039935   BABAEI A, TAGHAVI S A, MOHAMMADI A, MAHDIYAR M A, IRANPOUR P, EJTEHADI F, MOHAGHEGHZADEH A (Shiraz Univ of Medical Sciences, Iran, Email: ataghavi@yahoo. com) : Comparison of the efficacy of oral fenugreek seeds hydroalcoholic extract versus placebo in nonalcoholic fatty liver disease; a randomized, triple-blind controlled pilot clinical trial. Indian J Pharmacol 2020, 52(2), 86-93.

The aim of this study is to investigate the therapeutic property of hydroalcoholic extract of Fenugreek seeds in nonalcoholic fatty liver disease (NAFLD) in adult patients. This randomized, placebo‑controlled, parallel trial was conducted from November 2014 to June 2017. Patients aged between 18 and 70 years old with confirmed NAFLD were recruited from the Motahhari clinic, affiliated to Shiraz University of Medical Sciences, Iran. Participants either received 1 g hydroalcoholic extract of Fenugreek seeds or placebo daily for 3 months. The primary outcomes were changes in serum alanine transaminase and FibroScan controlled attenuation parameter score. Secondary outcome measures were changes in other laboratory data, liver stiffness measure, liver steatosis percent, and anthropometric variables. Participants were randomly assigned to the groups using blocked randomization method. Participants, investigators, and statistician were blinded to treatments allocation. After screening eighty patients, thirty patients met the inclusion criteria and were divided into two groups (1:1). After 3 months, two and four patients did not complete the trial in Fenugreek and placebo group, respectively. The changes in the anthropometrics, laboratories and FibroScan measurements were not statistically significant between the two groups. The evidence to prove the efficacy of the Fenugreek seeds’ hydroalcoholic extract in NAFLD was not strong enough; hence, further experiments are still needed to assess the possible efficacy of Fenugreek on the treatment of NAFLD. 

2 illus, 2 tables, 29 ref

039934   KADAM R L, SONTAKKE S D, TIPLE P, MOTGHARE V M, BAJAIT C S, KALIKAR M V (Psychiatry Dep, Government Medical Coll, Nagpur- 440 022, Maharashtra, Email: smitaavanti@ yahoo.co.in) : Comparative evaluation of efficacy and tolerability of vilazodone, escitalopram, and amitriptyline in patients of major depressive disorder: A randomized, parallel, open-label clinical study. Indian J Pharmacol 2020, 52(2), 79-85.

To evaluate and compare efficacy and tolerability of Vilazodone with Escitalopram and Amitriptyline in patients of major depressive disorder(MDD). This was a randomized, prospective, parallel-group, open label clinical study in which newly diagnosed patients of MDD were randomized to receive Tab Vilazodone 20 mg daily or Tab Escitalopram 20mg daily or Tab Amitriptyline 75mg daily for 12 weeks. Antidepressant activity was assessed by change in score from baseline to week 12 on HAMD-17 and MADRS scales while change in score on HAM-A scale was used to assess antianxiety effect. Change in scores on the three scales was also compared between the three treatment groups. Severity and causality of adverse events were assessed by the modified Hartwig & Siegel scale and Naranjo scale respectively. Data was analyzed in accordance with per protocol analysis. Reduction in HAMD-17 and MADRS scores was significantly more in vilazodone group compared to the other two drugs indicating that vilazodone is more efficacious antidepressant. Number of remitters were also significantly more in the vilazodone group (n=11) compared to escitalopram (n=4) (p<0.05) and amitriptyline (n=0) (p<0.001) at 12 weeks. Similar results were also obtained with HAM-A score. Number of patients showing MADRS sustained response at 12 weeks was statistically significantly more in vilazodone (n=12) and escitalopram (n=12) groups compared to amitriptyline (n=01) (p<0.001). Reported adverse events were constipation and sedation(amitriptyline group); nausea and headache(escitalopram and vilazodone groups).These adverse events were of mild severity. Most adverse events belonged to probable category. Vilazodone is more efficacious and well tolerated antidepressant compared to escitalopram and amitriptyline.

5 tables, 30 ref

039933   SACHDEVA J, MAHESH K V, SHREE R, JAIN G, KAPILA A T, SHASHIKALA T P, GOYAL M K, MODI M, LAL V (Neurology Dep, Postgraduate Institute of Medical and Research, Chandigarh- 160 012, Email: shashjules@ yahoo.co.in) : Use of rituximab in muscle-specific tyrosine kinase antibody-positive myasthenia gravis: Preliminary observations from a tertiary care center in Northern India. Indian J Pharmacol 2020, 52(1), 49-52.

