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  Table of Contents    
Year : 2019  |  Volume : 62  |  Issue : 5  |  Page : 22-42
Oral Presentations

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Date of Web Publication13-Dec-2019

How to cite this article:
. Oral Presentations. Indian J Pathol Microbiol 2019;62, Suppl S1:22-42

How to cite this URL:
. Oral Presentations. Indian J Pathol Microbiol [serial online] 2019 [cited 2022 Aug 16];62, Suppl S1:22-42. Available from: https://www.ijpmonline.org/text.asp?2019/62/5/22/272761

   OP1: Multisystemic pediatric Langerhans cell histiocytosis: A comprehensive clinico pathological and BRAF V600E mutation study at autopsy Top

Gargi Kapatia, Prateek Bhatia, Minu Singh1, Richa Jain1, Deepak Bansal1, Kirti Gupta

Departments of Pathology and1Pediatrics, Post Graduate Institute of Medical Education and Research, Chandigarh, India. E-mail: [email protected]

Introduction: Langerhans cell histiocytosis (LCH), a disorder of antigen- presenting cells, is the commonest disorder of the mononuclear phagocytic system. Diagnosis is always challenging due to heterogeneous clinical presentation. However, with the evolution and better understanding of its biology, many of these children are being diagnosed early and offered appropriate therapy. Despite these advances, in developing countries, an early diagnosis is still challenging due to resource constraints for specialized tests. As a result, many patients succumb to their disease. Autopsy data on LCH is notably lacking in the literature. Aim: We sought to analyze the clinical (including mutational) and morphologic features at autopsy in six proven cases of LCH. Methods: Study includes a detailed clinico-pathological and mutational analysis of 6 proven cases of LCH. Presence of BRAF V600E mutation was assessed by both Real Time PCR and Sanger sequencing. Results: A varied spectrum of organ involvement was noted with some rare and novel morphological findings, like nodular bronchiolocentric infiltration of LCH cells, lymphovascular emboli of LCH cells, and paucity of eosinophils within the infiltrate; these features have not been described earlier. Surprisingly, all cases were negative for BRAF V600E mutation on both RQ-PCR and Sanger sequencing. Conclusions: The present study is perhaps the first autopsy series on LCH. This extensive autopsy analysis represents a correlation of pathological features with clinical symptoms which provides clues for a timely diagnosis and appropriate therapeutic intervention. Also, our findings hint at the low frequency of BRAF V600E mutation in our LCH patients.

   OP2: Functional autopsy: A new paradigm Top

Bal Chander

Dr. Rajendra Prasad Government Medical College, Kangra, Himachal Pradesh, India. E-mail: [email protected]

Background: In autopsy 'normal/abnormal' is decided by gross and microscopic examination further aided by special stains which generally yields if not qualitative then semiquantitative data. Quantitative functionality of organs is never assessed and as a consequence the organs that might be morphologically 'normal' but functionally compromised do not come to attention. With the underlying assumption that normal morphology implies normal functionality we end up ignoring complexity of molecular dynamics at the time of death. In this preliminary study we decided to explore the feasibility of measuring one of the principal molecules as a surogate for functionality of the organ. Design: 20 adrenals were harvested from intrauterine death cases fetuses. All the organs were weiged and homogenized in triple distilled water. The supernatent was subjected to ELISA for Aldosterone. Results: There was wide variation in Aldosterone concentration per unit weight of organ with no linear correlation between concentration and gestational age or weight. Aldosterone was detected even in second trimester fetuses. Conclusion: Measurment of Aldosterone concentration in Adrenal glands demonstrated that morphologically 'normal' organs displays hitherto unexamined complexity and exposed significant limitations of conventional post mortem examination. Furthermore detection of Aldosterone even in second trimester fetuses which is contrary to conventional wisdom shows the importance of proper context and difference between results obtained via controlled and natural conditions.

   OP3: A study of histomorphological, immunohistochemical and molecular analysis of Ewing sarcoma and prognostic impact of p53 and ki67 Top

Kaniyappan Nambiyar, Nandita Kakkar, Bishan Dass Radotra, Rakesh Kapoor1, Deepak Bansal2, Naveen Kalra3

Departments of Histopathology and1Radiotherapy, Postgraduate Institute of Medical Education and Research,2Department of Pediatrics, Hemato-Oncology Division, Postgraduate Institute of Medical Education and Research,3Department of Radiodiagnosis and Imaging, Postgraduate Institute of Medical Education and Research, Chandigarh, India. E-mail: [email protected]

Background: Ewing sarcoma family of tumor (EWSFT) is a molecularly defined, morphologically heterogenous small-blue-round-cell neoplasia. The present study intends to study the EWSFTs with the help of histomorphological, immunohistochemical and molecular analyses and, further evaluates the utility of p53 and ki67 immunostains as prognostic markers. Design: A total of 32 cases of EWSFTs diagnosed between 2016 and 2017 were included in the study and were followed up till December 2018. EWSR1 translocation were studied by FISH technique using EWSR1-FLI11 Tri-colour Fusion/Translocation Probe Kit (Cytotest). Factors like tumor size, site, morphology, chemotherapy response, metastasis, localization of disease, and survival were assessed. In addition, p53 and ki67 immunostains were analysed using ImageJ software, and percentage nuclear positivity was correlated with the above-mentioned factors. Results and Discussion: The mean age of presentation in our study was found to be 24.3 years. Apart from classic morphology, variants like large cell, vascular, sclerosing, PNET and mixed type were found. CD99 was positive in all the cases. EWSR break apart was noted in 30 cases. FLI1 as a fusion partner was noted in 23 cases. The mean overall and disease-free survival were 24.5 and 9.7 months, respectively. Positive p53 expression showed significant association with development of metastasis later and outcome. Ki67 showed significant association with overall metastasis. Conclusion: Positive p53 and higher Ki67 seem to be related to poorer prognosis. Establishing p53 and ki67 as prognostic marker in EWSFTs needs further analysis including larger number of cases and longer follow up.

   OP4: Selecting the right immunohistochemical panel for diagnosis of synovial sarcoma Top

Manoj Gopal Madakshira, Uma Nahar Saikia, Bishan Dass Radotra, Lileswar Kaman

Post Graduate Institute of Medical Education and Research, Chandigarh, India. E-mail: [email protected]

Background: Synovial Sarcoma (SS) is a rare soft tissue sarcoma. We investigated the expression of TLE1, β-catenin, Calponin, SYT, INI1, CK7, CK19 and Claudin1 in SS. Design: 75 cases were reviewed. Initial IHC panel included positivity for EMA and/or PanCK, BCL2 and CD99 with absence of CD34. Cases which did not meet criteria were subjected to FISH (SS18 probe). TMA was constructed including selected cases, differential diagnosis, and controls. IHC was carried out using antibodies against TLE1, β-Catenin, INI1, CK7, CK19, SYT, Calponin, and Claudin1; and their expression was scored. Results and Discussion: 40 cases satisfied the inclusion criteria. TLE1 was seen to be a robust diagnostic marker, by virtue of its nuclear staining pattern. Cytoplasmic accumulation and nuclear translocation of β-catenin were seen in select cases. Calponin showed consistent cytoplasmic staining including poorly differentiated SS. SYT is a promising maker, exhibiting a nuclear staining pattern. INI1 showed consistent partial to complete loss of nuclear staining pattern. CK7 proved to be better in comparison with CK19. Claudin1 faithfully delineated the epithelial component. Conclusion: An initial panel CK7, CD99, BCL2, CD34, TLE1/SYT is recommended for diagnosis of SS. INI1 is useful to distinguish from tumors having a non-epithelioid/rhabdoid morphology. Calponin is useful to exclude poorly differentiated SS from its mimics. However, in equivocal cases confirmation by FISH or RT-PCR still remains the gold standard.

   OP5: Histopathological spectrum of vascular anomalies with special reference to immunohistochemistry Top

Swathi Chilukuri, Uma Pallivela, Vahini Gudeli, Asha Thota

Department of Pathology, ASRAM Medical College, Eluru, Andhra Pradesh, India. E-mail: [email protected]

Introduction: Vascular anomalies comprise a diverse group of tumors and malformations. Despite of the differences in evolution, clinical course, morphology and treatment “hemangioma” commonly referred for vascular anomalies. Multidisciplinary approach along with histopathological confirmation and definitive diagnosis by immuno profiling is necessary for the differentiation and diagnosis. Objectives: The aim of the study was to classify the vascular anomalies according to revised classification of ISSVA – 2018 and for definitive diagnosis of infantile hemangiomas (IH) with immunohisto- chemistry by Glucose transporter protein isoform 1 (GULT1). Materials and Methods: This is a retrospective study comprising of 47 cases which includes vascular tumors (n=22) and malformations (n=25). Complete clinico-demographic details was collected from Jan 2017 to June 2019. Hemangiomas and vascular malformations were distinguished by Verhoff van-Gieson stain wherever necessary and further confirmed by GLUTI. Results: Vascular malformations were common in age group of 0-20 years comprising 53% compared to hemangiomas 47% and predominantly above 40 years age. The common site is being Head & Neck region in both groups. All the infantile hemangiomas cases were positive for GLUT1 marker. Conclusion: Consistency in terminology of vascular lesions is necessary for the proper diagnosis and management. Verhoff van-Gieson stain helps in confirming AVM's. GLUT1 is an important diagnostic marker for the definitive diagnosis of infantile hemangiomas.

   OP6: A comparative study of a novel marker, EZH2, in different immunophenotypes of carcinoma breast and its overexpression in triple negative Top

Sukanya Ghosh

R. G. Kar Medical College, Kolkata, West Bengal, India. E-mail: [email protected]

Bakground: Enhancer of zeste homolog 2(EZH2), is a histone methyl transferase and works cooperatively with histone deacetylases (HDACs) in the same protein complex to mediate gene transcription repression by increasing histone H3Lyzs 27 methylation (H3K27me3). EZH2 regulates two important processes for metastasis: epithelial to mesenchymal transition and migration; increasing breast cancer stem cell numbers. EZH2 is overexpressed in numerous cancers, including triple negative breast cancer. Thus inhibition of EZH2 may be harnessed for targeted therapy for this disease. The aims and objectives are:

  1. To study the magnitude of EZH2 expression in breast carcinoma
  2. To compare EZH2 expression in different immunophenotyes of breast carcinoma
  3. To study the overexpression of EZH2 in triple negative breast carcinoma.

Design: A cross sectional study was done on 41 cases with epithelial breast tumour, in collaboration with Department of Pathology and General Surgery over a period of one year. Results and Discussion: Inclusion criteria consisted of estrogen, progesterone and her2neu receptor and Ki 67 expression in them. Immunohistochemistry for EZH2 in formalin preserved paraffin embedded tissue specimen was done and characterization of expression patterns of EZH2 transcripts were done. ER positive breast cancers with low proliferative index (Ki67< 15%) showed lowest expression of EZH2 and triple negative breast carcinomas showed highest expression of EZH2. Conclusion: Our study shows a strong association between EZH2 expression and triple negative breast carcinoma. Extensive studies need to be done in deciphering the role of EZH2 in the prognostication of breast carcinomas.

   OP7: Expression of programmed death ligand-1 in carcinoma breast and its correlation with prognostic parameters Top

Pallavi Punhani, Charanjeet Ahluwalia, Rashmi Arora

Vardhaman Mahavir Medical College and Safdarjung Hospital, New Delhi, India. E-mail: [email protected]

Background: PD-L1 (programmed death ligand 1), is a T cell inhibitory molecule that leads to T cell anergy and/or apoptosis. PD-L1 association with prognosis of various cancers has been a research topic of considerable interest. Several studies have been conducted to evaluate expression of PD-L1 in breast cancer. However, there is lack of literature for such studies in Indian patients. We aim to study the expression of PD-L1 in carcinoma breast and find out its correlation with prognostic parameters. Design: A cross sectional study to evaluate expression of PD-L1 in carcinoma breast and its correlation with prognostic parameters such as histological grade and surrogate molecular classification.15 cases (n=15) of Invasive Carcinoma Breast were taken and every case was evaluated for histological type and grade. Surrogate Molecular Classification using immunohistochemistry for ER, PR, Her-2 neu and Ki-67 was done for all the cases along with PD-L1. Results: All 15 cases were diagnosed to be Infiltrating Ductal Carcinoma, NOS type.

IHC performed in all 15 cases was found to be negative for PD-L1. A positive control for PD-L1 was done on tonsil which showed membranous positivity in the epithelial crypt lining. Conclusion: PD-L1 expression was negative in all 15 cases having different grades and different receptor status. PD-L1 expression may not be associated with a specific grade or hormonal status in breast cancer cases.

   OP8: MCM2 expression in triple negative breast cancers and its relationship to histologic grade, stage and Ki-67 expression Top

Meghadipa Mandal

PGT in R.G. Kar Medical College and Hospital, The West Bengal University of Health Sciences, Kolkata, West Bengal, India. E-mail: [email protected]

Background: Triple negative breast cancers being unresponsive to conventional therapy, identification of molecular pathways associated with increased aggressiveness is important. Both Ki-67 and MCM2 are cell proliferation markers. We studied the relationship between MCM2 expression & Ki-67 expression, histologic grade and stage of tumor. Design: The study included a cohort of 17 breast cancer patients. FFPE tumor tissue samples were collected after mastectomy. BR Grading and TNM staging done. IHC assay done on representative blocks for ER, PR, HER2, Ki-67 & MCM2. 12 blocks were triple negative, 1 each being Luminal A, Luminal B & HER 2 enriched. The slides were assessed by light microscopy and analysis done by SPSS-25. Results and Discussion:

  1. The median age of study population is 51 yrs.
  2. 59% cases are BR Grade 3 & 70% cases are Locally Advanced Breast Carcinomas (LABC).
  3. The median expression of Ki-67 is 20% & MCM2 is 70%, taken as cut-off.
  4. MCM2 expression is higher (≥70%) in Grade 3 (80% versus 43% in Grade 2).
  5. 58% of LABCs have high MCM2 expression.
  6. No difference in MCM2 expression with nodal staging was seen.
  7. The expression of Ki-67 (92%) & MCM2 (75%) was high in triple negative cases. They have positive correlation (rho= 0.364).

Conclusion: Median expression of MCM2 was higher than Ki-67 in triple negative cases. Advanced disease is associated with higher MCM2 expression. MCM2 may be an attractive alternative to Ki-67. Small sample size is the main limitation.

   OP9: Comparative analysis of mucins in infiltrating micropapillary carcinoma and different types of mucinous carcinomas of breast Top

Arshi Beg, Tanuja Shet1

Departments of Oncopathology and1Pathology, Tata Memorial Hospital, Mumbai, Maharashtra, India. E-mail: [email protected]

Background: 86 % of mucinous carcinoma in our population have a micropapillary pattern with propensity to produce IMPC recurrences with reduction in survival rate. Design: 59 mucinous carcinomas were divided into a) MUMPC( n=30) :- mucinous variants of micropapillary carcinoma b) MU,MPC (n=16) :- Mucinous carcinoma of micropapillary type mixed with IMPC (c) SVPCMUMPC (n=9):- Mixed tumors composed of MUMPC and solid variant of papillary carcinoma with mucinand (d) MUS(n= 4):- mucinous carcinomas with signet ring cells. MUC1, MUC1core, MUC2, MUC3, MUC4, MUC5AC, MUC6 were studied by immunohistochemistry (IHC) in all the cases and compared with 21 (IMPC). Results: The subtype of mucinous carcinoma significantly affected disease free survival (p value =0.007) but not the nodal status or overall survival. MUMPC:- All tumors were positive for MUC2. A cytoplasmic staining pattern with MUC1/MUC1core was significantly associated with a negative nodal status (p value=0.032). MU, MPC:- The IMPC component in these tumors showed MUC2 and MUC3 in 72% and MUC6 in 36% cases. Absence of MUC6 was associated with recurrence and a negative hormone receptor status. SVPC/MUMPC: - All tumors showed low expression of MUC1. MUC2 was seen in all tumors. MUC3 was associated with a negative nodal status. IMPC:- MUC3, MUC 2 and MUC6 was seen in 71%, 9% and 19%. Presence of MUC6 was associated with lack of recurrence. Conclusion: Mucin IHC reflects and affirms the histological subtypes of mucinous carcinoma. For the first time we have documented that the angioinvasive IMPC also express MUC2 and MUC6 and this may affect prognosis.

   OP10: Prognostic significance of serum vascular endothelial growth factor and hypoxia-inducible factor - 1 alpha immunoexpression in breast cancer Top

Sonam Sharma, Saumil Garg

Department of Pathology, Kalpana Chawla Government Medical College, Karnal, Haryana, India. E-mail: [email protected]

Background: Understanding the development and progression of breast cancer is still critical in the quest for successful therapeutic intervention. During tumorigenesis, under the influence of intratumoral hypoxia there is activation of a hypoxia-dependent gene expression program, which in turn upregulates hypoxia-inducible factors (HIF) and induce the expression of multiple genes which are involved in angiogenesis, thereby activating transcription of various potent proangiogenic cytokines like vascular endothelial growth factor (VEGF) which further causes tumor evolvement. The present study was conducted on breast cancer patients with an aim to (1) estimate their pre-operative serum VEGF (sVEGF) levels (2) determine the HIF-1alpha immunoexpression in post-operative specimens (3) find correlation between them and with other clinicopathological parameters. Design: A total of 50 female breast cancer cases proved on fine needle aspiration cytology were considered for the study. Their serum samples were subjected to ELISA for VEGF level estimation. Histopathological typing and grading was done for the resected specimens followed by immunohistochemistry for HIF-1alpha. Results: Increased sVEGF levels and HIF-1alpha expression was seen in 42% and 82% of the cases respectively. 19/21 cases showing increased levels of sVEGF, also exhibited high HIF-1alpha expression. A statistically significant correlation was observed between the increased sVEGF levels and lymph node metastasis as well as lymphovascular invasion. Conclusion: Both the pre-operative sVEGF levels and post-operative HIF-1alpha immunoexpression of the breast cancer patients can be useful for predicting the prognosis. Nevertheless, these tumor markers can also act as novel therapeutic targets in future for the better management of such cases.

   OP11: Expression of PD-L1 and Ki-67 proliferation index in invasive breast cancer and correlation with TILs and clinico-pathological characteristics Top

Rashim Sharma, Meenakshi Rao, Sudeep Khera, Ramkaran Chaudhary, Jeewan Ram Vishnoi, Puneet Pareek, Poonam Abhay Elhence

All India Institute of Medical Sciences, Jodhpur, Rajasthan, India. E-mail: [email protected]

Background: Breast cancer is most common cancer accounting for approximately one fourth of all cancers. Breast cancer ranking is number one amongst all Indian females with age adjusted rate being as high as 25.8 per 100,000 women. This study is done to ascertain and correlate the PD-L1 (Programmed Death Ligand-1) expression along with Ki-67 labelling index and TILs (Tumour Infiltrating Lymphocytes). Design: Study type: Descriptive, observational, comprising of 114 cases from 2016-2018. All patients diagnosed with invasive breast carcinoma with available hormone receptor status were included. Core biopsy, wedge biopsy, lumpectomy and MRM specimens were examined. Results and Discussion: Out of 114 cases, 86 cases received no chemotherapy. 75 cases (87%) showed high Ki-67 labelling index. Of these 33.33% cases showed positive PD-L1 expression in tumour and 45% in sTILs. Out of 86 cases 25% concordance was noted in PD-L1 expression in both tumour and sTILs. 28 cases were post chemotherapy, out of which 21(75%) cases showed high Ki-67 index. Out of these, 38.09% cases showed high PD-L1 expression in tumour and 66.66% in sTILs. Of these 28 cases, 32.14% concordance was noted in PD-L1 expression in both tumour and sTILs. Conclusion: Our study results showed correlation of Ki-67 with PD-L1 expression in tumour cells and sTILs. PD-L1 positivity in tumour cells and sTILs also showed concordance. Thus, it can be predicted that PD-L1 expression might be related to a TIL-mediated anti-tumour inflammatory reaction.

