most cases which showed PD-L1 positivity and/or high intratumoral CD3+ and CD8+ TILs were from older age group (>50 years). There was no significant difference in PD-L1 expression and presence of TILs between males and females. Conclusion: This was a preliminary study of a larger study including various carcinomas. PD-L1 positivity correlated with high stromal TILs and with presence of intratumoral TILs.

Keywords: Carcinoma, programmed death ligand-1, tumor-infiltrating lymphocytes

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Electron microscopy of juvenile nasopharyngeal angiofibroma: Clinical and histopathological correlates

Himadri Shukla, Atin Singhai, Suresh Babu, Anupam Mishra

King George's Medical University, Lucknow, Uttar Pradesh, India

Introduction: The histological and electron-microscopic (EM) characteristics of juvenile nasopharyngeal angiofibroma (JNA) have been described but no study has compared them with one another or with clinical features. Objectives: To compare ultrastructural characteristics of JNA with clinical parameters. Materials and Methods: This prospective study included histology of 21 samples of which only 13 underwent transmission-EM. Four clinical parameters (age, duration, epistaxis, recurrence), three radiological-staging, 13 histological and 15 EM characteristics were considered. A descriptive analysis for association of these characteristics and also with clinical parameters was attempted. Results: Dense-intranuclear-inclusions (DNI) and peripheral-nuclear-irregularities were universally encountered while other very prominent features were nuclear-blebs-and-pockets, myoid-features, thin-vessel-wall (TnVW), irregular-vascular-contour (IVC) and fibrous-stroma (FS). Statistical significance was obtained between recurrence with histology (p = 0.04) and Fisch staging with EM (p = 0.001). While muscle-in-vessel-wall, thick-vessel-wall, mast-cells, stellate-stromal-cells and cellular stroma predominated in recurrent cases, the upfront disease showed predominance of scar-like-stroma, fusiform-stromal-cells, IVC, TnVW, FS, organised-collagen-bundles and less-cellular-stroma. A very unique Rod-like-structures were appreciated in the cytoplasm of the fibroblast for the first time. Conclusion: While histological parameters of recurrence need further validation, a larger sample may better define histopathological surrogate for predicting intracellular dynamics that may further correlate with underlying cellular stresses. Hence an 'ultrastructural staging' may better customise treatment protocol and prognosis. Furthermore 'characteristic' unique rods need to be further investigated along with validation of viral aetiology for DNI.

Keywords: Electron microscopy, juvenile nasopharyngeal angiofibroma nasopharynx

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Cross sectional descriptive study of histopathological spectrum of nasal, paranasal and nasopharyngeal masses

Annu Khokhar, Manjiri Karandikar

B.V.D.U Medical College, Pune, Maharashtra, India

Introduction: Nasal, paranasal and nasopharyngeal regions harbour diverse lesions including many non-neoplastic and neoplastic lesions presenting with varied spectrum of symptoms. Histopathology with IHC of these excised masses is the mainstay for the definitive diagnosis. Hence a study was undertaken to study the spectrum of these masses histopathologically. Objectives: 1. Study the histopathological spectrum of benign and malignant neoplasms and correlate clinically and radiologically. 2. Determine diagnostic accuracy of IHC in malignant tumors in applicable cases. 3. To determine incidence of fungal infections of this region. Materials and Methods: A descriptive cross-sectional study conducted for nasal, paranasal and nasopharyngeal masses in department of Pathology of a tertiary care center of Maharashtra with clinical and radiological data of 4 years. H&E stain along with PAS stain, GMS stain and IHC were performed wherever required. Followed by statistical analysis. Results: 150 nasal, paranasal and nasopharyngeal masses- 46 neoplastic lesions, 104 inflammatory. Most common: benign -Inverted Papilloma (10 cases), malignant- Non-Hodgkin Lymphoma (7 cases), inflammatory lesion - Inflammatory Polyp (35 cases), fungal infection - Mucormycosis (35 cases) of which 26 patients had Covid-19 infection. IHC were used for definitive diagnosis: Squamous cell carcinoma p63, p16, ki67, Hodgkins and Non-Hodgkins lymphoma- lymphoma panel, Neuroendocrine carcinoma- Synaptophysin and Chromogrannin A, Undifferentiated Nasopharyngeal Carcinoma -CK, CD3, CD20, Synaptophysin, Kki67, Neuroblastoma-S100, Synaptophysin, Meningioma- EMA, S100, CD99, TLE1, Malignant melanoma- S100, MelanA. Conclusion: Most common: fungal infection- Mucormycosis, benign lesion- Inverted Papilloma, malignant- Non-Hodgkins lymphoma.

Keywords: IHC, lymphoma, nasophayngeal carcinoma

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Polarised microscopy of sinonasal tissue in post COVID fungal infections

Neha, Garg, Dayananda S Biligi

Department of Pathology, Bangalore Medical College and Research Institute, Bengaluru, Karnataka, India

Introduction: COVID-19 pandemic has seen surge in several opportunistic fungal infections with unprecedented increase in Rhino Orbito Cerebral Mucormycosis (ROCM). A consistent finding in COVID fungal infections is histopathological presence of refractile and birefringent crystals which were also examined under Scanning Electron Microscope- Energy Dispersive X-ray Analysis (SEM-EDXA). Objectives: 1. To determine presence or absence of crystals and their frequency in COVID fungal infections. 2. To analyse pattern of birefringence. 3. To correlate crystal deposition with inflammatory pattern. 4. To determine elemental composition of crystals by SEM-EDXA. Materials and Methods: Samples from 120 COVID patients were recieved over 2 months. H & E and special stained sections were examined for fungal elements, inflammation and refractile crystals. The slides were viewed under polarised microscope for presence and pattern of birefringence. One representative block was subjected to SEM -EDXA for elemental composition of crystals. Findings correlated by suitable statistical tests. Results: There is significant association between fungal positive cases and needle shaped birefringent crystals (p value:0.00). Birefringence is strongly associated with Aspergillus (p value:0.002) followed by Mucor (p value:0.00). It is also significantly associated with Inflammatory pattern (mostly: acute), ischaemic necrosis, granuloma and comorbidities. On SEM EDXA, the highest percentage of element found in birefringent areas: Calcium followed by Sodium, Magnesium, Zinc and Iron. Conclusion: Since most of the post COVID fungal infections did show significant refractile precipitates, analysing birefringence pattern and their chemical recognition might give us clue towards pathogenesis and prognosis of the disease process.

Keywords: Birefringence, COVID, mucormycosis

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Histopathological prognostic markers of oral squamous cell carcinoma and their correlation with regional lymph node metastasis

Garima Singh, K Sujatha, Pretty Singh

Apollomedics Super Speciality Hospital, Lucknow, Uttar Pradesh, India

Introduction: Oral cancer has become one of the most clamant conditions today with cases rising all around the world. India is no exception to it and is responsible for a quarter of the cases of the global load. The northern part of country with abysmal penetration of oral health awareness and greater fondness for chewable tobacco like gutka and betel quid, has been particularly at a higher risk. In view of minimal studies available on the subject in this region, this study can prove to be seminal in understanding the prognostic factors of oral cancer. Objectives: To estimate the association of defined histopathological parameters with regional lymph node metastasis. Materials and Methods: An observational retrospective study of 50 biopsy-proven Oral Squamous Cell Carcinoma patients who underwent radical resection with lymph node dissection over a period of 1 year in our hospital. Patient details like age, gender and habits will be noted. Gross and histopathological microscopic features which will be included in the study are primary tumor site, tumor size, differentiation, tumor-stroma ratio, depth of invasion, worst pattern of invasion, perineural invasion, lymphovascular invasion, tumor infiltrating lymphocytes, surgical margin involvement and underlying bone involvement, and will be further correlated with regional lymph node metastasis. Results: The study is on-going and its results will throw light on the prognostic potential of various histopathological parameters on their correlation with regional lymph node metastasis. Conclusion: A tremendous incidence but a handful of research on prognostic factors of oral cancer in north India has made this study a need of hour.

Keywords: Oral cancer, oral squamous cell carcinoma

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A female carrying her twin for 47 years: A medical mystery

Neha Jain, Pooja Nagayach, Harendra Kumar, Lalit Kumar, Chandrakanta, Juhi Singhal

Sarojini Naidu Medical College, Agra, Uttar Pradesh, India

Introduction: Parasitic twin (Heteropagus) is a very rare anomaly. It refers to a type of conjoined twin in which tissues of an incompletely formed twin (parasite) are attached to and dependent upon a fully developed twin (autosite). The incidence is less than 1 in 1 million live births. The parasite has grossly recognizable fetal parts and is usually attached to the autosite by a pedicle. Objectives: Describe rare entity heteropagus twin. Materials and Methods: We present a 47 years old multiparous woman presented with abdominal lump since birth. On examination, swelling with stalk arising from epigastric region with evidence of rudimentary face was seen. β-hCG, α-fetoprotein, and routine blood investigation were within normal limits. On CECT, well differentiated viscera like liver, unilateral chest cage, scapula and vertebrae and fluid attenuating structure are seen on posterior aspect of lesion. Exploratory laparotomy and excision of swelling was done & sent for Histopathology. Grossly, one skin covered smooth multiglobular piece of tissue of variable consistency measuring 20X18X14 cm was received. External surface showed rudimentary face with hair, single ear, limb buds and a nipple & areola complex. Diagnosis of fetus in fetu (FIF) was confirmed. Results: FIF is development of high organotypic type with formation of vertebrae and arrangement of tissues around it. Male predominance of 78% in epigastric heteropagus (our case is female). In our case site of attachment of lesion was, epigastric region (only 45 cases reported). Conclusion: Main differentiating feature of FIF from teratoma is identification of vertebral bodies or limbs.

Keywords: Fetus in fetu, parasitic twins, teratoma

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Role of RBC indices and peripheral blood smear in diagnosis of Beta-Thalassemia major (TM)

Raveena Yallur, S S Hiremath, Deepa Masur

S. Nijalingappa Medical College and HSK Hospital, Navanagar, Bagalkot, Karnataka, India

Introduction: Thalassemia's are Heterogeneous Autosomal Recessive Hereditary Anemias characterized by reduced or absent Beta-globin chain synthesis. Around 68,000 children are born with various Thalassemia syndromes. Beta Thalassemia Major is a severe, transfusion-dependent, Inherited Anemia. Objectives: To study the RBC indices and Peripheral Blood Smear (PS) for establishing Thalassemia Major cases. Materials and Methods: Total 30 paediatric cases from july 2022 to September 2022 were evaluated in S. Nijalingappa medical college and HSK hospital, Bagalkot for a study period of 60 days. RBC indices were calculated in Mindray BC-6800 machine and Leishman stained Peripheral Blood Smears were evaluated. Results: Hematological parameters showed moderate to severe anemia with reduced Hb (less then 9 gm/dl), MCV (range 50-70 fl), MCH (12-30 pg) levels. RDW (12-14%) range. Peripheral Blood Smear showed RBC morphology as Microcytic Hypochromic cells, Anisocytosis, Poikilocytosis, Target cells, Polychromatophils, Basophilic stippling and Nucleated RBC's are seen. Conclusion: Diagnosis of Thalassemia Major is established in a proband younger age of 12 months by detecting a complete absence of HbA. Definitive diagnosis of beta thalassemia by Hb electrophosis and HPLC is not possible because of the dimished amount of HbA which overlaps the range. Thalassemia may be suspected based on Microcytic Hypochromic Anemia with Nucleated RBC's on Peripheral Blood Smear and confirmed by Molecular Analysis of Hemoglobin.

Keywords: PS, RBC indices, TM

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Efficacy of rapid, economical, acetic acid, papanicolaou stain in cervical smears as an alternative to conventional papanicolaou stain

Avinash Sahu, R K Chandrakar, Avishesh Singh

SSIMS, Bhilai, Chhattisgarh, India

Introduction: Papanicolaou (PAP) staining used as screening test which requires a large amount of alcohol. In India it has relevance because a laboratory needs a license from a government for supply of alcohol. Dighe et al used 1% acetic acid replacing alcohol in most of the steps. Acetic acid act as mild dehydrating agent is cheaper and easily available. This modified method refered as rapid economical acetic acid papanicolaou (REAP) stain method. Objectives: Since only two study has done on REAP stain, this study was undertaken to compare Conventional Pap stain with REAP stain for staining cervical smears. Materials and Methods: The study was comducted in cytology section of department of pathology, Tertiary care hospital for a period of 18 months. The cervical smears taken from gynecology outpatient departments as part of routeen screening as well as various indications. The two slide was preapared from each patient, were encoded and were immediately wet fixed in 95% ethyl alcohol. One silde smear with conventional pap stain and another one was smeared with REAP pap stain. The smear were numerically evaluated (Score) by three different observer using the modification of parameter given by Ng et al in four categories. Results: A total of 737 cases were included in the study. Out of the toatal 737 conventional pap smear, 675 showed excellent cytomorphology with a score of 4. The average score of 3.9 was observed with this stain. In REAP stain smear, excellent cytoplam and nuclear details was obsered in only 247 (33.6%) of study. The averager score with REAP stain was 3.13. Conclusion: Rapid, Economical, acetic acid, Papanicolaou stain is a simple cast effective and fast staining technique for cervical smears with less turn-around time and can be used in resource limited setting were alcohol availability is limited and cost factor is an issue. It can not ne utilized for routine staining in tertiary setup or for research purpose due to poor preservation of the staining quality.

Keywords: rapid economical acetic acid papanicolaou stain

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Unravelling the impacts of additional sex chromosomes in young adult female: Two rare presentations

Sigin George, Vikas Dagar, Barun Kumar Chakrabarty, Arijit Sen

Armed Forces Medical College, Pune, Maharashtra, India

Introduction: We present two rare cases of sex chromosomal aneuploidies in adult female: Triple X Syndrome and Turner variants - mosaicism with a Y Chromosome. Incidence of triple X syndrome is around 1:1,000 live female births. Only half of Turner syndrome (Incidence 1: 2500 live born female) show mosaicism. 6-9 % of Turner mosaic exhibit presence of Y chromosome or Y derived sequences. Objectives: The present study was planned to evaluate the patients using cytogenomics techniques. Materials and Methods: Both the patients were evaluated by peripheral blood conventional karyotyping using standard cytogenetic protocols. Interphase Fluorescence in situ hybridization (FISH) was performed. Chromosomal Microarray Analysis was performed only in Triple X syndrome case. Results: Case 1: Karyotype analysis of 34 year old female with history of recurrent pregnancy loss and decreased ovarian reserve showed presence of triple X syndrome with modal karyotype 47, XXX/46, XX. FISH studies reconfirmed the diagnosis and presence of mosaicism. Chromosomal microarray support the diagnosis but failed to detect low level mosaicism. Case 2: Karyotype analysis in a case of 17 year unmarried female presented with complaint of amenorrhea confirmed presence of mosaic Turner syndrome. with modal karyotype 45, X0/46, XY. The same finding was reconfirmed by aneuploidy FISH probe. The risk of gonadoblastoma is increased in females with 45, X/46, XY mosaicism. Conclusion: Our study emphasized on the recommendation of performing cytogenomic evaluation of all cases of decrease ovarian reserve and phenotypic Turner syndrome so that we should not miss out the diagnosis of the rare causes of sex chromosomal aneuploidies having prognostic significance.

Keywords: Mosiacism, Triple X syndrome, turner variants

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Immunohistochemical expression of IDH1, ATRX, P53 and Ki67 in diffuse gliomas: An analysis of 32 patients in a single center

Lokesh Kumar, N S Mani, Karandikar Manjiri, R C Nimbargi

Bharati Vidyapeeth Medical College, Pune, Maharashtra, India

Introduction: Gliomas are the commonest primary brain tumours. The 2016 WHO classification of CNS tumours introduced the role of molecular diagnosis with histology and immunohistochemistry (IHC) as it has a prognostic impact on treatment. Objectives: To study IHC expression of IDH1, ATRX, p53 and Ki67 in gliomas and classify them according to 2016 WHO CNS tumours classification Materials and Methods: IHC expression of IDH1, ATRX, p53 and Ki67 was studied in 32 diagnosed glioma cases. Representative tumour regions were selected, IHC performed, results recorded and statistically analysed. Results: The total 32 cases consisted of 16 astrocytomas [Grade II (11) and III (5)], 12 glioblastomas (GB, Grade IV) and 4 oligodendrogliomas (ODG, Grade II-III). Of the 11 diffuse astrocytomas, 10 showed IDH mutation (IDHm) and ATRX mutation (ATRXm), while p53 mutation (p53 m) was identified in only 4 of them. Amongst 5 cases of anaplastic astrocytoma, 4 revealed IDHm, 3 of which showed ATRXm and 1 showed p53 mutation. All 4 ODGs including one anaplastic type, showed IDHm, of which 3 showed ATRX retention as well as p53 wild type. Out of 12 GBs, 5 showed IDHm, 4 ATRXm and 2 p53m. 3 of GBs showed IDH wild type, ATRX retained while 2 were p53m. 4 of the GB cases were categorised as NOS type. Ki67 index was low in most of glial tumours and high in GB. Conclusion: Majority of astrocytomas are IDH mutant and can be opined on IHC. Molecular confirmation is required in ODGs and IDH wild type glioblastomas especially in less than 50 years age group.

Keywords: Astrocytomas, diffuse gliomas, IDH mutant

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[TAG:2]Miscellaneous [/TAG:2]

Role of morphometry in diagnosing premalignant and malignant lesion in liquid based cytology cervical (Pap smear)

Jaspal Singhsaini, Permeet Kaur Bagga, Jaspreet Singh

Government Medical College, Amritsar, Punjab, India

Introduction: Cervical cancer is the second most common cancer in India. The pathogenesis from low grade CIN to cervical cancer takes from 10 to 20 years, during which timely screening for premalignant lesions and early treatment is highly effective in preventing overt disease. Objectives: To study the morphometric parameters of the LBC Pap smear and correlate the lbc Pap smear cytology with morphometric parameters, exploring the possible role of morphometric analysis to improve the sensitivity and specificity for detection of premalignant and malignant conditions. Materials and Methods: The study was conducted on 200 lbc cervical samples which included normal and abnormal case findings, 20 squamous cells per slide were evaluated using Nikon instrument software. Cases screened and reported as squamous cell abnormality, cases showing reactive cellular changes and cases having normal cytology were included. Unsatisfactory samples, patients on intravaginal drugs and already diagnosed scc cases on radiotherapy were excluded. The parameters evaluated were cell area, cell perimeter, nuclear area, nuclear diameter, nuclear perimeter and N:C ratio. The lbc technique used was Surepath test. Results: It was observed that there was a gradual increase in n:c ratio, nuclear area, nuclear diameter, nuclear perimeter from normal to squamous epithelial lesions. Mean cell area and mean cell perimeter was more in normal cells compared to abnormal squamous epithelial lesions. Conclusion: Morphometry can be used as a diagnostic tool in differentiating between premalignant and malignant lesions of cervix.

Keywords: LBC, morphometry, squamous epithelial lesions

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Clinicopathological study and prognostic utility of Her2/neu expression in colorectal carcinoma

Annakishore Yadav, N Mohan Rao, V Mahidhar Reddy

Narayana Medical College, Nellore, Andhra Pradesh, India

Introduction: Colorectal carcinoma is 4th common in men and 3rd common in women Her2/neu belongs to epidermal growth factor receptor family and is a transmembrane protein with tyrosine kinase activity. It has been observed that HER2/neu over -expression in colorectal carcinoma gives good prognostic results. Objectives: To evaluate the expression of Her2/neu in colorectal carcinoma. To correlate the Immunohistocompatibility expression of Her2/neu in colorectal carcinoma by comparing with histopathological and prognostic factors like tumor grade, vascular invasion, perineural invasion, lymphatic invasion, distant metastasis, and stage of the tumor. Materials and Methods: Present study is conducted in Department of pathology, Narayana medical college and general hospital, Nellore It is conducted for a period of 1 year june2021 to may 2022. All the Specimens sent by surgical gastrentology and General surgery departments with colorectal carcinoma specimens are included in the study. Her2/neu immunohistochemistry is done and its expression is studied and prognosis of the patient is evaluated. Results: It has been observed that HER2/neu over -expression in colorectal carcinoma gives good prognostic. Conclusion: It has been observed that HER2/neu over -expression in colorectal carcinoma gives good prognostic.

Keywords: Her2 colorectal prognosis

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Post COVID-19 mucormycosis – Histopathology and associated factors

Shobhana Agrawal, Suvarna Patil, Manjiri Khade, Ajay Jungare, Dilip Sarate

Government Medical College, Akola, Maharashtra, India

Introduction: SARS-COV-2 infection (Covid-19) may be associated with wide range of bacterial and fungal infections. Mucormycosis is a common and potentially life threatening opportunistic fungal infection responsible for morbidity and mortality. Many factors like diabetes mellitus, hypertension and corticosteroid therapy might have been a role in the immunocompromised state of the patients. Objectives: To know the predisposing factors and role of histopathology in diagnosis and assessing the prognosis of post covid-19 mucormycosis cases. Materials and Methods: It is a prospective observational study conducted in tertiary care hospital over a period of 6 months from 1st April 2021 to 30th September 2021. FESS and Maxillectomy samples from 157 post covid-19 mucormycosis suspected cases were studied and details regarding history of diabetes mellitus, hypertension and corticosteroid therapy were retrieved. All tissue samples were examined under H&E stain and special fungal stain (PAS). Results: On histopathological examination, out of total 157 cases, 94 cases were found to be positive for mucormycosis. Of these 94 cases, 63 were males and 31 were females. Age range was from 23 to 75 years. 5 cases showed mixed mucormycosis and aspergillosis infection. Also out of 94 post covid-19 mucormycosis cases, 72 were diabetic, 21 were hypertensive and 68 had a history of corticosteroid intake for treatment of covid-19 infection. Conclusion: Histopathology plays a pivotal role in accurate diagnosis and assessing the severity and invasiveness of mucormycosis. Diabetes mellitus and corticosteroid use are the important associated factors.

Keywords: Corticosteroid therapy, diabetes mellitus, mucormycosis

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Multiple primary malignancies: A comprehensive analysis from a cancer institute in southern India

S Sivaranjani, Usha Amirtham, Akkamahadevi Patil, D Priya, G Champaka, M N Suma, Ashwini Nargund, Geetha V Patil Okaly

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Multiple primary malignancies (MPMs) have always created inquisitiveness amongst clinicians, radiologists and pathologists. The diagnosis invariably poses a challenge to all diagnosticians. The recent advances in diagnostic modalities, treatment and cancer screening programs have led to a substantial rise in incidence of MPMs. Objectives: To study the clinical and pathologic challenges in diagnosis and management of multiple primary malignancies. Materials and Methods: A retrospective analysis of cases with MPMs diagnosed from 2017-2022 was done. Patients with MPMs are defined as synchronous and metachronous based on 'Warren and Gates criteria'. 6 months (180 days) is considered to be the maximum period for the occurrence of synchronous tumors. Results: A total of 36,230 patients with malignant tumors were diagnosed at our institution from 2017 to 2022. 53 (0.14%) patients were histologically proven to have MPMs as defined above. The mean age of patients was 57.4 years (SD=12.3). The male to female ratio was 1:4.8. 39 cases (73.5%) were synchronous and 14 (26.5%) were metachronous. The mean time period between metachronous tumors was 5.14 years. Majority of them had double malignancies with solitary case having 3 synchronous primaries. Both the malignancies were solid tumors in most of the cases (86.7%), of which breast was the most common; while hematological malignancy was one of the neoplasms in 7 cases (13.2%). Conclusion: In the event of MPMs, a hereditary predisposition and syndromic association should be considered. Appropriate surveillance of primary malignancy and meticulous use of ancillary studies will help in early detection and proper management of these tumors.

Keywords: Metachronous, multiple primary malignancies, synchronous

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Evaluation of micrometastasis in head and neck squamous cell carcinomas using modified Papanicolaou stain and pan-cytokeratin immunostain

Ankita Garg, R P S Punia, Reetu Kundu, Anand Gupta, Nitin Gupta, Uma Handa

Government Medical College and Hospital, Sector-32, Chandigarh, India

Introduction: Lymph node metastasis is the most important prognostic factor for head and neck squamous cell carcinoma (SCC). Macrometastasis (>2 mm) can be routinely assessed by Hematoxylin and Eosin (H&E) staining however micrometastasis (0.2-2 mm) is frequently missed. Objectives: To detect micrometastasis in node negative SCC and to compare the diagnostic utility of modified Papanicolaou (PAP) stain with pan-cytokeratin immunostain (Pan-CK). Materials and Methods: Excised tumor and neck dissection specimens from 30 patients were assessed and their gross and other details were recorded. Specimens with all lymph nodes negative for metastasis on routine H&E stain were subsequently stained with Pan-CK and modified PAP stain and evaluated for micrometastasis. Results: Of the 382 lymph nodes examined 4 lymph nodes from 4 (13.3%) cases showed micrometastasis on Pan-CK which was taken as the gold standard for detection. These 4 cases were upstaged from pN0 to pN1mi. Micrometastasis were observed in 3 of these cases (10.0%) on modified PAP staining. Three of the four cases positive for micrometastasis had a primary tumor of the tongue which can be attributed to the rich lymphatic supply of the tongue, one case had primary tumor of the buccal mucosa. No statistically significant correlation could be derived with the other clinical and histopathological finding. Conclusion: Modified PAP stain was found to be less sensitive than Pan-CK in diagnosis of micrometastasis. Larger trials are however required to ascertain the significance of these small tumor deposits and the reproducibility and role of special staining as compared to the more sensitive techniques.