Approximately 10 %–15 % of patients with myasthenia gravis (MG) are refractory to standard treatment. A sizable chunk of these patients is due to muscle‑specific tyrosine kinase (MuSK) antibody‑positive MG which often runs a severe course with frequent relapses and poor response to conventional treatment. We report six patients with refractory MuSK‑positive MG who responded well to the treatment with rituximab. In this prospective institute‑based observational study, we report six MuSK antibody‑positive MG patients, who did not achieve remission with standard treatment and were later started on rituximab infusion. There was a significant clinical improvement in all patients after starting rituximab.  Rituximab is an effective immunomodulatory therapy in MuSK antibody‑positive MG patients who are not responding to the standard treatment.

1 table, 11 ref

039932   KOLOKAS K, KOUFAKIS T, AVRAMIDIS I, GEROU S, CHATZIDIMITRIOU M, KAZAKOS K, KOTSA K (Internal Medicine Dep, AHEPA Univ Hospital, Thessaloniki- 54636, Greece, Email: kalmanthou@ yahoo.gr) : Fasting insulin levels correlate with the frequency of hypoglycemic events in people with type 2 diabetes on treatment with sulfonylureas: A pilot study. Indian J Pharmacol 2020, 52(1), 44-8.

We aimed to explore whether fasting insulin levels correlate with the risk of hypoglycemia in people with Type 2 diabetes (T2D) receiving sulfonylureas (SUs). Our study included 58 individuals with T2D who had been on treatment with SUs, but not insulin, for more than 2 years. Confirmed hypoglycemic episodes during the past year were self‑reported by the patients, and a potential relationship of hypoglycemic event frequency with fasting insulin levels was investigated. Fasting insulin concentrations were found to have a low positive and statistically significant correlation with the number of cases of mild hypoglycemia per year (ρ = 0.279/P = 0.034) and a moderately positive and statistically significant correlation with the number of severe hypoglycemic events per month (ρ = 0.349/P = 0.007) and per year (ρ = 0.39/P = 0.002). Our results suggest that fasting insulin levels might be a predictor of the risk of hypoglycemia in people with T2D on treatment with SUs.

2 tables, 20 ref

039931   ZHANG W, CHEN H, DING Y, XIANG Q, ZHAO J, FENG W, YANG L (Jiangsu Univ, Jiangsu, China, Email: fwwujs@126.com) : Effect of chromium citrate on the mechanism of glucose transport and insulin resistance in buffalo rat liver cells. Indian J Pharmacol 2020, 52(1), 31-8.

Our published literature indicated that chromium citrate could regulate the glycemic index in alloxaninduced diabetic mice. The present study investigated the mechanism of chromium citrate in insulin resistance (IR) buffalo rat liver (BRL) cells. Chromium citrate was synthesized in our laboratory. BRL cells were purchased from the Chinese Academy of Sciences Cell Bank. The glucose transport and IR affected by chromium citrate in BRL cells were examined. The Thiazolyl Blue Tetrazolium Bromide (MTT) and glucose assay experiments were measured by microplate ELISA reader. The protein kinase B (Akt), glucose transporter‑4 (Glut4), and phosphor‑AMP‑activated protein kinase β1 levels were tested by Western blot, and the mRNA expression of glucose transport proteins (Akt2, Glut4, and AMPactivated protein kinase α2 (AMPKα2)) and insulin sensitivity proteins (insulin receptor substrate1 (IRS-1), phosphatidylinositol 3 kinase (PI3K), and peroxisome proliferator-activated receptor γ (PPARγ)) was measured by reverse transcription–polymerase chain reaction. The results indicated that the glucose absorption level of chromium citrate groups was higher than model group significantly. It demonstrated that chromium citrate could significantly improve glucose absorption in IR BRL cells. The Akt, Glut4, and phosphor‑AMPKβ1 levels in chromium citrate groups (at doses of 0.4, 0.2, and 0.1 μg Cr/mL) were markedly improved when compared with the other experiment groups, and chromium citrate could more effectively increase the Akt level than chromium trichloride. In addition, the mRNA expression of Akt2, Glut4, and AMPKα2 in chromium citrate groups was significantly improved when contrasted with model group. The consequences illustrated that chromium citrate can affect the IR BRL cells’ ameliorating glucose transport and IR.