   OP12: Mathematical pathological approach for the assessment of diffusion penetration length in breast carcinoma Top

Neha Singh

VMMS and Safdarjung Hospital, New Delhi, India. E-mail: [email protected]

Background: Mathematical Pathology is a research branch of pathology in which mathematics and physical principles are applied to study the nature of various diseases. In breast cancer there is a need for novel prognostic factors in view of individualized management of patients. Diffusion penetration length (DPL) calculated using mathematical pathological approach may emerge as one such novel parameter. Design: A total of 15 cases of breast carcinoma were studied with the aim to calculate diffusion penetration length of tumor in breast carcinoma using mathematical pathological approach and correlated with histopathological type, grade, stage and surrogate molecular classification. H&E stained sections were examined for the histological type and grade. IHC for ER, PR, HER-2 neu and Ki67 were performed for surrogate molecular classification. Ki-67 was also used to assess proliferation index. Additionally, IHC for cleaved caspase-3 was performed to measure apoptotic index. Geometric mean radius of tumor was obtained mammographically. DPL was calculated and correlated with existing prognostic parameters. Results and Discussion: Age range of the cases was 46.6 ± 13.5 yrs. Out of 15 cases, 12 were of grade II and 3 of grade III. Surrogate molecular classification of 9 cases were Luminal B, 3 cases were of Triple negative, 1 case of Luminal A and 2 cases of Her-2 neu enriched. Overall DPL was found to be 0.81 μm. DPL is a measure of how far molecules (e.g., chemotherapy, drugs, nutrients) diffuse through a medium (e.g., the tumor tissue). Conclusion: Diffusion penetration length may emerge as a novel parameter to assess breast cancer. It may prove to be of great utility in individualized management of these patients.

   OP13: The clinical, histomorphological and immunohistochemical characteristics of metaplastic carcinoma of breast: A descriptive study Top

Norton Stephen

JIPMER, Puducherry, India. E-mail: [email protected]

Background: Metaplastic carcinoma (MC) of the breast are a rare heterogeneous group of tumors, incidence of which is less than 1%. These are an unique set of tumours with varying subtypes with a poor prognosis and an increased chance of distant metastasis. Aim: To study the clinical, histomorphological, immunohistochemical features of Metaplastic carcinoma. Materials and Methods [Design]: This is a Descriptive study. Data was collected from 2015 till 2019. A total of 20 cases were available whose clinical, histomorphological and immunohistochemical were studied. Results: The median age of presentation was 50 years. 70% [14/20] cases were postmenopausal women. Trucut biopsy detected the possibility of Metaplastic carcinoma in 50% cases [10/20]. On Resection, 75% [15/20] showed Mixed typed Metaplastic carcinoma, the remainder showing epithelial type Metaplastic carcinoma. Axillary lymph node involvement was seen only in 10% [2/20] of the cases. ER, PR were negative in all the 20 cases [100%], Her2neu was positive [score3+] in 5% of the cases [1/20]. Ki67 >14% in 100% of the cases [20/20]. Two of the twenty cases received Neo-adjuvant Chemotherapy. Conclusion: Metaplastic carcinoma of the Breast are tumours known in elderly postmenopausal age group, larger tumour size, lesser chances of lymphnode metastasis, higher chance of recurrence and hematogenous spread with absence of Estrogen receptor (ER), Progesterone receptor (PR), Human epidermal growth factor -2 [Her-2 neu] and a high Ki67 index [>14%].

   OP14: Programmed cell death ligand-1 and tumor infiltrating lymphocytes, in patients with invasive ductal carcinoma breast pre and post neoadjuvant chemotherapy Top

Jhansi Jayachandran, Pampa Ch Toi, J Sree Rekha, Smita Kayal, D Kadambari

JIPMER, Puducherry, India. E-mail: [email protected]

Background: Programmed cell death protein 1 (PD-1) is an immune check-point on lymphocytes, activation of which promotes their apoptosis. By upregulating ligands for PD-1 (PDL1), cancer cells block responses by CD8+ Tumor infiltrating Lymphocytes and thereby immunosurveillance. Chemotherapy reduces actively dividing tumour load and could possibly reduce its protein expression, including PDL1. Design: 73 Patients diagnosed with IDC breast who have undergone NACT and Modified Radical Mastectomy were included. Immunohistochemistry was performed, for PDL-1 and CD8+T cells on the pre-chemotherapy biopsy and post NACT mastectomy specimens. Results and Discussion: Study population comprised of 40 Luminal A/B, 18 Triple Negative and 13 HER2neu positive cases. 6 cases had pathological complete response, postNACT. The mean Peritumoral TIL in post-chemotherapy Specimens (30.7%) was significantly higher than that in pre-chemotherapy biopsies (22.01%) (p=0.02), the difference being significantly higher in Luminal type A/B (p=0.009). 41(56.1%) of Pre-chemotherapy biopsies were PDL1 positive, compared to 10 (13.6%) of post NACT specimens, difference of which was significant (p=0.0001). Cases that had attained pathological complete response showed no significant difference in both TIL count as well as PDL1 positivity, pre and post chemotherapy (p=0.4). Conclusion: NACT decreases expression of PDL1 and causes immune activation. Targetting PDL1 in Breast cancer could pave way for better immunosurveillance and hence response.

   OP15: Clinico-pathological study of triple negative breast cancer and its association with androgen receptor in a tertiary care centre Top

Shubham Varshney

Kasturba Medical College, Manipal Academy of Higher Education, Manipal, Karnataka, India. E-mail: [email protected]

Objective: To evaluate the association of androgen receptor(AR) with various clinicopathological parameters in triple negative breast cancer (TNBC) including overall survival (OS) and disease free survival (DFS). Materials and Methods: Total 78 cases of IHC proven TNBC were included in 2-year ambispective study in tertiary care center. Clinicopathologic features of AR positive and negative TNBC were analyzed and compared. OS and DFS were evaluated for 51 patients (follow up available). Results: A 1% cut off was confirmed as appropriate threshold for AR positivity. The mean age for AR(+) group was 51.3 years, slightly higher compared to AR(-) group. There was no significant association of AR status with other parameters like, tumor size, laterality, tumor site, metastasis, skin & nipple involvement, DCIS, LVI and MBR grade. The Ki 67% median was 60% for AR(-) group which was significantly higher as compared to AR(+) group (10%). In univariate analysis, high Ki 67 (≥ 10) was an independent prognostic factor for DFS in TNBC (hazard ratio = 1.86, 95% CI = 1.12-2.81). In terms of disease outcome, AR(+) tumors showed improved OS and DFS as compared to AR(-) tumors. Conclusion: From present study, we concluded that AR(+) TNBC has significantly low Ki -67 index, better DFS & OS as compared to AR(-) TNBC. Also, high Ki-67 significantly predicts a higher incidence (1.86 fold higher) of lower DFS as compared to low Ki -67 group (< 10) in TNBC group and can be used for further classification of TNBC into 2 clinically divergent subtypes.

   OP16: Assessment of PD-L1 expression and tumour infiltrating immune cells across molecular subtypes of triple negative breast cancers Top

Ridhi Sood, Sandeep Kumar, Ashim Das, Gurpreet Singh1, Amanjit Bal

Departments of Histopathology and1General Surgery, PGIMER, Chandigarh, India. E-mail: [email protected]

Background: Triple-negative breast cancer (TNBC) accounting for 20-25% breast cancers has no approved therapeutic target due to lack of hormone receptors and Her-2 expression. Recently PD-1/PD-L1 targeted immunotherapy, has been investigated in locally advanced/metastatic TNBC. However, there is a lacuna on technical and prognostic aspects of PD-L1 biomarker testing. The aim of this study is to evaluate the tumor infiltrating immune cells (ICs) and PD-L1 expression across molecular subtypes of TNBC and assess prognostic significance. Design: TNBC tumour tissue microarrays from 197 treatment naïve cases were analysed for PD-L1 (22C3 Dako antibody) expression and ICs (as per the International TILs Working Group 2014). Results and Discussion: Based on IHC, the 197 cases, were subtyped as; Androgen Receptor positive (16%), Mesenchymal Stem cell Like (MSL, 14.8%), basal and MSL (14.5%), basal (14.1%), claudin low (12.5%) and unclassifiable (23.8%). Overall 88.8% cases had ICs less than <50%. PD-L1 positivity was seen in 36% and 11.2% cases in ICs and both ICs and TCs respectively. Highest PD-L1 positivity was seen in basal and unclassifiable group. PD-L1 expression in ICs correlated significantly with molecular subtypes (p value 0.017). Also there was correlation between >50% ICs and PD-L1 positivity (p value 0.000864). However, there was no correlation with ICs and PDL-1 expression in TCs. PD-L1 expression (TCs/ICs) did not correlate with tumor size and nodal status. Conclusion: This study highlights the heterogeneity within the TNBC group. The high PD-L1 expression in ICs of basal and unclassifiable group suggests that these can be targeted by immunotherapy.

   OP17: Evaluation of morphologic spectrum and PI3K/AKT mutations in metaplastic carcinomas of the breast Top

Saumya Gupta, Amanjit Bal1, Sandeep Kumar1, Ashim Das1, Gurpreet Singh2

Departments of Pathology,1Histopathology and2General Surgery, PGIMER, Chandigarh, India. E-mail: [email protected]

Background: Metaplastic carcinomas of the breast are comparatively rare, aggressive tumours of the breast with heterogenous morphology. They often contain conventional ductal invasive carcinoma component along with mesenchymal or squamous component and are typically triple-negative for estrogen receptor(ER), progesterone receptor(PR), and HER-2 overexpression, thus having no targeted therapy option. Aims and Objects: To study the morphologic spectrum of metaplastic carcinomas and look for prevalence of PI3K/AKT hot spot mutations in this cohort of breast cancer patients as possible target for therapy. Design: All cases of metaplastic carcinoma breast were retrieved and morphologic subtyping was done. The PI3K/AKT hot spot mutations were evaluated by Sanger Sequencing. The results were correlated with tumour size, and lymph node status. Results: A total of 39 cases of metaplastic carcinomas were diagnosed over a period of 5 years. The mean age of the patients was 51 years (range 27-82). Mean tumour size was 5.2 cm (range 1 to 15 cm). Axillary lymph node clearance was performed in 27 cases, of which 7 cases had positive nodes. Cases were categorized as spindle cell carcinomas (8), squamous cell carcinomas (12), carcinosarcomas (4), matrix producing metaplastic carcinomas (10) and fibromatosis like variants (5). All cases were triple negative on immunohistochemistry (ER-/PR-/Her2-). PI3K/AKT mutations were done in 10 cases (shall be done in more cases), however no hotspot mutations were noted. Conclusion: The results of this, highlight morphologic heterogeneity in metaplastic breast cancers. Mutations in PI3K/AKT gene, if seen, may provide options for targeted therapy in this aggressive tumour subset.

   OP18: Hematolymphoid neoplasms in effusion cytology subclassification, distribution and role of ancillary techniques: A 3-year retrospective study of 75 cases Top

Renu Sukumaran, Nileena Nayak, K Jayasree, Priya Mary Jacob, Rekha A Nair

Division of Pathology, Regional Cancer Centre, Thiruvananthapuram, Kerala, India. E-mail: [email protected]

Background: Involvement of body fluids can occur at the time of diagnosis or during the course of hematopoietic malignancies. Aims: 1) To study the morphology, subtypes and pattern of distribution of various hematolymphoid neoplasms in effusions, 2) To assess the role of ancillary techniques in the diagnosis. Materials and Methods: In this retrospective study, all cases of hematolymphoid neoplasms involving body fluids diagnosed from January 2016 to December 2018 were analyzed. Results: Total number of cases-75. It included 35 cases of B cell neoplasms, 33 cases of T cell neoplasms and 7 cases of myeloid neoplasms. Pleural fluid was involved in 56 cases (74.67 %), ascitic fluid in 17 cases (22.67 %) and pericardial fluid in 2 cases (2.67 %). B cell neoplasms included Burkitt lymphoma, diffuse large B cell lymphoma, follicular lymphoma, plasma cell myeloma, hairy cell leukemia and plasmablastic lymphoma. T cell neoplasms included T lymphoblastic lymphoma, anaplastic large cell lymphoma and adult T cell leukemia/lymphoma. Myeloid neoplasms included acute myeloid leukemia and myeloid blast crisis of CML. T lymphoblastic lymphoma was the most common subtype (29 cases- 38.7%), followed by DLBCL (12 cases- 16%). In 53 cases (70.67%), effusion was present in the initial presentation itself. Initial diagnosis of hematopoietic malignancy was made in effusion fluids in 25 cases (33.33% of the total) - with the help of flow cytometry in 20 cases and immunohistochemistry in cell blocks in five cases. Conclusion: Combining cytomorphology and ancillary techniques enable the accurate diagnosis of various hematolymphoid neoplasms in effusion cytology specimens.

   OP19: Utility of cell block from fine needle aspiration cytology for sub-categorisation of diffuse large B-cell lymphoma using Han's algorithm Top

Sumit Garg, Radhika Srinivasan, Manish Rohilla, Arvind Rajwanshi, Ashim Das1, Amanjit Bal1

Departments of Cytology and Gynaecological Pathology and1Histopathology, Post Graduate Institute of Medical Education and Research, Chandigarh, India. E-mail: [email protected]

Introduction: Diffuse Large B-Cell Lymphoma (DLBCL) is the most common NHL. DLBCL is further sub-classified as per Han's algorithm using surrogate immunohistochemistry(IHC) markers CD10, BCL6 and MUM1 as GCB type and Non-GCB type in accordance with the current WHO classification. Aim and Objective: To perform immunohistochemistry for CD10, BCL6 and MUM1 on cell blocks from fine needle aspiration cytology(FNAC) for sub-classifying cases of DLBCL and correlate it to histopathology thereby evaluating their utility. Materials and Methods: Retrospective analysis of all Non Hodgkin lymphoma cases diagnosed on cytology was performed to identify cases of DLBCL between Jan 2017 and June 2019 with available cell block for further immunocytochemistry and with histopathological follow-up excision lymph node biopsy. The cytomorphology and CB-IHC was compared with the final histopathology sign-out. Results: A total of 43 cases of DLBCL with histopathological follow-up were evaluated out of which 13 cases had to be excluded due to low cellularity in CB. Thus DLBCL sub-typing was carried out in 30 cases, out of which 10 cases were diagnosed as GCB type and twenty cases as Non-GCB type. Overall, histopathological-concordance was found in 90.5% cases (19/21). Out of two discordant cases one was of GCB type with CD10+ and another was unclassified type with MUM1-/CD10-/BCL6- on CB-IHC but were MUM1+ and therefore of ABC type on histopathology. Discussion and Conclusion: In cases of DLBCL with adequate material on cell blocks, DLBCL can be reliably sub-typed into GCB and ABCtypes based on CD10, MUM1 and BCL6 IHC and should be incorporated into routine clinical practice.

   OP20: Clinico histopathological correlation of granulomatous lesions of skin with special emphasis on leprosy Top

Raju Bhaisare, M Ravindranath

J.L.N. Hospital and Research Centre, Bhilai, Chhattisgarh, India. E-mail: [email protected]

Objective: Granulomatous inflammation of skin tissue is a commonly encountered in histopathology have challenging to diagnose due to amidst of histological features of various disease. The present study examined the histopathological profile of Granulomatous lesion of skin which comes in department of pathology of J L N hospital research and centre sector -9 Bhilai. Methods: The present study was observational study. It comprising of various granulomatous lesion specimen received over a period of three years (197 cases). All specimens were included, studied grossly and microscopically with routine, special stains and ancillary finding if available. Results: Total of 197 skin biopsies were received by the Department of Pathology. Among these, 47(23.85%) were histologically diagnosed as Granulomatous. Out of 47 Granulomatous skin lesions, 24 (12.18%) were histopathologically diagnosed as Non Lepromatous and 23 (11.67%) as Lepromatous. Infectious granulomatous lesions constituted of 30 (15.22% of total) cases. The most common etiology of granuloma was leprosy 23 (11.67%) cases, followed by cutaneous tuberculosis 7 (3.55%) cases. Conclusion: Granulomatous lesions of skin have varied spectrum, so histopathology is the key to differentiate one lesion from another which is beneficial for the management of lesion.

   OP21: Spectrum of skin lesions in pediatric age group Top

Sachin Uttamrao Raut, Vandana Sanklecha, Ashwin Natekar, Naresh Deonikar

Department of Pathology, Grant Government Medical College and JJ Group of Hospitals, Mumbai, Maharashtra, India. E-mail: [email protected]

Background: Skin diseases are common in children and surveys indicate that they are common reasons for pediatric visits to the outpatient department. The pathologist who wishes to interpret histopathological sections of skin diseases must familiarize himself with the clinical features of at least the common diseases, as the gross appearance of the lesions and their distribution is an important diagnostic aid to him. Objectives: 1. To analyze the spectrum of dermatopathological lesions in the pediatric population in tertiary care hospital over a two years period 2. To study the age and sex and anatomic distribution of the lesions. 3. To correlate the clinical diagnosis offered by the dermatologist with the histopathological diagnosis. 5. To assess the contribution of histopathology to the final diagnosis. Results and Conclusions: Pediatric dermatopathological biopsies constituted 6.15% of the total number of biopsy load. The age distribution pattern indicated high percentage of children in the 13-18 year age range (60.48%) and majority of cases were males compared to females. The anatomic distribution of the lesions revealed that the highest number of cases occurred in the extremities 54.02%, this was followed by the trunk and the head, neck and face. The most frequently encountered lesion was Borderline Tuberculoid Hansen's disease-4.03% followed by Lupus Vulgaris-2.41 % both belonging to the broad category of Infectious disease. Other lesions like Lichen planus- 3.22%, Vasculitis-3.22%, Non-Specific Dermatitis-3.22%, atopic dermatitis—1.61% and Psoriasis-1.61% were also encountered. Thus in a significant number of biopsies (77.41%), histopathology contributed to the diagnosis.

