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Background Indeterminate thyroid lesions have always been a grey zone in the field of thyroid cytopathology. Immunocytochemistry (ICC) has emerged as a promising tool to correctly classify these indeterminate thyroid lesions into benign and malignant. Hence we planned to assess a panel of immune markers in the diagnosis of indeterminate thyroid lesions consisting of Galectin-3, considered positive for malignancy and CD117 which is positive in benign follicular epithelial cells and negative in malignant lesions. Methods All the thyroid aspirates reported as indeterminate lesions over a period of 3 years were evaluated. Galectin-3 and CD117 immunocytochemistry was done in 50 alcohol fixed Pap stained smears of AUS/FLUS, FN/SFN and SM category lesions. The expression of both immune markers was assessed by semi-quantitative method and ICC score was calculated. Result Of 50 indeterminate lesions, 29 were positive for Galectin-3 and 21 were negative. CD117 was positive in 19 cases and rests 31 were negative. With the use of this ICC panel 29/30 indeterminate lesions in which histopathological correlation was available could be recategorized correctly into benign and malignant. The combined sensitivity and specificity of Galectin-3 and CD117 for categorising the indeterminate lesions into malignant category was 100%. Conclusion The combined use of positive and negative immune markers for thyroid malignancy increases the sensitivity and specificity of ICC to categorise the indeterminate thyroid lesions into benign and malignant. In cases with discordant ICC results we propose that inclusion of one additional positive and/or negative marker may resolve the diagnostic dilemma.

BACKGROUND: Bone allografts can elicit immune responses which is correlated with the presence of Human Leukocyte Antigen (HLA) and cellular DNA. It also has risk of causing occult infection arising out of contamination during its processing and storage. The presence of immunogenic materials like cells, cellular remnants and DNA in a decalcified bone allograft during different phases of processing has never been studied. Present study was conducted to explore- the cell viability using routine Hematoxylin and Eosin, presence of DNA using Feulgen staining and etiology of contamination in decalcified bone allograft during procurement, demineralization and ethanol preservation. METHODS: The harvested bones from patients undergoing hemireplacement/THR/TKR were processed to prepare decalcified bone allografts. The samples during procurement (A), HCL treatment (B) and ethanol preservation (C) were sent for histopathological analysis (number of osteocytes in the maximum density field under 40x and the cells demonstrating presence of DNA on feulgen stain) and microbiological assessment (aerobic/anaerobic/fungal cultures). RESULTS: Histopathological study demonstrated the presence of osteocytes and other cells like bone marrow, adipocytes, endothelial cells in the decal bone allograft. The average number of osteocytes gradually decreased from 55.47, 9.6, 0.86 in sample A, B, C, respectively. Feulgen staining confirmed the presence of DNA in osteocytes and other cells which decreased both qualitatively and quantitatively in subsequent stages of processing. Rate of contamination demonstrated at the procurement was 6.67% (Staphylococcus aureus). After treatment with HCl (demineralisation), 7.14% of non-contaminated allografts were found contaminated (Staphylococcus epidermidis). None of the remaining 13 non-contaminated allografts showed contamination after storage in ethanol. Overall 13% of the patients had positive cultures on microbiological assessment. CONCLUSION: The population of osteocytes in the harvested bone reduced significantly after processing with HCl and ethanol preservation. Presence of DNA, demonstrated by using Feulgen staining, was observed in bone marrow cells, adipocytes along with osteocytes which showed quantitative reduction on processing. Hence, antigenicity, conferred by cells and their DNA, reduced significantly after processing of decal bone. Contamination rate of banked decalcified allograft was 13%. Thus, culture and sensitivity tests should be carried out at each step of processing of decal bone allograft.

BACKGROUND: There is a paucity of data on the role of molecular methods in the diagnosis of osteoarticular tuberculosis. The present study was conducted to define the role of molecular (CBNAAT, LPA), phenotypic (AFB smear and culture) and histopathological evaluation in the diagnosis of osteoarticular TB. METHODS: Seventy-seven consecutive cases of osteoarticular tuberculosis were grouped into presumptive TB cases (group A) and presumptive drug-resistant cases (group B). Tissue samples obtained were submitted for CBNAAT, LPA, AFB smear, liquid culture and histological examinations. The diagnostic accuracy of each test was reported against histologically diagnosed cases and in all tests in tandem. RESULTS: Group A and group B had 65 and 12 cases, respectively. The diagnostic accuracy for tuberculosis was 84.62% by CBNAAT, 70.77% by LPA, 86.15% by molecular tests (combined), 47.69% by AFB smear, 50.77% by liquid culture and 87.69% by histology in group A, and 91.67% for CBNAAT, 83.33% for LPA, 91.67% for molecular tests (combined), 25% for AFB smear, 16.67% for liquid culture and 83.33% for histology in group B. The drug resistance detection rate was 4.62% on CBNAAT, 3.08% on LPA, 6.15% on molecular tests (combined) and 1.54% on DST in group A, while it was 33.33% on CBNAAT, 58.33% on LPA, 58.33% on molecular tests (combined) and 16.67% on DST among group B cases. Similar sensitivity rates for the various tests were obtained among both the groups on comparison with histology (taken as denominator). The addition of molecular methods increased the overall diagnostic accuracy (all tests in tandem) from 93.8 to 100% in group A and from 83.3 to 100% in group B cases. CONCLUSION: No single tests could diagnose tuberculosis in all cases; hence, samples should be evaluated by molecular tests (CBNAAT and LPA), AFB smear, culture and histological examinations simultaneously. The molecular tests have better demonstration of drug resistance from mycobacterial culture. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s43465-020-00326-w.