Keywords: Head/neck, micrometastasis, squamous cell carcinoma

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Intraoperative accuracy of frozen section diagnosis in ovarian masses: Two-years experience at a tertiary care hospital

M Rasheeda, R P S Punia, Ranjeev Bhagat, Phiza Aggarwal, *Poonam Goel, Uma Handa

Government Medical College and Hospital, Chandigarh, India

Introduction: Intra-operative frozen section (FS) aids in diagnosing and categorising the ovarian masses and guides the surgeon to optimise the management protocol. Objectives: To evaluate the diagnostic accuracy of FS in ovarian masses and analyse the discrepancy between FS diagnosis and final paraffin-embedded histopathological diagnosis. Materials and Methods: A retrospective comparative analysis between intraoperative FS diagnosis of clinically suspected ovarian masses and their corresponding final histopathological report were assessed over a period of two years. The discrepant cases were reviewed and re-evaluated and thus the overall accuracy was determined. Results: The study included 54 patients and the overall accuracy of frozen section in categorizing the ovarian neoplasm was 96.3%. Major discrepancy was noted in two cases, one of which was reported as benign surface epithelial tumor on FS but was finally diagnosed as leiomyoma and the other case was given as sex- cord stromal tumor which was later diagnosed as serous carcinoma with neuroendocrine carcinoma. With respect to surface epithelial tumors, eight cases (14.8%) were given a diagnosis of benign/borderline tumors, out of which four cases (7.4%) turned out to be benign and the other four turned out to be borderline. Conclusion: FS diagnosis has a high accuracy and is a crucial diagnostic tool in guiding intra-operative surgical management of ovarian neoplasms. The limitations of FSD includes the exact sub-categorization of the tumors especially in cases of benign and borderline surface epithelial tumors.

Keywords: Epithelial tumors, frozen section, ovarian neoplasm

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Clinicopathologic features of salivary duct carcinoma: Retrospective analysis from a tertiary cancer centre of North India

Zikki Fatima, Chowdhury, Shashikant Patne, Ipsita Dhal, Sadaf Hayat

HBCH and RC, Muzaffarpur, Bihar, India

Introduction: Salivary Duct Carcinoma (SDC) is a rare and aggressive high grade salivary gland malignancy, accounting for 0.2% of epithelial salivary gland neoplasms and 0.5% of salivary gland carcinomas. Objectives: To evaluate the clinicopathologic characteristics of SDC patients treated at a tertiary cancer centre. Materials and Methods: A retrospective analysis of SDC patients treated at tertiary cancer care centre of North India between January 2019 and August 2022 was undertaken. Results: A total of 20 histologically confirmed cases of SDCs were studied. Mean age of the patients was 59.85 years with a male predominance. Most common site involved was the parotid gland (75%). Most of the cases showed conventional histology of high grade mammary ductal carcinoma. Surgical resection was possible in 8 patients. Perineural and lymphovascular invasion was identified in 62.5% and 50% cases, respectively. Androgen receptor (AR) positivity and Her2/neu expression was detected in 80% and 41.17% cases, respectively. Regional lymph node involvement was noted in 70% of the cases and distant metastases in 50% cases, with the most common site being lung. Majority of the patients were administered palliative radiotherapy and chemotherapy. Median survival was 16 months and median time for distant metastasis was 3 months. Conclusion: Distinction of SDC from other salivary neoplasms is of prognostic and therapeutic significance, since it has a dismal prognosis. Ancillary studies for AR and Her2/neu amplification should be mandated on all SDC patients, as the benefits of androgen deprivation therapy and targeted Her2/neu therapy are becoming more evident.

Keywords: Parotid, salivary duct carcinoma, targeted therapy

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Clinicopathological features of 10 cases of phosphaturic mesenchymal tumor diagnosed at a tertiary cancer referral centre, India

Ashwini Duggad, P Choodamani, M Ramadwar, B Rekhi

Tata Memorial Hospital, Mumbai, Maharashtra, India

Introduction: Phosphaturic Mesenchymal Tumors (PMTs) are rare mesenchymal tumors with long standing but often debilitating symptoms. Nonetheless, they have characteristics histological features, and if diagnosed, patients may benefit because surgical excision is completely curable. Objectives: To study histopathological features and correlate with clinical findings and outcomes of 10 cases of PMTs, diagnosed at our institution. Materials and Methods: This was retrospective study, wherein 10 cases of PMTs, diagnosed between January 2013 and July 2022 were included. Results: The average age at time of diagnosis was 40 years with a M: F ratio of 4:1. Clinical symptoms included pathologic fractures, weakness, bone pain, and tumor site swelling. The results of biochemical investigation revealed elevated serum FGF23 levels, low serum phosphorus (average-1.51 mg/dl), and normal serum calcium levels (average-9.4 mg/dl) (in all the cases where available). On histopathology, all cases revealed tumor cells that were oval to short spindle-shaped and arranged in a hemangiopericytomatous pattern. Nine cases included multinucleate giant cells, with 8 of those cases exhibiting the typical “grungy calcifications.” In 8 out of 10 cases, prominent pseudo cystic areas were observed. All patients had complete post-operative symptom relief with no recurrence or metastasis, and in all cases where follow-up was available, all patients were disease-free at present. Conclusion: This constitutes one of the largest series of PMTs from our country. Histopathological characteristics, clinicoradiological, and biochemical data are all correlated with diagnosis of PMTs. It is crucial to get precise diagnosis of these tumors because they are benign neoplasms that can be totally treated with sufficient surgical excision.

Keywords: Grungy, hemangiopericytomatous, phosphaturic mesenchymal tumors

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Minimal residual disease in B cell acute lymphoblastic leukemia: Our experience

Gourang Paliwal, S Venkatesan, Deepti Mutreja, Harshit Khurana, Arijit Sen

Department of Pathology, Armed Forces Medical College, Pune, Maharashtra, India

Introduction: B – Acute Lymphoblastic Leukemia (B – ALL) constitutes around 85% of all acute leukemias. Minimal residual disease (MRD) has emerged as the holy grail of prognostication of Leukemia patients as well in monitoring response to treatment. It also helps to guide treatment decisions. MRD is defined as <0.01% of leukemic cells in bone marrow and can be assessed quantitatively either by flow cytometry or by detecting unique molecular signatures by polymerase chain reaction. Objectives: To study the clinicopathological profile of patients of B cell ALL and monitor the minimal residual disease by flow cytometry. Materials and Methods: This observational study was done at a tertiary care hospital in western Maharashtra over a period of 2 years. All the cases diagnosed as B-cell ALL were evaluated by complete blood count, bone marrow aspirate and biopsy, immunophenotyping by flow cytometry, immunohistochemistry and cytogenetics. MRD assessment was done by flow cytometry at the end of BFM Induction protocol and correlated with the clinicopathological parameters. Results: A total of 30 cases were included in the study, the mean age was 14 years. 6 patients were found MRD positive (>0.01%) post initiation of BFM protocol. MRD monitoring helped in switch over to 2nd line chemotherapy in one case and allogeneic stem cell transplantation in one patient. Conclusion: MRD measurement is an essential requirement for follow up of B cell ALL patients and can help treatment decisions. Flow cytometry is an effective and widely available technique with proven sensitivity in MRD detection. Besides MRD, it can also help identify aberrant phenotype expression.

Keywords: B – acute lymphoblastic leukemia, flow cytometry, minimal residual disease

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Insights into blood donors iron status: Current practices and future directions

S Susmitha, Basavaraj P Bommanahalli

Gadag Institute of Medical Sciences, Malasamudra, Karnataka, India

Introduction: Iron deficiency anemia is most common nutritional deficiency anemia and a potential problem among the blood donors. Two new parameters such as low hemoglobin density (LHD) and microcytic anemia factor (MAF) can be used as a new screening tool for early detection of anemia. Objectives: To access the utility of LHD and MAF in early detection of anemia among voluntary blood donors. Materials and Methods: It includes 110 voluntary blood donors. Complete blood count along with serum ferritin was estimated for all donors. Donors were classified into four groups based on serum ferritin. Group 1 being iron deficient donors, group IV being normal donors. Data was analysed and LHD and MAF were calculated from the above parameters. Results: LHD and MAF values were able to differentiate between iron deficient donors from the normal donors. MAF with a cut off value of 13.55 and 14.15 was found to be a good screening tool to differentiate group I and group II from the group IV donors. Similarly, LHD% cut off of 36.25 and 18.6 was able to differentiate group I and group II from the group IV donors respectively. Conclusion: Estimation of LHD and MAF among blood donors aid in the early detection of iron deficiency anemia. Though assessment of serum ferritin is not feasible in all setting, estimation of LHD and MAF helps in screening blood donors for iron deficiency without extra cost and additional blood sampling.

Keywords: Blood donors, ferritin, iron deficiency anemia

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Tumour budding in preoperative biopsies an independent prognostic factor to predict the risk of lymph node metastasis in oral cancers. A retrospective tertiary care centre study

Nugala Sindhura, M Vijayashree

Guntur Medical College, Guntur, Andhra Pradesh, India

Introduction: The enormous advancement in understanding of oral squamous cell carcinomas (OSCC) has not been accompanied by a significant reduction in the high morbidity and mortality rates associated primarily with disease recurrence and lymph node metastases. Objectives: To evaluate the tumour budding (TB) as an independent prognostic marker to predict lymph node metastasis in pre-operative biopsies. Materials and Methods: All patients of preoperative diagnostic oral cavity biopsies that also underwent resection with cervical LN dissection in our institute were included. The degree of differentiation, preoperative tumour budding, post-operative tumour budding, and intratumoral budding in preoperative biopsies along with other pathological parameters were all assessed histologically in each case. Retrospective study. Results: 32 OSCC cases were examined by haematoxylin and eosin-stained slides. By using univariate analysis, the histological factors depth of invasion (P = 0.04), pattern of invasion (P = 0.004), presence of preoperative tumour budding (P = 0.008), postoperative tumour budding (P = 0.004), intratumoral tumour budding (P = 0.019), and sex (P = 0.03) all significantly correlated with risk of lymph node metastasis. Other clinical and histological factors, including age, the largest tumour size, histological grade, lymphovascular invasion, perineural invasion, and stromal response, did not significantly correlate with the lymph node metastasis. Conclusion: The potential of morphological features, such as TB evaluated in OSCC diagnostic pre-operative biopsies aids in identifying patients who may benefit from more aggressive treatments.

Keywords: Budding, independent prognostic marker, oral

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Diagnosis of metastatic carcinoma of unknown primary: Our experience for past 1 year

Swagatika Samal, Debahuti Mohapatra, Pranita Mohanty, Soumya Surath Panda, Debirupa Dey

Department of Pathology, IMS and SUM Hospitals, Siksha O Anusandhan University, Bhubaneswar, Odisha, India

Introduction: Diagnosis of unknown primary is rare in the routine practice in histopathology. Although histopathology is considered to be the gold standard, often the primary site remains elusive in spite of extensive workup. The advancement of immunohistochemical (IHC) analysis has significantly increased diagnostic accuracy. In the present study, we have shared our experience in diagnosing tumours with unknown primary. Objectives: To know the occurrence carcinoma of unknown primary in our institution. Materials and Methods: It is a retrospective cohort study conducted in the Department of Pathology, IMS & SUM Hospital, from October 2021 to September 2022. A total number of 132 conjugative cases of CUP were included in the study. The HE slides, along with the IHC markers, were reviewed to find out the primary site of malignancy. Results: Of the total 132 patients, 63 were males and 69 were females. The average age of the study population was 52 years (range 24-84 years). The omental metastasis was the most common (22/132), followed by liver (20/132), pleural fluid (12/132), lymph node metastasis (27/132), ascitic fluid (18/132), bone (6/132), brain (6/132). The most common variety was adenocarcinoma (85/132), followed by serous carcinoma (12/132), squamous cell carcinoma (6/132), poorly differentiated carcinoma (6/132) and renal cell carcinoma. Conclusion: Besides routine histopathology, the judicious use of IHC can ensure the primary site of origin in most cases and later reduces the financial stress by the commencement of early palliative therapy.

Keywords: Carcinoma of unknown primary, diagnosis, immunohistochemistry

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Demographic study of ABO and Rh blood groups among blood donors in western Maharashtra

Prasad Chaudhari, Nikhil S Deshpande, Bharati P Baviskar, Ravindra R Karle, Suryakant D Dongre

Dr. Balasaheb Vikhe Patil Rural Medical College, Pravara Institute of Medical Sciences (DU), Loni, Maharashtra, India

Introduction: Blood group determination is most important for cross matching before blood or blood component transfusion. The demographic study can help to prevent shortfall of required blood group by recruiting voluntary donors for such required blood groups. Objectives: To find blood group wise population distribution in a tertiary care hospital in western Maharashtra region. Materials and Methods: This was a retrospective study conducted for a duration of a year. Medically fit blood donors according to the blood bank guidelines were accepted at blood donation camps and hospital blood bank. The blood samples were collected in Ethylene Diamine Tetra Acetic acid (EDTA) tube by the method of venepuncture. The blood group detection was done with the help of Ortho Vision blood group analyser using gelcard technology. Results: A total of 4536 donors included of which 98.18% (n=4453) were males & 1.82% (n=83) were females. The most common blood group was B Rh Positive – 31.65% (n=1436). Least common blood group was AB Rh Negative - 0.37% (n=17). The blood groups were found in decreasing frequency of B>O>A>AB. We also got 3 donors of Bombay blood group. Rh positive donors were 94.57% (n=4290) while Rh negative donors were 5.35% (n=243). Conclusion: The commonest blood group found is B Rh Positive. Maximum donors were male but female donors need to be encouraged. The panels of rare blood group voluntary donors should always be ready for emergency situation.

Keywords: ABO, Bombay blood group, Rh

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Clinicopathological study of Mucormycosis in resected specimens of COVID-19 patients: An institutional study

K Manjula, R V Sowjanya Lakshmi

Chikkaballapura Institute of Medical Sciences, Chikkaballapur, Karnataka, India

Introduction: Coronavirus disease 2019 (Covid-19) is an infection caused by severe acute respiratory syndrome coronavirus -2 (SARS-CoV-2). Covid19 has been associated with a wide range of opportunistic bacterial and fungal infections. Aspergillus and Candida have been reported as the main fungal pathogens for co-infection in people with COVID-19. Recently, several cases of mucormycosis in people with COVID-19 have been increasingly reported world-wide, in particular from India. Objectives: (1) To study the clinicopathological characteristics of Mucormycosis in covid 19 patients. Materials and Methods: This is an observational study done in the department of pathology. All the covid positive specimens sent for histopathological examinations were included in the study. Patient's presentation details, laboratory investigation details, radiological findings and other available details of investigation were collected. Also details regarding co morbidities, clinical management, and operative details will be collected. Gross deails of the resected specimens were noted. Routine processing of the tissue was done. Histopathological examinations of the sections were done. Special stains (PAS and GMS) were used to confirm the mucormycosis. Microscopic examination of sections (Hematoxylin and Eosin) was done, findings were recorded (type of inflammation, angioinvasion, perineural invasion, haemorrhage, necrosis, ete) and analysied. Results: A total of 85 cases of mucormycosis were studied in the present study. Male to female ratio was 2.86:1. The most common age group affected was 40- 49 years, median age was 44.5 years, accounting to 29.41%. The most common histopathological finding associated with mucormycosiswas acutesuppurative inflammation and coagulative necrosis. Conclusion: Mucormycosisia an highly invasive fungal whichcannot be diagnosed by rapid tests. In many patients affected with mucormycosis, the outcome is poor. Early Diagnosis and reversing the predisposing factors are most important. Controlling the infection requires increased awareness, better tests to diagnose at the earliest, controlling the diabetes and usage of corticosteroids wisely.

Keywords: Angioinvasion, mucormycosis, necrosis

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Pathologist and patient: One on one

Rajatha Anand, H Chaithra, J P Geetha

Department of Pathology, Sri Siddhartha Medical College, Tumakuru, Karnataka, India

Introduction: Pathologists have always been an integral part of the healthcare system striving to deliver quality care to patients. However the speciality has mostly been invisible, with only certain specialities receiving attention from the public population. Forensic pathology has been highlighted extensively by the media in many cases, yet the public knows very little about what pathologists actually do in general. Even though a pathologist is known as a consultant's consultant, the missing recognition affects pathologists' reputation, makes recruiting very challenging, and does not help the pathologist community's lobbying. Objectives: To analyse the public perception of a Pathologist among general population. Materials and Methods: In our survey with 200 respondents the questionnaire used was developed and approved by Manitoba Research Ethics Board, USA. We used Pearson's X2 test to evaluate the association between correct responses and if respondent had undergone the procedure. Two-sided p values < 0.05 were considered statistically significant. Results: In our study Pathology was identified as a medical speciality by only 25 %, 33% assumed that is takes 4 years to become a pathologist, only 8% identified that it would take more than 8 year to become a Pathologist. The most commonly undertaken test with the identified specialist were Pap test (gynaecologist), breast biopsies or surgical excisions (surgeons / oncosurgeons), blood tests (medical laboratory technicians). The number of patients who identified pathologist as ultimate decision maker remained low 15%. Conclusion: Implementing patient-pathologist discussions would certainly improve the quality of patient care, help patients to understand disease process better and make more informed decisions. It would be worth investing our time into this process as it would provide an opportunity to change the image our speciality in the society.

Keywords: Manitoba research, pathologist, patient

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Placental thrombosis in women with prothrombotic coagulation profile: A 5-year retrospective study

Mary Mathew

Department of Pathology, KMC, Manipal, Karnataka, India

Introduction: Placenta is one of the most under-utilized organs that can provide vital clues for the cause of fetal mortality and morbidity. Women with abnormal prothrombotic coagulation profile present with recurrent fetal loss or still birth. The examination of the placenta can aid in establishing the cause of death in the fetus and provide guidance for further investigations and management. Objectives: The aim of this study is to present the clinical and histopathological findings in placentas of women with abnormal prothrombotic coagulation profile. Materials and Methods: This was a retrospective 5-year study of placentas sent to the Department of Pathology, KMC, Manipal, in women with bad obstetric history. The clinical history and investigations were retrieved from the hospital records and the placentas of women with abnormal coagulation profile were reviewed. Results: Out of a total of 70 placentas, 17 placentas showed features of fetal vascular malperfusion with multiple thrombi within the chorionic plate, basal plate and/or stem vessels. Conclusion: Placentas provide useful clues in women with bad obstetric history and should be submitted for histopathological examination. The evidence of thrombi within the vessels of the placenta provides important information to advise women to undergo further testing to elucidate the cause of the prothrombotic state and prevent adverse fetal outcomes.

Keywords: Coagulation, placenta, thrombus

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Evaluation of clinical, histomorphological and immunohistochemical markers (EGFR, OCT 3/4 and PDL1) in oral squamous cell carcinoma

T N Suresh, Swaroop Raj, S M Azeem Mohiyuddin

Department of Pathology, Sri Devaraj Urs Medical colleg, Kolar, Karnataka, India

Introduction: Oral squamous cell carcinoma (OSCC) is considered the most common malignant tumor of the head and neck region in India. The PDL1- induced suppression of immune surveillance is considered as poor prognostic factor. Studies done outside India have reported association between stem cell biomarker OCT ¾ expression to the metastatic potential of cancer. EGFRtargeted therapy is in clinical trial for Head and neck cancer. Studies on these theranostic biomarkers are not well documented in Indian literature hence needs to be studied in local population. Objectives: To study clinical, histopathological parameters and expression of EGFR, OCT 3/4 and PDL1 IHC marker in oral squamous cell carcinoma. Materials and Methods: 103 Oral cancer operated in our institute during 2018- 2020 were studied. PDL1 expression in >5% of tumor, EGFR, OCT 3/4 expression in > 10% of tumour considered as positive. Results: Oral cancer was predominant in Female (M:F=1:4.4). Majority of the cases (61%) are in the age group of 40-60 years. Buccal mucosa (65%) is the commonest site. Lymph node positivity is seen in 36% cases. Majority of cases presented in TNM stage II (35%) followed by stage III (24%). EGFR expression in tumor area is seen in 42% cases. There was significant association between EGFR and OCT 3/4 expression with lymph node metastasis and TNM staging. PDL1 expression in tumor area is seen in 70% cases. However there was no significant association between PDL1 expression with lymph node metastasis and TNM staging. Conclusion: Study of EGFR, OCT 3/4 and PDL1in oral cancer helps in prognosis and treatment.

Keywords: EGFR, OCT3/4, oral cancer, PDL1

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Multicentre analysis of the prognostic significance of worst pattern of invasion – Type 4 and proposal for inclusion as a mandatory minimum dataset reporting criteria

Paromita Roy, Daphne Fonseca, Ritu Saxena, L M Chandrasekar Rao, Hemant Kumar Nemade, Indranil Mallick

Tata Medical Center, Kolkata, West Bengal, India

Introduction: Worst pattern of invasion (WPOI) is a novel prognostic marker and mandatory reporting of WPOI-5 has been recommended by AJCC. Objectives: We aimed to look at the prognostic significance of WPOI-4. Materials and Methods: In this retrospective multicenter study, we re-reviewed the slides for consecutive oral cavity cancers to assess presence of WPOI-4 and WPOI-5 and correlated with survival. Results: 293 patients from Kolkata and 135 patients from Hyderabad, were analysed. WPOI4 was present in 240 (56.1%) and WPOI5 in 88 (20.3%). Median FU of 25 months was available for 388 patients. Median survival was 94.0 months and 2-year and 3 year survival 81.1% and 77.7% respectively. For WPOI-4 negative 2 year DFS 85.2% vs 77.9% for positive, 3 year DFS 84.3% vs 72.3% (log rank test, p = 0.04). For WPOI-5 negative 2 year DFS 83.8% vs 69.5% for positive, 3 year DFS 81.4% vs 62.7% (log rank test, p < 0.005) We found that 54 patients had features of both WPOI-4 and WPOI-5. On reclassification of patients to consider this group to be a separate category, this cohort had the worst outcomes with 2 -year and 3-year DFS of 59.8% and 48.3% (log rank test, p <0.005). Conclusion: The presence of features of WPOI-4 and WPOI-5 impact DFS in oral cancers. Presence of features of both has the poorest prognosis and may merit a separate WPOI group.

Keywords: Squamous carcinoma, worst pattern of invasion

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Granulomatous inflammation in malignancies: An interesting coincidence or association?

Nupur Trivedi, Deepti Gupta, Shefali Agarwal, Anjali Tewari, Abhimanyu Kapoor

Regency Hospital Ltd, Kanpur, Uttar Pradesh, India

Introduction: Granulomatous inflammation is a group of disorders sharing the common histological evidence of granuloma formation. Granulomas develop in a number of immunologically mediated infectious (mycobacteria, fungi, parasites, etc.) and non-infectious (sarcoidosis, foreign bodies, Wegener's granulomatosis, inflammatory bowel disease, etc.) conditions. Granulomatous reactions occurring within the lymph nodes draining the carcinoma or within the stroma of the malignancies are a well known but uncommon occurrence. Objectives: To evaluate the occurrence of granulomatous reactions in malignancies, their incidence, probable pathogenesis and diagnostic modalities to differentiate between co-existing infection and tumour induced granuloma. Materials and Methods: We hereby report a series of cases studied over a period of one year in a tertiary care hospital in Kanpur to evaluate the occurrence of granulomatous reactions in malignancies, their incidence, probable pathogenesis and diagnostic modalities to differentiate between co-existing infection and tumour induced granuloma. Results: Of the total 365 specimens of carcinomas with lymph node dissection, 42 cases showed coexisting granulomas in the tumour stroma/ draining lymph nodes. Out of 42 cases, 3 were diagnosed as coexisting Sarcoidosis, 6 as coexisting tuberculosis and majority; 33 as having sarcoid like granulomas. Conclusion: Of the total 365 specimens of carcinomas with lymph node dissection, 42 cases showed coexisting granulomas in the tumour stroma/ draining lymph nodes. Out of 42 cases, 3 were diagnosed as coexisting Sarcoidosis, 6 as coexisting tuberculosis and majority; 33 as having sarcoid like granulomas.