   OP22: Expression of osteopontin in psoriatic lesions Top

Rohini Garg, Meena Harsh, Nadia Shirazi

Himalayan Institute of Medical Sciences, Dehradun, Uttarakhand, India. E-mail: [email protected]

Background: Osteopontin (OPN) is a secreted phosphorylated glycoprotein first isolated in 1986 from osteoblast. It is also known as bone sialoprotein. The regulation of OPN expression has been identified in various carcinomas and autoimmune diseases including psoriasis. The aims of the study were to study the expression of OPN in psoriatic lesions and to correlate it with the severity of disease using “Psoriasis area severity index (PASI)” Materials and Methods: Observational study was conducted on 35 cases of psoriasis over a period of one year. The clinical history was recorded in detail along with PASI score, pathological and immunohistochemical findings were evaluated. Results and Discussion: Study showed that maximum number of cases (n=23:65.71%) showed expression of OPN upto lower half of epidermis (2+) while 17.14% (n=6) cases showed expression in whole epidermis. For dermal inflammatory infiltrate, 51.4% (n=18) cases showed moderate expression (2+) while 17.1% (n=6) showed marked expression. While comparing the severity with PASI score, it was seen that with a high mean PASI score of (26.45+_6.14) expression was in whole epidermis. Similar results were in dermis. With a mean PASI score of (29.26+_5.04), expression of OPN was severe in dermal inflammatory infiltrate while with a mean score of (11.01+_6.0), OPN expression was mild. Thus showing a statistically significant result with p value<0.05. Conclusion: This study shows that OPN expression varies with severity of disease and therefore can be used as a marker of disease severity.

   OP23: Clinico-cytopathological evaluation of skin lesions with special reference to bullous lesions: Institutional study Top

Priyanka Dhingra, Parul Singhal, Ajay Kansal

Sarawathi Institute of Medical Sciences, Chaudhary Charan Singh University, Uttar Pradesh, India. E-mail: [email protected]

Background: Cytopathology of skin has been documented to be useful in the diagnosis of several skin lesions. The present study was conducted to correlate the clinical diagnosis with cytology and histopathology for the diagnosis of various nodular inflammatory lesions. Aims: Evaluate cytopathology as a quick non-invasive method for early diagnosis of bullous lesions, neoplastic and preneoplastic skin lesions and to correlate the clinical, cytological and histopathological findings of various skin lesions. Materials and Methods: The study was retrospective carried out over a period of 1 year in which 100 patients of skin lesions were included. Skin scraping, Tzanck smears, slit smears and fine needle aspiration cytology, were done to obtain material for cytological examination. Excisional biopsy, incisional biopsy and punch biopsy was done to obtain tissue for histopathological examination. The slides were stained with routine and special stains as and when required. Results: Out of the 100 patients,60 were males and 40 females. The most common non-neoplastic lesions observed were vesico-bullous lesions which comprised of 48 cases followed by neoplastic lesions which consisted of 28 cases, ofwhich 07 were benign and 21 malignant. Concordant results between cytology and histopathology was seen in majority (91.7%) of lesions studied. Conclusion: Cytology performed skillfully and with perfection, leads to an early diagnosis in majority of the lesions, as the observed cytomorphological features of various skin lesions were fairly distinctive making cytology a fairly sensitive 'patient compliant' technique for rapid diagnosis of skin lesions.

   OP24: Mitotic activity >15/ 50 high power field, rather than weiss criteria is a reliable indicator of outcome in adrenocortical carcinoma in children Top

Divya Aggarwal, Rimesh Pal1, Debajyoti Chatterjee, Uma Nahar Saikia, Bishan Dass Radotra, Rama Walia1, Arunanshu Behera2

Departments of Histopathology,1Endocrinology and2General Surgery, PGIMER, Chandigarh, India. E-mail: [email protected]

Introduction: Adrenocortical carcinoma (ACC) is a rare and aggressive malignancy in children. Due to rarity of the disease, few studies have validated the criteria helpful in predicting the long term outcome. Materials and Methods: Institutional histopathology database was searched for all cases of adrenocortical carcinomas in children (age <18 years) diagnosed from January 2010 to December 2018 (n=13). All demographic, clinical and pathologic data was collected including retrieval of slides and clinical and radiological follow-up. All cases were screened and modified Weiss score was calculated. Results: Of a total of 13 cases, mean age was 50±53 months with male: female ratio of 1.17:1. Follow-up was available for 9 patients, of which 4 cases had recurrence, 1 developed contralateral ACC, 1 died and 3 were free of disease at last follow-up. Mean duration of symptoms was 7.9±6.4 months and mean duration of follow-up was 6.7±2.6 months. Most common symptom was excessive pubic hair growth (56%), followed by weight gain (33.3%). Hypertension was noted in 67% (6/9) of the cases. Mitotic figures more than 15 per 50 hpf was constantly associated with an adverse outcome, while other pathologic criteria including tumor size, weight, capsular or vascular invasion, extension to extra-adrenal tissue and necrosis showed poor correlation with outcome. Conclusion: Mitotic activity (>15/ 50 hpf), rather than Weiss criteria is a reliable indicator of the outcome in ACC in children. However, studies with larger sample size and longer follow-up are required for further validation of these criteria as independent prognostic indicators.

   OP25: Diagnostic utility of CD56, CK19 and P63 in differentiation of papillary carcinoma thyroid from follicular neoplasms Top

Ruby Sahu, Minakshi Bhardwaj, Arvind Ahuja, D S Chauhan, Purnima Malhotra, C K Durga

ABVIMS and DR. RML Hospital, New Delhi, India. E-mail: [email protected]

Background: Thyroid diseases are commonest endocrine disorders worldwide. Majority of the thyroid nodules are papillary thyroid carcinoma followed by follicular carcinoma. Diagnostic dilemma may arise when an encapsulated nodule with a follicular pattern of growth exhibits clear nuclei with grooves and so distinguishing follicular adenoma from encapsulated FVPTC becomes difficult. Aim of this study was to evaluate the individual as well as combined expression of CD56, CK19 and P63 in the differentiation of papillary carcinoma from follicular neoplasms. Design: A total of thirty cases of histologically proven papillary carcinoma, follicular carcinoma and FVPTC lesions over a period of one and half years were included in this study. Detailed histomorphological and immunohistochemical analysis of all three markers CD56, CK19 and P63 were performed in all the cases. Results and Discussion: Specificity of CK19 and CD56 were 100%, 84.62% respectively in differentiating PTC from Follicular carcinoma when they were used individually while specificity were 84.62% when used in combination. Combination of CD 56 and p63 was most specific (100%) for differentiation of FVPTC from follicular carcinoma. Combinations of all the three markers were 100% sensitive. Conclusion: Panel of all three IHC markers when used together were more useful to reach definitive diagnosis than individual markers. These markers are useful in cases with morphological overlap specially to diagnose PTC and FVPTC.

   OP26: Study of apoptosis, proliferation and BRAF in colorectal carcinoma in Indian patients Top

Shilpi Saxena, V Srinivas, Rajat Jagani, Deep Kumar Raman1, Prabal Deb, M S Brar

Military Hospital,1Department of Pathology, Military Hospital, Allahabad, Uttar Pradesh, India. E-mail: [email protected]

Background: Colorectal Carcinoma (CRC) is a common cause of death worldwide with rising incidence in India. Many biomarkers have been studied in CRC patients in various populations and have been correlated with tumour differentiation, prognosis and targeted therapies. However similar studies in patients of CRC in India are lacking and no study has been performed till date to study apoptosis, proliferation and BRAF mutation on same tumour in Indian patients, which we aim to do. Design: Cross sectional study. The aim was to study distribution of p53, Bcl2, MIB-1 and BRAF in colorectal carcinoma in Indian patients and correlation with tumour grade and American Joint Committee on Cancer (AJCC) stage of CRC using Immunohistochemistry (IHC) in 65 consecutive cases of CRC in tertiary care centre. The results were statistically analysed using SPSS version 20. Results and Discussion: Out of 65 cases 36.92% (24/65) cases were positive for p53 (p=0.001). Bcl-2, MIB-1 and BRAF positivity was seen in 32.31%, 49.23% and 4.62% respectively. Expression of BRAF was significantly higher in higher stages (P=0.029). No significant correlation was found in between expression of other three markers with grade and AJCC stage. No significant association was found between apoptosis, proliferation and BRAF markers in this study. Conclusions: The study concluded that BRAF mutation is not very common in Indian CRC patients but is predictor of higher stage of CRC. However larger population based studies need to be done to find out prevalence, significance and association of these biomarkers in Indian patients.

   OP27: Histological assessment and use of immunohistochemical markers for detection of dysplasia in Barrett's esophageal mucosa Top

Prateek Kinra, Gaurav Gahlot1, Prasenjit Das2, Rajni Yadav2, Siddhartha Datta Gupta2

Armed Forces Medical College, Pune, Maharashtra,1Army Hospital, R and R, New Delhi,2AIIMS, New Delhi, India. E-mail: [email protected]

Background: Histological assessment of dysplasia in Barrett's esophagus (BE) has high inter-observer variability. Hence, use of ancillary markers for early detection of dysplasia in BE is an important clinical question. Design: In this retrospective study consecutive cases of BE (n= 59), over a period of 4 years were included. Sections were reviewed independently by 3 senior qualified pathologists, who graded the dysplasia according to the Vienna Classification system and inter-observer agreement was analysed using the Kappa statistics. Subsequently Alpha-Methyl Acyl-CoA Racemase (AMACR), p53, CyclinD1, β-catenin, H2AX and M30 immunohistochemical (IHC) stains were examined on the following disease categories: BE with no dysplasia [NFD] (45), BE with indefinite for dysplasia (IFD) (4), low grade dysplasia (LGD) (3), high grade dysplasia (HGD) (2) and in adenocarcinomas (5). H score was calculated by adding up products of different grades of stain distribution and stain intensities (range of scores 0-300). Results: Among the 3 pathologists, overall agreement was poor (k 0.06; 95% CI -0.089 to 0.145), with highest disagreement noted for differentiating the LGD and IFDs (k= 0.21). After revising the histological criteria, the kappa improved to 0.53. Among the IHC stains performed, p53, β-catenin, H2AX and M30 stains were significantly useful to differentiate between IFD and LGD (P values: 0.04, 0.004, 0.05 & 0.04, respectively). AMACR and β-catenin stains though were up-regulated in HGD/ adenocarcinomas than in other categories, their expression were not statistically different between the IFD and LGDs. Conclusions: Detailed Histology and IHC can accurately grade BE associated dysplasia.

   OP28: Immuno-histochemical expression of CD44 and Ki67 in gastric adenocarcinoma and its correlation with clinico-pathologic parameters Top

Taiba Farooque, Parul Maheshwari, Amit V Varma

Sri Aurobindo Medical College and P.G. Institute, Indore, Madhya Pradesh, India. E-mail: [email protected]

Background: Gastric carcinoma is a leading cause of deaths due to cancer in the world. Symptomatic gastric carcinomas are typically detected at advanced stages, with poor 5-year survival rates despite modern multimodal treatment strategies. There is a need for novel biomarkers to predict patient outcomes. In this study, we have studied expression of biomarkers related to stem cells i.e.CD44 and proliferative index marker, Ki67. The aim was to evaluate the expression of CD44 and Ki67 in Gastric adenocarcinoma (GAC) in relation to clinical and histo-morphological parameters. Design: All histopathologically confirmed cases of GAC, including biopsies and gastrectomy specimens, received in the Department of Pathology at Sri Aurobindo Medical College & PG Institute, Indore from January 2018 to June 2019 were studied and biomarkers were applied. Results: Of total 53 cases of GAC, mean age was 55 years with M:F ratio of 2.78:1. Most common site was pyloro-antral region (35.84%). As per the WHO Histological Classification, most cases were of Tubular subtype (47.16%), with least cases of Mucinous (13.21%) subtype. According to Lauren's Histological Classification, the Intestinal (35.66%) subtype was appreciated more than Diffuse (34%) subtype. Biomarker study showed that both Ki67 and CD44 expression was significantly more in grade III tumours compared to grade-I & II, and diffuse tumors showed more Ki67 & CD44 positivity compared to the tubular variant of Lauren's classification of GAC. Conclusion: Both markers can have a role in prognosticating GAC, since their expression shows increased positivity with increasing grade of tumor.

   OP29: Study of survivin biomarker in carcinoma colon Top

Pratibha Chandra

Armed Forces Medical College, Pune, Maharashtra, India. E-mail: [email protected]

Background: Survivin is a newly detected protein which has been researched in various carcinomas. Survivin suppress both extrinsic and intrinsic pathways of apoptosis and regulates cell division. This study correlates the presence of Survivin in carcinoma colon with known prognostic variables. Design: Paraffin blocks of 102 patients with carcinoma colon were retrieved from archives. The cases were reviewed as per the existing CAP protocol and the IHC was carried out using the polymer-based detection system. The immuno-histochemistry expression was semi-quantified using the IRS Scoring System which encompasses the stain intensity and percentage positive cells. Survivin expression was correlated with age at diagnosis, gender, grade, nodal status, lymphovascular/ perineural invasion, resected margin status and tumor stage. Result and Discussion: 26/102 cases showed Survivin positivity. The mean age of the study population=45 yrs (range 21-84). The gender distribution was 74:28 (M:F). High survivin expression was seen in cases with higher grade (p=0.002), high stage (p=0.002), lymph node positivity status (p=0.002), LVI/PNI status (p=0.02 / 0.001 respectively). However, no significant correlation was found between Survivin and gender. No statistical test could be applied for resection margin status as the number of positive cases in category were few. Conclusion: Survivin expression increases as the tumor grade worsens in carcinoma colon and is maintained in approximately all high group staged tumors along with tumors showing lymphovascular and perineural invasion. The study concludes that Survivin can be used a surrogate maker for worse prognosis in initial biopsy of cases having carcinoma colon.

Keywords: Survivin, carcinoma colon

   OP30: Study of KRAS, MLH1, MSH2 and MSH6 in colorectal adenocarcinomas using immunohistochemistry Top

Soumia Burada, N Rukmangadha, V Venkatarami Reddy1

Departments of Pathology and1Surgical Gastroenterology, SVIMS, Tirupati, Andhra Pradesh, India. E-mail: [email protected]

Background: Colorectal cancer(CRC) is one of the most common cancer across the Globe. Majority of the CRC's are sporadic and <6% are familial or hereditary. Chromosomal instability (CIN) is major pathway in CRC, caused due to mutations in APC, KRAS, P53. Microsatellite instability (MSI) is major pathway in lynch syndrome and in 15% of the sporadic CRC caused by mutations in mismatch repair (MMR) genes such as MutL homolog1(MLH1), MutS homolog2 (MSH2), postmeiotic segregation (PMS2) and MutS homolog6(MSH6). Design: A total of 150 cases were studied prospectively in the department of pathology, SVIMS Tirupati, during the period of January 2018 to September 2019. IHC with KRAS, MLH1, MSH2 and MSH6 is done on morphologically diagnosed cases of colorectal adenocarcinomas. Results and Discussion: Out of 150 patients 93 are male and 57 are female, between 28 to 82years of age, among 150 cases 123 are conventional adenocarcinomas, 22 were mucinous and 5 signet ring cell carcinomas. Of the 150 cases, 109 were moderately differentiated, 24 were well differentiated and17 poorly differentiated. Most of the tumors are from colon. Microsatellite stability/instability status and KRAS mutation status were studied using IHC and evaluated the associations between MMR and KRAS mutations with clinicopathological parameters and compared with similar other studies, including the benefits of IHC over MSI testing. Conclusion: This is a prospective analysis of correlations between KRAS mutation and MMR status with clinicopathological characteristics in CRC patients. They are helpful to predict prognosis and treatment response and hence can be used as prognostic and predictive markers.

   OP31: Evaluation of tumor budding, tumor infiltrating lymphocytes and tumor eosinophilic infiltration with respect to pathologic stage in colorectal adenocarcinoma Top

Priya Sahu, Arvind Ahuja, Purnima Malhotra, D S Chauhan, Amrita Talwar, Minakshi Bhardwaj

Department of Pathology, ABVIMS and Dr Ram Manohar Lohia Hospital, New Delhi, India. E-mail: [email protected]

Background: In Colorectal carcinomas (CRC), TNM system is regarded as the only standard for prognostication and staging. However, additional prognostic markers are needed to predict the disease outcome and improved patient management. In this study, we assessed tumor budding, tumor infiltrating lymphocytes (TIL) and tumor eosinophilic infiltration at invasive front of colorectal adenocarcinoma (NOS) and its correlation with Pathologic Stage. Design: It was a cross-sectional observational study. We enrolled 78 cases of CRC over the period of 4 years. Cases diagnosed as Adenocarcinoma NOS were further evaluated. Mucinous and signet ring carcinomas were excluded. Histomorphologic parameters: tumor budding, TILs and tumor eosinophilic infiltration were stratified in three categories. Tumor budding: 0-4, 5-9, ≥10 buds/20x field; TILs: <10%, 11-49%, ≥50%; tumor eosinophilic infiltration: 1-9, 10-50, >50/HPF. Correlation among individual parameter and its impact on the pathologic (pTN) stage were evaluated by using chi square test. A p value of <0.05 was considered statistically significant. The analysis was done using SPSS version 21.0. Results: Out of 78 cases of colorectal carcinoma, 57 (73%) were Adenocarcinoma NOS. Mean age of presentation was 50.9 years with male to female ratio 2:1. Our data supported significant association of tumor budding with pT category (p-value 0.01) and lymph node metastasis (p-value 0.01). Among individual parameters significant association was found between tumor budding and TILs (p-value 0.03).

Conclusion: Tumor budding is a robust and reproducible prognostic marker that can predict nodal metastasis. Thus, it may be useful in recognising the high risk cases and their management decisions.

   OP32: PD-L1 in colorectal carcinomas Top

Ekta Sethi1,2, Jyoti R Kini1,2, Saraswathy Sreeram1,2, Hema Kini1,2

1Kasturba Medical College, Mangalore,2Manipal Academy of Higher Education, Manipal, Karnataka, India. E-mail: [email protected]

Background: Colorectal carcinoma is the third most commonly diagnosed cancer in the world in men and second in women. It is the most common malignancy affecting the gastrointestinal tract. PD-L1, an immune checkpoint inhibitor, has been widely studied in lung non-small cell carcinomas, melanomas, urothelial carcinomas. Currently expression of PD-L1 in colorectal carcinoma is of interest. It has importance as targeted FDA approved therapy is now available for PD-L1 positive tumors, improving survival rate and prognosis. Design: Fifty nine colorectal carcinoma cases were taken from Department of Pathology, Kasturba Medical College, Mangalore archives from January 2017- December 2018. Male to female ratio was 1.19:1. Pre-operative treated cases were excluded. PD-L1 (CAL 10 clone) staining was done on paraffin embedded formalin fixed blocks. Results and Discussion: Out of 59 colorectal carcinoma specimens, 39 (66.1%) displayed positive membranous and/or cytoplasmic positivity while 20 (33.9%) had negative staining. PD-L1 staining was quantified by intensity of staining and percentage of tumor positive cells. Intensity of more than 2+ was taken as positive. PD-L1 intensity of staining had a significant relationship with peritumoral lymphocytic response. Percentage of PD-L1 positive tumor cells was significantly associated with number of lymph nodes positive for metastasis. Conclusion: PD-L1 positivity was seen in more than half of the colorectal carcinomas cases studied. It was significantly associated with higher lymph node status. This study implies that there is scope for immunotherapy to be considered in our subset of Indian population.