Cataract is the second leading cause of preventable blindness on the globe. Several programs across the country have been running efficiently to increase the cataract surgical rates and decrease blindness due to cataract. The current COVID-19 pandemic has led to a complete halt of these programs and thus accumulating all the elective cataract procedures. At present with the better understanding of the safety precautions among the health care workers and general population the Government of India (GoI) has given clearance for functioning of eye care facilities. In order to facilitate smooth functioning of every clinic, in this paper, we prepared preferred practice pattern based on consensus discussions between leading ophthalmologists in India including representatives from major governmental and private institutions as well as the All India Ophthalmological Society leadership. These guidelines will be applicable to all practice settings including tertiary institutions, corporate and group practices and individual eye clinics. The guidelines include triage, use of personal protective equipment, precautions to be taken in the OPD and operating room as well for elective cataract screening and surgery. These guidelines have been prepared based on current situation but are expected to evolve over a period of time based on the ongoing pandemic and guidelines from GoI.

Evidence from current studies show that squamous cell carcinomas at oral and oropharyngeal sites are distinct and unique, with their own separate etiopathogenesis, treatment, and prognosis. The aim of this work is to correlate p16 immunohistochemical expression with histomorphological features suggestive of HPV infection in oral and oropharyngeal squamous cell carcinoma. A total of 50 consecutive biopsy cases of oral squamous cell carcinoma (OSCC) and 50 consecutive biopsy cases of oropharyngeal squamous cell carcinoma (OPSCC) were evaluated for features suggestive of HPV infection like focal basaloid appearance, nests, and lobules of tumor cells with pushing borders, absence of stromal reaction, central necrosis, focal lymphoepithelial morphology, presence of koilocytes, and non-keratinizing or hybrid morphology. Immunostaining was performed using p16 monoclonal antibody (clone mouse 16P04). Only cases showing a moderate (2+) to high intensity (3+) staining in more than 75% cells were taken as p16 immunopositive. The histological features were correlated with p16 immunopositivity. A total of 18/50 (36%) cases of oral squamous cell carcinoma and 27/50 (54%) cases of oropharyngeal squamous cell carcinoma were p16 immunopositive. On statistical analysis, only nests/lobules with pushing borders were found to have a significant correlation with p16 immunopositivity (P value = 0.0012) for OSCC cases. For OPSCC cases, four histological features namely nests and lobules with pushing borders (P value = 0.0001), focal basaloid appearance (P value = 0.0041), lymphoepithelial morphology (P value = 0.0029), and non-keratinizing/hybrid morphology (P value = 0.0141) had a significant correlation with p16 immunopositivity. Histomorphological features are more helpful in predicting p16 immunopositivity in OPSCC than OSCC.

Juvenile xanthogranuloma (JXG) is a rare type of non-Langerhan cell histiocytic disorder, which is mostly confined to skin of head and neck. It is a self-limiting benign condition, which does not require surgery. We present a case of 8-month-old girl child with multiple yellowish brown colored papules over scalp, face, and neck. A clinical diagnosis of cutaneous mastocytosis was made. Fine-needle aspiration cytology (FNAC) smears showed foamy macrophages along with mixed inflammatory infiltrate and few touton giant cells. A diagnosis of JXG was rendered which was confirmed on histopathology and immunohistochemistry.Juvenile xanthogranuloma can be diagnosed on FNAC based on its characteristic cytologic features; however, it requires a high index of suspicion by cytopathologist. Cytological diagnosis of JXG can save the patient from unnecessary surgical biopsy or excision.