Keywords: Granulomatous reactions, multimodality approach, sarcoidosis, tuberculosis

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Orbital meningioma in Asian Indians: A retrospective study of 56 specimens

Dilip Mishra, Swathi Kaliki

LV Prasad Eye Institute, Hyderabad, Telangana, India

Introduction: Meningiomas constitute 4% of intraorbital tumors and 20% of symptomatic intracranial tumors. Orbital meningiomas most commonly arise from the optic nerve and extend to the orbit through the adjacent structures mainly the greater wing of sphenoid. Objectives: To examine the cohort of intraorbital meningiomas according to the current World Health Organization (WHO) classification and evaluate demographics and histopathology of orbital meningiomas. Materials and Methods: This was a retrospective study approved by Institutional Ethics Committee and included 56 cases of orbital meningioma which were incised between 2001 and 2019 at our institute. The data were reviewed for clinical presentation, demography and histopathological findings. Results: The mean age at presentation was 38.8 ± 18 years (range, 3 to 71 years). Three tumors arose in children. Median duration of clinical presentation was 8.7 months (inter-quartile range, 2.4 months to 2 years). There were 22 (39.3%) males and 34 (60.7%) females. Left eye was more commonly affected (n = 30, 53.6%) and right eye was affected in 26 (46.4%) cases. Most common clinical presentation was proptosis found in 51 (91.1%) cases. The commonest clinico-radiological diagnosis was sphenoid wing meningioma (n = 17, 30.4%) followed by optic nerve meningioma (n = 15, 26.8%) and fungal granuloma in (n = 4, 7.1%). Metastasis was suspected in 3 (5.4%) cases. Most of the tumors were WHO grade 1 (n = 52; 92.9%). Four tumors (7.1%) were atypical meningioma of grade II with nuclear atypia and more mitotic figures. Psammomatous calcification was found in (n=20; 35.7%) tumors. Conclusion: Orbital meningioma is a rare tumor of orbit and more commonly arises in middle aged females. Proptosis is the most common symptom. Histopathologically, meningiothelial meningioma of WHO grade 1 is frequently found in Asian Indians.

Keywords: Meningioma, sphenoid wing

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Tumour associated macrophages in breast cancer and their role in response to neoadjuvant chemotherapy – A pilot study

Anugnya, Ranjoalkar, Lohita Krishna, Sudipta Naskar, Rekha V Kumar, B S Srinath, Aruna Korlimarla

Shri Shankara Cancer Hospital and Research Centre, Bengaluru, Karnataka, India

Introduction: M2 tumour associated macrophages (TAMs) are implicated in tumour progression and therapy resistance in breast cancer (BC). We have characterized M2-TAMs and their correlation with neoadjuvant chemotherapy (NACT) response in matched pre and post-treatment tissues. Objectives: To calculate combined H-score (CHS) for CD68 and CD163 and Macrophage polarisation score (MPS) in pre and post-treatment tissues and to correlate them individually and together (TAM score) with clinicopathological characteristics and chemotherapy response. Materials and Methods: Pre and post-treatment matched tissues (n=45) of operable BC treated between 2018-2020 were studied. CHS using immunohistochemistry (IHC) for CD68 and CD163 and MPS for gene expression using q-RT-PCR was calculated. A TAM score was established using CHS and MPS. Residual Cancer Burden scores divided patients into: complete responders (CR)(20%); partial-responders (PR)(51%) and non-responders (NR)(29%). Univariate and multivariate analysis was performed between scores and clinicopathological parameters. Results: A positive correlation was observed between high MPS and high H-score (p=0.083). The average H-score and the average high MPS in pre (52.11) and post- treatment (12.16) tissues was highest in PRs. There was a statistically significant correlation between CHS on post-treatment tissues and pathological response (p=0.017). TAM score in residual tumours was associated with increased pT size and significantly with LVI (p=0.03). 11% of patients with high TAM score in resected specimen were non-responders. There was no statistical correlation of MPS and TAM score with pathological response in pre -treatment tissues. Conclusion: It is postulated herein that there is utility in examining TAM score in residual tumours for choice of subsequent adjuvant chemotherapy.

Keywords: Breast cancer, chemoresistance, tumour associated macrophages

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Single multipurpose magnetic automated ready-to-use extraction kit for different clinical samples

Kavita Khadke, Ashwini Jadhav, Shivani Singh, Sneha Thakur, Sujata Hajra, Rajas Warke

Himedia Laboratories Pvt Ltd., Thane, Maharashtra, India

Introduction: In order to reduce the hassles of ordering multiple purification kits, we have come up with the Multipurpose magnetic bead-based nucleic acid automated extraction kit. Objectives: To develop a kit to carry out efficient extraction of nucleic acid from wide range of samples such as whole blood, plasma, serum, saliva, buccal cells, buffy coat, cultured cells, urine, FFPE, tissue. Materials and Methods: Himedia's MB583MPF HiPurA® Multi- purpose Magnetic Nucleic Acid Purification Kit, Insta Nx® Mag96, InstaNx® Mag32 Pro extractor, Insta Q96® plus real time PCR, MagMAX DNA Multi-Sample Ultra 2.0 Kit, KingfisherFlex, MBPCR268 Hi-PCR® Human Housekeeping Gene Detection Probe PCR Kit, Nanodrop. Results: 20 plasma, serum, buccal swabs, urine samples each, 5 FFPE tissue samples were extracted and efficiency was checked by PCR assays. 120 blood, 15 buffy coat, 25 mixed cell suspensions, 5 tissue, 40 saliva samples were extracted. Spectral measurement and real time PCR was performed to demonstrate nucleic acid purity, an average concentration of more than 40 ng/ul and A260/A280 ratio between 1.6 to 1.9 was recorded. Conclusion: A single prefilled magnetic automated kit used for extraction of Nucleic Acid from various types of samples showed good results as compared to competitor. The magnetic beads bind to nucleic acid efficiently providing high-quality nucleic acid from various biological samples. The prefilled plates provided in the kit makes the kit user friendly by saving time for filling reagents for samples to be processed and minimizes the risk of contamination.

Keywords: Automated magnetic extraction

   AOP266 Top

   Miscellaneous Top

Challenges in assessment of medullary bone invasion in oral cavity cancers and its prognostic significance

Badrinath Venkatesh, Paromita Roy, Indranil Mallick

Tata Medical Centre, Newtown Action Area III, Kolkata, West Bengal, India

Introduction: Presence of bone invasion 'through cortical bone' upstages oral cavity squamous carcinoma (OSCC) to pT4a. This medullary invasion needs to be differentiated from superficial erosion which does not change stage. Objectives: To assess the difficulties in differentiating cortical erosion from medullary invasion and evaluate the prognostic significance of the different patterns of bone involvement. Materials and Methods: We retrospectively reviewed OSCC cases with bone resection treated at our center for last 10 years to assess bone invasion in 3 categories – no invasion / erosion / medullary invasion. Results: 210/368 patients (57.1%) had no bone involvement; 136 (36.9%) had erosion and 22 (5.9%) had medullary invasion. The median follow up was 26.5 months. The disease-free survival (DSS) estimates at 3 years is 63.1%; 3 year DFS for no involvement/erosion was 64.4% vs 42.7% for medullary invasion (log rank test, p = 0.12). The 3 year loco regional control for no involvement /erosion was 75.9 % vs 65.6% for invasion (log rank test, p = 0.22). In multivariable analysis bone invasion was not an independent prognostic factor (hazard ration 1.4, 95% CI 0.72 to 2.69, p = 0.34). The difficulties in assessing bone invasion include – bone fragmentation, unclear junction between cortex and medulla especially near tooth socket. Conclusion: Our study shows while there is a difference in DFS and locoregional control, it is not statistically significant. This may be due to less number of cases with bone invasion and larger multicenter study is proposed to find out the true significance of bone invasion.

Keywords: Cortical erosion, medullary invasion, tooth socket

   AOP267 Top

   Miscellaneous Top

Intra-operative grossing to achieve R-0 resection in oral cancer surgery

Sudipta Naskar, M S Sulakshana, M P Sreeram, Narayana Subramaniam, Rekha V Kumar

Sri Shankara Cancer Hospital and Research Centre, Bengaluru, Karnataka, India

Introduction: Consistently achieving clear margins in oral cancer is challenging in view of the complex anatomy, unpredictable patterns of tumour spread and lack of anatomical compartments in the oral cavity. Frozen section, except in specific situations, is of limited value to achieve this. Objectives: To perform immediate grossing during surgery to assess margin adequacy using a standardized protocol, and to correlate with final margin status. Materials and Methods: Fifty patients undergoing curative intent oral cancer were included. After resection aimed at 1-cm gross margins, the margins were tagged & the specimen were then taken down to the pathology lab, where it was inked and grossed in the presence of one of the ablative surgeons. Gross margins < 7 mm were revised. Specimen driven frozen section was performed for suspected areas. Analysis was performed to determine final margin status and necessity for margin revision based on the current technique. Results: 21/50 (42%) patients were found to have close margins during intra-operative grossing, in spite of the fact that the surgeon was satisfied with the adequacy of resection. In spite of revision of margins, 3 patients (6%) still had positive or close margins (<5 mm) in the final histopathology. 29 patients (58%) who had free margins during grossing did not have close margins. Conclusion: This technique was useful to identify submucosal spread. For those requiring revision, the standardization of technique allowed for accurate identification of representative margins. It also allows good communication between the surgeon and pathologist to ensure there was no ambiguity in grossing complex specimens.

Keywords: Intra-operative grossing, oral cancer, R0 resection

   AOP268 Top

   Neuropathology Top

Immunohistochemical evaluation of S-100 protein, neurofilament protein (NFP) and glial-fibrillary acidic-protein (GFAP) immunoreactivity in spindle cell tumors of CNS: A study of 50 cases

Priya Patel, Shikha Ghanghoria

MGM Medical College, Indore, Madhya Pradesh, India

Introduction: The collective expression of antigens produced in immature or mature myelin-producing glia was evaluated in spindle cell tumors arising from CNS and cranial and paraspinal nerves. In S-100 protein, NFP, and, Glial-fibrillary acidic-protein (GFAP) immunoreactivity were evaluated immunohistochemically using commercially available antibodies. Objectives: Immunohistochemical evaluation of S-100 protein, Neurofilament protein (NFP) & Glial-fibrillary acidic-protein (GFAP) immunoreactivity in spindle cell tumors. Materials and Methods: The 40 cases of tumors arising from cranial and paraspinal nerves and 10 cases of spindle cell tumors of CNS are studied which showed histologic features resembling that of schwannomas. A panel of above markers were applied on each case to make the final diagnosis. Results: All cases of tumors that arises from nerves showed strongly and diffusely positive for S-100 protein (nuclear and cytoplasmic staining), while none of the CNS tumor showed S-100 immunoreactivity. This make our diagnosis that the tumors that arose from cranial and paraspinal nerves are Schwannoma. The NFP-positive axons were identified in 11 of 50 (22%) cases of tumors, arising from nerves but the immunoreactivity with NFP was variable. None of the cases of CNS tumors showed immunoreactivity with NFP and GFAP, which suggest that none of tumor are of glial in origin. Conclusion: Thus, it is concluded that S-100 is the better and important marker of Schwann cell neoplasms. The combined immunohistochemical evaluation of potential Schwann cell markers such as GFAP, NFP and S-100 offers extra advantage in making the diagnosis more accurate.

Keywords: CNS tumor, Glial-fibrillary acidic-protein, neurofilament protein, S-100

   AOP269 Top

[TAG:2]Neuropathology [/TAG:2]

Isocitrate dehydrogenase (IDH) in CNS tumors – A study of 100 cases

Venus Khan, Shikha Ghanghoria

M.G.M. Medical College, Indore, Madhya Pradesh, India

Introduction: The annual global age-standardized incidence of primary malignant brain tumors is 3.7 per 100,000 for males and 2.6 per 100,000 for females. Mutations of isocitrate dehydrogenase 1 genes have been identified as early molecular events in the development of gliomas. Objectives: Study the role of IDH mutations in prognosis of CNS tumors & correlation with WHO grade and immunohistochemical expression of Ki-67. Materials and Methods: Herein we report data from major medical center of MP (M Y Hospital & MGM M College). IDH1 mutation was analysed by IHC in 100 patients of CNS tumors and their markers applied for diagnostic accuracy were GFAP, S-100, EMA & synaptophysin. Results: Out of 100 CNS tumors 42 cases were gliomas and IDH1 mutation were absent in grade I glioma (out of 02 cases), in grade II gliomas IDH1 mutations were observed in 12/14 cases, in grade III gliomas all cases are showing IDH1 mutation (04 cases) In grade IV glioma, 08/22 cases show IDH mutation. Increased glioma grades are associated with increased age and Ki67 index. Other 58 tumors include meningioma, medulloblastoma, metastatic and others. Conclusion: IDH1 mutations are associated with the progression of grade of gliomas and the presence or absence of IDH mutations can help to predict the prognosis.

Keywords: CNS tumors, IDH (Isocitrate dehydrogenase)

   AOP270 Top

   Neuropathology Top

Expression of programmed death ligand 1 in gliomas

Havarala Keerthana, Deepti Mutreja

Armed Forces Medical College, Pune, Maharashtra, India

Introduction: The ability of gliomas to induce immunosuppression limits the innate defense against tumor growth and thus poses a significant challenge to the development of new therapies. Physiologically, PD-L1 is expressed to protect tissues from excessive inflammation, but malignant tumours express PD-L1 to escape from host immune surveillance. The PD-1/PD-L1 axis is believed to play a key role in the immune evasion mechanism of tumors. Objectives: To study the expression of PDL-1 marker by IHC in patients diagnosed with gliomas. Materials and Methods: Descriptive study includes 60 cases of glioma diagnosed at tertiary care center in span of 3 years. H&E and IHC stained section of brain biopsies will be taken for the diagnosis of Gliomas in prospective cases and of preserved tissue blocks in retrospective cases. Representative sections will be subjected to immunostaining by PD-L1. Results: The staining of cytoplasm or membranes was deemed to be positive for PD-L1 expression in over 5% tumor cells. Conclusion: Studies have reported that the PD-1/PD-L1 pathway plays a key role in glioma progression and in the efficacy of immunotherapies. Therefore, we focused our work to investigate the PD-L1 expression in gliomas and its association with WHO grading of Gliomas by immunohistochemistry but our study showed no significancy.

Keywords: Glioma, programmed death ligand 1

   AOP271 Top

   Neuropathology Top

Detection and differentiation of IDH mutations in 20 cases of gliomas through RT-PCR

Vartika Mishra, Rajesh Gaur, Sudha Iyengar, Gajendra Pal

Gajraraja Medical College, Gwalior, Madhya Pradesh, India

Introduction: Presence of IDH mutation is an indicator of better prognosis as it increases chemotherapeutic and radiotherapeutic sensitivity of the gliomas. It has accurate ability to detect even a single neoplastic astrocyte. This can provide ground for treatment strategies targeting IDH mutations like IDH inhibitors. Real -time polymerase chain reaction is used in this study to detect and differentiate IDH Mutations in Formalin- Fixed Paraffin embedded tissues of glioma patients. Objectives: To detect common mutations in Gliomas and thus determine the prognosis and direct further therapy. Materials and Methods: 20 diagnosed cases of gliomas were selected. DNA was extracted from Formalin- Fixed Paraffin embedded tissue and quantification was done. DNA detection was done for IDH1 and IDH2 mutations and differentiation for IDH1 R132H, IDH1 R132C, IDH1 R100Q, IDH2 R140X and IDH2 R172X. Results: 5 cases (25%) out of 20 showed IDH1 R132H mutation. These gliomas were located in Frontal region and patients belonged to age group of 31-40 years. Conclusion: We can conclude IDH 1 R132H to be the most common IDH mutation in our study. This finding can be further used for targeted therapies.

Keywords: DNA, glioma, IDH

   AOP272 Top

   Neuropathology Top

Study of histopathological spectrum of meningiomas and correlation of grade with clinical and radiological features

Apurva Ramteke, Rajendra Chaudhari, Preeti Bajaj, Jyoti Sonawane, Duhita Kodare

Dr. Vasantrao Pawar Medical College, Hospital and Research Centre, Nashik, Maharashtra, India

Introduction: Meningiomas comprise 36.6% of all primary CNS tumors reported histologically. Female to male ratio in intracranial meningiomas is approximately 2:1 and 9:1 in the spine. Objectives: 1. To study histopathological spectrum of meningiomas. 2. To study correlation between histopathological grading and tumor enhancement. Materials and Methods: Histologically diagnosed 22 cases of meningiomas were studied from January 2021 to July 2022. All patients operated for intracranial or extracranial meningiomas during the study period who had clinical follow up and radiological imaging were included. The exclusion criteria was patients who had undergone previous radiotherapy. Statistical analysis was done. Results: Out of 22 cases, 16 were females and 6 males, age ranging from 32 to 70 years. Majority of the patients complained of headache. Nineteen cases (86.36%) showed intracranial involvement, out of which thirteen were supratentorial and six infratentorial and three (13.64%) cases showed spinal involvement. One case (4.55%) out of 22 was found in intraventricular region. The most common grade was meningioma WHO grade 1 (14 cases, 63.64%). Seven cases (31.82%) were diagnosed with WHO grade 2 and one case (4.54%) with WHO grade 3. Eight cases (57.14%) of grade 1 and three cases (42.85%) of grade 2 meningioma showed heterogenous tumor enhancement. One case of grade 3 meningioma showed homogenous tumor enhancement. Three cases (13.64%) showed discrepancy in radiological and histopathological diagnosis. Conclusion: Meningioma has female predominance and grade 1 occurs at early age than others. Radiological findings may be useful in preoperative characterization of lesion, but histopathological examination is essential for definitive diagnosis of meningioma.

Keywords: Meningioma, radiological features, WHO grade

   AOP273 Top

   Neuropathology Top

Immunohistochemical expression of EMT markers twist and snail + slug in adult diffuse gliomas and its correlation with known prognostic factors

Sherrin Jacob, Charanjeet Ahluwalia, K B Shankar, Sunil Ranga

VMMC and Safdarjung Hospital, New Delhi, India

Introduction: Gliomas are the most common primary malignant tumors of the brain with a median age of 39.3 years. Glioblastoma, carrying the worst prognosis among gliomas is the most common tumor affecting this population. Here, we contemplate to study the expression Epithelial Mesenchymal Transition (EMT) markers in adult diffuse gliomas. Objectives: 1. Evaluate the expression of EMT markers, Twist and Snail + Slug in adult diffuse glioma and score based on intensity and extent of staining. 2. To correlate the score of EMT expression with prognostic factors namely, WHO grade, Ki 67 proliferation index, ATRX, p53. Materials and Methods: • H&E-stained sections of the tumor along with Immunohistochemical sections of GFAP, IDH, p53, ATRX, Ki67 was evaluated for diagnosis, categorization and grading of glioma. • IHC of Twist and Snail +Slug at 1:100 and 1:1000 dilution respectively was done to evaluate and score the intensity and extent of staining. Results: We have analyzed 80 cases of adult diffuse glioma over a period of 2 years and found a statistically significant correlation between Snail + Slug score and WHO grade of tumour (p value =0.003). A similar statistically significant correlation was found between Twist score and WHO grade of tumour (p value =0.002). The result also highlights a correlation of Twist and Snail+Slug with Ki67, ATRX and p53. Conclusion: Our study suggests the use of EMT markers to predict the prognosis of the tumor. Advent of targeted therapy against these proteins may impact the prognosis in the near future.

Keywords: Gliomas, snail+slug, twist

   AOP274 Top

   Neuropathology Top

Differential expression of CD34, EMA and Inhibin in differentiating meningioma, hemangioblastoma and solitary fibrous tumour

Nuneno Nakhro, Meetu Agrawal, K B Shankar*, Snigdha Goel, Sunil Ranga

VMMC and Safdarjung Hospital, New Delhi, India

Introduction: Meningioma, hemangioblastoma and solitary fibrous tumour of the central nervous system are generally low grade, slow-growing tumours. Differentiating them can be challenging in cases when they show similar histomorphology. Although the conventional haematoxylin and eosin (H&E) staining remains the mainstay of histopathological diagnosis, additional use of appropriate immunohistochemistry (IHC) panel helps to substantiate the diagnosis. Objectives: To evaluate the differential expression of CD34, Epithelial membrane antigen (EMA) and Inhibin as important markers in segregating meningioma, hemangioblastoma and solitary fibrous tumour in central nervous system. Materials and Methods: A retrospective study was carried out in our institution from January, 2022 to August, 2022. During this period, a total of 40 intracranial tumours, morphologically classified as meningioma, hemangioblastoma and solitary fibrous tumours were included in the study. Immunohistochemistry was performed using CD34, EMA, and Inhibin antibody with the available clones and as per the manufacturer's instructions. Their expressions were analysed and interpreted according to standard literature. Results: All meningioma subtypes with the exception of atypical meningiomas showed immunopositivity for EMA while being negative for Inhibin and CD34, except fibroblastic meningiomas which showed CD34 positive. On contrary, all hemangioblastomas showed strong immunopositivity for Inhibin and CD34 while being negative for EMA. Solitary fibrous tumour also showed immunopositivity for CD34 and focal positive for EMA while being negative for Inhibin. Conclusion: The current study results suggest that the IHC panel (CD34, EMA, Inhibin) helps to differentiate between meningioma, hemangioblastoma and solitary fibrous tumour in cases where morphological features are overlapping.

Keywords: Hemangioblastoma, meningioma, solitary fibrous tumour

   AOP275 Top

   Neuropathology Top

Diagnostic utility of Intraoperative squash cytology in central nervous sytem tumors and tumor like lesions with histopathological correlation

Akanksha Yadav, Varsha Kumar, Divya Singh, Pankaj Gupta, Swati Tyagi, Vatsala Misra

Department of Pathology, Moti Lal Nehru Medical College, Prayagraj, Uttar Pradesh, India

Introduction: Squash smear is the most widely used cytological technique in intraoperative neuropathology. This technique is simple, rapid, inexpensive and accurate intraoperative diagnostic modality which provides a rapid pathological diagnosis. Objectives: The aim of this study was to compare the diagnostic accuracy of intraoperative squash cytology with Histopathological examination. Immunocytochemistry was applied as and when required. Materials and Methods: An observational cross sectional study was conducted in the department of Pathology from January 2020 to September 2022. Total of 110 cases of CNS space occupying lesions were included in our study. 40 tissue specimens were received for squash cytology. Smears were prepared from the tissue sent and the remaining tissue was submitted for histopathological analysis. Squash cytology was compared with histopathological diagnosis. Immunocytochemistry was applied on cell block preparations when needed. Results: Gliomas (44%) were most frequently encountered tumour followed by Meningioma (29%) and Schwannoma (09%). Amongst the Gliomas, majority were grade 4 (73%). Diagnostic accuracy of squash cytology was 100% with sensitivity 100%, specificity 100%, positive predictive value 94.11% and negative predictive value 100%. Cell block followed by ICC helped in differentiating reactive gliosis and glioma, CNS Lymphoma from high grade glioma. Conclusion: Squash cytology is an indispensable diagnostic tool for estimating the grade of tumor, differentiating primary from secondary tumours. Immunocytochemistry provides accurate insights with respect to Ki67 index for grading of gliomas, differentiating gliomas from reactive gliosis and differentiating CNS lymphoma from high grade gliomas.

Keywords: CNS tumors, immunocytochemistry, squash cytology

   AOP276 Top

   Neuropathology Top

Clinicopathological analysis of aggressive meningiomas in a tertiary care centre

Nitika Yadav, Surabhi Tyagi, B S Sharma

Mahtama Gandhi Medical College and Hospital, Jaipur, Rajasthan, India

Introduction: Meningiomas are derived from meningiothelial cells of the arachnoid layer. Meningiomas are categorized into 3 grades according to the WHO 2016 Classification. Aggressive meningiomas includes WHO Grade II & III, which have been included in our study. Objectives: Clinicopathological analysis of aggressive meningiomas. Materials and Methods: Total 31 cases of aggressive meningiomas were included. The specimens were received and processed as per the standard protocols & appropriate IHC guidelines. Results: Total Grade II cases were 20 out of which 15 cases were of atypical meningiomas with high MIB -1 activity, 2 cases were of clear cell meningiomas and 3 cases were of chordoid meningiomas. Total Grade III cases were 10 cases (Rhabdoid-5 cases, Anaplastic- 3 cases, Papillary- 2 cases). 1 case was of lymphoplasmocytic rich meningioma with brain invasion and marginally increased MIB 1. In our study male gender, middle age & cerebral convexity had higher predilection for aggressive meningiomas. Conclusion: The aggressive WHO Grade II & III meningiomas have a poor prognosis and the post operative treatment and follow up is affected by the grade. Therefore it is important to classify meningiomas according to their grades even if they may look benign on first look careful inspection of the sections is advised.

Keywords: Agressive, meningioma, MIB-1

   AOP277 Top

   Neuropathology Top

Study of single nucleotide polymorphism in MGMT gene promoter and correlating MGMT methylation with prognosis in glioblastoma

Krachi Agarwal, Shivanjali Raghuvanshi, Ajay Singh, Chhitij Srivastava, Sudhir Singh

King George's Medical University, Lucknow, Uttar Pradesh, India

Introduction: Glioblastoma (GBM) is the most common and aggressive brain tumor with median survival of 15 months. MGMT gene (O6-methylguanine-DNA methyl-transferase) promoter methylation is the strong predictive biomarker for response to alkylating chemotherapy (temozolomide) in glioblastoma. Several single-nucleotide polymorphisms (SNPs) exist in the MGMT promoter region of which rs1625649 downregulates MGMT protein expression, providing better chemotherapeutic response. Objectives: To study MGMT gene methylation and SNPrs1625649 in Glioblastoma and correlating with patient survival. Materials and Methods: A tertiary care hospital study, including 66 histologically and immunohistochemically (IDH-1, p53) proven glioblastoma cases. Genomic DNA isolated from paraffin embedded tissue was PCR (polymerase chain reaction) amplified to study MGMT methylation and RFLP (restriction fragment length polymorphism) to determine SNPrs1625649. Patient was followed up for progression free survival (PFS). Results: The mean patient age was 43 years with male to female ratio1.6:1. Headache was the most common presenting complain (87%). MGMT methylation (MGMT+) identified in 37 (56.1%) cases while 29 (43.9%) were unmethylated. SNPrs1625649 with genotype AA, CA, CC was 10 (15.2%), 15 (22.7%) and 41 (62.1%) respectively. MGMT methylation was associated with sex (p-value 0.042) and not with age (p-value 0.764). Mean PFS in MGMT+ was 2 months (p-value<0.049) as compared to 1.1 months in unmethylated MGMT. Mean PFS of 1.8 months in MGMT methylated GBM with AA genotype was longer than those with genotype CA and CC. Conclusion: MGMT promoter methylation and homozygous SNPrs1625649 (AA genotype) in promoter region correlated with better PFS. The study highlights the prognostic value of MGMT in glioblastoma patient treated with temozolomide.