   OP33: Importance of P16ink4a marker in oral neoplasms Top

Umarani Pallivilla, Swathi Chilukuri, Vahini Gudeli, Asha Thota

Department of Pathology, ASRAM Medical College, Eluru, Andhra Pradesh, India. E-mail: [email protected]

Introduction: Human Papilloma viruses (HPV) association and risk of oral cancers development reported dates back in 1983, when HPV16 was detected in oral squamous cell carcinoma (OSCC) tumors. HPV 16 regulates cell cycles arrest and apoptosis by regulating the Tumor suppressor p16INK4a protein and increased or decreased expression was found in HPV 16 associated oral SCC. Aims and Objectives: The aim of this study is to evaluate the role of p16INK4A protein in OSCC and epithelial dysplasia's by immunohistochemistry. Materials and Methods: This is a retrospective study, 29 OSCC cases from January 2017 to July 2019 were included. Histopathological studies were done by standard Hematoxylin & Eosin staining on tissue section followed by immune profiling with p16INK4a collected from punch biopsies and surgically resected tumors. Tumor classification was done according to the criteria proposed by WHO. Results: In this present study male predominant were seen and age group was 40 to 60 years and common site of the tumor was tongue. Histopathologically well differentiated SCC were more in number. The immunoexpression of p16 stain was present at nuclear and cytoplasmic level in basal dysplastic epithelium. P16 had a weak intensity at level of tumor proper and also at invasion front. Conclusion: P16 is useful marker in identifying dysplastic lesions and decreased immune expression is a predictive factor for neoplastic transformation.

   OP34: Analysis of PDL1, Her2NEU and p53 in gastric and gastroesophageal junction carcinoma Top

Animesh Saurabh, Vandana Raphael1, Biswajit Dey1, Caleb Harris2, Vikas Kantilal Jagtap3, Umesh Das4

NEIGRIHMS, Departments of1Pathology,2Surgical Oncology,3Radiation Oncology and4Medical Oncology, NEIGRIHMS, Shillong, Meghalaya, India. E-mail: [email protected]

Background: The process of tumorigenesis in gastric carcinoma involves multiple genetic alterations including overexpression of PDL1, amplification of Her2neu and mutation of p53 among others. In the present study, the expressions of PDL1, Her2neu and p53 were analyzed in relation to clinicopathological parameters in gastric and gastroesophageal junction carcinoma. Methods: We examined 100 biopsy and resection samples of gastric and gastroesophageal carcinomas for PDL1, Her2neu and p53 protein expressions using immunohistochemistry. Scorings of were done based on intensity and percentage of tumour cells expressing the markers. Results: PDL1, Her2neu and p53 positivity was seen in 37%, 38% and 40% respectively. The analysis showed PDL1 expression was significantly associated with depth of invasion (p=0.0007), nodal metastasis (p=0.0003) and AJCC staging (p=0.0085). Her2neu expression was significantly associated with with histological grading (p=0.0043), Lauren Classification (p=0.0042), depth of invasion (p=0.04) and nodal metastasis (p=0.017). p53 expression was significantly associated with depth of invasion(p=0.01), nodal metastasis(p=0.025) and distant metastsis (p=0.005). Combined analysis of PDL1 and Her2neu showed significant association with histological grading (p=0.017), Lauren Classification (p=0.005), depth of invasion(p=0.0035) and nodal metastasis (p=0.00073). Univariate Cox regression analysis showed that depth of invasion, nodal metastasis, distant metastasis, AJCC staging and p53 were negative prognostic factors for patients' overall survival. In multivariate analysis, distant metastasis and Her2neu expression were independent prognostic factors. Conclusion: PDL1, Her2neu and p53 expression were significantly correlated with advanced clinicopathological features and play an important role in the development of gastric cancer, Prognostication, patients' survival. Targeted therapy against these markers may be beneficial.

   OP35: Assessing microsatellite instability in colon cancer with the help of immunohistochemical markers in a tertiary care center of South India Top

Rushreeta Anjali Deuri, Debasis Gochhait, Biswajit Dubashi

Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India. E-mail: [email protected]

Background: Mismatch repair defect in Colonic carcinoma accounts for approximately 15% of all sporadic carcinomas and has an evident molecular phenotypic presentation, easily identifiable with the help of IHC markers. We wanted to study the morphological and immunohistochemical features of Colon cancers with the help of microsatellite markers such as MLH1, MSH2, MSH6, PMS2 and to correlate clinicopathologically the outcome. Design: Following Institute's Ethics committee approval, 60 cases were collected with complete clinical details from the year Jan'17- Dec'18. Blocks were collected and markers were standardized. The site of the tumor, biochemical, morphological and histological patterns were analyzed, and IHC was performed in 15 cases using the standard protocol. Results: Study population comprised 41 males (68%) and 19 females (32%) with an average of 54.7 years. ALP for most of the patients was elevated (86%) with a mean of 239 U/L. At the time of diagnosis, 34(56%) cases were diagnosed to be stage I/II and 26(44%) cases were diagnosed to be stage III. IHCs were done in 15 cases. 2 cases (13%) showed loss of MLH1 with one case showing concurrent loss of MLH1 and PMS2. 2 cases showed loss of MSH6 (13%) and 4 cases showed loss of PMS2 (26%). Of the 15 cases analyzed, the male to female ratio was 1:1. 67% of cases underwent right colectomy and were diagnosed as stage III. Six cases are still under follow up, rest were lost to follow up. Conclusion: Further markers and correlation with the outcome of the disease are still under progress.

   OP36: Validation of newly introduced, G3 NET in the WHO (2017) classification of gastroentero-pancreastic neuroendocrine neoplasms Top

A Divya, Roopa Paulose

Amrita Institute of Medical Sciences, Kochi, Kerala, India. E-mail: [email protected]

Background: The classification and terminologies of gastroenteropancreatic neuroendocrine neoplasms(GEP-NEN) have evolved over the last two decades and continue to. WHO (2017) classification of pancreatic NEN introduced “G3 well differentiated NEN”, separating it from poorly differentiated neuroendocrine carcinoma(NEC). The genomics of GEP-NEN and NEC are distinctly different, with MEN1, DAXX, and ATRX being drivers in the former and TP53 and RB1 inactivation in the latter. Aim of the Study: To assess the prognostic significance between “well differentiated G3 NEN” and NEC and to determine the association of ATRX and p53 expression with tumor grade and patient survival. Methods: This is a retrospective study of 68 GEP-NENs wherein all G3 cases are reviewed and reclassified based on morphology into G3 NEN and NEC. Immunohistochemical expression of p53 and ATRX is correlated with patient demographics, tumor grade, stage, treatment response and patient survival. Results: The mean age was 50 years and there was male preponderance. Tumour grade and differentiation of the study population are shown in Table 1. Eight (42.1%) of 19 Grade 3 tumors were reclassified as G3 NEN of WHO (2017). There was no statistical difference in the patient survival between the G3 NEN and NEC. IHC for p53 and ATRX is being performed on Grades 2 and 3 NENs and will be correlated with patient survival; Results to follow. Conclusion: Our study did not show significant difference in the survival between G3 NEN and NEC. We hypothesize that p53 and ATRX will help in substratifying G2 and G3 NENs with different outcomes.

   OP37: Expression of Her -2/NEU in primary gastric and gastro-esophageal adenocarcinoma: An experience from a tertiary centre in S. India Top

Aditi Damle, Roopa Paulose, Divya Ail, Wesley Jose1

Amrita Institute of Medical Sciences, Kochi,1Department of Medical Oncology, Amrita Institute of Medical Sciences, Kochi, Kerala, India. E-mail: [email protected]

Introduction: Gastric carcinoma is a common cause of cancer-related mortality. Expression of Her2/neu in advanced gastric / gastroesophageal adenocarcinoma has been shown to have an improved outcome with targeted therapy (trastuzumab). There is limited Indian data on correlation of expression of Her2 neu in gastric cancers with survival. Aims and Objective: To determine frequency of immunohistochemical expression of HER-2/neu in primary gastric / gastro-oesophageal adenocarcinoma, its correlation with various clinicopathological parameters and outcome. Methodology: This was a retrospective study of Her 2/Neu expression in 114 primary gastric/GEJ adenocarcinoma with 5 year follow up. Details regarding patient demographics, tumour site, histological type of adenocarcinoma (Lauren's classification) and tumour stage were obtained from hospital information systen. HER2/neu immunohistochemistry performed in all cases was objectively scored as per current recommendations into 0/1+/2+/3. Data was analyzed using SPSS software. Results: There was strong (3+) Her2/neu immunostaining in 13 (11.4%), equivocal (2+) staining in 9 (7.89%) and negative (1+/0) in 92 patients. Among the Her2/Neu positive (3+) patients, 92.3% were males, mean age at diagnosis was 64.7 years, most common site was GE junction (69.23%), 84.6% were intestinal type of well/moderately differentiation. 76.9% of Her2/Neu positive (3+) had advanced tumor stage and 84.61 % had metastases [Table 1]. The overall survival was 3.7 years in 3+ group and 5.4 years in the other group [Figure 1]. None of the patients were treated with Trastuzumab. Conclusion: Expression of Her2/neu in gastric/GEJ adenocarcinomas though low in our population, was associated with lower survival and can be considered as an independent poor prognosticator.

   OP38: Deregulation of epigenetic modifier Enhancer zeste homolog 2 in gall bladder carcinoma: A clinicopathological study Top

Gayatri Behera, Susama Patra, Tushar S Mishra1, Dilip K Muduly2, Suvradeep Mitra, Suvendu Purkait

Departments of Pathology,1Surgery and2Surgical Oncology, All India Institute of Medical Science, Bhubaneswar, Odisha, India. E-mail: [email protected]

Background: Gallbladder carcinoma (GBC) is the most aggressive malignant tumor of the gastrointestinal tract. Recently, key molecules involves in histone modification especially Enhancer zeste homolog 2 (EZH2) and tri-methyl lysin 27 residue of histone H3 (H3K27me3) have been implicated in the biological behavior of various malignancies. However, there is only limited data regarding their role in GBC. Methods: A total of 31 cases of GBC, 6 cases of dysplasia and 20 cases of chronic cholecystitis (CC) were included. IHC for EZH2 and H3K27me3 was performed in tissue microarray. A scoring system including staining intensity and proportion of positively stained neoplastic cells was used for assessment. Results and Discussion: EZH2 expression was absent in CC and the adjacent non-neoplastic epithelium while half of the cases of dysplasia and 93.5% GBC showed immunoreactivity. The immunohistochemical score was also significantly higher in GBC as compared to dysplasia. The cases with high proliferative activity and advanced stage showed a higher EZH2 expression. In contrast to EZH2, strong and diffuse expression of H3K27me3 was detected in all cases of CC and dysplasia. Although immunopositivity was present in all cases of GBC also, 12.5% of cases showed low expression. Conclusion: The epigenetic modifier EZH2 possibly has an important role in the malignant progression of GBC. As the expression is consistently absent in the non-neoplastic epithelium and rate of immunopositivity is very high in preneoplastic/neoplastic lesions, EZH2 may be an important diagnostic biomarker. Given the reversible property of epigenetic alterations, EZH2 may be a novel target for GBC.

   OP39: Flow cytometric analysis of CD34+ hematopoetic stem cells: A comparison between single and dual platform methodology using ISHAGE protocol at a Top

Meena Mittal, Ashok Panchonia, Shikha Ghanghoria, Astha Dawani, Sachin Sharma

MGM Medical College, Indore, Madhya Pradesh, India. E-mail: [email protected]

Background: ISHAGE set guidelines for detection of CD34 cells , using that an absolute CD34 count is generated by incorporating the leukocyte count from an automated hematology analyser (two-plat method). In this study, we modified the basic ISHAGE method with addition of known number of fluorospheres, to reduce errors inherent to sample washing/centrifugation. Design: CD34+ cell counting results is compared with and without using separate hematology analyzer (single-plat.vs dual-plat.). A 68 samples of peripheral blood, apheresis packs & bone marrow aspiration were analysed and compared using ISHAGE protocol with and without fluorescent microspheres. Dual-platform CD34+ cell counting incorporated data from navious modal 2 L&8 Results and Discussion: Subtle differences in CD34+ cell counting between 2 systems and 2 methods did not achieve statistical significance. ISHAGE guidelines for detection of CD34+ cells are based on a four-parameter FC (CD34PE/CD45PerCP staining, side and forward angle light scatter) thus employing multiparameter gating strategy. With two-platform ISHAGE protocol, an absolute CD34+ count is generated by incorporating the leukocyte count from an hematology analyser. The single-platform ISHAGE method to determine the absolute CD34+ count directly from a FC includes the use of analyzing tubes with a known number of fluorescent beads. Discrepancies of up to 2.5% (two-plat.)--2.6% (single-plat.) in enumeration of CD34+ cells in leukapheresis product and 4.8% (two-plat.)--4.9% (single-plat.) in bone marrow is noted. Conclusion: It is the accurate determination CD34+ cell enumeration, properly validated, can support care for patients undergoing transplants setting. These modifications may improve the inter laboratory reproducibility.

   OP40: CD66c expression in pediatric B-cell aute lymphoblastic leukemia and its role in minimal residual disease Top

Santosh Suman, Sangeeta Pahuja, Sunita Sharma, Shailaja Shukla, Jagdish Chandra

Lady Hardinge Medical College, New Delhi, India. E-mail: [email protected]

Background: Acute Lymphoblastic Leukemia (ALL) is the most common childhood malignancy with majority being B-cell ALL. Minimal Residual Disease (MRD) assessment is one of the powerful prognostic indicator which helps to detect leukemia relapse. The aim of this study was to evaluate the expression of CD66c in pediatric B-cell ALL patients and its role in Minimal Residual Disease (MRD). Design: This hospital based follow up analytical study was conducted on 30 newly diagnosed pediatric B-cell ALL cases. Peripheral blood sample and/ or bone marrow samples were collected at day 0 and day 35 after written informed consent. Flowcytometric analysis was performed on the provided sample on day0 (Basic Panel & LAIPs) and day35 (only LAIPs). Results and Discussion: Mean age of study was 4.8 years with male:female ratio of 2:1. At day 0, expression of CD66c was seen in 50% cases. At day 35, 40% of the total cases were MRD positive and out of these, CD66c was helpful in MRD detection in 58.3% cases. Sensitivity of CD38 underexpression, CD58 overexpression, CD123 in MRD detection was 91.7% for each. Statistically significant (p 0.000) strong positive correlation was seen between CD66c/CD123 and CD66c/CD81. Conclusion: In our study, CD66c was expressed in 50% cases at day 0 and was helpful in predicting MRD in 58.3% of the positive cases which is in concordance with other studies. However, combination of LAIPs (CD38 underexpression, CD58 overexpression, CD66c, CD123 and CD81 expression, etc) is required for better detection of MRD in B cell ALL cases.

   OP41: Primary extranodal lymphoma: A cryptic enemy tertiary care hospital experience of 3 years Top

G P S Gahlot

Army Hospital Research and Referral, Delhi, India, E-mail:[email protected]

Background: Primary extranodal Non-Hodgkin lymphoma (pENL) is a lymphoma with a dominant extranodal compartment with no or minor nodal involvement. This study was aimed to access epidemiological trend, anatomical distribution and histomorphological patterns of pENL in a Tertiary Care Superspeciality Hospital of North India. Design: It was a retrospective study conducted in Department of Laboratory Sciences & Molecular Medicine over a period of 36 months. Clinical details available on requisition form and haematoxylin & eosin stained slides with relevant immunohistochemistry slides were retrieved from archive and analyzed. All cases were classified on the basis of their morphological and immunohistochemical profile according to WHO 2016 Classification. Results and Discussion: pENL cases outnumbered NHL involving lymph nodes & constituted 41.45% (131/316) among all lymphoma and 53.91% (131/243) NHL cases respectively. The mean age in pENL (51.96 years) was slightly lower than corresponding nodal NHL (53.10 years) cases with overall male predominance. Most common sites involved gastrointestinal tract 25.96% followed by head & neck 20.61% with least involvement of urinary bladder 0.77%. Cases of B cell origin 87.79% were dominated over T cell origin 11.46%. Commonest histomorphological type among B and T cell cases were diffuse large B cell lymphoma 67.94% and PTCL, NOS 5.34% respectively. Conclusion: Diagnosis of pENL can be challenging due to their variable clinical presentation, morphological diversities, molecular alteration and lack of uniformity in histopathological classification system. The incidence of pENL is increasing due to better diagnostic immunophenotyping and imaging modalities thus enabling the timely effective management.

   OP42: Review of cases of histiocytic and dendritic cell neoplasms in a referral laboratory over a 5 year period Top

Subhalakshmi Sengupta, Debdeep Dey1

Narayana Superspeciality Hospital, Howrah,1Tata Meducal Center, Kolkata, West Bengal, India. E-mail: [email protected]

Background: Histiocytic and dendritic cell neoplasms are rare malignant tumours and account for about less than 1 % of all tumours arising in lymph nodes and soft tissues. Since these tumours are rare, their true incidence, sex, racial and geographical predilection is yet to be described. They are distinctive diseases with unique biology, clinical presentations, pathology, and treatment options. Morphology and immunohistochemistry evaluation remain key for diagnosing these neoplasms. Design: The computer records of a referral laboratory were searched over a period of 5 years (2013-2018) to detect the incidence of histiocytic and dendritic cell neoplasms. Results: There was one case of histiocytic sarcoma in an eight year-old girl who presented with a thigh mass. Three cases of follicular dendritic cell sarcomas were diagnosed with slight male predilection, two of them had presented with intraabdominal masses. The average age of the patients was 40 years. A 64 years male with a swelling in the axillary region was detected to have interdigitating cell sarcoma. These cases were unifocal and extranodal except in one case of follicular dendritic cell sarcoma which involved the tonsillar lymph node. Morphology together with an appropriate immunohistochemistry panel helped to diagnose these cases. Conclusions: Histiocytic and dendritic cell neoplasms resemble other nonhematopoietic neoplasms, and even reactive processes. Therefore, it needs a high index of suspicion to diagnose them. CD 45 should be included in the immunohistochemistry panel when dealing with a soft tissue sarcoma. Larger studies are required for defining their pathogenesis, treatment protocols and their outcome.

   OP43: Analysis of Epstein-Barr virus and PD-L1 expression in extranodal lymphomas of head and neck region Top

Chandana Rajbongshi, Vandana Raphael, Zachariah Chowdhury, N Brain Shunyu1, Umesh Das2, Purnima Laishram3

Departments of Pathology,1ENT,2Medical Oncology and3Radiation Oncology, North Eastern Indira Gandhi Regional Institute of Health and Medical Sciences, Shillong, Meghalaya, India. E-mail: [email protected]

Introduction: Malignant lymphomas represent 5% of all malignant neoplasms of the head and neck and involve nodal or extranodal sites. Association of extranodal lymphomas of head and neck region with oncogenic virus like Epstein-Barr virus and with immunodeficiency, these tumours are attractive targets for immune-based therapies. Aim and Objectives: This study was undertaken to evaluate if the immunopanel of EBV-LMP1 and PD-L1 is associated with extranodal head and neck lymphomas, and analyse the interaction of PD-L1 with EBV in these tumours. Materials and Methods: 40 patients of extranodal lymphomas were included. Histological typing/immunophenotyping, biopsy sites in head and neck region with clinical presentation, EBV status by IHC expression of EBV-LMP1 and PD-L1 expression in T-cell and B-cell lymphomas and association of PD-L1 with EBV-LMP1 were evaluated. Results: Average age of the patients included was 46.26 years, the male to female ratio being 1.6:1. Histologically, 21 cases were diagnosed as T-cell lymphomas and 19 cases were diagnosed as B-cell lymphomas. PD-L1 immunostain positivity was seen in 33 cases and 7 cases were negative. EBV-LMP1 immunostain positivity was seen in 5 cases and 35 cases were negative. Cases with EBV-LMP-1 expression upregulated PD-L1 in the tumor cells, especially in extranodal head and neck lymphomas, while the reverse could not be said to be true significant. Conclusion: Relatively high incidence of ENKTCL compared to B-cell lymphomas in the north eastern region showed positive correlation between EBV-LMP-1 and PD-L1 expression, which can become an attractive target for immunotherapy.