Background: Rheumatoid Arthritis (RA) is a devastating disease characterized by continual addition of leukocytes and T cells within the articular cavity causing inflammation and cartilage destruction. Withania somnifera is one of the most precious medicinal herbs, reported to have antioxidant, anti‐inflammatory, and immunomodulatory properties. Objective: The purpose of this study was to evaluate anti-inflammatory activity of aqueous extract of Withania somnifera roots (WSAq) in Collagen Induced Arthritic (CIA) rats. Methods: To achieve this, we assessed the level of inflammatory cytokines such as Tumor Necrosis Factor (TNF)-α, IL-1β, IL-6 and IL-10 in CIA rats. Further, transcription factor, oxidative stress parameters and CD+8 expressions were also analyzed in CIA rats. Results: Arthritic rats showed a greater increase in the levels of pro inflammatory cytokines such as TNF-α, IL-1β, IL-6, transcription factor NF-κB and a decrease in IL-10 concentration than controls rats. Oral administration of WSAq at a dose of 300mg/kg.wt. (WSAq300) appreciably attenuated the production of these pro inflammatory cytokines. This anti-inflammatory activity of WSAq300 might be partly mediated through an increase in the secretion of IL-10 and inhibition of NF-κB activity. Further, arthritic rats also show increased oxidative stress as compared to control rats. This increased oxidative stress in the arthritic rats appears to be the outcome of both an activated pro-oxidant and a poor antioxidant defense system. Treatment with WSAq300 strongly ameliorates all these ROS parameters significantly to near normal. Additional, metalloproteinase MMP-8 levels were also measured and found to be increased in CIA rats, which after treatment with WSAq300 came down to near normal. Conclusion: From the above results, it can be concluded that the use of WSAq300 may be a valuable supplement which can improve human arthritis.

Objective: Evaluation of endometrial scratching on pregnancy rates after IUI and comparing levels and distribution of IL-6 (inflammatory molecule) and E-Cadherin (adhesion molecule) by immunohistochemistry. Study design: Randomized case control study. Setting: Tertiary care hospital of Delhi, India. Material and methods: 240 women (120 in each group) with previous failed IUI cycles were enrolled. Study group patients were subjected to endometrial scratch by “Pipelle” on postovulatory day 6–8 in the preceding cycle. IUI was performed after controlled ovarian stimulation with gonadotropins. 20 patients of study group in whom IUI was cancelled underwent repeat scratch on postovulatory day 6–8. Levels and distribution of IL-6 and E-Cadherin were evaluated by Immunohistochemistry before and after the scratch. Main outcome: Comparison of pregnancy rates in women undergoing IUI after endometrial scratching with women undergoing IUI alone. Results and conclusion: There were 19 (19%) clinical pregnancies in the study group and 8 (7.6%) in the control group. There was a significant difference in between both the groups (p = 0.022). Number of pregnancies in unexplained infertility patients was 22.66% in study and 9.5% in control group (p = 0.0291). There was significant increase in score for IL-6 and E-Cadherin in both glands and stroma after scratching. Endometrial scratch is a simple, practical, cost effective and outpatient procedure which may increase pregnancy rates in previous failed IUI cycles. Increased intensity of staining; shift of major part of staining from gland or stroma in pre scratch samples to both glands and stroma in post scratch samples; and increased stromal decidualization point towards increased endometrial receptivity. Keywords: Endometrial scratching, IUI, Pregnancy rates, IL-6, E-Cadherin

To study the accuracy of frozen section biopsy for endometrial pathology in high-risk women with abnormal uterine bleeding (AUB).A case-control study was conducted between November 2017 to April 2019, a total of 150 women with postmenopausal bleeding, perimenopausal AUB, and high-risk women of age 40 years with AUB were recruited. All women underwent transvaginal sonography and Doppler, based on age-appropriate endometrial thickness cut-offs 80 women then underwent hysteroscopy. Based on hysteroscopy, women suspicious of malignancy were taken as cases (n = 40) and those with benign findings as controls (n = 40). All cases and controls underwent dilatation and curettage (DC) with frozen section (FS) and routine histopathology.The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (LR), negative LR, and overall test accuracy of FS were 90.9%, 93.19%, 83.33%, 96.19%, 13.8, 0.1 and 86.25% respectively for diagnosing endometrial hyperplasia and cancer taking histopathology as the gold standard. Correlation between frozen section biopsy and histopathology was highly significant (p 0.001) on DC specimens and the level of agreement was good (K = 0.778).In women suspicious of malignancy on hysteroscopy, frozen section has high accuracy on DC specimen and can be used to diagnose endometrial hyperplasia and cancer in an effort to fast-track investigations and work-up for definitive treatment while awaiting final histopathology.