Keywords: Glioblastoma, MGMT methylation, prognosis

   AOP278 Top

   Neuropathology Top

A retroprospective study of IDH1 and p53 mutations in gliomas in a tertiary care hospital

Nissy Louis, M C Sapna Patel

JSS Academy of Higher Education and Research, Mysuru, Karnataka, India

Introduction: Gliomas account for more than 70% of brain tumors and glioblastoma is the most frequent malignant histologic type. The diagnosis of glioma is based primarily on the WHO criteria that incorporates both histology and molecular parameters in defining tumor entities which is based on the presence or absence of isocitrate dehydrogenase (IDH1R132H) mutation. Objectives: 1. To study the expression of IDH1 and p53 mutation in gliomas. 2. To evaluate the association of IDH1 and p53 expressions with known clinicopathological variables. Materials and Methods: We studied all histopathologically diagnosed cases of gliomas who underwent surgery at JSS Hospital, Mysuru, Karnataka from October 2018 till September 2022. All the cases were appropriately reviewed and graded using the World Health Organization (WHO) criteria published in 2016. Immunohistochemistry (IHC) was performed for detection of IDH1 mutation and p53 expression. The tumor is classified as IDH1 positive with cytoplasmic staining and p53 positive when nuclear staining is observed in more than 5% of tumor cells. p53 expression is further quantified in scores as low- < 8 and high > 8 score. Results: A total of 35 cases were analysed and out of them 20 cases showed IDH1 positivity, 12 cases showed high p53 expression and 6 cases demonstrated both IDH1 positivity and high expression of p53. Conclusion: In gliomas, IDH1R132H mutations are associated with younger age at diagnosis, p53 mutations and favorable survival. Hence the present study of detection of IDH1 and p53 mutations could provide a reliable information for improved tailoring of patient therapy.

Keywords: Gliomas, IDH1, p53

   AOP279 Top

   Neuropathology Top

Study of vascular endothelial growth factor and microvessel density as angiogenic factor in adult diffuse gliomas: Their correlation with tumour grade

Soni, Vaishali Walke, Deepti Joshi, Tanya Sharma, Adesh Shrivastava, Ashwani Tandon, Neelkamal Kapoor

AIIMS, Bhopal, Madhya Pradesh, India

Introduction: High grade gliomas are vascular, aggressive primary brain tumours with worst outcome. They are characterized by abundant and aberrant vasculature which are leaky in nature and cause vasogenic oedema. As tumour angiogenesis is a necessary factor for growth and invasive properties of malignant neoplasm, microvessel density (MVD) and microvascular proliferation may correlate with tumour grade of gliomas and have prognosis. Objectives: The present study was conducted to evaluate Vascular Endothelial Growth Factor (VEGF) expression and MVD using CD34 in adult diffuse glioma; and to examine the microvascular patterns in glioblastoma on dual staining of CD34 and PAS. Materials and Methods: An ambispective, cross sectional, study was conducted on fifty histologically proven cases of adult diffuse glioma (WHO CNS Grade 2-4) diagnosed during a period of 3 years. CD34 and VEGF Immunohistochemistry was performed in each case. MVD was assessed after CD34 staining by hotspot method. VEGF expression scored was based on staining intensity and percentage positivity of tumour cells. Results: VEGF expression score 0 and 1+ was noted in low grade (diffuse) glioma whereas 2+ and 3+ scores in high grade tumours. Similarly low MVD count was evident in lower grade glioma while higher-grade gliomas revealed high MVD. Complex microvasculature patterns like glomeruli tufting and vascular garland are commonly observed in grade 4 gliomas. Conclusion: Heightened expression of VEGF, increased MVD and complex microvasculature patterns a hallmark of high-grade adult diffuse glioma. Over expression of VEGF also provide an opportunity for targeted therapy in high grade adult diffuse glioma.

Keywords: Angiogenesis, diffuse-glioma, microvessel density

   AOP280 Top

   Neuropathology Top

Role of intraoperative squash cytology in diagnosis of central nervous system lesions – A 2 years study

Sneha Sheelwanth, Shivaraj Hanchinal

ESIC Medical College, Kalaburagi, Karnataka, India

Introduction: Lesion within the Central Nervous System are diverse entities, ranging from inflammatory conditions to neoplasms. The annual incidence of tumours of CNS ranges from 10 to 17 per 100,000 persons for intracranial tumours, nearly 20% all of childhood cancers. Squash cytology is an indispensable diagnostic aid to CNS lesions. The definitive lesions is confirmed by histopathologcal examination. Objectives: The present study was carried out to study the squash cytology of CNS lesions and correlate with histopathology. Materials and Methods: A total of 30 cases of CNS lesions were studied over a period of 2 years from 2016 to 2018. Intraoperative squash smears were prepared stained with Heamotoxylin & Eosin, Giemsa and Toludiene blue. Histopathological examination was carried out from remaining sample for confirmative diagnosis. Results: Out of 30 cases, neoplastic cases were 22 and non-neoplastic were 8. Among neoplastic lesions, Meningiomas (5), Schwannoma (2), Fibrillary astrocytoma (2), Grade III Astrocytoma (1), Gliosarcoma (1), Glioblastoma Multiforme (1), Dysembryoplastic Neuroepithelial Tumour (1), Pleomorphic xanthastrocytoma (1), Clear cell ependymoma (1), Medulloblastoma (1), Central Primitive neuroectodermal tumour (1), Hemangioblastoma (1), Adamantinomatous Craniopharyngioma (1), Metastatic Follicular carcinoma thyroid (2), Metastatic poorly differentiated carcinoma (1). Among 8 non-neoplastic lesions, Dermoid cyst (2), Epidermoid cyst (2), Granulomatous inflammation (1), Tubercular inflammation (1), Inflammatory lesion (1), Arachnoid cyst (1). Conclusion: Advantage of squash cytology is that very tiny specimens can also be subjected to smear preparation which is extremely important in intracranial lesions. Squash cytology findings if interpretated with clinical and radiological finding will help to reach an reliable, accurate and rapid diagnosis of intracranial tumors.

Keywords: Central nervous system, meningioma, squash cytology

   AOP281 Top

   Neuropathology Top

Ultrastructural changes in leprous neuritis – Lessons learnt

A V Hema, Rashmi Santhoshkumar, Madhu Nagappa, Anita Mahadevan

NIMHANS, Bengaluru, Karnataka, India

Introduction: Leprosy continues to be a public health challenge in India. Peripheral neuropathy is common as M. leprae have affinity to Schwann cells. Although pathological changes are well known, ultrastructural studies are scarce. Objectives: Elucidate alterations in Schwann cells, axons, myelin, perineurium, fibroblasts, and blood vessels on ultrastructure. Materials and Methods: Leprous neuritis (n= 56) diagnosed between January 2021 to May 2022 were reviewed [H&E, Masson trichome for collagen, Kulchitsky Pal for myelin, Fite Farraco stain for lepra bacilli], subtyped as per Ridley Jopling criteria. Available biopsies were subjected to transmission electron microscopy (EM). Results: Histological subtyping showed Borderline tuberculoid (BT, n=16), borderline lepromatous (BL, n=10), Mid borderline (BB, n=4) and borderline lepromatous (BL) in 3. Chronic leprous neuritis (CLN, n=19) with endoneurial collagenization, scant inflammation and no bacilli was most frequent. EM performed in 16 [BL- 6, BB-4, CLN-4, BT- 2] revealed lepra bacilli within foam cells, unmyelinated Schwann cells, axons and fibroblasts. Only in BB, bacilli were detected in endothelium, and vascular smooth muscle cells. Myelin alterations included extensive fiber loss, separation of myelin lamellae and degradation. Prominent reduction in unmyelinated axons, reduced microtubule density with neurofilament accumulation was evident but Schwann cells appeared normal. Fibroblasts had irregular nuclei, mitochondria with loss of cristae. Hyperplastic endothelium occluding lumen with reduced pinocytic vesicles was evident in endoneurial vessels. Endoneurial collagen accumulation was more prominent in presence of higher bacillary density (BL). Conclusion: Neurofilament accumulation, microtubule reduction in unmyelinated axons, mitochondrial alterations in fibroblasts were unique findings in the study. Intriguingly, Schwann cells remained unaltered. These provide pathophysiologic insights into the mechanisms of nerve injury in this chronic infection.

Keywords: Leprosy, Schwann cell, ultrastructure

   AOP282 Top

   Neuropathology Top

Spectrum of phenotypic alterations in astrocytes in fungal infections of central nervous system

Aditi Goyal, Shilpa Rao, M Netravathi, A Arivazhagan, J Saini, Anita Mahadevan

NIMHANS, Bengaluru, Karnataka, India

Introduction: Astrocytes, most abundant cells in CNS, are diverse in morphology and function. As a response to cellular injury, astrocytes undergo proliferation, hypertrophy, extension and interdigitation of processes (astrogliosis). Astrocytic response in CNS infections has not been systematically evaluated. We sought to study astrocytic changes in fungal infections, incidence of which is rising due to increasing immunosuppressive states, ecological changes etc. Objectives: To evaluate morphological patterns of astrocytic response in CNS fungal infections. Materials and Methods: Clinical, demographic and neuropathological changes of twenty patients with CNS fungal infections were reviewed. H&E, special stains (Periodic-acid-Schiff, Gomori-methenamine-silver-stain) for fungal hyphae and immunohistochemistry for GFAP performed to evaluate astrocytic response. Results: Twenty cases including mucormycosis-3, invasive aspergillosis-3, aspergilloma-4, cryptococcoma-3, cryptococcal meningitis-3 and chromoblastomycosis-3, and candida-1 infection were studied. Granulomatous response was prominent in aspergilloma, cryptococcoma and chromoblastomycosis. Invasive aspergillosis and mucormycosis showed suppurative inflammation and microabscesses were seen in Candida. Cryptococcal meningitis showed minimal inflammation. Astrocytes showed varied morphologies dictated by histological patterns. Granulomatous inflammation showed total loss of astrocytes within necrotic centres. Surrounding granulomas, astrocytes were dysmorphic with thick, irregular clumped processes. Suppurative lesions showed hypertrophic astrocytes with stunted processes. Subpial stellate astrocytes with long slender processes were seen in cryptococcal meningitis. In acute lesions, clasmatodendrosis was prominent with fragmentation and/or beading of astrocytic processes. Conclusion: Varied tissue response of CNS fungal infections were associated with distinct astrocytic phenotypes. Dysmorphic changes can impact astrocytic function leading to altered blood-brain-barrier and neuronal dysfunction causing morbidity. Understanding glial pathophysiological changes may help abrogate neurological sequelae in survivors.

Keywords: Astrocytes, CNS, fungal infection

   AOP283 Top

   Neuropathology Top

Posterior fossa ependymoma: H3 K27me3 immunohistochemistry-based classification

P Rajalakshmi, Deepti Narasimhaiah, Jayanand Sudhir, Bejoy Thomas

Department of Pathology, Sree Chitra Tirunal Institute for Medical Sciences and Technology (SCTIMST), Thiruvananthapuram, Kerala, India

Introduction: Posterior fossa ependymomas are now histopathologically classified into posterior fossa group A (PFA) ependymoma and posterior fossa group B (PFB) ependymoma with prognostic implications. Immunohistochemistry for H3 p.K28me3 (K27me3) has emerged as a surrogate marker for this classification. Objectives: To classify posterior fossa ependymomas into PFA and PFB groups based on immunohistochemistry with clinical correlation. Materials and Methods: Retrospective study of 26 posterior fossa ependymomas from January 2016 to December 2021. Immunohistochemistry with H3 K27me3 and H3 K27M were performed. Clinical data were retrieved from archives. Results: Age ranged from 1 to 66 years (median: 11 years). With typical histomorphology of ependymoma, they showed EMA paranuclear dot positivity and variable GFAP positivity. PFA ependymoma: 14 cases characterized by loss of nuclear expression of H3 K27me3. Age ranged from 1 to 21 years (median: 6.5 years). All the cases were morphologically grade 3 and MIB-1 labelling index ranged from 10-40%. One case was positive for K27M-mutant H3. Of the 13 cases with follow-up, six had recurrence (46.1%). PFB ependymoma: 12 cases defined by intact nuclear expression of H3 K27me3. Age ranged from 11 to 66 years (median: 23 years). Morphologically, eight (66.6%) cases corresponded to grade 2 and MIB-1 labelling index ranged from 1-20%. None of the 10 cases with follow-up data had recurrence. Conclusion: PFA ependymomas manifest frequently in the first decade of life, commonly have high grade morphological features and higher chance of recurrence. Rare tumors can harbor H3 K27M mutation.

Keywords: H3K27me3, K27M-mutant H3, posterior fossa ependymoma

   AOP284 Top

   Pediatric Pathology Top

Clinico-pathological correlation and review of mesenchymal hamartomas of liver

Purvi Bansal, Poonam Panjwani, Girish Chinnaswamy, Sajid Shafique Qureshi, Vasundhara Rishikesh Patil, Maya Prasad, Mukta Ramadwar

Tata Memorial Centre, Mumbai, Maharashtra, India

Introduction: Hepatic mesenchymal hamartomas (HMH) are the second most common pediatric benign hepatic tumors with non-specific clinic-radiological presentation. Objectives: To provide a comprehensive overview of the clinical and radio-pathological correlation of the patients diagnosed with HMH in a tertiary cancer hospital. Materials and Methods: In this retrospective study, hematoxylin and eosin slides of patients diagnosed with HMH between 2011 to 2022 were studied. Clinical, paraclinical, histopathological and treatment data and follow up information were recorded. Results: Of the total 12 patients studied, (3 biopsies, 4 resections and 5 patients with both biopsy and resection specimens) median age was 1.5 years (Range: 1-7 years) with male: female ratio of 1:2. Eleven patients presented with abdominal distention and palpable lump. One patient of Wilms tumor with incidental HMH was identified. Serum AFP levels were raised in 4 patients (Range: 0.99-2638). Right: left lobe involvement ratio was 6:1. On radiology, 10 patients presented with a large heterogeneously enhancing lesion. The resection specimen showed large solid-cystic lesions. Histology revealed predominantly hyalinized stroma (with focal myxoid component in 5 patients), dilated blood vessels admixed with abnormal bile duct proliferation (all patients) and hepatocyte clusters. Histological correlation between biopsy and resection was noted. Follow up was available in 6 patients with no recurrence in them after complete resection, indicating excellent prognosis. Conclusion: HMH are benign pediatric hepatic tumors. Treatment with surgery alone has excellent prognosis. It requires a careful integrated clinical, radiological and pathological assessment for definitive diagnosis.

Keywords: Duct proliferation, fibro-myxoid stroma, mesenchymal hamartoma

   AOP285 Top

   Pediatric Pathology Top

Histopathological spectrum of non-Wilms pediatric renal tumors from a tertiary care center

Aishwarya Karthikeyan, Ipsita Panda, Nandita Kakkar, Manish Rohilla, Kirti Gupta, Radhika Srinivasan, Srinivasan Peyan, Richa Jain, Deepak Bansal, Amita Trehan

PGIMER, Chandigarh, India

Introduction: Pediatric renal tumors (PRT) encompass a highly heterogenous group of entities, with Wilms tumors (WT) accounting for 80-90% of cases. Non-Wilms pediatric renal tumors (NWPRT) are less common and have overlapping histo-morphological features, hence pose a diagnostic challenge. Objectives: This study aims to characterize the histological spectrum of NWPRT diagnosed at our tertiary care institute. Materials and Methods: A retrospective study was conducted on archival material received in our department of histopathology between 2012 to august 2022. Results: A total of 221 cases of PRT were diagnosed over ten years, of which 156 (70.59%) were Wilms tumors. Sixty-five (29.41%) cases were NWPRT of which 17 (26.15 %) were renal cell carcinomas, 12 (18.46%) were clear cell sarcoma of the kidney (CCSK), 11 (16.92%) were malignant rhabdoid tumors, 9 (13.85%) were congenital mesoblastic nephroma (CMN) (cellular= 6, mixed= 2, classical = 1), 5 (7.69%) were Ewings sarcoma, 4 (6.15%) were angiomyolipoma, 4 (6.15%) were multicystic nephroma, and 3 (4.61%) were neuroblastoma. Conclusion: NWPRT is a rare group of heterogeneous entities which often pose a diagnostic challenge to histopathologists. Awareness of the varied morphology and appropriate usage of ancillary testing aids in the diagnosis of these uncommon entities.

Keywords: Non-wilms, pediatric renal tumors, wilms tumors

   AOP286 Top

   Pediatric Pathology Top

Role of light microscopic examination of skin biopsy in diagnosis of neuronal ceroid lipofuscinoses

Aashita Agarwal, Divya Aggarwal, Lokesh Saini, Veena Laxmi, Poonam Elhence, Meenakshi Rao, Sudeep Khera, Deepak Vedant, Vikrant Verma, Kuldeep Singh, Sarbesh Tiwari

All India Institute of Medical Sciences, Jodhpur, Rajasthan, India

Introduction: Neuronal ceroid lipofuscinoses (NCL) are a group of rare neurodegenerative disorders characterised by visual and intellectual deterioration and seizures. Intralysosomal accumulation of autofluorescent lipopigments is demonstrable in neurons at autopsy, however, it also occurs in extracerebral tissues including skin, rectum and skeletal muscle. Extracerebral storage cytosomes (similar to neurons) have been reported in smooth and striated muscle, secretory epithelial cells from eccrine sweat glands, vascular endothelial and smooth muscle cells and ganglion cells of enteric nervous system. Objectives: To elucidate the role of light microscopic and fluorescent microscopic examination of skin biopsy in genetically proven cases of NCL. Materials and Methods: Five cases of NCL were identified, who had undergone axillary skin biopsy as a part of diagnostic work-up and were subsequently confirmed genetically. The biopsies were evaluated to identify light microscopy changes which are diagnostic of NCL. Five axillary skin biopsies (performed for other non-genetic skin conditions) were used as control. Results: Light microscopy changes were identified in the form of vacuolation of eccrine gland cells, with a brownish tinge of cytoplasm. These lipopigments stained with Periodic-acid Schiff (PAS) as PAS positive granules. On luxol fast blue staining with cresyl violet counterstaining, these were seen as blue cytoplasmic granules. These granules showed yellowish autofluorescence on fluorescent microscopic examination of unstained slides, while the age related lipofuscin pigment showed orange autofluorescence. Conclusion: Light microscopic examination of skin biopsy is a useful and under-utilised modality for the diagnosis of cases of NCL.

Keywords: Extracerebral accumulations, neuronal ceroid lipofuscinoses, skin

   AOP287 Top

   Thoracic Pathology Top

Histopathological study and immunohistochemical expression of epidermal growth factor receptor in lung tumours – A hospital based study at KLES Dr. Prabhakar Kore Hospital and MRC, Belagavi

Pratik Bhat, Ranjeet Kangle, Dhairyasheel Salunkhe

JNMC, Belagavi, Karnataka, India

Introduction: Currently, lung cancer is the commonest aetiology for cancer mortality in world As most of the lung cancers are in advanced stage of disease at the time of diagnosis, screening attempts have to be developed in future so that the cancer can be identified for the better outcome and prognosis. Further, it was studied that both EGFR mutation and gene amplification status were essential to know which tumours will respond to TKIs. Hence, the present study is being conducted to study histopathological spectrum of lung cancer types and EGFR expression by IHC. Objectives: To study histopathological spectrum of lung cancer types and EGFR expression by IHC. Materials and Methods: A total of 40 cases of lung tumours were included from January 2019 to December 2020. Clinical details and gross findings were obtained from medical records and grossing notes using a structured proforma. Paraffin embedded blocks were archived and histological findings & IHC expression of EGFR were analysed. Statistical analysis was done using chi square test and 'p' value of less than 0.05 was considered statistically significant. Results: Peak incidence of 40% of lung tumours was noted in the age group of 51-60 years, with a clear male preponderance. 65% were smokers, 52.50% had mixed diet intake and 15% were positive for family history of lung cancer. 50% of tumours in the study were located in the upper lobe of right lung. 52.50% were adenocarcinomas and 32.50% were squamous cell carcinomas with a single case of lepidic variant of adenocarcinoma and keratinizing variant of SCC each. 85% cases showed positive staining intensity for EGFR, among which 10% were weak positive, 35% were intermediate and 40% were strong positive. 72.50% cases were EGFR IHC positive by H-score method. 87.50% had cough as the most common symptom, 62.50% had dyspnoea, 62.50 % showed loss of appetite, 67.50% presented with weight loss and 45% had haemoptysis. Conclusion: Increasing age and types of lung tumours like squamous cell carcinomas, adenocarcinomas and their variants, showed a significant association with intensity of EGFR staining. A significant association between H-score and types of lung tumours was established in this study. Thus, the expression of EGFR protein can be used as a marker for targeted therapies in patients suffering from these lung tumours.

Keywords: EGFR, immunohistochemical, lung cancer

   AOP288 Top

   Thoracic Pathology Top

Myocardial tuberculosis: A 22 year autopsy study

Vishal Rajput, Heena Desai, U Sujith, Pradeep Vaideeswar, Gayatri Amonkar

Department of Pathology, Seth G S Medical College and K.E.M Hospital, Mumbai, Maharashtra, India

Introduction: Tuberculosis is primarily a pulmonary disease but it can affect any organ in the body including the heart. Tuberculous pericarditis is the commonest manifestation of cardiac tuberculosis. Myocardial involvement (isolated or with concomitant pericardial involvement) is uncommon. The diagnosis is often made during autopsy. Objectives: To study clinicopathological features of myocardial tuberculosis at autopsy. Materials and Methods: The study is carried out over the span of 22 years (2000-2021). Cases of myocardial tuberculosis in all age were studied with correlation to the clinical manifestations and investigations. Myocardial tuberculosis was classified into nodular, diffuse and miliary type on the basis of histopathology. Results: There were 20 cases of myocardial tuberculosis in the study period affecting 2 males and 18 females (5 cases of maternal mortality). The age range was between 14 years to 82 years. Duration of symptoms was ranging from 12 hours to 3 months. The patients clinically presented with cardiac symptoms like chest pain, breathlessness, palpitation along with extra-cardiac symptoms in some. Diffuse type of myocardial tuberculosis observed in 8 cases and miliary type in 12 cases. The other organs affected by tuberculosis include lungs, lymph nodes, liver, kidney, spleen, thyroid and diaphragm; tuberculous dissemination was seen in 4 patients. Conclusion: Very few cases of myocardial tuberculosis were reported till date and most of them diagnosed at post mortem. It should be suspected in patients in tuberculosis who develop cardiac symptoms. Endomyocardial biopsy will be useful tool for diagnosis.

Keywords: Autopsy, myocardial, tuberculosis

   AOP289 Top

   Thoracic Pathology Top

Clinical and molecular correlates of PDL-1 in patients with non-small cell lung cancer (NSCLC)

Khalid AbdulMannan, Daphne Fonseca, Sahithi Shilpa Arya, K Vaishnavi, V Kranthi Kumar, K Suseela

Basavatarakam Indo American Cancer Hospital and Research Institute, Hyderabad, Telangana, India

Introduction: Non-small cell lung cancers (NSCLC) have shown an improved outcome with immune checkpoint inhibitor (ICI) therapy targeting programmed cell death-1 or its ligand (PDL-1). Objectives: To correlate clinicopathological features with molecular phenotype across PDL-1 subgroups. Materials and Methods: A cohort of 100 NSCLC cases was investigated for PDL-1 expression, sample site, gender, histological type, and oncogenic mutation status. PDL-1 testing was performed using clone SP263 on Ventana benchmark and targeted next generation sequencing using lung and colon cancer research panel v2 for DNA and RNA analysis on thermofisher platform. Results: Median age of our cohort was 60 years. Cases were divided into PDL-1 negative (<1%), intermediate (1-49%), and high (>50%) groups. 51% of the cases showed a positive PDL-1 expression. Squamous cell carcinoma (SCC) exhibited higher PDL-1 expression than adenocarcinoma (AC) (p value 0.005). The KRAS expression was more frequent in males (p value 0.048). There was no difference between tumor location or sample types. EGFR mutation was exclusively seen in AC compared with other NSCLC. The other mutations did not show a statistically significant difference with PDL-1 expression. Conclusion: The integration of clinical, molecular, and immunological data will enhance the predictive value of PDL-1 biomarkers in lung adenocarcinoma.