   OP44: Significance of CD68 positive macrophages and CD8 positive T-cells in clinicopathological profile of nodal diffuse large B-cell lymphoma Top

Soundarya Ravi, J Sreerekha, Debdatta Basu, Smita Kayal

JIPMER, Puducherry, India. E-mail: [email protected]

Background: The role of microenvironment in the behavior of the Diffuse Large B cell Lymphoma (DLBCL) and predicting its treatment outcome can be understood by studying the cytotoxic T cells and macrophages in the tumour. Our aim is to study the clinicopathological profile and significance of CD 68 positive macrophages and CD8 positive T cells in nodal diffuse large B cell lymphoma. Design: The clinicopathological data and treatment outcome of 63 DLBCL patients diagnosed and treated at our institute for the past three years were analyzed. Germinal Centre(GCB) and Activated B cell type(ABC) subtyping was done according to Han's algorithm. CD68 and CD8 immunostain were performed in 27 patients and its association with immunophenotype and treatment outcome is studied. Results: Study group (n=63) had a mean age of 50.34 years with male: female ratio of 2.3:1 out of which there were 23(36.5%) GCB and 40(63.5%) ABC patients. There were 37 (58.7%) patients with Stage III/IV Ann arbor stage and 6 (9.5%) patients had bone marrow involvement at diagnosis. 15(23.8%) patients had relapse in which 11 patients constituted the ABC type(73.3%). Out of the 24(42.6%) patients who attained complete remission (CR), 10(43.5%) were GCB and 14(41.9%) were ABC subtypes. The distribution of CD8 positive T cells and CD68 positive macrophages in DLBCL subtypes has been tabulated below. Conclusion: ABC type is associated with increased frequency of non-remission and relapse. Further immunohistochemistry has to be performed to attain significant correlation between CD8+ T cells and Macrophages with the immunophenotype and treatment outcome.

   OP45: Rapid detection of rifampicin resistance in Mycobacterium tuberculosis by cartridge-based nucleic acid amplification test in a tertiary care center, India Top

Sumit Kumar, Surinder Kumar, Pallavi Kumari1

BPS, GMC(W), Sonepat, Haryana,1PGIMER, RML, Delhi, India. E-mail: [email protected]

Background: India constitutes one tenth of global burden of multidrug resistant tuberculosis (MDR-TB). Detection of MDR-TB takes 6-8 weeks by conventional drug susceptibility Testing (DST). Cartridge-based nucleic acid amplification test (CBNAAT) is based on real time amplification and Mycobacterium tuberculosis resistance is detected within hours. Design: In the present study samples presumptive of tuberculosis since February 2018 to July 2019 were subjected to CBNAAT for the diagnosis of rifampicin resistance. Results: In the present study total number of 3281 presumptive tuberculosis samples were tested by CBNAAT.Out of 3281 presumptive tuberculosis samples 963(29.35%) were mycobacterium positive and rifampicin sensitive and 66(2.01%) were positive for mycobacterium tuberculosis and were rifampicin resistant. Discussion: According to RNTCP report 2018, in the year 2017 total 10,77,377 number of test were performed by 628 CBNAAT machines out of which 37,488(3.48%) were rifampicin resistant tuberculosis. In the study done by R. Tripathi et al. rifampicin sensitive tuberculosis were detected in 35.8% and rifampicin resistant tuberculosis were in 53% samples. However Kumar et al. and Sharma et al. reported 25.8% rifampicin sensitive and 22% MDR respectively. In the study done by Deepak Arora et al. 84.21% samples were rifampicin sensitive and 15.78% samples were rifampicin resistant. In a study done by John Z. Metcalfe et al.28% samples were rifampicin sensitive and 20% samples were rifampicin resistant. Conclusion: Detection of rifampicin resistant tuberculosis by CBNAAT is done within few hours. Consequently early diagnosis of tuberculosis patients helps in early and precise treatment and prevents transmission of MDR strains of tuberculosis in the community.

   OP46: Association of GSTM-1 and GSTT gene polymorphism with histopathological grading in carcinoma breast Top

Sanya Jain, Vijay Kumar Bodal, Medhavi Dhir, Kuldeep Singh Ahi, Mohanvir Kaur, Siddharth Sharma1

GMC,1Thapar University, Patiala, Punjab, India

Background: Breast cancer possess a serious health problem as it is the most common invasive cancer in females world-wide. Apart from BRCA-1 and BRCA-2, two other breast cancer susceptibility genes have been identified, i.e. GSTM-1 and GSTT gene polymorphism. These two belong to a family of glutathione S-transferase enzyme which are Phase II intra cellular enzymes that catalyze cell detoxification. These genes are highly polymorphic, and any genetic variation changes an individual's susceptibility to carcinogen and toxins , increasing the risk of neoplasia. Design: 5 ml of venous blood sample from 100 histopathologically confirmed cases of carcinoma breast was collected for DNA extraction. The extracted DNA was subjected to multiplex PCR amplification of GSTM-1 and GSTT gene polymorphism. Results and Discussion: Out of 24 Grade 1 tumors, 18 were positive for GSTM1 and 17 were positive for GSTT.

  1. Of 48 grade 2 tumors, 30 were positive for GSTM1 and 28 were positive for GSTT1
  2. Out of 28 grade 3 tumors, 8 were positive for GSTM1 and 8 were positive for GSTT1.

Conclusion: In the population based study,GSTM-1 and GSTT gene polymorphism significantly increased the incidence of carcinoma breast. It provides both prognostic & therapeutic information and can be used as a tool in making strategy and planning for targeted therapy in breast carcinoma.

   OP47: Association of HLA-A, B, DR alleles in breast cancer Top

Neeti Nagar, Mukul Singh, Rashmi Arora

Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India. E-mail: [email protected]

Background: Tumor specific markers has critical role in early diagnosis and treatment of cancer. HLA molecules plays an important role in immunity and tumor response of the body. Widespread geographical distribution of HLA pleomorphism warrants regional studies to demonstrate alleles found in breast cancer. Aim to this study is to see the association the HLA -A, B, DR alleles in breast cancer cases. This association can provide potential area for targeted chemotherapeutic intervention. Design: Cross sectional study with the objective of studying the association of HLA-A, B, DR alleles in breast cancer. Ethylenediaminetetraacitic acid (EDTA) blood samples were taken from 30 histologically diagnosed cases of breast cancer and 30 healthy control for the study. HLA typing was performed using PCR single specific primer (Fluogene-a flourescent based HLA gene amplification detection method). Result and Discussion: In our study breast cancer cases were associated with HLA-DRB3, HLA-DRB4, HLA-DRB5*, as compared to controls which showed no association.HLA-DRB3* also showed an association with triple negative breast cancer. Breast cancer has a tendency of familial accumulation, suggesting that genetic factors might have role in etiology. Many studies emphasizing the relationship between HLA system and breast cancer. Identification of HLA alleles having role in breast cancer can provide a potential area for targeted chemotherapeutic intervention. Conclusion: Early diagnosis and treatment of breast cancer is evolving rapidly with advancing technology including genetics. So HLA alleles frequencies of different populations should be investigated for assessing important alleles, specified as risk factors of development of breast cancer.

   OP48: Association of HLA Class I (A and B) and HLA Class II (DR) alleles with autoimmune thyroid diseases Top

Divya Sethi, Mukul Singh, Krishna Biswas, Rashmi Arora

Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India. E-mail: [email protected]

Background: In iodine-replete areas, most people affected with thyroid disorders have autoimmune thyroid diseases, which include Hashimoto's thyroiditis and Graves' disease. HLA alleles have been associated with multiple autoimmune disorders. Some HLA alleles have higher affinity for autoantigenic thyroidal peptide and are associated with autoimmune thyroid disease. Aberrant HLA expression leads to activation of thyroid auto-reactive T lymphocytes. Various studies have been conducted worldwide; however, there is paucity of literature from India. Objective: To compare frequency of HLA CLASS I (A, B) and HLA Class II (DR) alleles in autoimmune thyroid disease with controls (healthy). Design: 22 cases and 22 controls were recruited for comparative cross-sectional study. Cases were either anti –TPO positive or FNA positive for lymphocytic thyroiditis. DNA was extracted with Innotrain kit and HLA typing was performed with Fluogene. Data was analysed using SPSS software version 21.0. Quantitative data was summarized as mean and standard deviation while qualitative data was summarized as proportion. Results and Discussion: Among cases, HLA A*02 was found in 13 instances, HLA B*40 in 10 instances and HLA DRB1*15 in 13 instances. These HLA alleles were found to have higher frequencies than in controls. Hence, these HLA alleles may be risk factors for the development of autoimmune thyroid disease. Conclusion: This study helps us identify susceptible population in Indian setting.

   OP49: Association of HLA-A, HLA-B and HLA-DR alleles in oral squamous cell carcinomas in Indian population Top

Juhi Shahab

VMMC and Safdarjang Hospital, New Delhi, India. E-mail: [email protected]

Background: Oral squamous cell carcinoma (OSCC) is the most common malignant epithelial neoplasm affecting the oral cavity. It is the 3rd most common cancer in India. Several HLA types are associated with an increased risk of various immunologically mediated diseases.There is also evidence that the HLA gene complex may mediate susceptibility to or protection from malignancies. Various studies have been conducted worldwide but there is paucity of literature in India regarding association of HLA alleles in oral squamous cell carcinoma patients. Objective: To study the association HLA- A, HLA-B and HLA-DR alleles in oral squamous cell carcinomas in Indian population. Design: HLA class I (HLA-A and HLA-B) alleles and class II (HLA-DR) alleles were determined in 50 patients with OSCC and the equal number of healthy controls (50) were taken for comparison.. DNA was extracted with Innotrain kit and HLA typing was performed with Fluogene. Data was analysed using SPSS software version 21.0. Results and Discussion: All the cases were compared with the healthy controls and the data was correlated with clinical stage of the oral squamous cell carcinoma. HLA A*24 and HLADRB1*15 alleles were found to have higher frequency than in controls. Conclusion: Thus this study helps us to establish the possible relationship between HLA-A,B and DR alleles in Oral squamous cell carcinoma patients about which there is paucity in Indian literature to the best of our knowledge.

   OP50: Molecular typing of HLA in renal transplant donors and recipients and correlation with various parameters at a tertiary care centre Top

Indu Yadav, Anjali Sharma, Mukul Singh, Rashmi Arora

Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India. E-mail: [email protected]

Background: Kidney transplant is one of commonest transplantation occurring around the world. The aim of this study is to detect most common HLA genotypes seen in End stage chronic kidney disease (ESCKD) patients. The HLA matching between the renal transplant recipients and donors would be correlated with Panel reactive antibody estimation and various other clinical parameters. Design: 40 patients of ESCKD and 40 donors (healthy controls) were included in the study. HLA A, HLA B AND HLA DR genotyping was performed using PCR-SSP (Fluogene -a fluorescence based HLA gene amplification detection system). The clinical and demographical data was collected and correlated. Results and Discussion: The most prevalent genotypic expression found in ESCKD patients was HLA-A*11,*24; HLA-B *40,*15 and HLA-DR*15,*12. Genotyping expression of MHC class I and II genes were compared between donors and recipients and was statistically correlated with patient's pre transplantation panel reactive antibody estimation and the various other clinical parameters like CMV immune status, frequency of dialysis undertaken, history of previous blood transfusions. Conclusion: Susceptible association of HLA alleles and various other parameters with ESCKD in a tertiary care centre was noted which may help in prognostication.

   OP51: Role of C4d in diagnosis and prognosis of native renal diseases Top

Satyam, Varsha Kumar, Vatsala Misra, P A Singh

MLN Medical College, Prayagraj, Uttar Pradesh, Inida. E-mail: [email protected]

Background: C4d is a biomarker of the complement cascade and has a role in the diagnosis of antibody mediated rejection in solid organ transplantation. It's role in disease progression has been studied recently in few native renal diseases. Design: A cross-sectional study was done at our hospital which included 50 cases of native renal diseases. Biopsy was stained with hematoxylin and eosin and periodic acid Schiff. Other stains were applied as per requirement. Immunofluorescence was performed with FITC conjugated anti IgG & C3. Further immunohistochemical staining for C4d was done and interpreted. Semi quantitative scoring of chronicity and activity index was done as required. Results and Discussion: Membranous glomerulopathy, Lupus Nephritis, Proliferative GN demonstrated C4d positivity along the glomerular capillary wall whereas no such staining was observed in MCD. Mesangial deposits of C4d was observed in IgA nephropathy. Positive correlation was noted between glomerular C4d and IF deposits of IgG & C3. The result is found to be statistically significant (p<0.05). Positive correlation between total C4d score and total chronicity index, interstitial fibrosis, tubular atrophy, S. creatinine was seen and found statistically significant (p < 0.05). Similarly positive correlation was seen between tubular C4d score and serum creatinine, glomerular C4d and 24hr urinary protein. Conclusion: C4d is a promising marker in the diagnosis and prognosis of renal diseases. A better understanding of the role of complements is required to know the exact pathway of complement activation in glomerular disease and to aid in diagnosis.

   OP52: Pauci-immune crescentic glomerulonephritis: Experience from a tertiary care hospital in India Top

Pallav Gupta

Sir Ganga Ram Hospital, Delhi, India. E-mail: [email protected]

Background: Pauci-immune crescentic glomerulonephritis (PICN) is characterized clinically by rapidly progressive renal failure developing over a period of days to weeks. On serology 80-90% patients have ANCA positivity however ANCA negative serology has be seen in 30-40% cases in various studies. Materials and Methods: Renal biopsy electronic records were retrospectively analyzed from January 2014 to December 2018. Renal biopsy slides were reviewed. Immunofluorescence findings, biochemical findings, ANCA serology findings by both indirect immunofluoroscence (IIF) and ELISA were retrieved from hospital records. Results: Mean age of the patients was 48 years. The mean serum creatinine at presentation was 6.57±4.31. ANCA positivity was seen in 41 patients (64. All the positive cases also tested positive on ELISA. 23 patients (36%) had ANCA negative serology on repeated testing. On histopathology, 49 patients had cellular to fibrocellular crescents whereas 15 patients had predominantly fibrous crescents. All but 5 patients had diffuse crescents (>50% glomeruli with crescents). Periglomerular or interstitial granulomas were identified in 3 patients and 13 patients had fibrinoid necrosis accompanying crescents. Extrarenal manifestations were seen in 16 patients. Follow up was available in forty patients (follow up duration 6-60 months) of which 4 patients died and 4 developed end stage renal disease (ESRD). Conclusion: ANCA associated PICN is the most common cause of rapidly progressive glomerulonephritis. Granuloma and vasculitis are infrequently seen in patients with PICN. Renal biopsy along with immunofluorescence plays an essential and important diagnostic role and is helpful in guiding treatment and accessing prognosis.

   OP53: Vascular endothelial growth factor expression in diabetic nephropathy: A clinicopathological study Top

Sagarika Sharma

JSS Hospital, Mysuru, Karnataka, India. E-mail: [email protected]

Introduction: Diabetes is a disease of epidemic proportions, and the number of people developing the disease is growing every year. The evolution of DN occurs over a period of 10–20 years, beginning from microalbuminuria and progressing to end-stage renal disease (ESRD). Diabetic nephropathy is a significant medical problem because of its increasing incidence, morbidity, and mortality. New treatments targeting cytokine imbalances in a timely and specific manner could go a long way in the prevention of development and progression of DN. Methods: 56 patients were studied and classified according to “pathologic classification of DN” by Tervaert et al. Data on histological, clinical parameters were calculated and correlated. Glomerular and tubular staining of VEGF was recorded. P values of less than 0.05 were considered statistically significant. Results: Of the 55 patients studied, 8 were class II, 24 were class III, and 23 were class IV. VEGF was positive for 6(75%) of class II, 17(70.83%) of class III, 8 (34.7%) of class IV cases. P value was found to be significant with the class of DN and serum creatinine, hypertension. It was also significant between VEGF positivity and DN class. Conclusion: An accurate estimate of damage in DN can only be achieved by the histological analysis of tissue samples. It is important to identify and differentiate the various pathologies at an early stage in order to prevent progression and potential complications. VEGF expression are increased at the onset of diabetes, can help provide early diagnosis and prevent progression to ESRD.

   OP54: Microsatellite instability in pediatric glioblastomas Top

Sheena Alphones, Paromita Roy

Tata Medical Centre, Kolkata, West Bengal, India. E-mail: [email protected]

Background: Microsatellite instability (MSI) is implicated in various cancers for their increased sensitivity to checkpoint inhibitor drugs and hereditary cancer syndromes. There are very few reported studies on MSI in pediatric high grade gliomas (pHGG). Aim: To evaluate the prevalence of MSI in pHGG using tissue microarray based immunohistochemical techniques. Materials and Methods: Paraffin blocks of consecutively reviewed pHGG (< 18 years), from May 2011 - 2019 were retrieved and immunohistochemical staining of one 2mm core from each case was done for 4 MSI antibodies (MLH1, PMS2, MSH2 and MSH6). Whole sections were stained in cases with scant tissue. The study was approved by the institutional review board (2019/TMC/166/IRB45). Results: A total of 10 cases were studied. Male: female ratio was 7:3. Average age of incidence was 10.5 years (range 8-16 ). Five cases showed strong diffuse staining for all 4 markers. In 2 cases there was focal weak staining of tumour cells due to processing artefacts in outside blocks (interpreted as positive). In one case MSH2 and 6 were lost and in 2 cases MLH1 and PMS2 were lost, so 30% of the cases showed immunohistochemical evidence of MSI. In one case internal control was also negative and it was interpreted as Congenital Mismatch Repair Deficiency (cMMRD). Conclusion: This is a preliminary attempt of assessment of prevalence of MSI in this rare subgroup of pHGG. There is no prior reported literature in India and further multi-center collaborative studies are imperative.