Introduction/Background Circulating tumor DNA found in women with malignancy, enters plasma due to lysis of cells at the interface between the primary tumor and the circulation. The primary objective of this study was to isolate and quantify cell-free DNA (cfDNA) from peripheral blood, analyze p53 mutations and correlate with tumor burden in epithelial ovarian malignancy. Secondary objective was to study the degree of agreement between cfDNA p53 mutations and tissue p53 immunohistochemistry. Methodology This prospective case-control study was carried out over 18 months from November 2018 to April 2020 at a tertiary teaching institution. Considering the exploratory nature of the study, study group (n=20) comprised women with epithelial ovarian malignancy. Control groups were women with borderline tumors (n=10) and benign epithelial ovarian tumors (n=10). 58 women who were treatment naive and admitted for surgery entered the study but only those with a final histopathology of epithelial ovarian tumor (malignant, borderline and benign) were included. Peritoneal carcinomatosis index (PCI), surgical complexity score and cytoreductive score was calculated in women undergoing primary cytoreduction. Plasma samples for cfDNA was collected just before surgery and stored at -20oC. cfDNA was extracted from plasma serum using a DNA isolation kit and quantified with Nanodrop Spectrophotometer. ARMS PCR was used to detect a point mutation in Exon 8, codon 239 of p53 using primer pairs. p53 immunostaining was performed on tissue samples using monoclonal antibody directed against p53. Statistical analysis was done using SPSS version 21. Results In women with malignant ovarian cancer isolated cfDNA was highest (1330 ng/mL) in comparison to those with benign or borderline ovarian tumors (748.5 ng/mL and 448.5 ng/mL, respectively) reaching statistical significance, p=0.023. Quantity of cfDNA also correlated well with the histopathological grade of the tumor and stage of the disease, p Analysis of cfDNA p53 mutation in exon 8 showed that 55% of the women diagnosed with malignant ovarian tumor harboured this mutation (p=0.043). Correlation of tissue p53 with cfDNA p53 mutation was statistically significant, p=0.007. All women with malignant ovarian tumor in whom cfDNA p53 mutation was present at codon 239 of exon 8 stained positive for tissue p53 mutation. Conclusion cfDNA p53 mutation in exon 8 was detected at higher frequency in women with malignant epithelial ovarian cancer. Significant correlation was seen between tissue p53 and cfDNA p53 mutation suggesting that mutational analysis of cfDNA could act as biomarker for the diagnosis of ovarian tumors. Disclosures Shalini Rajaram – No conflict of interest Aanchal Verma - No conflict of interest Rajarshi Kar - No conflict of interest Vinod Arora - No conflict of interest Bindiya Gupta - No conflict of interest Sandhya Jain - No conflict of interest

Background Urine cytology is the corner-stone for the diagnosis of urothelial neoplasia; however, a substantial proportion of low-grade carcinomas are reported as inconclusive owing to scant cellularity and subtle cytological features. Biomarkers applied on urine sediment smears of such patients are likely to be clinically relevant. Access to Food and Drug Administration approved urinary biomarkers in resource limited setting is poor. Detection of cytokeratin 20 (CK20) in urine sediments, although still a research tool, is a promising marker as immunocytochemistry is performed regularly in several Indian laboratories. Objective We tested the clinical utility of CK20 immunocytochemistry as a potential low-cost adjunct to urine cytology in diagnosis of low-grade urothelial carcinoma. One hundred and fifty fresh, voided urine specimens from 42 cases of biopsy proven urothelial neoplasia (14 high grade, 28 combined low-grade [n=26]) and low malignant potential [n=2]), and 20 non-neoplastic lesions were included in the study sample. Results Confident diagnosis of malignancy was possible in five (17.8%) low-grade malignancies. Thirteen of 16 (81.3%) low-grade malignancies with inconclusive cytology showed positive CK20 expression. This reduced the proportion of low-grade cases with inconclusive cytology from 57.1% to 10.7% (P=.021). In addition, we could correctly classify one case of bladder lithiasis with false positive urine cytology. Discrepant CK20 staining (positive) was seen in one patient with acute cystitis. Conclusions CK20 expression in non-umbrella cells is a robust marker of urinary bladder carcinoma. It has potential clinical utility for identification of low-grade urothelial malignancy with inconclusive cytological diagnosis.

Introduction Extrapulmonary tuberculosis (EPTB) is one of the major diagnostic challenges in clinical practice. The aim of the study was to compare the utility of Polymerase Chain Reaction (PCR) with conventional techniques in the diagnosis of EPTB. Materials and methods Specimens from 150 patients with a presumptive diagnosis of EPTB were examined by Zeihl-Neelsen staining and mycobacterial culture on Lowenstein Jensen medium. Histopathology was done for lymph node aspirates and endometrial biopsies. DNA from all the samples was amplified by PCR targeting 123 bp and 240 bp fragments of IS6110 and MPT64 genes respectively. Results IS6110 PCR was more sensitive (82.1%) as compared to PCR for MPT64 gene (81%) though the difference was not found to be statistically significant. Four (2.6%) and 2 (1.3%) samples were positive by IS6110-PCR only and MPT64-PCR only respectively.Conclusion: PCR with multiple targets can have potentially important role in improving diagnostic accuracy of EPTB.