Keywords: Genetic alteration, lung cancer, PDL-1

   AOP290 Top

[TAG:2]Thoracic Pathology [/TAG:2]

Interstitial capillary congestion and blood stasis: Incidental or factual finding in lungs of COVID positive autopsy cases

Preeti Agarwal, Uma Shankar Singh, Shiuli Rathore, Anoop Kumar Verma, Ravindra Kumar Garg

Department of Pathology, King George's Medical University, Lucknow, Uttar Pradesh, India

Introduction: Diffuse alveolar damage was commonly seen in patients who succumbed to COVID-19. The question arises whether these are observed in COVID positive patients who do not have pulmonary symptoms. This can be answered by histological study of lung samples from patient's who do not die due to covid related complications or pulmonary involvement. Objectives: We conducted a study on postmortem lung samples from people who did not die of covid related acute lung injury. The aim was to study the morphological variations in lung samples and compare them with studied alterations in lung in covid. Materials and Methods: In this prospective postmortem study subjects who died within 24 hours due to acute injuries with positive RT-PCR throat swab, and had no signs and symptoms of covid pneumonia were included. Detailed morphological analysis of the lung samples was performed. Results: Total 20 subjects were enrolled. Morphology wise most frequent change seen in the lungs was emphysema followed by capillary congestion which were curvilinear in 85% (17/20), acute phase of diffuse alveolar damage was seen in 25% subjects, its organizing phase was seen in 10%. Acute bronchopneumonia was observed in 20% of study population. Another interesting finding was active proliferation of fibroblasts seen in the interalveolar septa and small collection of these cells in the interstitial spaces. Interestingly hyaline membrane and squamous metaplasia was readily seen in cases with DAD. Conclusion: Presence of curvilinear capillary congestion and red blood cell stacking backs role of cytokines which regulate vascular tone and congestion. Presence of fibrotic nodule supports early fibrosis ensues in patients suffering from covid even when clinical symptoms have not commenced.

Keywords: COVID-19, COVID lung, DAD

   AOP291 Top

   Thoracic Pathology Top

Clinico-pathological features of primary pulmonary hepatoid adenocarcinoma (PPHAC); An under-recognized and aggressive lung neoplasm with distinct immunophenotype (TTF1-neg/CK7-pos/HepPar1-pos)

Konil Varshney, Rajiv Kaushal, Trupti Pai, Neha Mittal, Kumar Prabhash, Amit Janu, C S Pramesh

SRL Daignostics, Mumbai, Maharashtra, India

Introduction: Primary pulmonary hepatoid adenocarcinoma (PPHAC) is a rare, aggressive neoplasm. Till date, it has failed to establish a distinct place in WHO classification for lung tumour. Objectives: We hereby, present the clinic-pathological spectrum of PPHAC. Materials and Methods: PPHAC diagnosed between 2012-2020 were retrieved from pathology archives. Cases with known extra-thoracic primary were excluded. Clinico-radiological details were recorded and histopathological features were reviewed. Results: A total of 45 cases of PPHAC were recognized. Exclusive predominance for males (43/45) and smokers (39/40) with median age of 55 years (range 43–78 years) was noted. Tumour was predominantly located in upper lobe [(N=32/45 (71%)] and ranged from 3 cm – 11 cm in size. AFP levels were raised in 1/3 cases. Histopathological examination, revealed predominantly solid (n=20) and trabecular pattern (n=12). Tumor cells were large polygonal with abundant eosinophilic to clear cytoplasm in 89% (40/45) cases. IHC showed a consistent and distinctive immunoprofile i.e. CK7+/HepPar-1+/TTF1- in all 45 cases. Molecular evaluation (n=25) revealed consistent negativity for EGFR mutations, EML4-ALK, ROS1 gene rearrangements and Met amplification. Chemo-radiotherapy (n=25) was the main treatment modality. Almost half of patients (22/45) had metastatic disease at presentation. Surgical resection was possible in only 4 cases. Median follow up was 12 months (range: 2 months – 5 years) and 8 patients (24%) succumbed within six months of diagnosis. Conclusion: PPHAC is emerging as a distinctive, highly aggressive, under-recognised entity which occurs exclusively in smoker males. In view of consistent Hep-Par1 positivity and TTF1 negativity, it may represent a potential diagnostic pitfall.

Keywords: Adenocarcinoma, aggressive, lung

   AOP292 Top

   Urogenital Pathology Top

Interobserver variability in the grading of prostatic adenocarcinomas according to the 2019 Gleason grading system

Mansi Agarwal, Hema Kini, Saraswathy Sreeram, Jyoti Kini

Kasturba Medical College, Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, India

Introduction: Gleason grading system is a histological grading system used for prognostication and management of prostatic adenocarcinomas (PCa). It has undergone significant revisions over the past years, but continues to have deficiencies that can potentially impact patient care. Objectives: We studied the reproducibility of the Gleason grading system as per the 2019 International Society of Urological Pathology (ISUP) guidelines, by determining interobserver variability and agreement. Materials and Methods: An institutional based retrospective study was conducted with fifty needle core biopsies confirmed to be PCa, received in the year 2021. Histological analysis was done by three observers (P1, P2 and P3) according to 2019 ISUP guidelines. The interobserver agreement was calculated using the interclass correlation coefficient (ICC). Results: Among the fifty cases, complete agreement was seen in 92% cases (n=46/50). Majority of the cases were of Gleason score (GS) 3+3 (18%, n=9) and least for GS 5+3 (n=1) in 2% cases. Disagreement was noted in 4 cases with a grade 4 or 5. Perfect agreement with an ICC (1.00) was seen in all cases for grade groups (GG) (n=50). Most discrepancy involved GG4 and GG5, with either over or under grading. Conclusion: The 2019 ISUP modified guidelines show less interobserver variability and good reproducibility. Standardized reporting can help in improving the results. However, specialized training programs for general pathologists ensure better results thus maximizing the diagnostic utility.

Keywords: Gleason score, grade group, prostatic adenocarcinoma

   AOP293 Top

   Urogenital Pathology Top

Study of CD10 expression pattern in prostatic adenocarcinoma and its correlation with Gleason's grade

Poulomi Biswas, Anup Kumar Roy, Tapan Kumar Mandal

Nil Ratan Sircar Medical College and Hospital, Kolkata, West Bengal, India

Introduction: Benign prostatic hyperplasia and carcinoma of the prostate are frequently increasing with advancing age. CD10 is a 100-kDa transmembrane glycoprotein involved in cleavage and inactivation of certain peptide hormones important for signal transduction. This is a zinc-dependent enzyme that is widely expressed in epithelial cells of kidney, breast, lung, intestine and prostate. Objectives: The aim of our study is to evaluate CD10 expression pattern in benign and malignant prostatic lesions by immunohistochemistry & correlate it with gleason grade group & pre treatment PSA value in prostatic adenocarcinoma. Materials and Methods: Immunohistochemical staining for CD10 is performed on 56 cases of paraffin-embedded tissue from transurethral resected and core biopsy specimen of prostate. Results: In our study we have found that CD10 positivity was more in prostatic adenocarcinoma patients rather than BPH patients with a significant p value of 0.003. We have also found that there is positive linear correlation between CD10 positivity & Gleason grade group & PSA value with significant p value of 0.0016 & 0.0083 respectively. Conclusion: In conclusion, a high CD10 expression may accurately diagnose prostatic adenocarcinoma from BPH, which aids in diagnostic accuracy.

Keywords: CD10, Gleason grade group, prostatic adenocarcinoma

   AOP294 Top

   Urogenital Pathology Top

Histopathological spectrum of ovarian neoplasms in tertiary care centre [1 year study]

K Leena, Naveen Kumar Verma, R K Chandrakar

Shri Shankara Institute of Medical Sciences, Bhilai, Chhattisgarh, India

Introduction: Ovarian neoplasms are 3rd most common site of primary malignancy in female genital tract after cervix and endometrium representing about 30% of all cancers of the female genital system. It is the seventh leading cause of cancer death. Objectives: The Study was undertaken to determine different histological types of ovarian tumours, in a tertiary care centre in Bhilai Chhattisgarh. Materials and Methods: The data pertaining to samples of ovarian tumour specimens received for histopathological evaluation at Central Laboratory of SHRI SHANKARA INSTITUTE OF MEDICAL SCIENCES BHILAI, over a one year period (September 2020 to September 2021). Were analysed to determine the histological pattern and the age distribution of all lesions. The study included 60 histopathologically proven and reported cases. These were classified according to the WHO classification of ovarian tumours. Results: The majority of ovarian lesions received for evaluation were benign and unilateral. Most patients were in the third to sixth decades of life. Of 60 cases of ovarian tumours. Benign neoplastic lesions were 54, malignant were 6. Most commonly encountered benign ovarian tumour were Surface epithelial tumours followed by sex cord tumours -6 and germ cell tumours -4. Out of malignant tumours 2 were of borderline malignancy and 4 were malignant [serous and mucinous carcinoma]. Conclusion: Most common benign tumour encountered in this study was serous cystadenoma. Early diagnosis and appropriate treatment of ovarian neoplasms favour the good prognosis.

Keywords: Benign, borderline, epithelial tomours, malignant

   AOP295 Top

   Urogenital Pathology Top

Diagnostic accuracy of ROMA index in ovarian tumor – A study of 80 cases

Mrudula Yerawar, Shikha Ghanghoria, Ravi Jain, Venus, Priya

MGM Medical College, Indore, Madhya Pradesh, India

Introduction: Ovarian tumors are leading cause of cancer deaths amongst women. Deaths are mostly due to late detection of ovarian malignancy. Risk of Ovarian Malignancy Algorithm [ROMA] created by Moore et al. combines tumor markers CA125 and HE4 (preoperative levels) taking into account menopausal status of each patient, and is an index which aids in early detection of ovarian malignancy. Objectives: To know diagnostic accuracy of Roma index by correlating it with histopathological diagnosis of ovarian tumor. Materials and Methods: A study was conducted in M G M Medical College, Indore, MP. 80 cases of ovarian neoplasm were studied & ROMA index was evaluated. The histopathological diagnosis and ROMA index values were correlated to assess diagnostic accuracy of ROMA index. Results: Out of 80 cases 45% were benign, 15 % borderline and 40% were malignant. Mean ROMA index value in benign is 16.3%, in borderline 25.9% and in malignant is 84.9%. Higher values of CA125, HE4 & ROMA INDEX was found in malignant cases. Conclusion: In 92.7% cases ROMA index could correctly diagnose and match the histopathological diagnosis, indicating high positive predictive value therefore ROMA Index is more useful parameter than CA125 & HE4 alone to differentiate between malignant & benign tumors, also between epithelial ovarian cancer and non malignant adnexal tumors.

Keywords: Ovarian tumors, Risk of Ovarian Malignancy Algorithm Index

   AOP296 Top

   Urogenital Pathology Top

A study of histopathological spectrum of endometrium in patients of abnormal uterine bleeding

Sonal Chandrakar, Rabia Parveen Siddiqui, Varsha Pandey

PT. JNM Medical College, Raipur, Chhattisgarh, India

Introduction: Endometrial diseases ranked among the most common gynecological disorder globally. Abnormal uterine bleeding is defined as any alteration in the pattern/volume of menstrual blood flow. Objectives: Objective was to determine spectrum of endometrial pathologies in different age groups patients presenting with AUB. Materials and Methods: The study was done on biopsies with chief complaint of AUB (excluding the organic causes) in Department of Pathology, Pt. JNM Medical College Raipur (C.G) over the period of 2 years. The specimens were routinely processed. H&E stained slides were studied for endometrial pattern. A statistical analysis between age of presentation and specific endometrial cause was done using chi-square test. Results: Out of 159 cases, 46.5% women were perimenopausal (40-49 years) and the commonest pattern was normal cyclic endometrium (36.5%). 25.2% women were in reproductive age group (<40 years) with the commonest pattern of typical hyperplasia (55%). 28.3% woman were post menopausal (>49 years) with the commonest pattern of endometrioid carcinoma. Other Endometrial patterns included disordered proliferative, atrophic, endometritis, polyp, arias stella reaction, luteal phase defect and progesterone effect. Conclusion: Patients with AUB show varying spectrum of endometrial pattern, ranging from cyclic endometrium to carcinomas. Histopathological examination is the gold standard diagnostic tool in evaluation of AUB and there is an age specific association of endometrial lesions.

Keywords: Abnormal uterine bleeding (AUB), H&E (Hematoxyline & Eosin)

   AOP297 Top

   Urogenital Pathology Top

Evaluation of programmed death ligand 1 and p16 expression in penile tumors

Sabiha Tasneem, Hema Kini, Saraswathy Sreeram

Kasturba Medical College, Mangalore, Karnataka, India

Introduction: Penile malignancy is rare. High risk Human papilloma virus (hrHPV) infection is one of the risk factors. p16 is used as a surrogate marker for HPV. Programmed death ligand 1 (PD-L1) is an important immune checkpoint regulator. Thus, it is used in immunotherapy. Objectives: This study was conducted 1. To evaluate the morphology of all penile tumors and classify them into HPV positive and negative groups. 2. To correlate p16 expression with the histologic types of tumor. 3. To evaluate pattern of PD-L1 expression and its correlation with p16 positive and negative penile tumors. Materials and Methods: Retrospective study of 50 cases of penile carcinoma diagnosed between January 2016 to December 2021. Clinical data and pathological data of selected cases were collected from the archives. H&E slides were reviewed. Immunohistochemical staining for p16 and PD-L1 was performed, and the expression of these markers were evaluated. Results: All men in this study were aged 31 to 90 years. Usual squamous cell carcinoma was the most common histologic subtype (82%). p16 was positive in 24.4% cases. PD-L1 showed patchy positivity. PD-L1 was expressed in 6% cases and all these cases were negative for p16. Conclusion: Localized penile carcinoma usually has good prognosis (90%) after penectomy. However, prognosis worsens in advanced stage. Anti-PD-L1 therapy has shown promising results in advanced melanoma and non-small cell lung cancer. Thus, it is being evaluated in the treatment of advanced penile cancer.

Keywords: Human papilloma virus, p16, programmed death ligand 1, penile malignancy

   AOP298 Top

   Urogenital Pathology Top

Xanthogranulomatous and pseudoxanthomatous inflammation of the female genital tract

Parul Mehrotra, Gayatri Ravikumar

St. Johns Medical College, Bengaluru, Karnataka, India

Introduction: Xanthogranulomatous inflammation (XGI) of the female genital tract (FGT) is very rare. The clinical presentation and pathological associations are not established in large studies. Objectives: To study the various clinico-pathological presentations of XGI in FGT. Materials and Methods: Cases diagnosed as XGI of FGT at a tertiary care hospital over 10 years were reviewed with relevant clinical data. Results: There were 40 cases reported as XGI. Of these 29 cases were idiopathic/primary XGI (PXGI). In 11 cases, a pseudoxanthomatous inflammation was seen in association with other lesions (PXI). The mean age of presentation of PXGI was 46 years. Common clinical presentations were mass per abdomen or pain abdomen (24%). Amongst PXGI, 6 patients had an associated urinary tract infection and one patient had a coexisting fungal infection. Surgical procedures done in PXGI were: Hysterectomy with salphingo-ophorectomy (n=21), salpingo-ophorectomy (n=6), salpingectomy (n=1) and endometrial biopsy (n=1). Frozen section was done for 10 cases. Bilateral involvement was seen in 5 cases. The common sites of involvement were:ovary (7), Fallopian tube (6), tubo-ovarian (11), endometrium (2), myometrium along with adnexa involvement (3). The PXI was seen in ovary (6), Fallopian tube (4) and myometrium (1). The associated conditions were teratoma (4), high grade ovarian serous carcinoma (2), foreign body (1), dedifferentiated endometrioid carcinoma (1), ectopic tubal gestation (1), metastatic mucinous adenocarcinoma (1) and endosalpingiosis with fibroid (1). Conclusion: It is important to identify XGI as it clinically mimics malignancy and can be associated with increased Ca-125. PXGI in FGT is rare and often seen in adnexa, with rare occurrence in myometrium and endometrium. PXI had varied associations and should be differentiated from PXGI due to distinct pathogenesis and management.

Keywords: Pseudoxanthomatous, xanthogranulomatous endometritis, xanthogranulomatous inflammation

   AOP299 Top

   Urogenital Pathology Top

Placental correlates of spontaneous and indicated preterm births

Victoria Liza, Gayatri Ravikumar

St. Johns Medical College and Hospital, Bengaluru, Karnataka, India

Introduction: Preterm births (PTB) can be classified as Spontaneous PTB (SPTB) and Indicated PTB (IPTB). Does placental findings differ and will it help to understand the role of placental in SPTB and IPTB forms the basis of this study. Objectives: To study the difference in placental pathology between SPTB and IPTB. Materials and Methods: Retrospective study analyzing singleton preterm placentas from more than 25 weeks gestation, received over 3 years. PTBs were classified as SPTB and IPTB based on clinical details. Placentas from stillborn/intrauterine demise (IUD) and multifetal gestation were excluded. Results: SPTB (n=154) and IPTB (n=292) placentas were included. The clinical presentations of SPTB were PPROM (46.1%), preterm contraction (27.3%) and antepartum haemorrhage (26.6%). The most common indication in IPTB was severe pre-eclampsia (59.2%). The mean maternal age (yr) was higher in IPTB (25.9 ± 4.8, 27.1 ± 4.8, p=0.016) and mean birth weight (gm) was low (1.8 ± .54, 1.6 ±.57, p<0.001). Placental macroscopic findings showed lower mean placental weight (309 ± 81 vs 274 ± 102, p<0.001), length (15 ± 2.2 vs 14 ± 2.3, p=0.003), breadth (12 ± 1.9 vs 12 ± 1.9, p<0.001) and umbilical cord diameter (1.1 ± .29 vs 1 ± .31, p<0.001) in IPTB placentas. Microscopic analysis showed maternal (26.6% vs 72.9%) and fetal (37.7% vs 49.7%) vascular malperfusion lesions, thrombohematoma (3.9%vs13.4%) and membrane hypoxia (18.8% vs 31.8%) to be higher in IPTB (p<0.001), while maternal inflammatory response (38.3 vs 17.5%) and fetal inflammatory response (12.3% vs 1.4%) were significantly higher (both p<0.001) in SPTB. Conclusion: The acute inflammatory pathology was commonly seen in SPTB, thus proving the role of inflammation in activation of spontaneous parturition. Placentas from IPTB were smaller and weighed lesser with higher frequency of vascular malperfusion lesions.

Keywords: Indicated preterm, placental pathology, spontaneous preterm

   AOP300 Top

   Urogenital Pathology Top

Accuracy of intra-operative frozen section in the diagnosis of ovarian tumors

Mayuri Kute, Rajendra Chaudhari, Preeti Bajaj, Jyoti Sonawane, Duhita Kodare

Dr. Vasantrao Pawar Medical College, Hospital and Research Centre, Nashik, Maharashtra, India

Introduction: Discrimination of benign and malignant tumors during surgery in gynaecologic patients with adnexal masses is important for the management of the patient. The use of frozen section analysis has had a great impact on these patients and has become indispensable in diagnosing malignancies. The results of the frozen examination determine the course of surgery. Objectives: To compare frozen section results with definitive histopathological results of ovarian tumors diagnosed intra-operatively. Materials and Methods: Frozen section results of the patients with a final diagnosis of ovarian tumors operated at our institute between September 2019 to September 2022 were analyzed. The concordance of frozen section diagnosis was determined by comparing the frozen section result with the final pathological diagnosis. Results: Data of 39 patients was analyzed. The results of frozen section were 27 cases (69.2%) benign and 12 cases (30.8%) malignant. The results of paraffin block were 26 cases (66.7%) as benign and 13 cases (33.3%) as malignant. Sensitivity of frozen section in diagnosis of malignant lesions is 92.31% with specificity of 100%, positive predictive value of 100% and negative predictive value of 96.30%. Conclusion: Intraoperative frozen section is useful in situations where the nature of the ovarian tumor is uncertain whether benign or malignant. It is a highly sensitive and specific modality and thus helps the surgeon in planning further surgical management.

Keywords: Frozen section, histopathological results, ovarian tumors

   AOP301 Top

   Urogenital Pathology Top

Histopathological study of urinary bladder neoplasms

Sneha S Nayak, M N Suma, Usha Amirtham, S Balu

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: Urinary bladder cancer is the ninth most common cancer worldwide. Urothelial neoplasms represent about 90% of all bladder tumours. True sarcomas are uncommon in the bladder. Objectives: To assess urinary bladder neoplasms with regard to age, sex, histopathological features and categorize according to WHO 2022 classification of urinary tract tumours. Materials and Methods: A retrospective study was done on resection specimens received in our department between January 2020 and September 2022. Results: A total of 54 resected specimens were studied which included radical cystoprostatectomy (43), radical cystectomy (9) and partial cystectomy (2). Majority of patients were male (94%) and were in the age group 61-70 years. The histopathological spectrum showed predominantly high grade myoinvasive urothelial carcinoma, 5 cases of low grade urothelial carcinoma, 1 rhabdomyosarcoma and 1 leiomyosarcoma. Out of five specimens where patients had received neoadjuvant chemotherapy, 2 had no residual tumour. Among the urothelial carcinomas, one case with nested subtype and 21 cases of divergent differentiation were noted, squamous being most frequent. 15 cases had lymphovascular invasion out of which 46.67% cases showed divergent differentiation. Conclusion: Majority of urinary bladder neoplasms are of epithelial origin with male predominance. Invasion to the muscularis propria layer correlates with high grade tumour. Extensive sampling of tumour, quantification of divergent components and thorough sampling of lymph nodes is necessary in view of potential management implications.

Keywords: Divergent differentiation, myoinvasive, urinary bladder neoplasms

   AOP302 Top

[TAG:2]Urogenital Pathology [/TAG:2]

Evaluation of CD44 expression in prostatic adenocarcinoma: An institutional study

Priyanka Subhashish Das, Soumya Hadimani, Bodapati Sivaramakrishna

Sri Devaraj URS Medical College, Kolar, Karnataka, India

Introduction: Prostate adenocarcinoma (PCa) is the second most common cause of cancer and cancer-related deaths among men worldwide. CD44 plays an important role in mediating cell to cell and cell to matrix interaction, hence promoting the maintenance of tissue integrity and inhibiting tumor metastasis. Objectives: 1. To study the expression of CD44 in prostatic adenocarcinoma. 2. To know the association of CD44 expression with presence of metastases, tumor stage, with PSA level serum and gleason's score (GS). Materials and Methods: This is a cross sectional study done on Trans Urethral Resection of Prostate (TURP), prostatic core biopsy specimens received from Department of Urology from 2020-2022 are included in the study (50 PCa cases). Immunoreaction to CD44 antibodies is evaluated by calculating positively stained cell percentage and staining intensity. P value <0.05 is considered as significant. Results: Out of 50 Cases, 17 cases (34%) showed low GS with 12 cases (70%) score-5 and 5 cases (30%) showed score-6 expression for CD44 and 33 (66%) cases showed high GS with 22 (66.7%) cases score-4, 9 cases (27.3%) score-3 and 2 cases (6%) showed score-2 expression. Intensity of CD44 expression is stronger in 17 cases with low GS. Loss of CD44 expression is associated with greater tumor aggressiveness. The positive expression of CD44 expression on immunohistochemistry was strongly associated with lower S.PSA levels, GS. Conclusion: We conclude that once cancerous properties have been established, cells tend to lose the ability of CD44 expression. CD44 expression in low grade carcinomas suggest that CD44 maintains the tumor in a differentiated, gland forming state so it may not act as a cancer stem cell marker in prostate carcinogenesis.

Keywords: CD44, Gleason's score (GS), prostate adenocarcinoma (PCa)

   AOP303 Top

   Urogenital Pathology Top

Significance of apoptotic index and p16INK4A protein expression in cervical intraepithelial neoplasia and squamous cell carcinoma of cervix

Elizabeth Katepogu, Surekha B Hipparagi

Shri BM Patil Medical College, Vijayapura, Karnataka, India

Introduction: Cervical cancer is a leading cause of cancer related death among women. P16INK4A expression correlates excellently with grade of cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC) & can be used for screening cervical malignancies. Apoptosis play an important role in tumor progression and development of tumors and apoptotic bodies helps to grade dysplasia. Hence p16INK4A expression and apoptotic index (AI) in CIN and SCC cervix has been studied. Objectives: To study the apoptotic index (AI) & p16INK4A expression in cervical intraepithelial neoplasia and squamous cell carcinoma of cervix & to correlate with non-neoplastic lesions of the cervix. Materials and Methods: 73 cervical biopsy specimens diagnosed as CIN, invasive SCC and also non neoplastic cervical lesions as controls are studied. One section stained with H&E for histomorphologic diagnosis and apoptotic index. Other section is subjected to p16INK4A staining. Results: Apoptotic Index (AI) increased progressively from normal to carcinoma but decreased with decreased differentiation of tumor. Expression of P16INK4a noted in 100% cases of carcinoma cervix, 100% cases of HSIL and 25% cases of LSIL with p value 0.0001. Conclusion: Apoptotic index and p16INK4A can be used to screen cervical malignancies and to differentiate between normal, premalignant and malignant lesions of cervix.