   OP55: Histomorphological study of spectrum of diffuse astrocytomas and oligodendrogliomas with expression of isocitrate dehydrogenase-1 mutation Top

Pranshu Sisodia, Harendra Kumar, Garima Dundy, Chandrakanta, Rajni Bharti, Alok Kumar Gupta

S.N Medical College, Agra, Uttar Pradesh, India. E-mail: [email protected]

Background: Diffuse astrocytic tumors account for approximately 60% to 80% of all gliomas and about 30% of all CNS tumors in the adult population. The 2016 update of the 2007 WHO classification breaks old tradition of classification of brain tumors based on concepts of histogenesis and incorporates well-established molecular parameters into the classification of diffuse gliomas, which are now grouped together on the basis of isocitrate dehydrogenase (IDH1 and IDH2) genetic status. Design: The present study was cross-sectional type and aimed at histomorphological spectrum of diffuse gliomas in adult population, to assess expression of IDH-1 mutation and correlation with tumor type & grade. It included intracranial tumors with the clinical & radiological diagnosis of diffuse astrocytomas & oligodendrogliomas during the study period from January 2018 to June 2019. Total 50 cases of diffuse astrocytic and oligodendroglial tumors were studied. The histopathological confirmation of diffuse gliomas was made on routine H&E and processed for IDH-1 immunostain (Dako & Dianova). Results and Discussion: The mean and median age of patients was 43.8 years and 42 years respectively. Male to female ratio was 3.1:1. Frontal lobe was the commonest site of involvement (13 cases, 26%). Maximum cases were of glioblastoma (24 cases, 48%). IDH-1 mutation was found in 44 cases (88%). Oligodendrogliomas and anaplastic astrocytomas were having highest percentage of IDH-1 positive cases of each (100%). Conclusion: IDH status is important from the prognostic & therapeutic point of view, as it will presumably guide the treatment of biologically similar brain tumors.

   OP56: Study of expression and role of isocitrate dehydrogenase 1, ATRX and P53 mutation in gliomas Top

Susmita Sarma, Yookarin Khonglah, Jaya Mishra, Arindom Kakati1, Pranjal Phukan2

Departments of Pathology,1Neurosurgery and2Radiology, NEIGRIHMS, Shillong, Meghalaya, India. E-mail: [email protected]

Background: Gliomas account for 45% of all intracranial tumors. Newer technologies have allowed deeper analysis of many gliomas leading to the discovery of IDH mutations and their association with markers like ATRX, p53 for better diagnosis, prognosis in gliomas. The present study analyzes the expression of IDH1, ATRX and p53 in different types and grades of gliomas. Correlation with various clinico-pathological parameters was done. A follow up to prognosticate gliomas was also attempted. Materials and Methods: This is an ambispective study from last 5 years. Immunohistochemistry for IDH1, ATRX and p53 were done and reported based on percentage of tumor cells expressing the markers. Results: Total of 53 glioma cases were obtained fulfilling the inclusion and exclusion criteria. Patient's age ranged from 10 to 53 years. M:F ratio was 1.3:1. 88%: Diffuse Astrocytoma, 80%: Anaplastic Astrocytoma, 90%: Oligodendroglioma,60%: Anaplastic Oligodendroglioma and 54%: Glioblastoma showed IDH1 positivity. Significant association between positive IDH1 expression & low grade gliomas (p=0.028) and loss of ATRX expression with histological lineage (p=<0.0001) was found. Combined analysis of expression of IDH1, ATRX versus IDH1, ATRX and p53 with WHO grade showed significant association. Follow-up of 32 patients were obtained of which 24 were IDH1 positive. 22/24 were alive with median survival of 21.5 months (92%). IDH1- cases were 8 of which 5 were alive with median survival of 15.8 months (62%). Conclusion: Majority of the gliomas expressing IDH1 mutation are associated with improved survival. Combined analysis of IDH1, ATRX and p53 aids in diagnosis, prognosis and further therapy of gliomas.

   OP57: CAPN3 gene expression in limb girdle muscular dystrophy 2A with special emphasis on its mutations Top

Sukanya Banerjee, B D Radotra, Rakesh K Vashishta, Manoj K Goyal1, Manni L Guptasarma2

Departments of Histopathology,1Neurology and2Immunopathology, PGIMER, Chandigarh, India. E-mail: [email protected]

Background: Limb Girdle Muscular Dystrophy (LGMD) is a heterogeneous group of neuromuscular disorders which are autosomally inherited. Among 30 types of LGMD, Calpainopathy (LGMD2A) is an autosomal recessive muscle disease which occurs due to mutations in CAPN3 gene. Patients are clinically diagnosed with proximal muscle weakness in young age and usually accompanied by bilateral scapular winging. We aim to study expression of CAPN3 gene and its correlation with genetic analysis. Design: The study includes 15 patients with clinical and histopathological features of LGMD and 8 control subjects. The biopsies were evaluated using Enzyme histochemistry, Immunohistochemistry followed by Real time PCR, Sanger sequencing of 8 exons of CAPN3 gene (1,3,5,6,7,19,21,22). Results and Discussion: Among 15 patients with clinical and pathological features suggestive of Calpainopathy, all showed significantly reduced CAPN3 gene expression. Additionally one patient showed mutation in exon 21 (c.2260G>A; p.754A>T). Another patient showed splice site changes in exon 19 of CAPN3 gene (c.2051-1G>T). Mutation in exon 22 was also observed in two patients (c.2338G>C; p.780D>H). Conclusion: Reduced expression of CAPN3 gene indicates that the gene is down regulated in patients with LGMD2A. Alterations in exon 19, 21, 22 of CAPN3 gene further confirms the down regulation of this gene in LGMD2A patients.

   OP58: Profile of irregular antibodies in blood transfusion recipients Top

Abhilasha Yadav, Aparna Bhardwaj, Gaurav Raturi

Shri Guru Ram Rai Institute of Medical and Health Sciences, Dehradun, Uttarakhand, India. E-mail: [email protected]

Background: Several irregular red blood cell alloantibodies are produced by alloimmunization of antigens, are clinically important because they cause hemolysis in neonates and in transfused patients. According to the American Association of Blood Banks Standards, antibody screening plays a pivotal role in pre-transfusion testing to reduce incompatibility in most blood centres. Designs: This is a prospective study conducted at the Blood Bank of a tertiary care centre in Dehradun over a period of two years. Blood samples of 1000 individuals comprising exposed and non exposed individuals were collected and were submitted for ABO, RhD grouping, Direct and Indirect Coombs Test. Antibody screening and identification using gel technique was further performed in case of any unexpected reaction. Results and Discussion: Of 1000 samples, antibodies were identified in 57 (5.7%) cases. These cases were further subdivided into exposed (480 cases) and non-exposed cases (520 cases).Of the exposed group that included 480 cases comprising individuals with repeated transfusions and multiple pregnancies, antibodies were identified in 45 (9.3%) with multiple alloantibodies being observed in those who had received more than five transfusions. Of 520 non -exposed cases, antibodies were identified in 12(2.3%) and majority of them had single alloantibodies. Of 57antibody positive cases majority of them (50cases, 87.7%) were females with Anti D alloantibody being the commonest (31.5%). Conclusion: Since the risk of alloimmunization is more common in multitransfused patients it is advisable to screen at least those cases for irregular antibodies.

   OP59: Study of immunohistochemical expression of epidermal growth factor receptor and its association with epithelial mesenchymal transition markers (e-cadherin, vimentin) in primary head and neck squamous cell carcinoma Top

Nita Yadav, Kiran Agarwal, Sunil Kumar1

Departments of Pathology and1ENT, Lady Hardinge Medical College, New Delhi, India. E-mail: [email protected]

Objectives: To study immunohistochemical expression of epidermal growth factor receptor and its association with epithelial mesenchymal transition markers (e-cadherin, vimentin) in primary head and neck squamous cell carcinoma. Methods: Hospital based descriptive observational cross sectional study was performed in 50 newly diagnosed histopathologically proven cases of primary head and neck squamous cell carcinoma after bilingual written consent. The biopsies were collected and immunohistochemistry was performed for EGFR, E-cadherin and vimentin. Analysis of immunohistochemical scoring was done for the markers. Results: Out of 50 cases, 86% cases showed significant positive EGFR expression (p<0.01). High EGFR expression (score 3,4) was found in 24(55.81%) and low EGFR expression in 19(44.19%) cases. No significant association of EGFR expression was seen with histological grade of tumour. There were 11(84.61%) cases which were EGFR positive and showed EMT transition (E-cadherin negative, vimentin positive). 22(88%) EGFR positive showed E-cadherin positive and negative vimentin expression. However no significant association was seen between EGFR and EMT markers in our study (p=0.256). Conclusions: Significant positive EGFR expression was observed in cases of HNSCC. However EMT markers did not show any significant correlation with EGFR expression and clinicopathological parameters.

Keywords: Epidermal growth factor receptor, epithelial mesenchymal transition, head and neck squamous cell carcinoma, immunohistochemistry

   OP60: Morphology and morphometry of umbilical cord vessels in preeclampsia and gestational hypertension Top

Kavitha Kannan, Manjiri Phansalkar, Moses Ambroise, A Padma

Pondicherry Institute of Medical Sciences, Puducherry, India. E-mail: [email protected]

Background: The placenta plays a key role in the pathogenesis of preeclampsia (PE). There are numerous studies in the literature about the morphological and morphometrical alterations of the placenta in PE. But there is a lack of studies on these alterations in umbilical cord vessels. Hence we undertook this study on the significance of morphological and morphometrical changes in umbilical cord vessels. Objectives: To study the morphological and morphometrical changes in umbilical cord vessels in gestational hypertension and PE in comparison with the umbilical cord of normal pregnancy. Design: 60 cases (control – 30, case including PE and gestational Hypertension-30) were studied. Haematoxylin & Eosin and Verhoeff VanGiesonstained slides were examined for morphological changes. Morphometric parameters like vessel area, wall area, lumen area, wall lumen ratio of both arteries and vein were assessed by using Image J software. The severity of changes were correlated with gestational age, birth weight, and other clinical parameters. Results: In the case group, the umbilical vein shows duplication and fragmentation of internal elastic lamina (IEL). Most of the arteries of the case group show remnants of IEL. Additional microscopic findings like retraction spaces and perivascular hemorrhages are present in the case group. Morphometry variables like arteries wall lumen ratio (p-value -0.010) and vein wall lumen ratio (p-value -0.039) are reduced in the case group compared to controls. Conclusion: The Present study reveals various alterations in the umbilical cord vessels which could affect fetal development. Morphometry is more accurate in highlighting the subtle changes in blood vessels.

   OP61: Comparison of expression of PD-L1, Ki 67 and p53 in dysplasia and squamous cell carcinoma of the oral cavity Top

Monica Rajamuneeswaran, Chintamani Pathak, Rashmi Arora

VMMC and Safdarjung Hospital, New Delhi, India. E-mail: [email protected]

Background: Oral cancer is one of the most prevalent cancers worldwide. Squamous cell carcinoma represents >90% of oral cancers. PD-L1 is an immune check point molecule that attenuates the immune response. Its high expression is associated with lower immune response to tumor leading to poor prognosis. Ki-67 is a proliferation marker whose increased expression is associated with increasing grades of tumor. High expression of p53 (tumor suppressor) is associated with worse prognosis. Limited literature is available regarding the comparison of expression of PD-L1, Ki-67 and p53 in oral dysplastic lesions and squamous cell carcinoma. This study aims to fill that lacunae by using immunohistochemistry. Objectives: To compare the expression of PD-L1, Ki-67 and p53 in oral dysplastic lesions and squamous cell carcinoma. Design: 20 cases each of oral dysplasia and squamous cell carcinoma were taken in this study. Immunohistochemistry was performed for PD-L1, Ki-67 and p53 in all cases. Results: Ki-67 and p53 showed faint positivity in dysplasia while increased positivity was seen in SCC cases. 6 of the 20 cases of SCC were faintly positive for PD-L1 while all the dysplastic lesions were negative. Conclusion: Ki-67 and p53 expression increases as the lesion progresses from dysplasia to carcinoma. Though PD-L1 was faintly positive in few of the SCC cases, the results were not statistically significant. Considering the sample size taken for pilot study, further analysis has to be done to make the results more valid.

   OP62: Histomorphological study of lesions of nose and paranasal sinuses with special reference to status of human papilloma virus in neoplastic lesions Top

Akhilesh Singh, Vatsala Misra, Varsha Kumar1

Department of Pathology, MLN Medical College, Prayagraj, Uttar Pradesh, India. E-mail: [email protected]

Background: HPV has been found to be associated with various lesion of head and neck region. However its role has not been studied in detail in neoplastic lesions of nose and paranasal sinuses. Therefore present study has been done to study histomorphology of these lesions in detail and find out prevalence of HPV in neoplastic lesions. Design: It was combind study of prospective and retrospective type. Specimen (biopsy/surgical) was received in department of pathology from last 5 years including 2 year of prospective specimens. Slides and blocks of retrospective cases was taken out and re-evaluate in details. Prospective specimens were fixed in 10% formalin, processed by conventional methods. 3μ section taken and stained with hematoxylin and eosin. IHC for HPV was done for neoplastic lesions. Results and Discussion: Out of 140 cases, 77 (55%) were non-neoplastic and 63(45%) were neoplastic. The commonest site was nose, followed by PNS and nasopharynx with male predominace in all lesions. Nasal polyp was the commonest non-neoplastic lesion followed by angiofibroma in benign neoplasm (30%) and SCC in malignant neoplasm (15%). The commonest age group was 2nd and 3rd decades for non-neoplastic & benign lesions and 5th to 7th decade for malignant neoplasm. HPV positivity was seen in inverted papilloma (40%), SCC (20%) and ACC (75%). Conclusion: Nasal polyp was commonest non-neopastic lesion followed angifibroma in benign and SCC in malignant neoplasm. HPV may have a role in causation of benign papillomatous and neoplastic lesions of nose and PNS.

   OP63: Association of human papilloma virus and Epstein Barr virus with squamous cell carcinoma in upper aerodigestive tract Top

Lity Dhar

Maulana Azad Medical College, New Delhi, India. E-mail: [email protected]

Background: Squamous cell carcinoma (SCC) of upper aerodigestive tract (UADT) has been showing changing trends in its incidence showing an increasing trend, particularly in younger age group in the absence of risk factors like tobacco and alcohol. This shift has been attributed to the infection by viruses. Design: Immunohistochemistry was performed using p16, LMP1, p53, p63 in paraffin sections of 100 diagnosed cases of SCC of UADT. Results: The median age for both HPV & EBV was 49.5 years with M:F ratio for HPV 3.8:1, while for EBV 37:1. HPV & EBV positivity were found in 29% & 38% cases respectively. Oral cavity comprised of 32% & 41.9% for HPV & EBV respectively. 4% cases co-expressed both. 30.9% and 32.3% MDSCC cases were positive for HPV & EBV respectively. HPV involved stage 1 SCC while EBV involved stage 4a most commonly. HPV+ve cases showed decreased p53 expression. p63 expression of higher grade was noted in both HPV+ve and EBV+ve cases. Conclusion: Oral cavity was the commonest site involved with male predominance for both HPV & EBV. It was detected more in patients <50 years. MDSCC was the commonest grade involved. HPV involved lower stages, EBV was more seen in higher stages SCC. An inverse relation was noted between HPV and p53 expression while increased p63 expression was noted in both HPV & EBV positive cases. A co-infection of 4% was seen. A larger study with longer follow up is required to study the association of viruses and predict the prognosis better.

   OP64: Morphological spectrum and IDH 1/2 mutation status in sinonasal undifferentiated carcinoma Top

Balamurugan Thirunavukkarasu, Amanjit Bal, Parul Gupta, Sandeep Kumar, Ashim Das, Naresh Panda1, Sushmita Goshal2

Departments of Histopathology,1Otorhinolaryngology and2Radiotherapy, Postgraduate Institute of Medical Education and Research, Chandigarh, India. E-mail: [email protected]

Background: Sinonasal undifferentiated carcinoma (SNUC) is a high-grade malignancy with limited treatment options and poor outcome. Uniform diagnostic criteria are not well established for this variant. Also, there are no recurrent genetic alterations identified except for recently explored IDH 1/2 mutations. Aim: To decipher morphologic spectrum of SNUCs and look for prevalence of IDH1/2 mutations. Materials and Methods: Retrospective analyses of sinonasal carcinomas were done from a period of 2005 – 2019. Cases were analyzed by two histopathologists. Immunohistochemistry panel including p40, panCK, INI-1, CD56 and CK-7 were performed in all cases. IDH1/2 mutations were analyzed in sinonasal undifferentiated carcinoma by Sanger sequencing. Results: A total of 147 cases were identified. p40 positive cases with squamous differentiation (n = 109), adenocarcinoma (n=18) and one case of neuroendocrine carcinoma and 13 cases of undifferentiated carcinoma (p40 negative; CK positive). Six cases had inadequate tissue in paraffin block. For SNUCs, the mean age was 53.6 years (25-68 years). Most common site was nasal cavity (9) followed by ethmoid sinus (2) and maxilla (1) and extension to nasopharynx in one case. Twelve cases showed sheets of undifferentiated cells with tumour necrosis. One case showed focal rhabdoid morphology and was INI-1 deficient. IDH1 mutations was noted only in one case c.273 G>A on exon 4. Conclusion: The diagnosis of SNUC should be made after exclusion of other epithelial malignancy after an extensive panel of IHC. IDH1 mutation, though rare, should be looked for as they have significant implications for therapy with IDH inhibitors.

   OP65: Comparison between digital image analysis and visually assessed ki67 labelling index: A study of 120 cases Top

Kajal Angural

Dayanand Medical College and Hospital, Ludhiana, Punjab, India. E-mail: [email protected]

Introduction: The Ki67 staining is frequently used in oncology to estimate a tumor's proliferation index. Using immunohistochemistry of a biopsy, cells are scored as Ki67 positive or negative, and the biopsy is assigned a percent Ki67-positive value. Objectives: Immunohistochemical Ki67 labelling index (Ki67 LI) reflects proliferative activity and is a potential prognostic/predictive marker of carcinomas. However, its clinical utility is hindered by the lack of standardized measurement methodologies. Besides tissue heterogeneity aspects, the key element of methodology remains accurate estimation of Ki67-stained/counterstained tumour cell profiles. This study aims to compare visual method and digital image analysis (DIA) and to develop a methodology to ensure and improve accuracy of the DIA approach. Materials and Methods: Histopathologically diagnosed malignancies in the department of pathology, Dayanand medical college and hospital, Ludhiana, further subjected to ki67 IHC marker for final evaluation and categorisation of the type of tumor. This study includes 120 such cases starting from January 2019. In our study, we examined whether Ki67 LI in cancers calculated using image analysis software correlates with that measured on the basis of visually. Conclusion: The merits of calculating Ki67 LI using immunoratio DIA are: the staining intensity of the cells to be counted can be adjusted, the portion of a tumor including “hot spots” for counting can be chosen and also many cancer cells can be counted more rapidly using immunoratio DIA than by visual observation. This method is particularly important to categorise neuroendocrine tumors.

   OP66: Accuracy of intraoperative frozen section in assessing margins in oral cancer resection: A tertiary care hospital based study Top

Surabhi Pandey

SRMSIMS, Bareilly, Uttar Pradesh, India. E-mail: [email protected]

Introduction: To study the accuracy of intraoperative frozen section diagnosis in assessing margins in oral cancer resection. But reliability of frozen sections in predicting the ultimate final margin status is unknown. We compared frozen and permanent reads to identify risk factors for overall discrepancies between intraoperative and final margin status. Aims and Objective: To evaluate the role of frozen section diagnosis as a guide in resection of oral cancer lesions. Materials and Methods: Pathology reports of 112 surgical resections between 2018 and 2019 were retrospectively reviewed. A total of 54 cases, met inclusion criteria. The reports of frozen sections are compared with the results of histologic findings obtained by routine paraffin embedded Haemotoxylin and eosin method. Results: The total of 264 margins, from 54 patients were examined. 251 margins showed concordance with the permanent section of the same tissue, an accuracy rate of 95.07%. However, 4.93% of cases had a final positive margin not detected by frozen section.