Background & objectives: Psoriasis is a recurrent hyper-proliferative skin disease which is often associated with free radical generation, abnormal lipid metabolism and increased inflammatory secretion that induce cardiovascular risk in these patients. The present study was intended to evaluate serum lipids, lipoprotein and oxidants-antioxidants status and to establish their relationship with atherogenic risk markers [oxidized low-density lipoprotein (oxLDL) and high-sensitivity C-reactive protein (hsCRP)] in patients with psoriasis. Methods: The study was conducted on 150 psoriasis patients and 150 age- and sex-matched healthy controls. Overnight fasting blood samples were obtained for lipids, lipoproteins, lipid oxidation and peroxidation products [oxLDL, malondialdehyde (MDA)], antioxidant enzymes [reduced glutathione (GSH) and total antioxidant status] levels and hsCRP estimations. >Results: The mean levels of atherogenic lipids [total cholesterol (PInterpretation & conclusions: The study results suggest that LDL oxidation and reactive oxygen species in addition to inflammatory markers may play a pivotal role in inducing atherosclerosis in patients of psoriasis.

Context: In the context of competency-based medical education being advocated worldwide, fine needle aspiration biopsy (FNAB) is considered as an entrustable professional activity (EPA). There is no information regarding how much time and training are required to achieve a “competent level” for performing and documenting FNAB in the Indian context. Aim: To determine the time taken by an average postgraduate pathology trainee to become competent in performing FNAB with respect to history taking, clinical examination, and fine needle aspirate adequacy. Settings and Design: A descriptive, retrospective, chart-based audit was conducted in the Department of Pathology. Materials and Methods: FNAB chart records documented during 3 years of postgraduate training by a cohort of 13 postgraduate (PG) resident trainees admitted in 2010 were included in the study. Adequacy rates and criteria for adequacy were defined for the purpose of the study. Statistical analysis: Data was entered in MS Excel and analyzed using the Statistical Package for the Social Sciences version 20.0. The adequacy rates are presented as percentages and time taken to achieve adequacy rates as median values. Results: A total of 3272 charts were audited. Median time taken to achieve 85% adequacy rate for history taking, for clinical examination by the first and the second criteria, and for FNAB were 1 month, 1 month and 3.5 months, and 1 month, respectively. Conclusions: Although the mean time taken to achieve 85% adequacy rates for FNAB was 1 month, there was wide variation in the time durations between the residents to achieve this level.

Objectives This study was designed to correlate expression of epithelial mesenchymal transition (EMT) pathway markers i.e., E-cadherin and Vimentin with surgicopathological extent of EOC and to type the tumour using p53 immunohistochemistry staining. Methods Women with malignant and benign epithelial ovarian tumours were studied. Sample size was calculated with 80% power and 5% level of significance ;22 cases (EOC) and 22 controls (benign ovarian tumour) were recruited. m-RNA and protein expression of E-cadherin and vimentin was done by real time PCR and IHC staining and p53 by IHC. Peritoneal extent of disease was calculated by peritoneal carcinomatosis index (PCI) and tumour resection by completeness of cytoreductive score (CCS) and correlations derived. Results In advanced EOC, positive correlation was found between PCI and CCS with correlation coefficient of 0.495, p-value Conclusions Expression of E-cadherin decreased and Vimentin increased in EOC which is in synchrony with EMT pathway, however larger studies are needed to derive an association between these markers and extent of disease.

To test the clinical utility of biomarkers p16/Ki-67 expression in cervical cytology smears as a marker for transforming HPV infection. Experimental study at a tertiary care hospital. Women who were screened positive on Pap and visual inspection tests (n = 280) underwent colposcopy and biopsy. p16/Ki-67 immunostaining was performed in abnormal Pap smears (n = 86), and HPV DNA testing was also performed in the same women. Sensitivity, specificity, negative predictive value, positive predictive value and likelihood ratios were calculated for each biomarker separately and in combination. McNemar test and receiver operating characteristic (ROC) curves were used to compare sensitivity and specificity of biomarkers with HPV DNA. Areas under the ROC curve (AUC) were compared using the Chi-square test. Eighty-six women with abnormal cytology were evaluated with p16/Ki-67 immunocytochemistry; 20.9% (n = 18) and 18.6% (n = 16) were positive for each biomarker, while dual marker was positive in 15% (n = 13). In all smears, the sensitivity of p16INK4a/Ki-67 in detecting CIN 2+ lesion was 76.9% and specificity was 95.8%. For ASCUS (n = 42) and LSIL (n = 23) smears, specificity and negative predictive value of p16/Ki-67 for CIN 2+ were 100% with a likelihood ratio (LR+) of 27 and 25, respectively, suggesting good diagnostic accuracy. In comparison with HPV DNA testing, combined marker p16/Ki-67 was significantly more specific (p = 0.003); AUC was 0.734 and 0.635, respectively. p16/Ki-67 evaluation in cervical cytology is a valuable biomarker in triaging for CIN 2+ disease in ASCUS and LSIL smears.