5 illus, 1 table, 32 ref

039930   MAGAZINE R, VENKATACHALA S K, GONEPPANAVAR U, SURENDRA V U, GUDDATTU V, CHOGTU B (Pharmacology Dep, Kasturba Medical Coll, Manipal- 576 104, Karnataka, Email: bharti.magazine@ manipal.edu) : Comparison of midazolam and low-dose dexmedetomidine in flexible bronchoscopy: A prospective, randomized, double-blinded study. Indian J Pharmacol 2020, 52(1), 23-30.

Dexmedetomidine is a clinically useful drug for providing sedation, but concern regarding its cardiovascular side effect profile can limit its widespread use during routine diagnostic flexible bronchoscopy (FB). Patients between 18 and 65 years of age, who required diagnostic FB, were screened. Eligible patients were randomized to either receive 0.5 μg/kg intravenous dexmedetomidine over 10 min or intravenous midazolam 0.035 mg/kg over 1 min. If required, rescue medication (intravenous midazolam 0.5 mg bolus) was administered. The primary outcome measure was the composite score. Other parameters observed were numerical rating scale, hemodynamic variables, oxygen saturation, number of doses of rescue medication, visual analog scale score for cough, ease of bronchoscopy, Ramsay Sedation Score, and postprocedure patient response after 24 h of bronchoscopy.  A total of 54 patients were enrolled, 27 in each group. Total composite score (mean ± standard deviation) in dexmedetomidine and midazolam group at nasopharynx was 7.04 ± 2.19 and 6.59 ± 1.55 (P = 0.387), respectively. The corresponding values at the level of trachea were 9.22 ± 3.69 and 8.63 ± 2.13 (P = 0.475). In the dexmedetomidine group, patient response after 24 h of bronchoscopy showed the quality of sedation to be excellent in three patients, good in 10, fair in 11, and poor in 3 and discomfort to be nil in 14, mild 7, moderate in 3, and severe in 3. The corresponding values in the midazolam group for the quality of sedation were 0, 9, 18, 0 and for discomfort 10, 16, 1, 0. Other parameters did not reveal any statistically significant difference. Dexmedetomidine at a dose of 0.5 μg/kg may provide clinically useful conscious sedation, comparable to midazolam.

2 illus, 5 tables, 26 ref

039929   ABIDI A, RIZVI D A, SAXENA K, CHAUDHARY S, AHMAD A (Pharmacology Dep, Lucknow Medical Coll, Lucknow- 226 003, Uttar Pradesh, Email: afrozabidi@gmail. com) : The evaluation of efficacy and safety of methotrexate and pioglitazone in psoriasis patients: A randomized, open-labeled, active-controlled clinical trial. Indian J Pharmacol 2020, 52(1), 16-22.

Psoriasis is a chronic inflammatory disease showing co‑existence with metabolic syndrome (MS), as has been confirmed by numerous epidemiologic studies in recent times. In this study, the aim was to ascertain the beneficial effects of pioglitazone in psoriasis, simultaneously targeting the improvement of MS parameters. We conducted a prospective randomized open‑labeled parallel‑group interventional study in patients of moderate‑to‑severe chronic plaque psoriasis. A total of 90 patients were inducted in study and divided into three groups of standard treatment (methotrexate 7.5 mg/week for 12 weeks), active treatment (pioglitazone 15 mg tablets once daily for 12 weeks), and their combination. Primary outcome was taken as percentage Psoriasis Area and Severity Index (PASI) improvement from baseline; secondary outcomes were PASI‑75, safety profile, and MS parameters. Intergroup evaluation of PASI score showed that standard treatment methotrexate and active treatment pioglitazone were comparable. Combination of methotrexate and pioglitazone proved superior in efficacy from both standard and active treatment in 8 and 12 weeks. Adverse drug reactions were mild and treated symptomatically. Pioglitazone and combination group also demonstrated beneficial efficacy in parameter of MS hence establishing it as a potential therapy in psoriasis with MS. Pioglitazone alone or in combination with standard treatment may be a safe alternative drug for psoriasis coexisting with MS proving beneficial for both.

2 illus, 5 tables, 28 ref

039928   MAITI K, JAISWAL A, PAL D K (Urology Dep, Institute of Post Graduate Medical Education and Research, Kolkata- 700 020, West Bengal, Email: urologyipgmer@ gmail.com) : A comparative study of alpha-1a blockers (tamsulosin) versus estrogens in the treatment of lower urinary tract symptoms in perimenopausal females. Indian J Pharmacol 2020, 52(1), 6-9.