   OP67: Correlation of p16 and galectin 3 expression in squamous cell carcinomas of buccal mucosa Top

Shramana Mandal, Deepika, Meeta Singh, Nita Khurana, Vikas Malhotra

Department of Pathology and ENT, Maulana Azad Medical College, New Delhi, India. E-mail: [email protected]

Background: The incidence of head and neck squamous cell carcinoma (HNSCC) has been gradually increasing over the last three decades. Galectins are non-integrin β-galactoside -binding lectins. The galectin-3 expression is lost HNSCC p16INK4A is a gene associated with controlling the cell cycle in canceration and its expression increases from dysplasias to squamous cell carcinomas. Aim: To study the expression and correlation of p16 and galectin 3 in oral squamous cell carcinomas (buccal mucosa). Design: We evaluated 60 patients of buccal squamous cell carcinoma. Immunohistochemical (IHC) analysis was performed using avidin- biotin complex method. Tissue were stained for p16 and galectin 3. Results and Discussion: The age of the patients ranged from 27 to 70 years of age. There were 48 Male and 14 female. All the cases were of Squamous cell carcinoma with moderately differentiated being the most common type (40 out 62). The p16 expression was noted in 45% of the cases and correlated with the grade of tumour (P=0.05). The expression of galectin 3 did not correlate with grade and that of p16 expression. Conclusion: P16 is a surrogate marker of HPV and these carcinomas are associated with a favourable prognosis. It may used for early detection of cancer and may also help in therapy in patients of HNSCC.

   OP68: Benign vascular anomalies: An attempt for transition from morphological to etiological classification Top

Lavleen Singh

Chacha Nehru Bal Chikitsalaya, Delhi, India. E-mail: [email protected]

The International Society for the Study of Vascular Anomalies (ISSVA) devised a multidisciplinary etiopathogenesis based approach to classify benign vascular anomalies into tumors and malformations. This classification scheme has major therapeutic and prognostic implications as treatment modalities differ for both the categories. Inappropriate usage of term “hemangioma” for etiopathogenetically distinct entities is commonly seen in clinical practice leading to delivery of incorrect treatment to the patients. We aimed to study the histomorphological and immunohistochemical features of benign vascular anomalies for their precise histopathological classification. A total of 44 cases diagnosed over a period of 3.5 years were reviewed, and reclassified into 3 major groups i.e. infantile hemangioma, congenital hemangioma and vascular malformation based on ISSVA classification and prototypical histopathological features. Biopsies were reviewed based on 7 histopathological criteria viz. endothelial morphology, mitotic activity, intralesional nerve bundles, intralesional inflammation, prominent vessel type, variability in the size of vessels, and the plane of the lesion. A panel of GLUT-1, WT-1 and Ki-67 was performed in each case. Three cases each of infantile hemangioma and congenital hemangioma and 38 cases of vascular malformations were diagnosed. Endothelial cell morphology (p<0.001), mitotic activity (p<0.001), and intralesional nerve bundles (p<0.02) were found to be statistically significant in differentiating hemangiomas from malformations. GLUT-1 (p<0.05) and Ki-67 labeling index (p<0.001) were useful to distinguish infantile hemangiomas from vascular malformations. To conclude, ISSVA classification of benign vascular anomalies can be reliably done on histopathology. However, every case must be interpreted in the light of clinical and radiological features.

   OP69: Immunoexpression of PD-1 and PD-L1 in nonsmall cell lung carcinoma and its correlation with other clinicopathological parameters Top

Michael Leonard Anthony, Nilotpal Chowdhury, K Arathi, Mayank Mishra, Shalinee Rao

AIIMS, Rishikesh, Uttarakhand, India. E-mail: [email protected]

Background: This study attempted to study the prevalence of PD-1 and PD-L1 positive cases in Non-Small Cell Lung Carcinoma (NSCLC) and their association with other clinico-pathological parameters in a tertiary care setting in North India. Design: This was an observational cross-sectional study. One hundred consecutive histologically proven NSCLC cases having sufficient tumour material from July 2016 – July 2018 were examined and the prevalence studied by IHC. Results: The PD-1 positivity in lymphocytes was 29% (95% CI: 20.4% - 38.9%). Membranous positivity for PD-L1 in tumor cells was 27% (95% CI: 18.6% - 36.8%) and in tumor infiltrating lymphocytes was 22% (95% CI: 14.3% - 31.4%). There was no statistically significant association between PD-1 or PD-L1 status with age, gender, smoking , pleural effusion, clinical stage, histological type or infiltration by lymphocytes. Discussion: PD-L1 immunoexpression was seen in the tumour cell membrane and tumour infiltrating lymphocytes. Most targeted therapies require 1% membranous positivity for classifying a tumor as PD-L1 positive; additionally Atezolizumab requires staining of 1% immune cells as well. These PD-1 positive lymphocytes were observed within the epithelial tumour cell formations as well as within the tumour stroma. Additionally, PD-1 and PD-L1 expression were found to be independent of one another. Conclusion: The moderately high prevalence may justify routine testing for PD1 or PD-L1 in NSCLC, which should preferably be carried out in all cases, rather than any selected subsets.

   OP70: Clinicopathological study and classification of thymomas based on 2015 WHO classification of thymic tumors: A single center experience Top

Monalisa Hui, Shantveer G Uppin, Megha S Uppin, Tara Roshni Paul, RV Kumar, M Ambaresh Rao

Nizams Institute of Medical Sciences, Hyderabad, Telangana, India. E-mail: [email protected]

Background: The classification of thymic tumors has been revised several times over past few years. The recent WHO Classification has introduced certain conceptual changes to overcome unsatisfactory reproducibility encountered in the previous edition. So we attempted to study the clinicopathological findings of thymomas and classify according to the criteria laid down in 2015 WHO Classification. Design: All the consecutive cases of thymomas diagnosed on resected specimens between January 2011 to June 2019 were analyzed retrospectively. Cases diagnosed on core needle biopsies or fine needle aspiration cytology were excluded. The hematoxylin and eosin stained slides were reviewed. The thymomas were classified according to 2015 WHO classification. The histopathological parameters analyzed included transcapsular invasion, margin status, lymph nodal involvement and changes in the adjacent thymus. Immunohistochemistry results were reviewed where ever available. Results: Of total 100 cases, there was associated myasthenia gravis in 40 patients. The histopathological subtype included 34 cases of B2 followed by 21 AB,15 type A,10 type B1and 9 B3 thymoma. Combined histology was seen in 11 cases of which a minor B2 and B3 component were seen in 5 and 3 cases respectively. The B2 and B3 thymomas were more commonly associated with a higher stage. The staging of the patients did not differ significantly in patients with or without MG. Conclusion: The overall most common histological subtype was B2. Recognition of B2 and B3 areas in combined histological pattern would caution the clinician for close follow up and adjuvant therapy if required.

   OP71: The prevalence of PDL-1 expression in lung cancer: Real-world experience from a tertiary care oncology centre Top

Rajiv Kumar, Sathyanaraynan Rajaganesan, Amit Joshi, Kumar Prabhash

Tata Memorial Hospital, Mumbai, Maharashtra, India. E-mail: [email protected]

Background: Increasing demands for PD-LI (Programmed Death-Ligand 1) testing has been observed recently and its role is emerging as a potential biomarker for immunotherapy. However, there is paucity of the Indian data on prevalence of PDL1 expression. Design: We conducted a retrospective audit of lung cancer patients, tested with PDL-1 by using rabbit Anti-Human PD-L1 monoclonal antibody (clone SP263) on VENTANA BENCHMARK XT between January 2017–August 2019, to evaluate the prevalence of PD-L1 expression and record its correlation with clinic-pathological factors. PD-L1 expression of>=1% in the tumor cells was regarded as positive. Further, stratification of the positive cases was done, in subgroups of 1 – 24, 25-49 and>=50% PD-L1 expression respectively. Results: A total of 450 cases of lung cancer patients were tested for PDL-1. The median age was 56.95 years (range: 29-83years). The majority of patients were male (N=306, 68.1%), smokers (N=175, 39%) and stage III/IV (n=360, 80 %). PD-L1 positivity was seen in 28% (n=126) of all cases. On further stratification of positive cases, 82%, 12% and 6% PD-L1 positivity was noted in subgroups of 1 – 24, 25-49 and>=50% PD-L1 expression respectively. PDL-1 expression correlated with higher nuclear grade (p=0.001), smoking (p=0.025), solid histological type (p=0.045) and TTF1 positivity (p=0.028), but not with EGFR status. Conclusions: Although, the overall positivity rate of PD-L1 expression was 28%, only 6% of them had expression more than 50%. More studies with larger sample sizes will be necessary to find out the exact prevalence of PDL-1 expression in Indian population.

   OP72: Correlation of biomarkers and frozen section diagnosis with paraffin histopathological diagnosis in suspected ovarian cancer Top

Tanushri Mukherjee

Command Hospital, Chandimandir, Haryana, India. E-mail: [email protected]

Background: Ovarian cancers pose diagnostic dilemma and is problematic for decision making for the gynaecological oncologist as well as the pathologist. The use of intra operative frozen section can aid significantly in decision making and assist in choosing the correct operative path once a mass lesion of ovaries is discovered. Design: Over a two year period, 50 cases of Suspected Ovarian cancers were examined by intra operative frozen section as well as followed up with histopathology in paraffin sections. Results were categorized in two strata – benign and malignant. Results and Discussion: A comparison between frozen section diagnosis and findings on paraffin section showed that the sensitivity of frozen section in diagnosis of malignant lesions is 97.14%, with specificity 93.33%, positive predictive value 97.14% and negative predictive value 93.33%. Among 50 cases, 01 case was reported as false positive and 01 reported false negative. Conclusion: Intra operative frozen section is a highly sensitive and specific modality for diagnosis of malignant lesions of the ovary.

   OP73: Immunohistochemical and morphometric evaluation of angiogenesis in placentae of low birth weight babies using vascular endothelial growth factor and CD105 Top

Col Jasvinder Kaur Bhatia, Maj Manish Sharma1, D Boruah, Brig Ajay Malik

Department of Pathology, Armed Forces Medical College,1Graded Specialist Pathology, MH CTC, Pune, Maharashtra, India. E-mail: [email protected]

Background: VEGF is involved in the placental vascular development and immunohistochemical expression of VEGF may reflect hypoxic status of the placenta in hypoxic states. CD105 stains the blood vessels so may be used as a marker of vascularity in placenta. The objective of the study is to evaluate angiogenesis in placentae of low birth babies by VEGF & CD105. Methods: This is a prospective case-control study at a tertiary care centre in Western India. 60 placentae of LBW newborns were taken as cases and 40 placentae of babies weighing more than 2.5 kg were included as controls. Clinical history of mothers was retrieved from hospital records. The placentae were examined grossly and sections were taken. The clinical, gross and microscopic findings were studied in both the groups. Immunohistochemistry was performed by using markers CD 34, VEGF & CD105 and morphometric evaluation of parameters was calculated. Results: We found that the expression of VEGF was significantly higher in LBW placentae than controls (p<0.001). Morphometry was performed on CD105 stained placental sections and we found increased vascular parameters in LBW placentae. Conclusion: We found significant differences both in the expression of CD105 & VEGF in placentae of low birth weight babies. Increased placental expression of CD105 may result in vascular dysfunction leading to chronic fetal hypoxia, which may induce VEGF to stimulate angiogenesis.

   OP74: A study on association of the epithelial changes in the fallopian tube and the epithelial neoplasms of ovary using p53 Top

Vandana Maroo, Suchismita Chakrabarti, Anadi Roy Chowdhury

R.G. Kar Medical College and Hospital, Kolkata, West Bengal, India. E-mail: [email protected]

Background: The origin of ovarian carcinomas was considered to be de novo previously. But recent studies show that the type II Ovarian carcinomas, majority constituting High Grade Serous Carcinomas originates from the fimbrial end of the fallopian tubes. Design: Observational Cross sectional study. Results and Discussion: The fimbrial ends of the fallopian tubes in 20 cases with epithelial Carcinomas of the ovary (18- Serous Carcinoma and 2- Mucinous Carcinoma) were submitted. The sections were stained with Haematoxylin and Eosin and P53 tumour marker. A minimum of 12 Tubal Secretory epithelial cells staining for p53 were considered to be P53 positive. Out of the total 20 cases, it was found that the fallopian tubes of 2 cases of mucinous carcinomas did not stain for p53 while 10 cases (55.5%) of High Grade serous carcinomas were positive with p53,5 of them (27.7%) were dysplastic and remaining 3 (16.8%) were negative. Among the positive and dysplastic cases of serous carcinomas (10+5=15), the fimbrial ends of the fallopian tube was submitted in 13 of them (86.7%) while in the rest 2 (13.3%), the proximal ends of the fallopian tubes was submitted. Conclusion: There is correlation of the fimbrial ends of the fallopian tube with the High Grade Serous Carcinomas of the ovary. This knowledge can be used to remove the entire tube with Hysterectomies and Fimbriectomy instead of simple tubectomy should be done for sterilisation as a prophylactic measure to reduce the risk of ovarian cancers.

   OP75: Molecular characterization of malignant epithelial ovarian tumours with the help of immunohistochemical with KRAS, BRAF, PTEN, PIK3CA and P53 Top

Ejju Krishna Chaithanya, Amit Kumar Chowhan, H Narendra1, A Y Lakshmi2

Departments of Pathology,1Surgical Oncology and2Radiology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India. E-mail: [email protected]

Background: Ovarian cancer is the leading cause of gynaecologic cancer death in females and often presents at an advanced stage. Epithelial ovarian carcinoma (EOC) is a highly heterogenous disease characterized by multiple histological subtypes. Recent advances in the molecular characterization of EOCs have provided the basis for a simplified classification scheme in which these cancers are classified as either type I or type II tumours. These two categories have implications regarding disease pathogenesis, management and prognosis. Design: All oophorectomy specimens diagnosed with epithelial malignancy during the period from January 2018 to August 2019 were included in the study. IHC was performed with surrogate markers KRAS, BRAF, PTEN, PIK3CA and P53 to analyse mutation status of EOCs and hence classified as type I and type II. Results: Out of 52 cases of EOCs, 43 are serous carcinomas (34 high grade, 9 low grade), 6 mucinous carcinomas (all low grade), 3 endometrioid carcinomas (all low grade). IHC was performed on these to classify them as type I & type II according to the predicted mutation and will be discussed. Discussion: Type I and type II ovarian tumours develop independently along different molecular pathways. They differ also in terms of precursor lesions & stage at presentation, clinical behaviour, response to treatment and prognosis. So these should be considered as two distinct entities.

   OP76: Clinicopathological spectrum of nonurothelial bladder tumors and role of immunohistochemical in the diagnosis Top

Neha Sharma, Arvind Ahuja, A K Sen, D S Chauhan, Purnima Malhotra, Minakshi Bhardwaj

ABVIMS and DR RML Hospital, New Delhi, India. E-mail: [email protected]

Background: Non urothelial bladder tumors (NUBT) are relatively rare, accounting for approximately 5 % of carcinoma arising at this location. These tumors frequently pose a diagnostic and therapeutic challenge, therefore immunohistochemistry is a useful adjunct to arrive at the conclusive diagnosis. Design: The objectives of the study were to analyse the clinicopathological and immunohistochemical features of NUBTs. This is a retrospective study of NUBTs diagnosed over a period of five- and half-years. Patients' files were retrieved from the archives. Gross and microscopic features were recorded. Simple percentage and frequencies were used to interpret the data. A panel of IHC was applied in the morphologically challenging cases. Results and Discussion: A total 17 cases (2.1% of all bladder tumors) of NUBT were found. The age of the study population ranged from 18 to 80 years with a male: female ratio of 1.4:1. The most common presenting symptom was gross hematuria and the most common location was right lateral bladder wall. Muscle invasion was seen in 23.5 % of cases and large areas of necrosis were observed in 35% of the cases. The cases included squamous cell carcinoma (23.5%), adenocarcinoma (secondary and primary: 23.5%), signet ring carcinoma (5.8%), undifferentiated carcinoma (5.8%), neuroendocrine carcinoma (23.5%) and mesenchymal tumors (leiomyoma and leiomyosarcoma 17.6%). Conclusion: NUBTs present with similar clinicoradiological findings as urothelial carcinoma but have different prognostic and therapeutic implications. Combined approach including adequate clinical information, histomorphology and IHC are essential for definite diagnosis.

   OP77: The utility of evaluating mismatch repair proteins in endometrial carcinoma: An experience from a tertiary referral centre in North India Top

Ekta Jain, Lata Kini, Sarita Prasad, Aparna Dhar, Shivani Sharma, Aditi Dewan

Core Diagnostics Private Limited, Gurgaon, Haryana, India. E-mail: [email protected]

Background: Endometrial cancer (EC) is one of the common gynaecological malignancies worldwide. Around 25-30% of EC patients have Mismatch repair deficiency (MMRd). Lynch syndrome is caused by germline mutations in same MMR genes. Lynch-associated tumours appear to have better prognosis, however implications for prognosis and survival in these tumours are less known. Microsatellite insufficiency (MSI) is associated with high neoantigen loads and number of tumor infiltrating lymphocytes, which overexpresses PD-1 and PD-L1 and are thus excellent candidates for PD-1-targeted immunotherapies. In this study, we aim to evaluate utility of MMR in patients with EC and its clinico-pathological correlation. Methods: Seventy-five cases of EC which underwent MMR evaluation over a period of four years at our centre were included. Demographics, clinical details, histopathological and immunohistochemical (IHC) parameters were recorded. Tumors with loss-of at least one protein were considered MMR deficient (MMRd) and those with intact expression were MMR proficient (MMRp). Family history, post evaluation treatment given and survival data was evaluated. Results: Of 75 cases tested, 21(28%) were MMRd. Frequencies of IHC MMR loss of expression were: MLH1/PMS2: 16 (21%), MSH6 loss only: 2 (2.6%), MSH2/MSH6 loss:1 (1.3%), PMS2 loss: 1 (1.4%). In MMRd cases, most common histologic tumor type was endometrioid adenocarcinoma (70%).Further analysis in terms of pedigree evaluation and therapy outcomes are ongoing. Conclusions: MSI plays an important role in the progress of endometrial cancer. Routine testing of MMR proteins in endometrial cancer can contribute to screening of LS families and make immunotherapy available as a treatment option.

   OP78: To study the expression of p16, epidermal growth factor receptor and CD44 in squamous cell carcinoma uterine cervix Top

Kanika Gupta

Maulana Azad Medical College, New Delhi, India. E-mail: [email protected]

Objective: Cervical cancer is most common cancer in Indian women. Main cause is persistent human papilloma virus infection (HPV). P16 expression increases by pRb gene inactivation by high risk HPV.1 Epidermal Growth Factor Receptor (EGFR) overexpression leads to invasion, enhanced angiogenesis and metastasis. HPV prevents EGFR degradation.2 Cluster differentiation 44 (CD44) aberrant expression leads to tumour extension and metastasis.3. Aim of Study:

  1. To study expression of p16, EGFR and CD44 in squamous cell carcinoma (SCC) uterine cervix
  2. To compare different grades (WHO) and clinical stages (TNM) of SCC cervix using combined panel of immunohistochemical markers including p16, EGFR and CD44.