Background Fine needle aspiration cytology (FNAC) plays a pivotal role in evaluating salivary gland (SG) tumors. Several studies have shown diagnostic utility of MILAN system for reporting salivary gland cytopathology (MSRSGC) by examining risk of malignancy but only an occasional study has focused on interobserver variability. Hence, the present study was undertaken to assess the agreement among cytopathologists with varying experience in SG cytopathology using MSRSGC and to re-evaluate discordant cytohistological diagnoses for possible causes of misinterpretation. Methods All SG lesions subjected to FNAC over a period of 3½ years were studied. The cases were critically reviewed by 2 pathologists with variable experience in cytopathology using MSRSGC and concordance level among them was calculated. Cytohistological discordant diagnoses were reclassified and possible causes of misinterpretation during routine reporting were evaluated. Results Of 150 SG aspirates categorized according to MSRSGC, diagnostic disagreement between 2 pathologists was found in 10. Unweighted Cohen's Kappa score between consultant and resident was 0.812 (high). Among 55 cases with histological correlation, cytohistological discordance was seen in 12. True pitfalls constituted 50% of discordant cases while rest 50% were attributed to practical issues (turnaround time and heavy case load) during routine reporting. Conclusion MSRSGC can be used with good reproducibility between observers with variable cytopathology experience. Heterogeneous nature of SG neoplasm is a known pitfall in FNA diagnosis of SG neoplasms. During routine reporting turnaround time, heavy case load and reporting by cytopathologists with variable experience add on to challenges faced in reporting cytopathology of SG neoplasm.

Basal cell carcinoma (BCC) is the most common skin cancer worldwide. Surgical excision is considered to be the primary therapeutic modality wherever possible. For inoperable cases, 5% imiquimod seems to be a good alternative. We present two cases of nodular pigmented BCCs on the face in elderly women successfully treated with 5% imiquimod cream application resulting in complete clinical clearance of lesion as well as on histology and dermatoscopy. There was no recurrence of the lesion on 2 years follow-up for the first and 1.5 years for the second patient.

We report a 45-year-old, apparently healthy sero-negative man, presenting with multiple ulcers on the glans penis for a duration of three months. There was no significant inguinal lymphadenopathy. He showed no improvement on systemic antibiotics and acyclovir. Histopathology revealed the diagnosis of genital tuberculosis (TB), and polymerase chain reaction for Mycobacterium tuberculosis tested positive. The patient responded well to category I anti-tubercular treatment with complete resolution of lesions in six months. It is important to consider a differential diagnosis of penile TB in patients with non-healing genital ulcers.

e18012 Background: VEGF initiates endothelial cell proliferation and angiogenesis in tumour cells while CA IX has an important role in pH regulation and elimination of hypoxia generated acid load during glycolysis.The present study was designed to identify these biomarkers and their correlation with tissue spread and behavior and eventually could lead to development of novel target therapies. The objective of this study was to correlate serum VEGF levels and tissue CA IX expression with clinicopathological features of carcinoma cervix stage IA to IIA. Methods: This exploratory case-control study included 19 cases with carcinoma cervix FIGO 2009 stage IA to IIA and 19 age matched controls. Out of the 19 cases, 12 underwent surgery and 7 received chemoradiation. Pre-therapy blood samples were collected for serum VEGF estimation from both cases and controls. For CA IX expression estimation, cervical tissue was taken for analysis. For cases, post-operative / post chemoradiation blood samples were taken for VEGF level estimation after 6 weeks. Results: The difference in mean serum VEGF levels between cases 1235.84 pg/ml (SD± 433.67) and controls 153.74 pg/ml (SD± 108.11) was statistically significant (p = 0.001). Mean serum VEGF level between pre and post-operative cases was 1108 pg/ml (SD± 407.62) and 391.42 pg/ml (SD± 270.71) while in pre and post chemoradiation cases it was 1455 pg/ml (SD± 413.38) and 623 pg/ml (SD± 238.41). This decrease was statistically significant (p = 0.00 & p = 0.018). VEGF positively correlated with histopathologic type (p = 0.016), grade (p = 0.012), and lymphovascular space invasion (p = 0.042).CA IX expression was seen in all cases while in controls no expression was seen. CA IX expression had a positive association with lymphnode metastasis (p = 0.041). Conclusions: Elevated pre-therapeutic serum VEGF suggests spread of tumor and can be used as a prognostic marker for cancer cervix. CA IX immunostaining in cervical biopsy samples can be used as a surrogate marker for tumor aggressiveness and distant spread.