Lower urinary tract symptoms (LUTS) in perimenopausal females are very common. It can be treated with alpha‑blockers or application of topical oestrogen. The purpose of this study is to compare the efficacy of alpha‑blockers versus topical estrogen in the treatment of LUTS in perimenopausal females. All perimenopausal females between the age group of 45 and 60 years who present with the symptom of voiding LUTS were divided into two groups. Acute urinary retention patients were excluded from the study. The first group was given alpha‑blocker (tamsulosin) and other group was given topical estrogen application (0.5 %–1 %) in the periurethral region. Patients were followed up clinically by voiding components of the International Prostate Symptom Score and objectively by uroflowmetry and postvoid residual (PVR) urine estimation (ultrasonography). Alpha‑blocker group had 40 females and topical estrogen group had 40 females. During the 6‑week period, 8 patients of the first group and 6 patients of the estrogen group discontinued the treatment. In the first group, pretreatment mean Qmax (maximum flow rate) of patients was 7.2 ml/s and posttreatment Qmax was 18.4. In the second group, the values were 7.4 ml/s and 10.2, respectively. This difference was statistically significant (P < 0.0001). In the first group, pretreatment PVR urine was significant, which became insignificant after the treatment, whereas in the second group, PVR was significant posttreatment also. Alpha‑1a blockers should be used as the first‑line medical management in perimenopausal females with symptoms of LUTS, as they have a clear advantage over topical estrogens.

3 illus, 1 table, 7 ref

039927   GAUR K, PURI V, SHUKLA S, SHARMA S, SUMAN S, SINGH R (Pathology Dep, Lady Hardinge Medical Coll, New Delhi- 110 001, Email: drvandanapuri@gmail.com) : Finish before the start: Analyzing preanalytical sample errors in a tertiary care hematology laboratory. Indian J Pathol Microbiol 2020, 63(3), 435-40.

To evaluate the types and frequencies of preanalytical errors occurring in a tertiary care hematology diagnostic center and (b) To evaluate differences if any, across groups [outpatient data (OPD) vs inpatient data (IPD), type of test requested [complete blood count (CBC) vs coagulation] and laboratory (routine vs emergency). A prospective study was conducted over a period of nine months (August 2017–April 2018) to address the above objectives. All samples received in the clinical hematology division of our institute were included in the analysis. Categories of preanalytical errors were defined. This included insufficient, clotted, diluted, and lipemic samples. Clerical errors such as wrong/absent sample labeling, requisition form‑sample mismatch, and wrong vacutainer selection were also documented. IPD and OPD data, as well as data pertaining to samples sent for different tests [complete blood count (CBC)/coagulation] and in the routine and emergency laboratories, were segregated. All errors in each category were recorded as numbers and corresponding percentages (proportions). The two‑tailed z‑test was applied to assess the significance of the difference in proportions across all groups. Statistical significance was kept at P < 0.05. A total of 189,104 samples were received in the clinical hematology laboratory during the aforementioned period, out of which preanalytical errors were found in 4052 (2.14 %) samples. Inadequate sample quantity (ISQ) comprised the bulk of preanalytical errors in our laboratory (1.11 % of total samples) followed by sample clots (0.88 %). There was no significant difference in the error frequencies in OPD and IPD (P = 0.1031). The proportion of errors was higher in routine vis‑à‑vis emergency samples and also in samples sent for coagulation analysis vis‑à‑vis CBC.

3 illus, 5 tables, 22 ref

039926   REKHI B, KARMARKAR S, GUPTA C, DEODHAR K K, MENON S, PATHUTHARA S, MAHESHWARI A, SHYLASREE T S, GUPTA S (Surgical Pathology Dep, Tata Memorial Hospital, Mumbai- 400 012, Maharashtra, Email: rekhi.bharat@gmail.com) : Evaluation of cell blocks from effusion specimens in Gynecologic Oncopathology: An experience of 220 cases, diagnosed at a Tertiary Cancer Referral Center. Indian J Pathol Microbiol 2020, 63(3), 427-34.