Keywords: Apoptotic index, carcinoma cervix, P16INK4a

   AOP304 Top

   Urogenital Pathology Top

Assessment of endometrium using transvaginal sonographic endometrial thickness and endometrial biopsy in perimenopausal abnormal uterine bleeding

Esha Abdul Salam, Gopinath Rajesh, Anjali Rao

Department of Pathology, K.V.G. Medical College and Hospital, Sullia, Karnataka, India

Introduction: Abnormal uterine bleeding (AUB) is a significant clinical presentation among perimenopausal women. In developing countries like India, prevalence of AUB is around 17.9%. Objectives: Study is intended to investigate the association between endometrial thickness in transvaginal sonography (TVS), the results of endometrial biopsy in perimenopausal women with AUB and to evaluate the combined use in diagnosis of endometrial lesions. Materials and Methods: In this retrospective study, 249 perimenopausal women with AUB who underwent TVS and endometrial biopsy were enrolled. Findings of biopsy were compared with hysterectomy specimens whenever data was available. Data was analysed using SPSS V 16. Sensitivity and specificity was calculated for preoperative endometrial biopsy. Results: Menorrhagia was most common bleeding pattern seen in 61.8%. 159 samples were identified to be benign lesions, 85 were with Hyperplasia and 5 were Malignant. The mean endometrial thickness in subgroup without disease were 5.26±3.8 mm, benign were 8.57± 3.06 and Malignancy were 26.40 ±8.79 mm. On comparing hysterectomy specimens, the sensitivity of Endometrial biopsy to identify Hyperplasia with or without atypia was 89.79% and carcinoma was 100%. Conclusion: From this study, we concluded that the combined usage of endometrial thickness using TVS and endometrial biopsy is a reliable diagnostic protocol for detecting endometrial lesions like hyperplasia and carcinoma. We also inferred that if endometrial thickness is 5 mm or less, further investigations are not indicated. One of the drawbacks identified was that during sampling of endometrium with pipelle, endometrial polyps may sometimes be missed.

Keywords: Abnormal uterine bleeding, endometrial biopsy, transvaginal sonography

   AOP305 Top

   Urogenital Pathology Top

Panoramic view of histopathology of nephrectomy specimens

Shatarupa Das, Ranu Tiwari Mishra, Noopur Gupta, Vijay Kumar Shrivastava, Sanjay Kumar Totade

Netaji Subhash Chandra Bose Medical College, Jabalpur, Madhya Pradesh, India

Introduction: Kidneys are affected by various disease processes resulting in permanent deterioration in functions causing significant morbidity and mortality. Nephrectomy is the standard surgical procedure performed in cases of suspected malignant renal disease and End Stage Renal Disease (ESRD). Objectives: To evaluate histopathological features of various lesions in Nephrectomy specimens and classify renal neoplasms as per “WHO Histological Classification”and study their age and gender distribution. Materials and Methods: A retrospective study done for a period of 1 year from January to December 2021 which includes 40 specimens received in our department. Patient particulars and radiological details were noted. According to CAP protocol, representative sections processed, H&E staining and reporting was done. Results: Among 40 specimens, the affected age range was 1-78 years having male:female ratio 1:1, majority in 30-50 years with 20 cases (50%). Non-neoplastic cases were 30 (75%) and neoplastic were 10 (25%), all malignant. Among malignant lesions, 3 cases each of Papillary RCC and Chromophobe RCC (60%), 2 cases (20%) clear cell RCC, 1 case of post NACT Wilm's tumour were reported. A rare case with hidden malignancy of “Urothelial Carcinoma with glandular differentiation” with Xanthogranulomatous Pyelonephritis was reported in a nephrectomy specimen of nephrolithiasis. Among non-neoplastic lesions, most common diagnosis was Chronic Pyelonephritis with nephrolithiasis, 23 cases (76.6%) followed by 2 cases each of Necrotising granulomatous pyelonephritis and Xanthogranulomatous pyelonephritis (13.3%). Out of 23 cases of chronic pyelonephritis, 1 case showed squamous metaplasia of calyx. Another rare case of post chemotherapy myeloid sarcoma also showed chronic pyelonephritis. Conclusion: Meticulous histopathological examination of surgical specimens is necessary for proper diagnosis and further treatment as it not only confirms clinical diagnosis but may also detect unsuspected associated malignant lesions.

Keywords: Chronic pyelonephritis, nephrectomy, renal cell carcinoma, urothelial carcinoma

   AOP306 Top

   Urogenital Pathology Top

Histomorphological spectrum of non-neoplastic lesions in nephrectomies

Sneha Ravindra, Aneel Myageri

SDM College of Medical Sciences and Hospital, Dharwad, Karnataka, India

Introduction: Nephrectomy is a common procedure indicated for variety of non-neoplastic lesions (chronic infections, obstruction, calculus disease or severe traumatic injury) and for neoplasms (benign/malignant tumours). Most common non-neoplastic lesions encountered are chronic pyelonephritis and reflux/obstructive uropathy. Objectives: 1. To study histopathological features and enlist the non-neoplastic lesions in nephrectomies at our institute. 2. To study demographic pattern. Materials and Methods: Present study is a 4-year cross-sectional study from January 2018 to December 2021 conducted at department of pathology. All microscopically confirmed non-neoplastic lesions in nephrectomies were included in the study. Case details were retrieved from Medical Records. Results: A total of 51 non-neoplastic nephrectomies were received during study period. Most patients belonged to 5th decade. Of 51 cases, 54.90% (n=28) were females and 45.098% (n=23) were males. Right kidney was seen involved more frequently. Flank pain was the most common clinical presentation. Most common non-neoplastic lesion encountered was chronic pyelonephritis (49.01%) and least common being interstitial nephritis (1.96%), renal infarction (1.96%) and renal mucormycosis (1.96%). Conclusion: Wide range of lesions are encountered in these nephrectomies. In immunosuppression, renal fungal infection leading to nephrectomy is not unlikely.

Keywords: Chronic pyelonephritis, nephrectomy, non-functioning kidney

   AOP307 Top

   Urogenital Pathology Top

Morphology and immunohistochemistry based renal tumors classification – A 4- year retrospective study

Shifa Pathan, Aneel Myageri

SDM College of Medical Sciences and Hospital, Dharwad, Karnataka, India

Introduction: Classification of renal tumors has undergone marked changes (WHO 2022) and requires genetic analysis, however morphologic and immunohistochemistry features aid in diagnosis and classification. Objectives: To classify renal tumors according to WHO 2022 classification based on morphology and IHC. Materials and Methods: Retrospective 4-year study of nephrectomy patients done for renal tumor are included. Demographical, clinical, histopathological features and IHC were studied and reallotted the diagnoses according to WHO 2022. Results: Total of 35 nephrectomies included. Most patients belonged to 5th-7th decade and with male preponderance. 30 cases were classified directly based on clinico-morphologic features and diagnosis remained the same. 4 cases required IHC and changed the previous diagnosis. One Papillary RCC were subcategorized as Eosinophilic SolidCystic variant of Papillary RCC (ESC-PRCC). One Carcinosarcoma was renamed as High Grade Carcinoma. One Eosinophilic granular variant of CCRCC was changed to Clear cell RCC. One Oncocytic Tubo-papillary RCC was changed to Type 1 Papillary RCC. One case of Papillary RCC required further genetic analysis as IHC features couldn't conclude. Conclusion: Judicious use of IHC will help in subcategorizing most renal tumors as per WHO 2022. Genetic analysis is indispensable.

Keywords: Nephrectomy, renal tumors

   AOP308 Top

   Urogenital Pathology Top

Immunohistochemical expression of B7-H3 and VEGF in invasive squamous cell carcinoma of uterine cervix

Monica Mondal, Preeti Diwaker, Vinod Kumar Arora, Sonal Sharma

University College of Medical Sciences and Guru Teg Bahadur Hospital, New Delhi, India

Introduction: B7-H3, also known as CD276, has a potential role in tumor angiogenesis. Recently, many therapeutic approaches targeting B7-H3, are undergoing evaluation and have shown impressive therapeutic efficacy. B7-H3 and VEGF are aberrantly overexpressed in many cancers and has been associated with a poor clinical outcome. However, there is paucity of literature documenting any association between them in invasive cervical squamous cell carcinomas (CSCCs). Hence, we intend to study the association of these two markers on angiogenesis in CSCC. Objectives: To evaluate and compare the immunohistochemical expression of B7-H3 and VEGF in invasive squamous cell carcinoma of uterine cervix. Materials and Methods: A retrospective study was conducted on 30 histopathologically diagnosed cases (resected specimens) of invasive CSCCs. IHC was performed using Human B7-H3 antibody and Anti-VEGF antibody. IHC slides were semi-quantitatively graded on the basis of intensity of staining (negative 0; weak: 1; moderate: 2; strong: 3) and percentage cell positivity. H-score for B7-H3 and VEGF was calculated. p-value < 0.05 was considered as statistically significant. Results: Statistically significant positive correlation was found between immunoexpression of B7-H3 and VEGF (p = 0.000; τ = .601) as well as between B7-H3 and VEGF intensity score 2 (p = 0.050) and score 3 (p = 0.001). Conclusion: The positive correlation between immunoexpression of B7-H3 and VEGF in CSCC cases highlights the proposed role of B7-H3 in angiogenesis. Hence, targeted therapy against B7-H3 can be utilized for treatment and better prognostic outcomes as well as for overcoming the problem of chemoresistance in cervical squamous cell carcinomas.

Keywords: B7-H3, cervical squamous cell carcinoma, VEGF

   AOP309 Top

   Urogenital Pathology Top

Grade 3 endometrial adenocarcinoma: A comparative assessment of clinicopathological and immunohistochemical features of endometrioid and serous carcinoma

Lavina Manu, Saloni Bansal, B Vishakha, Julian Crasta

St. Johns Medical College and Research Institute, Bengaluru, Karnataka, India

Introduction: High Grade endometrial carcinomas include a heterogenous group of disorders which tend to present in an advanced stage with histological overlap and requires confirmatory immunohistochemistry. Objectives: To study the clinicopathological and immunohistochemical features of FIGO grade 3 endometrial carcinomas with special emphasis to endometrioid and serous morphologies. Materials and Methods: Retrospective analysis of clinical and pathological features of FIGO grade 3 carcinomas from 2016-2022 by record review. Categorical variables are analysed using Chi square test/ Fischer's exact test. Results: Total of 40 cases, including serous (n=16), endometrioid (n=15), carcinosarcomas (n=6) and clear cell (n=3) were reviewed. Mean age at presentation was 61 years and 80% were multiparous with risk factors and presented with postmenopausal bleeding. Endometrioid tumors exhibited more frequent myometrial invasion of >50% (p =0.20) and lymphovascular invasion (p=0.60) whereas advanced stage with locoregional spread (p=0.05) and lymphnode involvement (p= 0.968) was more common in serous carcinomas. Among immunohistochemical markers, difference in staining with PR (p=0.002) and mutant pattern of p53 staining (p=0.001) was statistically significant though higher extent and intensity ER staining was more frequent with endometrioid histology. Mean DFS was better in serous carcinoma (31.5 vs. 25 months), whereas overall survival was better in endometrioid carcinoma (38 vs. 36.5 months). Conclusion: High grade endometrial carcinomas are aggressive tumours of postmenopausal women which exhibit higher invasive tendencies with poor survival. An immunohistochemical panel of PR, ER and p53 can differentiate between the endometrioid and serous histologies.

Keywords: Endometrioid, immunohistochemical, serous

   AOP310 Top

   Urogenital Pathology Top

Histopathological spectrum of lesions IB nephrectomy specimens in a tertiary care hospital

Shabnum Makandar, Sateesh Chavan, Purushotham Reddy

Karnataka Institute of Medical Sciences, Hubballi, Karnataka, India

Introduction: The standard surgical procedure in urological practice that is performed in end stage/nonfunctioning kidney or in suspected malignant neoplasm is nephrectomy. It is indicated in patients who present with a wide range of clinical conditions that include obstruction, symptomatic chronic infections, calculus disease as well as severe traumatic injury to renal cell carcinomas. Objectives: To study histomorphological features of renal lesions encountered in nephrectomy specimens. Materials and Methods: This is a retrospective (Jan 2011 to Dec 2020) and prospective (Jan 2021 to Aug 2022) unicentre study on total 88 cases of nephrectomy over a period of 11.5 years. Formalin fixed specimens were grossed and stained with routine H&E. Further detailed histopathological examination was done. and for the retrospective cases details are retrieved from the archives of the departmental records. Results: A total of 88 cases were studied. The age of patient ranged from 2 to 74 years. Highest percentage of nephrectomy was seen in 4th to 5th decade of life. Non-neoplastic lesions constituted 68 cases and chronic pyelonephritis was the most common lesion. Out of 20 neoplastic lesions, 76% of cases were malignant. Clear cell renal cell carcinoma was the most common malignant tumour. Conclusion: A wide range of lesions are encountered on histopathology of nephrectomy specimens, some of which are incidental finding in histopathology may be misdiagnosed clinically and radiologically; therefore, it is mandatory that every nephrectomy specimen be subjected to detailed histopathological examination. In renal cell carcinoma, nuclear grade and staging of tumors are necessary for therapy and prognosis.

Keywords: Chronic pyelonephritis, nephrectomy, renal cell carcinoma

   AOP311 Top

   Urogenital Pathology Top

Study of histopathological spectrum of ovarian tumors

S Mahalakshmi, S Arathi, Purushotham Reddy


Introduction: Among female genital tract-related malignancies, ovarian tumors are the fourth leading cause of mortality in women. The present study was conducted to identify the various histopathological spectrums of ovarian neoplasm (ON) according to the World Health Organization (WHO) classification 2020 in our center. Objectives: 1. To study the incidence of different types of ovarian tumors in our institute. 2. To study the histopathological features of various tumors of the ovary. 3. To classify ovarian tumors according to the new WHO classification. Materials and Methods: This cross-sectional study was conducted over a period of 5 years (August 2017 to July 2022) in the Department of Pathology, Karnataka Institute Of Medical Sciences, Hubli, Karnataka. The study included 416 cases of ovarian neoplasms (ONs). After proper fixation and thorough gross examination, sections were routinely processed and examined. The distribution of the various histomorphological spectrum of ovarian tumors was studied according to the WHO classification. Results: Out of the 416 cases studied, 267 (64.18%) were benign, 12 (2.88%) borderline, 36 (8.65%) malignant, and 1 (0.24%) metastatic tumour. Surface epithelial tumors (SETs) (327 (78.60%)) were the most common of all, followed by germ cell tumors (GCTs) (57 (13.7%)) and sex cord-stromal tumors (SSTs) (18 (4.32%)). The largest percentage (65%) of ONs was seen in 25-45 years of age group. Conclusion: The present study shows various histopathological features of ONs. Benign tumors are more common than malignant tumors. Among the benign tumors, serous tumors were most common of all. Thus, an accurate histological diagnosis is important to initiate a proper management plan.

Keywords: Germ cell, histopathology, ovarian tumors

   AOP312 Top

   Urogenital Pathology Top

Prognostic significance of tumor stroma ratio and stromal reactions in muscle invasive bladder cancer

Ishu Ghiloria, Varuna Mallya, Shramana Mandal, Nita Khurana, Anubhav Vindal

Maulana Azad Medical College, New Delhi, India

Introduction: The stroma surrounding the tumor plays a prime role in supporting & nourishing the tumor parenchyma. Tumor cells interactions with its microenvironment leads to tumor progression & spread. Prognostic role of tumor stroma ratio (TSR) & stromal reaction patterns remains unexplored in Muscle invasive bladder cancer (MIBC). Objectives: The aim was to study the relationship of TSR & stromal reaction patterns in MIBC with clinicopathological parameters. Materials and Methods: 28 Patients who underwent radical cystectomy for MIBC were included after ethical clearance. Their tumor stroma ratio was calculated & they were classified as TSR > 50% & <50%. Results: A higher stroma content (TSR<50%) was found to have positive correlation with lymphovascular invasion (p=0.003) & edema type reaction pattern (p=0.001). A positive correlation was also noted between the reaction patterns & LVI (p=0.002). Conclusion: A high stromal content was significantly associated with higher presence of LVI, as these tumors tend to invade lymphatics & result in spread & poor outcome. Also, edema type reaction pattern was observed in tumors with higher stroma content again highlighting the aggressiveness of the tumor. The above methods are simple, easily reproducible & cost effective, should form a part of the routine reporting checklist for MIBC, thus helping indetify high risk patients warranting additional therapy.

Keywords: Bladder cancer, stromal reactions, tumor stroma

   AOP313 Top

   Urogenital Pathology Top

Role of PDL-1 in serous carcinoma ovary and its association with tumor infiltrating lymphocytes (TILs)

Anubhuti Chaturvedi, Reena Tomar, Shramana Mandal, Deepti Goswami, M Bharanidharan

Maulana Azad Medical College and Lok Nayak Hospital, New Delhi, India

Introduction: Ovarian malignancies despite having lesser incidence, have a high death-to-incidence ratio, attributed to unavailability of effective screening tools, and absence of early symptoms, hence presentation at advanced stages, when the prognosis is poor. Programmed death ligand 1 (PDL-1) is a 40 kDa transmembrane protein that has been speculated to play a role in the tumor microenvironment, suppressing the adaptive response of the immune system. Tumor Infiltrating Lymphocytes (TILs), can provide important prognostic information and predict response to treatments. Prominent TIL is associated with high PDL-1 expression in various histological types of ovarian carcinomas, despite both being independent prognostic markers. Objectives: To evaluate the expression of PDL-1 in Serous Carcinoma Ovary by IHC, and study its association with TILs. Materials and Methods: 30 diagnosed cases of Serous Carcinoma Ovary were taken retrospectively from the Records of Pathology department. Clinico-histomorphological features were noted. The immunohistochemical expression of PDL-1 was assessed using PathnSitu Biotechnologies (USA) Rabbit Monoclonal Antibody, Clone- PB7H1, RTU. Grading was done on the percentage of positive cells, taking a cut-off of 1% for positivity. Results: 33% of the cases in this study expressed PDL-1. All the cases expressing PDL-1 showed the presence of TILs, found statistically significant, with a p-value of 0.02. Conclusion: Mapping the expression of PD-L1 in Serous Carcinoma of the Ovary is clinically relevant because, apart from its prognostic value, it will provide information about response to immunotherapy/targeted therapy.

Keywords: Programmed death ligand 1, serous carcinoma ovary, tumor infiltrating lymphocytes

   AOP314 Top

   Urogenital Pathology Top

Prognostic significance of immunohistochemical expression of Enhancer of Zeste Homolog 2 and Endothelin-I and its correlation with clinicopathological variables in carcinoma cervix

Aditi Priya, Jai Kumar Chaurasia, K Pushpalatha, Hemlata Panwar, Neelkamal Kapoor

AIIMS, Bhopal, Madhya Pradesh, India

Introduction: Carcinoma cervix is the fourth most common cancer worldwide and is one of the leading causes of cancer death in women. Recently, immunohistochemical (IHC) expression of biomarkers have been utilised as indicators of disease progression, aggressiveness and can prediction of prognosis in various cancers. Here, we studied the IHC expression of Enhancer of Zeste Homolog 2 (EZH2) and Endothelin-I (ET-1) in carcinoma cervix. EZH2 is a histone methyltransferase and catalyses methylation of histone H3 and plays important role in tumor cell proliferation, invasion and metastasis. Similarly, engagement of ETAR receptor by ET-1 triggers activation of tumor proliferation, angiogenesis and invasiveness. Objectives: To study the IHC expression of EZH2 and ET-I in cervical carcinomas and correlate it with clinico-pathological features. Materials and Methods: Immunohistochemical expression of EZH2 and ET-1 was studied in 50 paraffin-embedded tissue specimens of carcinoma cervix in AIIMS, Bhopal between between January 2018 –June 2022. Their correlation to the clinicpathologic features were analyzed. Results: High EZH2 expression was noted in 82% of cervical carcinoma cases with a significant relation with histologic type, tumor grade, FIGO stage and pTNM stage (p<0.001). Endothelin-1 overexpression was detected in 40% of the studied cases with a significant association with tumor size, histologic type, FIGO stage and pTNM stage (p<0.001). Conclusion: Hence, EZH2 and ET-1 may act as biomarkers of aggressive cervical cancer and could potentially be used in targeted therapy.

Keywords: Endothelin-I, Enhancer of Zeste Homolog 2, immunohistochemistry

   AOP315 Top

   Urogenital Pathology Top

P53 Expression in endometrial carcinoma: Clinico-pathological correlation

Garvita Singh, Nita Khurana, Asmita Rathore, Shramana Mandal

Maulana Azad Medical College, New Delhi, India

Introduction: Endometrial carcinoma is 6th most common carcinoma of female genital tract. Objectives: P53 expression on endometrial carcinoma is an area of interest for differentiating type 1 from type 2 endometrial carcinoma. Materials and Methods: The study included 20 cases of type I endometrial carcinoma and 5 cases of Type 2 endometrial carcinoma. P53 immunohistochemistry was seen as wild type, mutant type including both null type and overexpression type. Microscopically parameters noted and immunohistochemical staining was done using primary antibodies-mouse monoclonal anti-p53 antibody (Pathn Situ). Results: 25 cases of endometrial carcinoma were included in the present study. 20 cases of Type 1 endometrial carcinoma had strong immuno-expression of estrogen and progesterone receptor with wild type p53 expression, corroborating with the endometrioid histology. Low ER & PR expression was found in solid areas in grade 2 and 3 tumours; these cases lacked p16 expression which was seen to be expressed in five cases of type II carcinomas which in addition had markedly reduced or complete lack of ER and PR expression and showed mutant type p53 pattern. Conclusion: P53 expression was seen in 20 cases of endometriod carcinoma wild type followed by 3 cases showed overexpression and 2 cases showed null/mutant type of p53 expression. Along with p53 expression ER and PR expression was also observed in the present study and on the basis of histo-morphological and strong diffuse ER and PR positivity 20 cases were immunohistochemically classified as type 1 endometrial carcinoma and 5 cases which were either negative or had low weak ER and PR they were classified as type 2 endometrial carcinoma.

Keywords: Clinicopathological correlation, endometrial carcinoma, P53

   AOP316 Top

   Urogenital Pathology Top

Clinico-pathological study of recurrent endometrial carcinomas

Padmavathi Kamath, Gayatri Ravikumar, Kiran Kulkarni

St. Johns Medical College and Hospital, Bengaluru, Karnataka, India

Introduction: Recurrence in endometrial carcinomas (EC) pose major problem in management of these patients. The histopathological features of primary tumour in patients with recurrences are studied. Objectives: To study the detailed histopathological features in recurrent EC. Materials and Methods: A 10-year retrospective clinico-pathological study of recurrent EC received in Department of Pathology, St. John's medical college. Results: There were 15 cases of recurrent EC. Adjuvant chemotherapy with or without radiotherapy was given for 8 patients and radiotherapy for 2. Recurrence interval was 1-7 years. The common sites of recurrence was vaginal (6), liver (2), abdominal wall (2), 1 case each of omentum, urinary bladder, peritoneum, paravertebral. No events of death due to recurrence/disease. The histological types of primary was endometrioid (10), serous (2), mixed endometrioid- serous (1), clear cell carcinoma (1) and MMMT (1). There were FIGO grade 1 (4), grade 2 (3) and grade 3 (3) endometrioid carcinomas. Mucinous differentiation was seen (3). The pattern of infiltration (n=11) was diffuse (5), broad pushing front (2), adenoma malignum like (2) and limited superficial invasion (2) and no MELF pattern. The stromal reaction pattern (n=11) was desmoplastic (3), inflammatory (3) and no stromal reaction (5). All high grade EC (8) had recurrence within 2 years. Two case were stage III, others were Stage I. LVSI (5), LUS involvement (3), pelvic- paraaortic node (4) involvement was seen. The recurrent tumors showed similar morphology as primary. Conclusion: Conventional histological features may play minimal role in predicting recurrence. The study highlights the need for molecular methods to understand tumour biology and microenvironment.

Keywords: Invasion pattern, LVSI, recurrent endometrial carcinoma

   AOP317 Top

   Urogenital Pathology Top

Title: Renal CD68+ macrophages in IgA nephropathy: Association with Oxford classification

Divya Waghray, Gouri Kulkarni, S Suchitha

JSSAHER, Mysuru, Karnataka, India

Introduction: IgA nephropathy (IgAN) is the most common primary glomerulonephritis, an important cause of end-stage kidney disease. The Oxford classification, using evidence from around the world, resulted in the MESTC (mesangial hypercellularity, endocapillary hypercellularity, segmental sclerosis and interstitial fibrosis/tubular atrophy and crescents) criteria. Endocapillary hypercellularity are treatment-responsive lesions and is important to recognize them objectively. Objectives: To quantify the glomerular macrophages in IgA nephropathy using CD68+ immunostain, evaluate its association with the variables of Oxford classification and known clinical and histopathological prognostic factors. Materials and Methods: Sixty biopsy proven cases of IgAN were studied. Clinical and laboratory parameters, histological and CD68+ immunohistochemical data were recorded. The findings were analyzed and correlated. A p value of <0.05 was considered statistically significant. Results: E1 score showed a significance with all the test parameters, highest with CD68+ max (p=0.008). T value also showed statistical significance, highest with total CD68+ cells (p=0.001). The sensitivity and specificity for all the parameters for CD68+ max, total CD68+ cells and CD68+/Glom were 84.6% and 81%, 100% and 37%, and 76.9% and 57.4% respectively, when 7, 8.5 and 2.45 (number of macrophages) were used as cut offs respectively. Conclusion: Recognition of endocapillary hypercellularity (E score) plays a vital role in the management and prognosis of IgAN patients. In view of its clinical importance and poor reproducibility, there is a strong need to improve scoring methodology. CD68+ immunostain can be used as an effective marker for recognition of such lesions.