Background: Majun Brahmi (MB) and Itrifal Muqawwi Dimagh (IMD) are polyherbal Unani formulations traditionally used for the treatment of mild cognitive impairment and memory enhancement. Although they are widely used, there is no particular scientific evidence regarding the toxicity of these medicines. Objective: The present study was undertaken to evaluate the safety of MB and IMD polyherbal Unani formulation. Materials and Methods: Wistar albino rats of both genders were treated with three doses of each drug – MB and IMD, i.e., 513.88, 1027.77, and 2055.54 mg/kg body weight (bwt) orally for 14 days for acute and 90 days for subchronic oral toxicity studies. At the end of the study, bwt gain, hematology, clinical biochemistry, and histopathological examination were performed. Although no significant changes in biochemical parameters occurred at the highest dose, mild damage to the liver, kidney, and spleen was observed. Results: At the lowest dose and at the dose of 1027.77 mg/kg bwt (therapeutic effective dose [TED]) of both the drugs, no abnormalities were observed in the biochemical and hematological parameters as well as histopathological findings. Conclusion: None of the rats exhibited apparent toxicity or mortality. The results of the present study provide evidence that both the formulations are not toxic at the TED dose and hence are clinically safe.

Background: Cancer of the uterine cervix is the second most common cancer among women worldwide. More than three fourths of these patients are diagnosed at advanced stages, leading to poor prospects of long term survival and cure. Introduction of Papanicolaou (Pap test) cytological screening for cervical precancerous lesions has significantly reduced the incidence and mortality of cervical cancer. Human papillomavirus (HPV) DNA testing has been recently demonstrated to be efficient to be integrated into screening programs, so it can be used to triage women with equivocal cytological abnormalities and also identifies women at risk of residual or recurrent disease after treatment of cervical intraepithelial neoplasia. However, it fails into triage low grade lesions. HPV DNA test confirms infection by the virus, present in 99% of cases. However, it does not discriminate between transient and persistent infection as it is crucial because it is the persistent infection which progresses to neoplasia. Histological assessment of cervical biopsy that is considered as “gold standard” can be hampered by intra and inter observer variability. A more specific triage marker is required to identify women who would need colposcopy. Hence, p16INK4a has emerged as a new diagnostic and prognostic biomarker. Aims and objectives: This study was conducted to evaluate the utility of staining of p16INK4a on conventional Pap smears and its comparison with corresponding biopsies. Material and Methods: 50 cases of conventional Papanicolaou stained cervical smears cases were randomly selected from the archive of cytopathology laboratory. The cervical smears were re-evaluated for adequacy, preservation of cells, cytomorphology and various lesions were categorized according to The Bethesda system 2001 (TBS) classification. Consecutive such cases were selected for which both cytological and histological material were available. Immunostaining of cervical cytological specimens for p16 was performed using monoclonal murine antibody, clone 16P04, JC2. Ready to use antibody was used for immunostaining as per manufacturer's protocol. Results: Out of the 50 smears of preneoplastic and invasive lesions, ASCUS was seen in 5 cases, LSIL in 10 cases, HSIL in 20 cases, SCC in 20 cases and AGUS in 5 cases. Of the 5 cases of ASCUS, histopathology showed 3 chronic cervicitis and 1 each in CIN-1 and CIN-2. Histopathological diagnosis of 10 cases of LSIL showed 6 CIN-1 and 2 each in CIN-2 and chronic cervicitis. Similarly, for 10 cases of HSIL, 3 were CIN-2, 5 CIN-3 and 2 SCC. All 20 cases of carcinoma showed SCC while 5 cases of AGUS showed 4 chronic cervicitis and one adenocarcinoma. Immunohistochemical staining of p16INK4A showed weak positivity in 3 cases of chronic cervicitis and 4 cases of CIN-1. In CIN-2 cases, 66.67% showed strong positivity, CIN-3 showed 80% while both carcinoma and adenocarcinoma showed 100% strong positivity. The sensitivity of immunohistochemical staining of p16INK4A was 77.5%, specificity was 100%, PPV 100% and NPV 52.6%. The relation between histological and cytological immunostaining of p16INK4A. Conclusion: p16INK4A is a reliable marker for dysplastic squamous and glandular cervical cells both in tissue sections and in cervical smears. p16 immunostaining can be easily performed on CPS, and there is high concordance of positivity on smears and tissue sections.