Materials and Methods: Sixty two cases of histopathologically diagnosed untreated SCC cervix were included in study and immunohistochemistry for p16, EGFR and CD44 was done. Results: CD44 expression increased with increasing stage of SCC cervix (p=0.007). A positive significant correlation between expression of EGFR and CD44 expression (p=0.001); and p16 and EGFR expression (p=0.015) was observed. No statistically significant association of clinical stage with p16 and EGFR respectively and of p16, EGFR and CD44 with histological types was observed. Conclusion: The study focused on correlation of p16, EGFR and CD44 expression with histological type and clinical stage of cervical SCC and clinical parameters. CD44 expression can aid in staging carcinoma. These results add to growing literature suggesting that HPV interaction changes biology of EGFR by preventing degradation. However, larger study is required to validate results, monitor cervical lesions and predict progression of lesion based on these markers.

   OP79: To study over expression of p53 and BCL2 in correlation to human papilloma virus infection in premalignant and malignant lesion of cervix Top

Vijayanand Choudhary, Sangeeta Pankaj1

Departments of Hematology and1Gynecooncology, Indira Gandhi Institute of Medical Sciences, Patna, Bihar, India. E-mail: [email protected]

Background: Cancer of the cervix is second most common cancer in Indian women after breast cancer. Persistent high risk human papilloma virus (HPV) infection is the best predictor for increased risk of cervical cancer, but expression of certain biomarkers of cell proliferation and apoptosis have been studied. This study was undertaken to study the significance of p53 and bcl2 overexpression in premalignant and malignant lesions of cervix and their correlation with HPV infection. HPV virus contains oncoprotein E6 which binds to and degrades p53, a tumor suppressor gene resulting in increased cell proliferation and decreased apoptosis. Bcl2 oncogene inhibits apoptosis thereby increasing cell proliferation. Design: 37 women patients were enrolled for cytology, HPV DNA, p53, bcl2 and histopathology. Aim was to evaluate the sensitivity and specificity of these tests considering histopathology as gold standard. Results and Discussion: Total of 37 cases of cervical neoplasia including 32 cases of invasive carcinoma and 5 cases of CIN were seen. HPV was positive in 24 cases of invasive carcinoma and negative in all 5 cases of CIN. In invasive carcinoma, p53 was positive in 28 cases and bcl2 was positive in 23 cases. In CIN, p53 was positive in 2 cases and bcl2 was positive in 1 case. Conclusion: In this study p53 showed higher positivity compared to bcl2 in invasive carcinoma. In CIN I both were negative thus behaving like benign lesion. In CIN II no correlation was seen. CIN III behaved like their invasive counterpart showing both p53 and bcl2 positivity.

   OP80: Molecular classification of urothelial carcinomas using immunohistochemical surrogates Top

Rishabh Sahai, Prashant Joshi, Sanjeev Kishore, Ankur Mittal

AIIMS, Rishikesh, Uttarakhand, India. E-mail: [email protected]

Background: Lacunae in optimizing treatment and predicting prognosis of Urothelial carcinoma has led to molecular classification broadly into luminal and basal subtypes based on molecular profile. Here, we evaluate the reproducibility of characteristics of molecular subtypes of Urothelial carcinoma using immunohistochemical surrogates as reported by Dadhania et al in 2016. Design: 144 TURBT (Transurethral resection of bladder tumor) specimens sent to Department of Pathology, AIIMS, Rishikesh from September 2017 to January 2019 were included. Immunohistochemical expression of markers GATA3, CK5/6, p53, Ki67 and Her2neu were evaluated. Hematoxylin and eosin slides were reviewed for morphological characteristics. Statistical analysis was done using software R. Results and Discussion: Out of 144 cases, 41(28.5%) were non muscle invasive tumors while 103(71.5%) were muscle invasive tumors. GATA3 was seen positive in 86(59.7%) cases while negative in 58(40.3%) cases. CK5/6 was positive in 13(9.03%) cases while negative in 72(50%) cases. Basal expression was seen in 59(40.97%) cases. Aberrant immunoexpression of p53 was noted in 88(61.1%) cases while 56(38.9%) cases demonstrated wild type p53 immunoexpression. Ki67 ranged from 1% to 90% and significant associations were seen with grade of tumor and presence of inflammation. Her2neu immunoexpression was seen in 20(13.9%) cases. Conclusion: Urothelial carcinomas can be classified on basis of GATA3 and CK5/6 expression. However in our study, 27(18.75%) cases were double negative which is higher than reported in literature (4%-5%) and 40.97% cases demonstrated expression of CK5/6 limited to basal layers. In addition, Her2neu overexpression was particularly noted in areas of tumor with high Ki67 expression.

   OP81: Finding bacteria with flow Top

S Kannan, Joy J Mammen, Sukesh C Nair, Rani Diana Sahni, K P P Abhilash, Nitty S Matthews, Gina Maryann Chandy, G Vinoth Kumar, Indra S M David, Josephine

Christian Medical College, Vellore, Tamil Nadu, India. E-mail: [email protected]

Background: Cost-effective and quick screening tool to diagnose Urinary tract infections (UTIs) would create an impact on laboratory economics and management of patients, especially in the Emergency Department. Sysmex urinanalyzer, UN-9000 analyses urine-sediment by flow cytometry resulting in rapid and cost-effective quantification of bacteria. Design: Prospective longitudinal observational study was planned with aims of 1) Determining a flowcytometric cut-off value of urine bacteriuria in automated SysmexUN9000 urinanalyzer to diagnose urinary tract infections. 2) Assess the machine's capacity in differentiating between gram-negative and gram-positive bacterial infection, compared to urine culture. 304 urine samples from consecutive patients admitted to the Emergency Medicine (June 2018 to May 2019) were tested in SysmexUN9000 urinanalyzer and were compared with corresponding culture results. Bacterial counts and scatter patterns were analysed to ascertain which would be suited to identify UTI the earliest. Results and Discussion: A flow cytometric bacteriuria count of 239/HPF was determined to be the best at 80% sensitivity and 63% specificity from the ROC-curve. Type III pattern in scattergram had a statistically significant association with gram-negative bacteria infection (p<0.001, specificity: 88.70%). Type II pattern was indicative of contaminants (p<0.001, specificity: 91.70%) due to improper collection. 'UTI flag' determined by the machine had 99% sensitivity (p<0.001) in detecting culture-positive samples and 'Gram-negative flag' had 94.70% specificity (p<0.001) in picking up culture-confirmed gram-negative bacteria. Conclusions: We have found that a rapid screening of samples on Sysmex urine analyzer can provide an early and reliable indication of gram-negative UTI providing guidance for the initiation of appropriate antibiotics in the emergency department.

   OP82: Spectrum of morphological lesions in 161 nephrectomies Top

Madhuri Singh, Piyusha Naragude, Akshi Raj, Sunita Bamanikar, Archana Buch, Shirish Chandanwale

Dr D Y Patil Medical College and Hospital, Pune, Maharashtra, India. E-mail: [email protected]

Background: Kidneys can be involved in variety of pathological processes. It is a common procedure in urological practice. The incidence of nephrectomies for non-neoplastic condition such as chronic pyelonephritis is decreasing and nephrectomy for malignant lesions is increasing in some developed countries The study is carried out to find out the frequency of various neoplastic and non-neoplastic lesions in nephrectomy specimens and to study histomorphological features in a tertiary care hospital. Design: The study include 161 nephrectomy specimens and their morphology was studied. Lesions were categorized into non neoplastic and neoplastic. In malignant lesions nuclear Grading System was used to grade clear cell renal cell carcinoma and papillary renal cell carcinoma. International society of Pediatric Oncology staging was used to stage Wilms, tumors. Staging of renal cell carcinoma was done by TNM classification as per WHO. Results: Out of 161 nephrectomy specimens the 81(50.31%) were males and 80(49.68%) were females. Mean age was 45.43 years. 115 (71.42%) nephrectomy specimens showed non neoplastic lesions and 46(28.57%) showed neoplastic lesions. Commonest non neoplastic lesions was chronic pyelonephritis in 106 (92.17%) patients. RCC (n=36) was the commonest neoplastic lesion. Conclusion: In developing country like India chronic pyelonephritis still remains to be the commonest cause of nephrectomy. The possible reason for change in trend can be early detection and better treatment modalities of inflammatory lesions of kidney in the developed countries as compared with developing countries.

   OP83: Comparative role of cytohistological techniques in diagnosis of upper gastrointestinal lesions Top

Kanika Kapoor, Kanika Kapoor, Prachi Singh, Seema Awasthi, Shyamoli Dutta, Ashutosh Kumar, Faiyaz Ahmad

Department of Pathology, Teerthanker Mahaveer Medical College and Research Centre, Moradabad, Uttar Pradesh, India. E-mail: [email protected]

Background: Early detection of various neoplastic and non-neoplastic lesions of upper gastrointestinal (GI) tract is crucial for correct and timely treatment. Histopathological examination (HPE) is gold standard but time consuming. Cytology offers a simple, inexpensive and rapid diagnostic method. Hence this study was planned to compare results of two cytological techniques [Imprint cytology (IC), Brush cytology (BC)] with the standard HPE for upper GI lesions. Design: Prospective study on 26 patients was performed with symptoms suggestive of upper GI pathology. Endoscopy was performed and brush smears along with imprint smears from endoscopic biopsies were prepared. Results of BC and IC were analysed, considering HPE as gold standard. Results: Our study included 17 males and 9 females (M:F=1.88:1) with age range- 20 to 85 years (mean age 48.6 years). Most common presentation was pain in abdomen (46.2%) followed by dysphagia (42.3%). Maximum lesions involved esophagus (42.3%) followed by duodenum (38.4%). Sensitivity and specificity of brush cytology was 85.7% & 87.5% and of imprint cytology was 88.8 % & 100% respectively. Discussion: Imprint cytology gave better results than brush cytology and was found superior as even few viable malignant cells in touch smear are sufficient for correct diagnosis within a short period. Conclusion: Imprint cytology is a simple, rapid, cost-effective and reliable procedure which can be adopted as a routine for immediate evaluation of various upper GI lesions. It is more diagnostic than BC whereas HPE remains gold standard. Combination of cytohistologic techniques facilitates exact tumour typing and assessment of tumour invasion.

   OP84: Cytomorphological spectrum of granulomatous mastitis: A study of 33 cases Top

Akshi Raj, Madhuri Singh, Piyusha Naragude, Manoj Swadhikar, Abhinav Shetty, Aniket Bhide, Shirish Chandanwale

Dr D Y Patil Medical College and Hospital, Pune, Maharashtra, India. E-mail: [email protected]

Background: Granulomatous mastitis is an uncommon benign breast disease. Varied aetiologies such as tuberculosis, foreign body reactions, sarcoidosis, fungal, parasitic infections and autoimmunity have been suggested. Pre-operative definitive diagnosis is essential for proper treatment. In developing countries like India, fine needle aspiration is widely used as a reliable technique for preoperative evaluation of palpable breast lumps. Design: The study includes a total of 33 cases of granulomatous mastitis diagnosed on fine needle aspiration. Cytomorphological features and clinical findings including radiological findings were reviewed. Cytomorphological features were correlated with histopathological features and clinical findings. Results and Discussion: A total of 33 patients showed varied cytomorphological features including epithelioid cells/granuloma with lymphocytes/plasma cells/polymorphs with or without necrosis/caseous necrosis and with or without giant cells. A total of 27 cases were diagnosed as tuberculous mastitis, 4 cases were diagnosed as idiopathic granulomatous lobular mastitis and 2 cases as foreign body granulomatous mastitis. ZN stain showed AFB in 13 smears. A total of 8 cases showed positive AFB culture. Fungal stain did not show fungi. FNA diagnosis was confirmed by histopathological diagnosis. Conclusion: Epithelioid granulomas with caseous necrosis with or without acid fast bacilli in cytology smears are diagnostic of tuberculosis. Cytology smears which show epithelioid granulomas with predominant polymorphs without necrosis and acid fast bacilli, in such cases the diagnosis of idiopathic granulomatous lobular mastitis must be considered. Histopathological examination is essential for definitive diagnosis in such cases.

   OP85: Expression of p16 and Ki67 in squamous intraepithelial lesions and squamous cell carcinoma of cervix Top

Ritika Hooda, Kulwant Singh, Swaran Kaur, Rajiv Mahendru, Parveen Rana Kundu, Ruchi Agarwal

BPS GMC (W), Sonepat, Haryana, India. E-mail: [email protected]

Background: Cervical cancer is a public health problem in India accounting for 17% of all cancer deaths among women aged 30-69 years. Invasive cervical cancer is preceded by precursor lesions hence cervical cancer has the greatest potential for prevention. p16 and Ki67 positivity in cervical smears is a marker of cervical dyskaryosis. The grade of positivity of both the markers is indicative of the higher grade of lesion hence predicting survival. Design: The aim of our study was to evaluate the expression of p16 and Ki67 in squamous intraepithelial lesions and squamous cell carcinoma of cervix. The study was conducted on 40 cervical biopsy and hysterectomy specimens received in the Department of Pathology, BPS GMC, KhanpurKalan. H&E stained sections were studied and classified. IHC assessment and grading for p16 and Ki67 was performed. Results and Discussion: Out of 40 cases, 36 cases were diagnosed as SCC and 4 cases as dysplasia. Staining for p16 was positive in all the cases of carcinoma (100%) and 50% of cases of dysplasia.(p<0.001) Ki67 was positive in all 40 cases (100%). A positive correlation between p16 and Ki67 was also found which is also statistically significant (p<0.001). Conclusion: The expression of p16 is upregulated with the increase in dysplastic cells to cervical carcinoma which indirectly indicates degree of malignancy. Ki67 which is usually positive in the lower third of the epithelium also covered almost entire thickness in cervical carcinoma. This finding could be useful to enhance diagnostic accuracy.

   OP86: Salivary gland lesions: Milan system and its diagnostic utility Top

Smriti Singh, Parveen Rana Kundu, Uma Garg, Swaran Kaur

BPS Government Medical College, Sonepat, Haryana, India. E-mail: [email protected]

Background: Salivary gland lesions represent about 3-6% of all head and neck lesions and at times warrant an urgent clinical management. Although an array of investigations aid in the diagnostic work up of the lesions but till date fine needle aspiration cytology (FNAC) has proved the most useful. However FNAC has its pitfalls and hence to improvise the reporting systems a new formulation was deviced named as the 'Milan System for Reporting the Salivary Gland Cytopathology'. Design: This study was undertaken to assess the utility of FNAC in reporting the salivary gland cytopathology and to assess the applicability of the Milan System. All the cases of salivary gland lesions, presenting to our department on whom FNA was done during the study period from January 2018 to September 2019, i.e. 85 cases were included in the study. The cytological diagnosis were categorized as per the Milan System into six categories. Results and Discussion: The various diagnostic values were calculated using the standard formula. The lesions showed a predominance in females (57.8%). Parotid glands were the most common site reported (61.4%). The distribution of various lesions as per Milan categorization were as follows- (1) Nondiagnostic (8.2%), (2) Non neoplastic (22.35%), (3) Atypia of undetermined significance (3.5%), (4a) Benign (48.5%), (4b) Uncertain Malignant Potential (4.7%), (5) Suspicious of Malignancy (7.0%), (6)Malignant (5.8%). Conclusion: The Milan system of categorization has proved as a useful tool in diagnostic assessment of salivary gland lesions. The new formulation has not only aided in guiding the clinical management of lesions but has also proved its efficiency in cytological diagnosis of difficult cases showing mixed features.

   OP87: Diagnostic utility of fine needle aspiration cytology in thyroid lesions Top

Deepti Agarwal, Swarn Kaur Saluja, Parveen Rana, Monika Gathwal, Shikha Goel

Department of Pathology, BPS Government Medical College for Women, Sonepat, Haryana, India. E-mail: [email protected]

Background: Fine-needle aspiration cytology (FNAC) is an inexpensive and highly accurate means in diagnosing neoplastic and non-neoplastic lesions of the thyroid. It is safe, quick, cost effective outpatient department procedure that helps in planning the management of the patient. Design: The present study was conducted in the Department of Pathology, BPSGMC (W), Khanpur Kalan, Sonipat over a period of 1 year to evaluate the diagnostic utility of FNAC in thyroid lesions and to calculate sensitivity, specificity and diagnostic accuracy of FNAC in diagnosis of thyroid lesions. A total of 278 cases of thyroid swelling were studied in whom FNAC of thyroid was done and diagnosis was given on conventional cytology. It was further categorized according to the Bethesda System. Cytological diagnosis was compared with the histopathology wherever available. Result: The commonest age group affected was 31-40yrs, female patients (92.0%) outnumbering the male patients (8.0%). On conventional cytology, the commonest lesion observed was colloid goiter. Maximum cases were observed in category II (61.86%), followed by category I (15.45%), category IV (10.8%), category VI (7.19%), category V (3.27%) & category III (1.43%). Histopathology was available in 77 cases. Statistical analysis of the data showed the diagnostic accuracy of fine needle aspiration cytology to be 92.2%. Fine needle aspiration cytology showed a sensitivity of 76% and a specificity of 100%. Conclusion: The simplicity, rapidity, high diagnostic accuracy, high sensitivity and high specificity of FNAC make it a valuable tool in the diagnosis and management of patients with thyroid lesions.

   OP88: Expression of p53 in benign and malignant prostate lesions and its correlation with serum prostate specific antigen level Top

Anu Sharan, Deepti Agarwal, Swaran Kaur, Ruchi Agarwal, Atul Khandelwal1

Departments of Pathology and1Surgery, Bhagat Phool Singh GMC for women, Sonepat, Haryana, India

Background: Prostate enlargement is a major health problem throughout the world. The most frequent prostate lesion in males is benign prostatic hyperplasia. Prostate cancer is the second most common cancer among men worldwide. Increased serum PSA has been observed in benign prostatic disorders and prostate cancer. Mutations of the p53 tumor suppressor gene have been implicated in numerous malignancies including prostate cancer. Design: The aim of our study was to evaluate the expression of p53 in benign and malignant prostate lesions and to correlate it with serum PSA level. The study was conducted on 54 prostate specimens received in the Department of Pathology, BPS GMC, Khanpur Kalan. Serum PSA levels were collected from requisition forms. H&E stained sections were studied and classified into benign and malignant lesions. Immunohistochemical assessment for p53 nuclear protein was performed. Results and Discussion: Out of 54 cases, 35 cases were diagnosed as BPH and 19 cases as adenocarcinoma prostate. An extremely significant increase (p<0.001) was observed in serum PSA concentration in adenocarcinoma prostate patients when compared to BPH patients. Among carcinoma cases, 11 (57.9%) were categorized as Gleason grade group 4. The p53 IHC staining was negative in all the 36 cases (100%) with benign disease and was positive in 16 cases (84.2%) of carcinoma prostate (p<0.001). Conclusion: It can be concluded that the IHC expression of p53 is significantly up-regulated in malignant lesions. Therefore, the application of immunohistochemistry along with serum PSA level becomes helpful in distinguishing prostate cancer from benign mimickers.

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