Keywords: CD68+ macrophages, endocapillary hypercellularity, IgA nephropathy

   AOP318 Top

   Urogenital Pathology Top

A study on adequacy of transurethral resection of prostate specimen sampling in accurately assessing pathological stage and Gleason score of incidentally detected prostatic carcinoma

Parvathy Ullas, Asila Puthalan, Renu Suresh Paul

VPS Lakeshore Hospital, Kochi, Kerala, India

Introduction: Incidental detection of prostate cancer in trans-urethral resection prostate (TURP) samples has increased. Carcinoma that is unsuspected clinically is referred to as stage T1 disease (subdivided into T1a and T1b). It is important for pathologist to accurately substage and grade these tumours since it mandates different patient care. Objectives: 1. To identify reasonable sampling approach to accurately evaluate TURP samples with incidentally detected prostate carcinoma without compromising the quality of the report. 2. To determine the cost implications for additional sampling. Materials and Methods: Thirty nine cases over a five year study period, in which incidental carcinoma was identified in the initial samples following College of American Pathologists (CAP) guidelines, and which were subsequently examined entirely formed the study group. A consensus opinion on Gleason score and tumor volume in the initial and additional samples were analysed. Cost analysis was done based on additional blocks used. Results: Out of 39 cases, 66.7% were stage T1a tumors (small volume tumours). A strong correlation between Gleason score and stage was found with majority (57.6%) of stage T1a tumours showing Gleason score 3+3, grade group 1. Tumor volume and Gleason score remained unchanged after complete sampling. An average loss of Rs 2592 per case was incurred after complete sampling. Conclusion: This study found that random sampling as per established CAP protocol is sufficient for accurate evaluation of incidentally detected prostatic carcinoma.

Keywords: Gleason score, prostatic carcinoma, trans-urethral resection prostate sampling

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   Urogenital Pathology Top

Histopathological spectrum of cervical biopsies in a Tertiary Care Center of Eastern Uttar Pradesh: A retrospective observational study

M Induparkavi, Ojas Gupta, Amrita G Kar, Neeraj Dhameja, Uma Pandey

Banaras Hindu University, Varanasi, Uttar Pradesh, India

Introduction: The major cervical lesions are due to infection, inflammation, and neoplastic conditions. Nonneoplastic lesions of the cervix are seen often in sexually active women. Inflammatory lesions include acute and chronic cervicitis and chronic granulomatous cervicitis. In India, cervical cancer is a significant health problem. Objectives: To study the demographic details. To study the incidence and prevalence of pre-malignant lesions. To study the incidence and prevalence of malignant lesions. Materials and Methods: Retrospective analysis of cervical biopsy samples submitted over 2 years from September 2020 to August 2022 were evaluated from our archival data. Results: A total of 668 cervical biopsy samples were received with a mean age of patients' 48.2 years, ranging from 18 years to 90 years. 40.4% (270) of the biopsies were non-neoplastic, with a predominance of chronic cervicitis (43.7%, 118), followed by benign endocervical polyp (31.8%, 86) out of total non-neoplastic entities. Pre-malignant lesions were 23.5% (77) out of all total neoplastic entities (398). Squamous cell carcinoma was the predominant malignant lesion (220, 32.9%) out of total cervical biopsies, and adenocarcinoma was 3.9% (9) out of total malignant biopsies. Conclusion: Chronic cervicitis and squamous cell carcinoma were the most common diagnosis among non-malignant and malignant cervical biopsies, respectively. Since the frequency of cervical cancer is high in our region, there is a need to have well-detailed national policies to be put in place to vaccinate younger females and increase the detection of preinvasive lesions to reduce the prevalence of cervical cancer.

Keywords: Biopsy, cervix, squamous cell carcinoma

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Comparing the diagnostic accuracy of combined serum biomarkers CA-125 and Apolipoprotein A1 over CA-125 alone in epithelial ovarian carcinoma

Vikas Kumar, Sachin Kolte, Saritha Shamsunder

Department of Pathology, VMMC and Safdarjung Hospital, New Delhi, India

Introduction: CA-125 is widely used classic biomarker for early diagnosis of epithelial ovarian carcinoma. Apart from that Apolipoprotein A1 is a newly studied serum biomarker that have anti-tumorigenic properties and appears to be decreased significantly in cases of epithelial ovarian carcinoma. Objectives: 1. To measure serum CA-125 & Apolipoprotein levels in epithelial ovarian tumor. 2. To compare serum CA-125 & Apolipoprotein levels in benign and malignant epithelial ovarian tumor. Materials and Methods: A total 60 cases were included in this cross-sectional study, done at Department of Pathology, VMMC and Safdarjung Hospital, New Delhi. Preoperative samples of patient which are diagnosed as adnexal mass on radiology were taken. Only patients diagnosed as epithelial ovarian tumor on Histopathology examination were taken for final evaluation. Results: Serum CA-125 levels were found to be significantly increased in malignant as compared to benign ovarian tumor. Serum Apolipoprotein A1 were found to be decreased in malignant as compared to benign ovarian tumor. Apolipoprotein A1 was found to augment the sensitivity & specificity, hence overall diagnostic accuracy of CA-125. Conclusion: All parameters consisting CA-125, Apolipoprotein A1 and CA-125 & ApoA1 combined significantly predicted HPE Impression: Malignant. CA-125 & ApoA1 combined comes out as best parameter in terms of diagnostic accuracy, Specificity & Positive Predictive Value (PPV).

Keywords: Apolipoprotein A1, CA-125, epithelial ovarian carcinoma

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CD117 immunoexpression in tumor cells and intratumoral and peritumoral mast cells in prostatic adenocarcinoma

Sandeep Kumar, Nadeem Tanveer, Sonal Sharma, Sanasam Jashmine Devi

University College of Medical Sciences, New Delhi, India

Introduction: Immunohistochemistry for CD117 has been studied as mast cell marker in several malignancies for its possible prognostic and therapeutic implications. Fewer studies on intratumoral and peritumoral mast cells density in prostatic adenocarcinoma have shown promising results. With the Gleason score modification in 2016, different Grade groups has been reassigned, hence, there is need to correlate the new Gleason score with the cells of tumor microenvironment. Objectives: To evaluate the immunoexpression of CD117 in tumor cells and intratumoral and peritumoral mast cells in prostatic adenocarcinoma cases. Materials and Methods: A retrospective study done on 40 prostatic adenocarcinoma cases, who had undergone TRUS- guided prostatic biopsy or radical prostatectomy in tertiary care hospital in North India. CD117 IHC on tumor cells and intratumoral and peritumoral mast cells was performed. The mast cells within and around the tumor were counted in three areas with highest mast cell density by scanning the entire section at low magnification and mean mast cells per HPF (×40) were calculated. Results: Intratumoral CD117 positive mast cells varied from 0-34 and mean was 8.03±8.40 while peritumoral CD117 positive mast cells (MCs) varied from 1-27 and mean was 7.80±6.28. Immunoexpression of CD117 in peritumoral MCs was found to be positively correlated with different grade groups of new Gleason scoring system (2016)(p= 0.02). However, CD117 positive intratumoral MCs density was not correlated with different grade groups of new Gleason scoring system (P=0.11). Conclusion: CD117 immunopositive mast cells can be used as a prognostic marker in prostatic adenocarcinoma cases.

Keywords: Intratumoral, mast cells, peritumoral

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Study of CD10 expression in prostatic adenocarcinoma

Spandan Nayak, K M Rao, S Agarwal, D P Mishra

Hitech Medical College and Hospital, Bhubaneswar, Odisha, India

Introduction: Adenocarcinoma of prostate is the second most common cause of cancer in men worldwide. Presently antibodies used in cocktail as triple stain are not 100% sensitive and specific. This calls for a new marker CD10, a transmembrane peptidase which plays an important role in pathogenesis of prostatic adenocarcinoma. Objectives: To study the expression of CD10 in prostatic adenocarcinoma and its correlation with histopathological grading (Gleason grading) alongwith serum PSA levels. Materials and Methods: This was an observational study conducted on 48 patients who were diagnosed as prostate cancer. Immunohistochemical profile was assessed for PSA and CD10 immunostaining. The intensity of CD10 expression and pattern of CD10 staining of tumor cells was evaluated. Results: The patients were in the age group of 45-95 years. As the grade group/Gleason score increased, the number of cases showing negative expression decreased and the pattern of expression changed from membranous to cytoplasmic to both types of expression. As the serum PSA levels increased, the intensity of expression changed from focally positive to diffusely positive. the pattern of expression also changed from membranous to cytoplasmic to both (membranous+cytoplasmic) types of expression with an increase in PSA levels. Conclusion: By immunohistochemical analysis we can identify CD10 positive tumors against which potential targeted therapies could be formulated.

Keywords: CD10, Gleason, prostate adenocarcinoma

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   Urogenital Pathology Top

Cancer stem cell markers in cervical cancer: Expression and effects on survival and radio-resistance

Chinmay Pendharkar, Surg Capt. Ritu Mehta, Col. P Sengupta, Air Cmde. Arijit Sen

Armed Forces Medical College, Pune, Maharashtra, India

Introduction: Cervical cancer is the second most common cancer of women in India. Multimodality treatment options are available. The cancer cells acquire radiation resistance by Cancer Stem Cells (CSCs). OCT4 is regulator in stem cell pluripotency and its expression is correlated with tumor recurrence and resistance to therapies. Sex-determining region Y-related high mobility group box (SOX2), is also the stem cell marker and is associated with tumor initiation and progression. Objectives: 1. To estimate the difference of expression of OCT4 and SOX2 histoscores in Cases of Cervical Cancer and Age-matched Controls 2. To estimate the association of OCT4 and SOX2 histoscores with Radio-resistance. Materials and Methods: Cases of cervical cancer (n = 31), and the age-matched controls were selected. Immunohistochemical staining for OCT4 and SOX2 was done. The Histoscores were calculated based on intensity of staining and percentage of cells showing positivity. The association of expression of OCT4 and SOX2 was tested. The association of histoscores was assessed with the FIGO stage, recurrence and radio-resistance. The survival analysis was performed to study association with the histoscores. Results: The OCT4 and SOX2 histoscores were significantly higher in cases than controls (p < 0.001). OCT4 and SOX4 histoscores did not show any association with the FIGO stage, recurrence, radio-resistance and survival. Conclusion: OCT4 and SOX2 expression has been previously described as the poor prognostic indicator, the present study failed to endorse their utility, which can be attributed to the small sample size. Larger sample size will be essential to ascertain their prognostic value.

Keywords: Cancer stem cells (CSC), OCT4, SOX2

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Histopathological spectrum of polypoidal lesion of cervix – A short case study

Milind Zade, Purwa R Patil, Sanjay R Bijwe

Grant Government Medical College, Mumbai, Maharashtra, India

Introduction: Cervical polyps are focal, hyperplastic protrusions of endocervical folds, including the epithelium and substansia propria. Endocervical polyp constitute the most common new growth of uterine cervix. Polypoidal lesion of cervix show various morphological spectrum ranging from inflammatory to neoplastic lesions. Objectives: To study histopathological spectrum of polypoidal lesion of cervix. Materials and Methods: Present study was conducted in the department of pathology in a tertiary care hospital over a period of one year. Women with polypoidal growth from the cervix visualized on per speculum, radiological examination were included in the study. Written informed consent was obtained. Patients were subjected to detail clinical, radiological examination. Specimens comprised of complete polyp or fragments of polyp were received in 10 % formalin. Specimens were processed, stained with H &amp; E stain. The sections were microscopically examined, findings were noted, data collected and analyzed. For diagnosis of malignant lesions, CAP protocol was applied. Results: Total 49 women were included in this study. Women were in the age group of 33 to 74 years. Endocervical polyp (22) was found to be most common lesion followed by leiomyoma (16), squamous metaplasia (03), inflammatory polyp (02), hyperplastic polyp (02), squamous cell carcinoma (02), focal dysplastic polyp (01), hemangioma (01), endometrial polyp (01), fibroepithelial polyp (01), flat condyloma (01), tuberculous inflammation (01), mixed mullerian tumor with epithelioid leiomyosarcomatous differentiation (01). Common associated clinical features were history of heavy menstrual bleeding or white discharge per vagina. Conclusion: Polypoidal lesion of cervix can be neoplastic or non-neoplastic in nature. It appears that polypoidal endocervicits, endocervical polyp, fibroepithelial polyp shows spectrum of similar pathogenesis with polypoidal endocervicits as initial lesion, endocervical polyp as intermediate lesion and fibroepithelial polyp as last stage in pathogenesis. Chances of malignancy presenting as cervical polyp is very low. Symptommatic polyp needs polypectomy. In asymptomatic cases, conservative management with follow up is expected.

Keywords: Cervix, heavy menstrual bleeding, leiomyoma

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Expression of programmed cell death ligand 1 (PD-L1) and mismatch repair status in squamous cell carcinomas of cervix

G Anju, Meenakshi Rao, Aasma Nalwa, Poonam Abhay Elhence

All India Institute of Medical Sciences, Jodhpur, Rajasthan, India

Introduction: Cervical cancer is the fourth most frequent cancer in women globally and the second most common cancer in India. (1) The co-regulatory molecule programmed death-ligand-1 (PD-L1), which is expressed on cancer cells and immune cells, reduces local tumor immunity, allowing tumor to metastasize. Mismatch repair deficiency has been shown to influence response to anti-PD-L1 therapy. Objectives: Expression of programmed cell death-ligand 1 (PD-L1) and MLH1, MSH2, PMS2 and MSH6 were assessed by immunohistochemistry in 50 cases of squamous cell carcinomas of the cervix. Materials and Methods: Expression of PD-L1 and mismatch repair status (MMR) was assessed by immunohistochemistry on 50 cases of SCCs of the cervix. Results: Mismatch repair deficiency was noted in 6% of cases. PD-L1 expression was noted in 80% and 84 % of tumor-infiltrating lymphocytes (TILs). A statistically significant association was noted between PD-L1 expression in the tumor and the grade of the tumor. All the deficient MMR SCCs of the cervix showed a CPS of <5 and one-third of the deficient MMR cases showed the absence of PD-L1 in the tumor. None of the nonkeratinizing SCCs had a CPS score of >50. Statistically, no significant association was noted between PD-L1, MMR with age, tumor grade, tumor necrosis, TILs. Conclusion: This study underscores the need for evaluation of the PD-L1 and mismatch repair status in malignancies as a routine practice for the management and prognostication.

Keywords: CPS, mismatch repair, programmed death-ligand-1

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Evaluation of apoptotic markers in the spectrum of morphological changes in endometrium in abnormal uterine bleeding

Nazia Walvir, Safia Rana, Zeeba S Jairajpuri, Sujata Jetley, Aruna Nigam

SKIMS, Srinagar, Jammu and Kashmir, India

Introduction: We aimed to histopathologically evaluate the morphological spectrum, apoptotic index (AI), and mitotic index (MI) of endometrial lesions in patients presenting with abnormal uterine bleeding (AUB). Objectives: 1. To estimate the apoptotic & mitotic index in the endometrial lesions. 2. To immunohistochemically assess the pattern of expression of the apoptotic markers (Bcl-2 and Bax) 3 To analyse the expression and balance between the apoptotic factors in the various endometrial pathologies. Materials and Methods: A cross-sectional study was done over a period of 18 months where a total of 60 newly diagnosed cases of perimenopausal women presenting with AUB were included. All H and E stained pathology slides from the specimens were reviewed for initial histopathological evaluation and diagnosis. Immunohistochemistry for Bcl-2 and Bax was done. The study subjects were divided into two groups: Group 1 included 30 cases of histologically proven proliferative endometrium (PE) and endometrial polyps and Group 2 included 30 cases of hyperplasia, endometrial epithelial neoplasia (EIN)/or carcinoma. For all cases, AI and MI were calculated and compared among the two groups. Results: Study included 60 patients of AUB, the mean age was 45.87 years. Most of the patients had heavy menstrual bleeding (66.67%). Endometrial carcinoma was diagnosed in 13 (21.67%) patients. There was a significant difference in the AI, MI, and ratio of AI/MI among various histopathological diagnoses. AI was highest for Endometrial carcinoma and lowest for hyperplasia. MI was highest for Endometrial carcinoma and lowest for hyperplasia. However, the ratio was incongruent as it was highest for hyperplasia without atypia and lowest for PE with endometrial carcinoma being midway. The Bcl-2 expression of relatively benign conditions (Group 1) was significantly higher than Group 2 (Endometrial epithelial neoplasia/carcinoma). Bax intensity showed an almost inverse pattern, being highest in endometrial carcinoma and lower in hyperplasia and polyps with the lowest expression in PE. Even the Bcl-2:Bax ratio was also highest for PE and lowest for endometrial carcinoma with others falling in between them. Conclusion: It can be concluded that the combination of the proliferative and apoptotic markers and the ratio will help as a tool in aiding the diagnosis of endometrial lesions for patients presenting with AUB.

Keywords: Apoptotic index, Bax-Bcl2, mitotic index

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Morphologic spectrum of prostate core needle biopsies with emphasis on Gleason score and Grade-A study from tertiary care cancer hospital in Southern India

Aparna Devi, G Champaka, Usha Amirtham, Akkamahadevi S Patil, M N Suma, Madhu

Kidwai Memorial Institute of Oncology, Bengaluru, Karnataka, India

Introduction: In India, prostate cancer incidence in elderly is 5 to 9/1 lakh population. Core needle biopsies aid in its definitive diagnosis. Objectives: 1. To study the clinicomorphologic spectrum of prostatic lesions detected in core needle biopsies and discuss the diagnostic challenges. 2. To assess the Gleason Score (GS) and Gleason Grade (GG) in acinar adenocarcinoma. Materials and Methods: This retrospective observational study included 119 cases consisting of 15 double sextant (DS), 68 bilateral (BLB) and 36 random biopsies (RB) from January 2019 to March 2022. Age, Prostate Specific Antigen (PSA) levels, percentage (%) of tumour, histological type, GS, and GG, perineural invasion were studied. Ancillary studies were performed in doubtful cases. Results: The spectrum included Acinar adenocarcinoma (72.2%), poorly differentiated malignancies (5.1%), small cell carcinoma (0.8%), non-neoplastic entities (17.7%) and normal biopsies (4.2%). Age range was 19-86 years. Most common GS was 4+5 and rate of detection of GS 3+3 was higher in DS biopsies (25%). Cases with high PSA levels had higher GS and GG in BLB (p= 0.0003 and 0.027 respectively), higher % of tumour in RB (p=0.048), and higher % of pattern 4 (p=0.022) in DS biopsies. Ancillary techniques aided in confirming and typing the malignancy in diagnostically challenging cases, such as prostate cancer mimics and poorly differentiated malignancies. Conclusion: Although, acinar adenocarcinoma is the dominant histology of prostate cancer, involvement of prostate by other malignancies should always be kept in the differential diagnosis. DS biopsies are superior for detection of acinar adenocarcinoma, especially with a GS 3+3.

Keywords: Double sextant, Gleason score, prostate biopsy

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   Urogenital Pathology Top

Expression of dendritic cells marker in microcystic, elongated and fragmented pattern of invasion in endometrial adenocarcinoma

Deepika Rana, Shramana Mandal, Varuna, Nita Khurana, Y M Mala

Maulana Azad Medical College, New Delhi, India

Introduction: Microcystic, elongated, and fragmented (MELF) glands are patterns of myometrial invasion associated with lymphovascular invasion, and lymph node metastasis and have a poor prognosis. Important components of the tumor microenvironment which play a central role in antitumor immunity are dendritic cells (DCs). DCS are antigen-presenting cells ability to activate T-lymphocytes to initiate immune responses. Thus, a large number of DCs in tumors suggest a favorable prognosis. Thus its role in the early detection of cases that may require additional radiotherapy for better prognosis. Objectives: Expression of Dendritic cells marker in the microcystic, elongated and fragmented pattern of invasion in Endometrial Adenocarcinoma. Materials and Methods: The retrospective study was conducted over 05 years. A total of 50 cases of Endometrioid carcinoma without adjuvant chemotherapy were included. Slides were reviewed for the presence of the MELF pattern of myometrial invasion and immunohistochemical staining was performed using the avidin-biotin complex method with antibodies to S-100 with DAB as the chromogen. Areas with positive dendritic cells will be counted under 400X and dendritic cells > 10 or greater per high power field taken as positive. Significance and Correlation between the MELF pattern and immunoexpression of S100 with clinicopathologic parameters. Results: All 50 cases were newly diagnosed and did not receive any therapy. The age ranged between 40 to 55 years and majority of patients were presented with bleeding per vagina. The tumor size ranged from 3 to 6 cm with negative lymph nodes. The MELF pattern had fewer number of S100 positive DC as compared to large number of DC in Non- MELF pattern. Conclusion: It is suggested that S-100 positive DC is inversely correlate with the MELF pattern and had a poor prognosis.

Keywords: Elongated, endometrial adenocarcinoma, fragmented, microcystic

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Clinicopathologic study of SET subtype of ovarian high-grade serous carcinoma and its correlation with immunoexpression of BRCA 1 and 2

Nimisha Dhankar, Gauri Kumari, Varuna Mallya, Shramana Mandal, Nita Khurana, Y M Mala

Maulana Azad Medical College, New Delhi, India

Introduction: Ovarian carcinomas with SET pattern have been classified under HGSCs in WHO 2020 classification. They harbor p53 mutation and express WT1. HGSC are frequently seen to have BRCA1 & BRCA2 loss, and they exhibit characteristic histopathologic features including solid, pseudoendometrioid/cribriform, transitional cell-like growth patterns (SET pattern), high mitotic index, nuclear pleomorphism & numerous TILs. HGSC-BRCA- have relatively better clinical outcome. Objectives: To investigate the clinicopathological characteristics and significance of solid, pseudoendometrioid and transitional (SET) ovarian high-grade serous carcinoma (HGSC) and to correlate the findings with BRCA 1 & 2 immunoexpression. Materials and Methods: 30 cases of HGSC (pre-chemotherapy) were reviewed for clinicopathological characteristics. They were assessed for SET pattern: solid pattern composed of diffuse sheets, pseudoendometrioid pattern with tumor in back to back nests with punched out microlumens and transitional comprising broad papillae with stratified layers of tumor cells. They were divided into three groups based on the amount of SET pattern: group A with SET<25% (HGSC classic). Group B: 25-59%, SET1 and group C: SET2 >50%. BRCA 1 and BRCA2 immunoexpression was also assessed, with nuclear expression in >10% cells considered positive. Results: Retrospectively analysis of 30 high grade serous carcinoma cases was done for SET pattern. Group A had 17/30 patients, group B had 9/30 patients and group C had 4/30 patients. Loss of BRCA1 and 2 expression was more commonly seen in patients with younger age, necrosis, increased TILs & SET morphology. Conclusion: HGSC with SET morphology more commonly had infiltrative borders, capsular breach, necrosis and loss of BRCA immunoexpression.

Keywords: BRCA1 & 2, high-grade serous carcinoma, SET pattern

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Spectrum of renal cell carcinoma-morphological features and select IHC findings – A one year data

Chitra Kalaivanan, Sandhya Sundram, J Natrajan1

Departments of Pathology and 1Urology, Sri Ramachandra Institute of Higher Education and Research, Chennai, Tamil Nadu, India

Introduction: The data from India is scarce and spectrum of a disease may differ across the globe. Hence this study was undertaken as an initiative to study the spectrum of morphological types of Renal Cell Carcinoma (RCC) in our institute, and its IHC pattern that aids in diagnosis and treatment. Objectives: 1. To study the spectrum of Renal Cell Carcinoma (RCC) over a period of 1 year. 2. To see the usefulness of IHC in subtyping RCC. Materials and Methods: This is a retrospective study which involved data analysis of RCC patients over a period of one year. Morphological types along with Immunohistochemistry (IHC) were also collected wherever available. Thus, the data were systematically analyzed and correlated with clinicopathological features. Results: A total of 52 Renal Cell Carcinoma (RCC) were considered for the study (Male - 38 and Female - 14). Most patients presented were in the fifth and seventh decades. The commonest histological type was Clear cell RCC (ccRCC) - 71.1%, followed by Papillary Type RCC - 13.4% and Chromophobe RCC - 5.7 %. The other rare morphological types were Leiomyomatous differentiation (1.9%), melanocytic differentiation (1.9%), RCC with papillary pattern (3.8%). 1 case of Rare Hybrid oncocytic tumor was also diagnosed. 60.4% of cases were Grade 2 types. The disease was confined in 67.4% of cases (pT1 and pT2), (26.6% - pT3) and (4.1% -.pT4). IHC was done in required cases for further categorization. Conclusion: Several subtypes of RCC are being increasingly reported and both morphological and IHC prove useful for categorization.