Lichen nitidus (LN) is a chronic inflammatory condition characterised by multiple asymptomatic shiny, flat-topped, pale to skin-coloured tiny papules. LN occurs most often in children and young adults with limbs, abdomen, chest, and penile shaft as common sites of predilection. Nail involvement is rare and includes irregular longitudinal grooving and ridging of the nail plate. We report two cases of generalised LN with nail dystrophy in 8- and 12-year-old boys along with the treatment response and review the pertinent literature.

Objectives The diagnosis of cutaneous tuberculosis is challenging due to its diverse clinical manifestations, paucibacillary state and lack of proper diagnostic tests. Clinico-pathological correlation is still frequently used for diagnosis. There is paucity of literature on cytomorphological features. Immunochemistry can help as an ancillary test. Methods Clinical diagnosis was made after thorough history and physical examination. Modified Fine Needle Aspiration technique was used to collect cytology samples and 3 mm punch biopsy for histological examination. Findings on histopathology were compared with cytomorphology. Immunochemical staining with anti-TB polyclonal antibody using standard Polymer-based-HRP immunochemistry technique and comparison of cytology and histology findings. Results The morphological spectrum of biopsy and cytology showed high correlation using nine parameters: necrosis, granulomas, giant cells, AFB, neutrophilic infiltrate, presence of lymphocytes, histiocytes, collagen bundles, and immunochemistry. Diagnostic correlation of FNA compared to biopsy was found to be 90.3%. On comparing cytomorphology of scrofuloderma and lupus vulgaris, all the parameters were found more frequently in scrofuloderma except for granulomas, giant cells and immunochemistry. Immunochemistry showed sensitivity and specificity of 90.3% and 70% on biopsy, respectively, compared to 67.7% and 60% on FNA, respectively. Combined sensitivity of IHC and ICC was 96.8%. Conclusions The cytomorphological spectrum of cutaneous tuberculosis is comparable to clinicohistopathology with a high correlation of 90.3%. However, sub classification on FNA is difficult on cytology alone. While FNAC is a better diagnostic tool for finding AFBs hence confirming the diagnosis, biopsy is better for immunochemistry. Thus, biopsy and FNA complement each other.

Malakoplakia is a rare chronic granulomatous inflammatory disease, related to immune deficiency and impaired bactericidal macrophage activity. Common sites of involvement include urinary bladder and kidney followed by gastrointestinal tract. We present an unusual case of abdominal malakoplakia diagnosed preoperatively on ultrasound-guided fine-needle aspiration cytology (FNAC). It presented as a large locally aggressive mass with extensive involvement of multiple bowel loops in an 18-year-old boy following blunt trauma to the abdomen. The demonstration of calcified laminated intracytoplasmic Michaelis-Gutman bodies in histiocytes is a reliable diagnostic feature of malakoplakia on FNAC smears, which can guide an appropriate medical management.

Background: Studies on ultrasonography (USG) guided fine needle aspiration cytology (FNAC) have been conducted in specialized settings such as thyroid, breast, and intra-abdominal aspirates. There is a paucity of literature on the practices of guided FNAC in a general cytopathology service. Aim: The aim was to determine prevailing practices of USG guided FNAC in a general cytopathology service of a teaching hospital. Settings and Design: Metropolitan hospital, clinical audit. Materials and Methods: Audit of 112 USG guided percutaneous FNAC done over 12 months. Statistical Analysis: Data were coded, entered in an excel spreadsheet and analyzed by translating into percentages and proportions. Results: The 112 guided FNACs included constituted 36 thyroid (32.14%), 45 intra-abdominal (40.17%), 11 breast (9.82%), 10 superficial lymph node (8.92%) and 10 soft tissue and miscellaneous (8.92%) lesions. Previous freehand FNAC was documented on the requisition forms in 14 cases. The reports were: Inadequate 33 (29.46%), nondiagnostic descriptive 35 (31.25%) or diagnostic 44 (39.28%). Inadequacy rates of aspirates from thyroid were 11 (30.56%) breast were 2 (18.18%), and intra-abdominal lesions were 13 (28.88%). Majority of the reports were nonstructured: 108 (96.42%) and nonrecommendatory: 101 (90.17%). Conclusions: Reporting practices varied and did not conform to a uniform structure. The inadequacy rates of breast and thyroid aspirates were comparable to the rates in the literature. Comparable studies were not available for intra-abdominal aspirates.