Keywords: Immunohistochemistry, morphology, renal cell carcinoma

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Clinicopathological spectrum of diabetic and non-diabetic renal lesions in patients with diabetes mellitus: An experience from tertiary care center

Netra Kori, Ranjana Ranade, Hephzibah Rani, Vidisha S Athanikar

SDMCMS and H, Dharwad, Karnataka, India

Introduction: Distinguishing diabetic and non -diabetic renal lesions in diabetic patients is essential on renal biopsies as their therapeutic and prognostic factors are different. We aimed to analyze the clinical, laboratory and renal biopsy features of diabetic and non-diabetic renal lesions in patients with Diabetes Mellitus. Objectives: 1. To estimate clinical and laboratory characteristics of Diabetic patients presenting with renal involvement. 2. To classify these patients as Diabetic renal disease and Non-diabetic ranal disease (NDRD) or an overlap based on renal biopsy findings. Materials and Methods: Renal biopsies from diabetic patients with renal involvement between January 2018 to 31st December 2021 were included. Light microscopy and Immunofluorescence findings were noted and cases were categorized into three groups namely Group-I (isolated NDRD); Group-II (NDRD superimposed on underlying DN); and Group-III (isolated DN) and further classified into categories as per Tervaert et al. Results: 133 cases were included in the study. Group I, II, III were represented by 9%, 43.6% and 46.6% respectively. The most common lesion in Group 1 being acute tubulointerstial nephritis (53%) and in group II had associated infection related glomerulonephritis (22% cases). Mean serum creatinine in group 1 and 3 respectively are 3.92 and 4.90. Proteinuria of 4+ was noted in 12.5% in group 1 and 30.95% cases of group 3. Diabetic retinopathy was present 45.16% in group 3. Conclusion: Shorter duration of disease, absence of retinopathy usually with nephrotic proteinuria strongly predict NDRD. Renal biopsy should be considered even in the absence of diabetic retinopathy to avoid delay in the diagnosis.

Keywords: Albuminuria, nodular glomerulosclerosis, retinopathy

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   Urogenital Pathology Top

Utility and applicability of direct immunofluorescence on formalin fixed paraffin embedded sections in diagnostic evaluation of renal biopsies

Tanya Sharma, Deepti Joshi, Erukkambattu Jayashankar, Mahendra Atlani, Girish Chandra Bhatt, Rajnish Joshi, Neelkamal Kapoor

Department of Pathology and Lab Medicine, All India Institute of Medical Sciences, Bhopal, Madhya Pradesh, India

Introduction: Direct Immunofluorescence on frozen tissue (DIF-F) is the gold standard technique, integral to diagnostic renal pathology. Immunofluorescence on paraffin embedded renal biopsies (IF-P) after enzyme treatment has been described in literature, however, has not found widespread use in renal pathology laboratories. In addition to cases which lack representative tissue on frozen sections, this technique also holds relevance in COVID-19 pandemic era, as the fresh tissue may be potentially infectious. Objectives: To analyse the utility of proteinase K digestion and DIF on paraffin embedded sections in diagnostic evaluation of medical renal diseases. Materials and Methods: A diagnostic accuracy study was conducted in the Department of Pathology and Lab Medicine, AIIMS Bhopal from March 2021 to August 2022. 30 consecutive renal biopsies received for diagnosis of medical renal disease were included in the study. A confirmed case of diffuse lupus nephritis was used for standardization of proteinase K enzymatic digestion with variation in timing of exposure at room temperature and stained with FITC-IgG. The FITC labelled antibodies (IgG, IgA, IgM, C3, C1q, kappa and lambda) were applied as per the light microscopic differential diagnoses and cases were graded independently in a blinded manner by two pathologists and compared to the grading of DIF-F results. Results: A high concordance was observed between the staining intensity using DIF-P as compared to DIF-F. Conclusion: Immunofluorescence on formalin fixed paraffin embedded tissue is a reliable salvage technique for renal diagnostic pathology, which can be useful in case of non-availability of representative fresh frozen tissue.

Keywords: Direct immunofluorescence, FFPE tissue, proteinase, renal biopsy

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   Urogenital Pathology Top

Pathological classification of endometrial carcinomas

R Nirosha, Indu R Nair, R Anupama, Prasanth S Ariyannur

Amrita Institute of Medical Sciences, Kochi, Kerala, India

Introduction: Integration of molecular features into the already existing pathological classification of ECs will offer significant prognostic information, chiefly for younger women struggling with tough choices regarding their reproductive healthiness. Objectives: The aim of the study was to classify ECs into 4 subtypes based on their molecular and IHC features and to find out association of each of these molecular subtypes with other pathological parameters-pathological subtype, grade, stage, LVE, MELF and node positivity. Materials and Methods: Prospective study done on 37 consecutive cases of fresh hysterectomy specimens (biopsy-proven) as ECs. 3-panel IHC p53, MSH6, PMS2, and DNAsequencing of selected regions of POLE gene was performed in each of 37 cases. ECs were subclassified into 4 subtypes, based on molecular and IHC features and association of each of these 4 subtypes with other pathological parameters were explored. Results: 37 cases studied. 29 patients-Type 1ECs and 8-Type 2ECs. 27 cases-low grade type and 10 cases-high grade type. 31 patients were low stage, 6 cases were high stage. Only 1 case (2.7%) showed mutation of POLE. 21 cases were assembled under CNL and 16 cases showed positivity for p53, MSI / POLE mutation. Type 2 EC subtype showed higher p53 positivity. Higher grade ECs showed more p53 positivity. All CNL cases were-Type 1 EC subtype and LG type. Conclusion: Our study confirms the type of carcinoma and grade correlates with p53 expression and CNL, p53 being associated with higher grade and type 2 ECs, whereas CNL is associated with low grade and type 1 ECs. There was only one case of POLE subtype identifiable in our study.

Keywords: Endometrial carcinoma, molecular classification, POLE mutation

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PDL1 expression in correlation with HPV status in cervical cancer: A one-year retrospective study

Kaumudi Konkay, C Padmavathi Devi, M Radhika, V Ann Mary

Department of Pathology, Siddhartha Medical College, Tumakuru, Karnataka, India

Introduction: Programmed cell death -ligand 1 (PDL-1) is an immune suppressive molecule expressed on tumour cells, antigen presenting cells and tumour infiltrating lymphocytes which regulates T- cell activation and belongs to B7-CD28 family. It binds to PD-1 on T cells and it helps in immune evasion by down regulating anti-tumour T cell activity. Immunomodulatory therapies using PD-1/PDL-1 inhibitors have been proven advantageous in malignant melanoma, advanced non-small cell lung carcinoma and urothelial carcinoma. Its utility in cervical carcinoma is still under research. The reported rate of expression of PDL-1 in cervical carcinoma ranges between 34.4% to 96%. Objectives: To enumerate the PDL-1 expression profile of cervical carcinoma patients in corelation with HPV status. Materials and Methods: A hospital based retrospective study of one year duration between January and December 2020 in Dept of pathology, GMC, Guntur after obtaining approval from institutional ethics committee. The specimens received in formalin, after routine processing, paraffin embedded tissue blocks prepared and sections cut were stained with Haematoxylin and eosin. The slides reviewed and appropriate areas devoid of necrosis and haemorrhage marked. Forty cases were selected and corresponding areas cores were taken and tissue microarray blocks were prepared and IHC with PDL1 done using a anti PDL1 rabbit monoclonal antibody, clone CAL10 using appropriate controls as per manufacturers guidelines. PDL-1 scoring system adopted by Reddy et al was adopted. The cells were scored as negative, low positive and positive based on intensity of positivity and percentage of positive cells. Statistical analysis: details entered in Microsoft excel sheet. Multivariate analysis done with Fisher exact test and p value of <0.05 considered significant. Results: PDL1 was positive (P) in 2 (Score 2b (P cells: ≥50%; Intensity: 2+ and/or 3 +), low positive (LP) in 14 (score 1a in 10 (P cells: <50%; Intensity: 1+), 1b in 4 (P cells: <50%; Intensity: 2+ and/or 3 +), and negative (N) (score 0) in 24 cases. Squamous cell carcinoma (SCC) were 36, of which PDL1 was P in 2, LP in 12, N in 22. Well differentiated SCC were 13, of which 5 were LP (score 1a – 4 (all p16 positive, HPV associated (HPVA)), 1b- 1, p16 N, HPV independent (HPVI)), 8 were N (7 HPVA, 1 HPVI). Moderately differentiated SCC were 23, of which 2 were P (score 2b), both HPVA, 7 were LP, all HPVA, 14 were N (13 HPVA, 1 HPVI). Adenocarcinoma case was one, LP (score 1a), HPVA. Adenosquamous carcinoma were 2, both N, HPVA. Small cell neuroendocrine carcinoma was one, N (1b), HPVA. Conclusion: The rate of PDL-1 expression in cervical carcinoma was 40% in this study. Strong positivity (2b) was associated with moderately differentiated SCC, HPV associated.

Keywords: Cervical carcinoma HPV associated, programmed cell death -ligand 1

   AOP336 Top

   Urogenital Pathology Top

Is C4d the magical marker in diagnosis of native renal diseases?

Varsha Kumar, Arun Yadav, Satyam, Vatsala Misra, Arvind Gupta, Sana Shadab

Department of Pathology, M.L.N. Medical College, Prayagraj, Uttar Pradesh, India

Introduction: C4d is a biomarker of the complement cascade and has a role in the of antibody mediated rejection in solid organ transplantation. It's role in disease progression has been studied recently in few native renal diseases. Objectives: The objective of the study was to evaluate the role of C4d in the diagnosis of native renal diseases. Materials and Methods: An observational cross-sectional study was conducted in the Department of Pathology from September 2020 to September 2021. A total of 93 renal biopsies of patients presenting with proteinuria/azotemia/isolated glomerular hematuria were included in the study. The semiquantitative intensity of Immunoglobulins and complements obtained on IF were correlated with the semiquantitative intensity of C4d by IHC. Statistical analysis was done by Pearson correlation coefficient to determine the P value. Results: Positive correlation was found between the IF deposits of immunoglobulins and C3 in the glomerulus with the C4d deposits demonstrated by immunohistochemistry, in minimal change disease and membranous nephropathy (R2= 1, 0.769). Weak positive correlation was obtained in case of MPGN and Lupus nephritis.(R2=0.33, 0.06). Conclusion: Pattern of glomerular C4d deposition can be a useful for diagnosing various diseases like MCD, MN, Lupus Nephritis, Membranoproliferative glomerulonephritis, IgA nephropathy. Given its diagnostic importance, staining for C4d might be incorporated into the routine analysis of renal biopsies.

Keywords: C4d, diagnosis, native renal diseases

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   Urogenital Pathology Top

Fetal growth in pregnancy with diabetes and its correlation with placental pathology

Shalini Bhalla, Akansha Sharma, Amita Pandey, Preeti Agarwal

Department of Pathology, King George's Medical University, Lucknow, Uttar Pradesh, India

Introduction: Diabetes in pregnant women is associated with increased incidence of macrosomia and fetal growth restriction. The possible cause in diabetic mothers' could be either maternal vasculopathy, placental dysfunction or overzealous glycemic control causing maternal hypoglycemia. Objectives: To assess fetal growth pattern in pregnancy complicated with diabetes and morphological and histopathological changes in the placenta. Materials and Methods: One year prospective Observational study on pregnant women who were known diabetics and those diagnosed with Gestational diabetes during the ante-natal visits. Fasting and post-prandial blood sugar estimation was done and glycemic control maintained for each trimester. After delivery placenta of both diabetic and non-diabetic women were sent in 10% formalin. Gross and Histopathological examination was done. Results: 110 women were registered of which 22 were lost to follow-up. Of the 88 women, 83 had gestational diabetes mellitus and 5 were pre-gestational diabetics. 69 women were less than 30 years of age. 22 gestational diabetics were diagnosed in first trimester, 46 in second and 15 in the third trimester. Majority of women with live birth (89.4%) had good glycemic control. Histopathological features of maternal vasculopathic changes and abnormal fetal placental circulation were significantly more in the placenta of diabetic women, however no significant association was seen with different extent of glycemic control. Incidence of Small for gestational age was dominant fetal growth pattern with 54.1% <10th percentile of birth weight. Conclusion: Placenta of diabetic women was larger and placental-fetal weight ratio was higher suggesting placental hypertrophy, however histopathological changes were not related to the fetal growth pattern.

Keywords: Fetal growth, gestational diabetes mellitus, placenta

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   Urogenital Pathology Top

A clinico-pathological study of epithelial tumours of ovary with emphasis on immunohistochemical expression of p53 and overexpression of Her2/neu in serous tumours of ovary

G V Neethu, Vardendra Kulkarni, Nisarga Rani

JJM Medical College, Davanagere, Karnataka, India

Introduction: The utility of biomarker in ovarian cancer show strong association between tumour marker and patient outcome. Varieties of biomarker like TP53, HER2/neu, PAX8, WT1, CDKN2A, ER, PR, EGFR have been evaluated for their utility in epithelial tumours of ovary. Various studies have shown association between p53 expression and grade of serous carcinoma of ovary. HER2/neu oncogene has been studied extensively for it's prognostic implications in ovarian cancer. Objectives: 1. To evaluate expression of p53 and overexpression of Her2/neu in serous borderline and serous carcinoma of ovary. 2. To correlate expression of p53 and overexpression of HER2/neu with grade of serous borderline and serous carcinoma of ovary. Materials and Methods: A total of 240 epithelial tumours of ovary diagnosed in department of Pathology J.J.M Medical College Davangere from June 2019 to May 2021 are included. Specimens sent in 10% formalin fixed, examined grossly and processed according to standard protocol. Immunohistochemical analysis for expression of p53 and overexpression of HER2/neu were performed on 30 cases of serous borderline tumours and serous carcinoma of ovary. Results: The following statistical tests performed for descriptive study -Percentage, Ratio and Proportion. Immuno-histochemical expression of p53 and HER2/neu in both group was compared using two tailed Fischers Exact test. Conclusion: In the present study of epithelial tumours of ovary, P53 expression showed higher association with high grade serous carcinoma followed by low grade serous carcinoma and borderline serous tumour. HER2/neu expression noted only in high grade serous carcinoma.

Keywords: HER2/neu study, p53 study, serous tumours

   AOP339 Top

   Urogenital Pathology Top

Adenomatoid tumor of the female genital tract: What the mind doesn't know, the eyes won't see

Neha Mittal, Santosh Menon, Bharat Rekhi, Kedar Deodhar

Tata Medical Center, Mumbai, India

Introduction: Adenomatoid tumor (AT) is a benign neoplasm of mesothelial origin, most commonly involving the epididymis. Its occurrence in the female genital tract (FGT) is an exceedingly rare and under-recognized phenomenon. Objectives: A retrospective observational study of cases of AT involving FGT diagnosed over 5 years (2018-2022). Materials and Methods: A total of 9 women, aged 39-65 years (mean: 52.6 years) were diagnosed. All but two (7/9) were incidental diagnoses on resected gynecological specimens (5 in myometrium and 3 in Fallopian tubes). Results: Two cases, clinically, mimicked malignancy; one each presenting with an inguinal and fallopian tube mass. The underlying histology warranting surgery were Endometrial endometrioid adenocarcinoma in 3 cases, and one case each of ovarian clear cell carcinoma, high grade serous carcinoma, mature cystic teratoma, endometriotic cyst, and one case of HPV-associated squamous carcinoma. On histology, microcystic angiomatoid histology was seen in 62.5% (5/8), microcystic trabecular in 25% (2/8), and one case each of oncocytoid (12.5%), and Leiomyoadenomatoid (12.5%) subtype. Salient histological findings include microcystic change in 8/9 (86.7%), epithelioid cells in 3/9 (33.3%), flattened cells in 5/9 (55.6%), oncocytoid cells in 1/9 (11.1%), signet ring-like cells in 3/9 (33.3%), fibrous stroma in 5/9 (55.6%), and absence of mitotic activity and necrosis in all (100%). On Immunohistochemistry, all cases were positive for keratins and Calretinin (nuclear and cytoplasmic), and negative for CD31. Conclusion: AT is rare, underdiagnosed, and misdiagnosed as it may mimic malignancy leading to inaccurate staging or management of gynecological tumors. A clonally driven origin due to TRAF mutations, and proposed morphological similarities with mesotheliomas necessitate accurate identification for larger multicentric studies.

Keywords: Adenomatoid tumor, female genital tract

   AOP340 Top

   Urogenital Pathology Top

Histomorphological spectrum of ovarian tumours in a tertiary care Hospital in North India

Anu Gupta, Subhash Bhardwaj

Government Medical College, Jammu and Kashmir, India

Introduction: Ovarian tumours are heterogenous group of neoplasms which account for one of the top five gynaecological malignancies in Indian women. The WHO classification of ovarian tumours is based on morphology with the recent molecular studies supporting the morphology based classification system. Molecular pathology has improved the knowledge of ovarian carcinomas identifying peculiar alterations for every histologic subtype. Ovarian neoplasms manifest at a late stage as the symptoms are delayed and non specific and even imaging modalities can be misleading and cytology has its own limitations, hence histopathology diagnosis remains the main stay. Objectives: To study histomorphological patterns of ovarian tumours and related molecular features essential for individualized clinical decision making. Materials and Methods: This is a two year retrospective study. A total of 82 cases of ovarian tumours were studied. Non neoplastic lesions were excluded from the study. Gross examination of the specimen along with clinical details were also taken into account for the final diagnosis given on H&E stained slides. All the ovarian tumours were classified based on WHO Classification. Results: Out of the total 82 ovarian tumours majority 64 (78.04%) were benign, 13 (15.85%) were malignant and 5 (6.09%) borderline. Age ranged from 13 to 70 years. Epithelial tumours were the commonest 52 (63.41%) followed by Germ cell tumours 21 (29.26%). Serouscystadenoma was the commonest ovarian tumour among the epithelial tumours and mature cystic teratoma in the germ cell tumours. In the malignant category High grade Serous carcinoma was commonest followed by Mucinous carcinoma. Conclusion: Ovarian tumours are silent killers, correct histomorphological diagnosis help gynaecologist in proper treatment and in institutions with limited provision of resources, histopathology remains the gold standard.

Keywords: Histopathology, ovarian, tumour

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   Urogenital Pathology Top

Comprehensive clinicopathogic study on a contemporary cohort of perivascular epithelioid cell tumor (PEComa) of the gynaecologic tract

Deepika Jain, Sayali Shinde, Samriti Arora, Mallika Dixit, Hena, Ekta Jain, Vipra Malik, Shivani Sharma, Juhi Varshney, Sambit K. Mohanty

Core Diagnostics, Gurugram, Haryana, India

Introduction: Perivascular epithelioid cell tumours (PEComa) of the gynecologic tract are rare. Molecularly these tumours are either TSC1/2-mutated and TFE3-rearranged. Recognition of the first group is imperative as these patients may benefit from targeted therapy with mTOR inhibitors. Objectives: Because of its rarity, we comprehend the overarching clinicopathologic features of a series of 11 patients with PEComas of the gynaecological tract. Materials and Methods: The clinical presentation, imaging, histopathology, immunohistochemistry (IHC) and management and follow-up data were analysed. Results: The patient's age ranged from 36 to 68 years (mean: 49.3 years). Stage tumours were located in the uterine corpus (6), cervix (2) and 1 each in the para-ovarian soft tissue and ovary. The tumors ranged from 3 mm to 16 cm in size. Epithelioid histology was present in 8 and spindled in 3 tumours. Nuclear atypia was mild in 2 and moderate to marked in 9 tumors. Mitoses ranged from 1 to 15 per 2.5 square millimetre. Seven and 4 tumours had necrosis and lymphovascular invasion. The tumours had a HMB45+(11)/S100+(5)/Melan A+(3)/SMA+(7)/Desmin+(2)/h-Caldesmon+(4)/panCK-.TFE3-/CD10- phenotype. Surgical resection was performed in all and none had adjuvant therapy. Six of seven patients with follow-up were alive and one died of extensive metastasis (follow-up: 1 months to 23 months). Application of gynaecologic-specific criteria, 4 tumours satisfied the criteria for malignancy. Conclusion: This is the largest series of PEComa of the gynaecologic tract from India. Four patients are categorised as malignant. None of our patients expressed TFE3 by IHC and they possibly fall into the TSC1/2-mutated PEComas. A multidisciplinary approach should be the key in treatment decision-making regarding gynaecological PEComas.

Keywords: Gynaecologic organs, PEComa, uterus

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   Urogenital Pathology Top

Evaluation of the prognostic significance of tumor-stroma ratio in radical prostatectomy specimens for prostatic adenocarcinoma

V Divya, M Archana, N Srivatsa, Rekha V Kumar

Sri Shankara Cancer Hospital and Research Centre, Bengaluru, Karnataka, India

Introduction: Prostate cancer (PCa) represents the second most- frequently diagnosed malignancy in men 1. Gleason grade group, pathological stage and surgical margins are independent predictors of recurrenceafter radical prostatectomy (RP) 2. In recent years, tumor to-stroma ratio (TSR) has been reported as a prognostic factor (3-6). Low TSR is associated with aggressive features in colon, lung and breast cancers 7, but play protective role in PCa.8 Despite its clinical potential, routine TSR scoring has not been implemented due to lack of standardisation and interobserver variability. There are very few studies on prognostic significance of TSR in PCa. Objectives: 1. To assess TSR in RP specimens for PCa. 2. To correlate TSR with known prognostic factors, Capra S score and time to biochemical recurrence (BR), if any. Materials and Methods: A retrospective study of histopathologically confirmed PCa who underwent RP at SSCHRC from January 2017 -August 2022 was conducted. Cases with inadequate clinical information were excluded. TSR was scored independently by two pathologists and statistically correlated with known prognostic factors, Capra S score and time to BR. Results: 145 patients who underwent RP were included. Multivariate analysis of prognosticfactors was done using Kendall Tau rank correlation and time to BR was plotted on Kaplan Meier' plot. TSRs ranged from 0.1 to 1.5 and were stratified to arrive at cutoff values that significantly correlate with CAPRA S score and time to BR. Conclusion: TSR quantification will help in identifying patients at high risk of progression.

Keywords: Microenvironment, prostate cancer, tumor-stroma ratio

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   Urogenital Pathology Top

Pathologic ultrastaging of sentinel lymph nodes in endometrial carcinoma for detection of micrometastases

K Shabnam, Indu R Nair, Anupama Rajanbabu, Viral Patel

Amrita Institute of Medical Sciences, Kochi, Kerala, India

Introduction: Lymph node status is an important prognostic factor and criterion for adjuvant therapy in endometrial cancers. Ultrastaging involves additional sectioning and staining of sentinel lymph node (SLN) with H&E and immunohistochemistry (IHC) to detect micrometastases. Objectives: To assess the value of ultrastaging of SLN of endometrial carcinoma and correlate it with other histopathological parameters and follow up. Materials and Methods: All patients who underwent staging surgery for endometrial carcinoma with negative mapped SLN from 2018 to 2021 were reviewed for ultrastaging. Protocol involved cutting an additional two 5-μm sections at each of two levels, 50-μm apart, from each block lacking metastasis on routine H&E. At each level, one slide was stained with H&E and with IHC using pan-cytokeratin (AE1:AE3). Results: Out of the 75 cases of endometrial carcinomas with negative SLN at routine H&E, ultrastaging detected an additional 6 cases (8%) with metastatic disease who would otherwise have been missed. Amongst these, 50 % were detected only in the second level and 50% only by IHC. These cases were of mostly type 1 (66%) with higher grade and showed lymphovascular invasion (66%) and outer myometrial invasion (83%) in comparison with the other 69 cases. Follow up period ranged from 6 mths to 4 yrs with relatively good prognosis. Conclusion: Pathological ultrastaging of SLN in endometrial carcinoma helps detect micrometastases which may be missed in usual evaluation. This study highlights the need for incorporation of ultrastaging in routine practice primarily for cases with higher grade, outer myometrial invasion and lymphovascular invasion for adequate treatment.

Keywords: Endometrial carcinoma, sentinel lymph node, ultrastaging

Evaluation of MUC1 as a prognostic marker in carcinoma breast: A swimmer's fleet

Shashwat Verma, G D Katyar, Mithila Bisht, Cheene Garg, Vibhuti Goal, Nitesh Mohan

Rohilkhand Medical College, Bareilly, Uttar Pradesh, India

Background: Immunohistochemical evaluation and classification of breast cancer into the fundamental molecular subtypes provide a far better understanding the biology of the disease as well as in individualised treatment. Some of the traditional marker such as estrogen, progesterone and HER-2 receptors have been typically evaluated. MUC1 a transmembrane mucin shown to have potential as a metastatic and prognostic marker in breast cancer. There has not been many research publications on the same in Indian population. Aim: The goal of the current study was to evaluate the importance of the MUC1 as a potential target marker in breast carcinoma. Methods: For microscopy 100 H&E slides of breast cancer patients were prepared followed by tissue microarray (TMA) and immunohistochemistry using ER, PR, HER2, Ki-67, and MUC1 were utilised. Results: 73% of breast carcinoma shows MUC1. MUC1 positive cases were predominately of higher Nottingham grades, i.e. Grade 2 and Grade 3, 80 and 76 % respectively and showed high significant association with MUC1 with p value of 0.0. Also, there was significant association of MUC1 expression with Luminal B with p value of 0.01, but mainly cases are of MUC1 positivity fall in HER2 enriched (88.2%) and triple negative (86.3%). Conclusion: MUC1 play a significant role in the biology of breast cancer and it was expressed in maximum number of cases and associated with high histological grade with significant association. MUC1 was mainly expressed in HER2 Enriched and Triple Negative Breast carcinomas, most aggressive molecular classes with poor prognosis.

Keywords: Breast carcinoma, immunohistochemistry, MUC1

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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0377-4929.362056

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