One of the common indications of ascitic fluid examination in gynecological oncopathology is the detection and classification of malignant cells, especially in cases of clinically suspicious tubo‑ovarian masses. The present study was undertaken to assess and validate the diagnostic utility of cell blocks (CBs) and compare its results with the corresponding conventional smears, prepared from effusion samples. CBs were prepared by thromboplastin technique in 220 cases. In 208 cases, diagnostic concordance between results obtained from smears and corresponding CBs was evaluated. Various antibody markers were tested, as per individual case. The average age of patients was 52.2 years. Positive immunohistochemical (IHC) staining for various markers was observed in 182 cases (82.7 %) The most frequently positive antibody marker was PAX8 (101/134), followed by p53 (85/92) [mutation type (either diffusely positive or completely negative)], WT1 (tumor cells) (80/112), calretinin (2/87) (diffuse), BerEP4 (21/49), CA125 (21/24), CK7 (31/39) and CK20 and CDX2, together (5/16). Various other IHC markers utilized, including their positive expression, were TTF1 (1/10), p40 (3/3), p63 (2/4), ER (21/29), HBME1 (1/7), GATA3 (1/4), and MIC2 (1/1). Complete diagnostic concordance between CBs and smears was observed in 170/208 cases (81.7 %). There were 20 major discordances, 10 minor and 8 cases with sampling errors. IHC was useful in classifying 158/182 (86.8 %) cases, including serous or Müllerian adenocarcinoma (n = 123), mostly high‑grade (121); metastatic squamous carcinoma (3); gastrointestinal‑type adenocarcinoma (8); pulmonary adenocarcinoma (1); breast adenocarcinoma (1); Ewing sarcoma (1); and mesothelioma (2). CBs are complementary to smears in the detection of gynecological malignancies, mostly high‑grade serous adenocarcinomas. These provide an opportunity for testing several IHC markers, for a precise diagnosis, including in various uncommon case scenarios, associated with significant therapeutic implications.

7 illus, 2 tables, 32 ref

039925   KOLEVA M S, DIKOV D I, BELOVEZHDOV V T, SARAFIAN V (General and Clinical Pathology Dep, Medical Univ, Plovdiv, Bulgaria, Email: mariya.kolevaivanova@gmail.com) : Eosinophilic metaplasia in transurethral resection of the prostate. Indian J Pathol Microbiol 2020, 63(3), 423-6.

To investigate prostatic eosinophilic metaplasia (EM) in a large series of cases and their relationship with the basic prostate pathology in TURP‑material: benign prostatic hyperplasia (BPH), National Institutes of Health category IV prostatitis (also called histologic prostatitis or HP), and prostatic adenocarcinoma (PCa). The relation between EM and basic prostate pathology: BPH, PCa, and HP. Around 61 consecutive TURP‑specimens were reviewed for the presence of EM. The tissue sections were stained routinely with hematoxylin‑eosin (HE), hematoxylin‑phloxine‑saffron (HPS), and periodic acid‑Schiff’s procedure. Simultaneously BPH, HP, and PCa were evaluated. We found EM in 55.7 % of TURP‑specimens. EM is located more often in the ductal epithelium (58.8 %) and is usually focal (73.5 %) and in small groups (88.2 %) of secretory luminal cells. They are associated with BPH and with a variable degree of HP in all cases. However, there is no association with PCa. Eosinophilic cytoplasmic granules in EM are better visualized with HPS. Zones induced by tissue electrocoagulation which mimic EM, are seen in the periphery of TURP‑fragments. EM in prostate is presented by the presence of eosinophilic cytoplasmic granules in benign secretory epithelium. The study presents the first attempt to investigate EM in a large series of patients. Our results enrich the available information about the histoepidemiology of prostatic EM. Moreover, EM is more common in a focal lesion, found in small groups of ductal secretory epithelial cells while EM in TURP‑specimens is associated with BPH and HP in all the cases.

4 illus, 18 ref

039924   ÇELIK S Y, ÇELIK O (Pathology Dep, Sitki Kocman Univ, Turkey, Email: oilhancelik@gmail.com) : Can TROP2 be used as a prognostic marker in endometrioid endometrial carcinoma?. Indian J Pathol Microbiol 2020, 63(3), 418-22.

Endometrioid‑type endometrial carcinoma is the most common malignancy of the female genital tract in developed countries. The prognosis greatly depends on the grade and stage of the disease. In some patients, the disease recurs in a short time after the surgical/medical therapy. Hence, it is important to predict the patients who will have worse prognosis at the beginning, to choose the appropriate treatment; resuming the search of new prognostic markers. Therefore, our study aimed to detect trophoblast cell surface antigen 2 (TROP2) as a new prognostic marker. The patients who underwent a hysterectomy and diagnosed with endometrioid‑type endometrial carcinoma were evaluated retrospectively and TROP2 immunostain was performed to their tumoral slides. We evaluated TROP2 expressions in 102 patients immunohistochemically who underwent hysterectomy with the diagnosis of endometrioid‑type endometrial carcinoma histopathologically and correlated them with the other generally accepted prognostic parameters. The Kolmogorov‑Smirnov test and Q‑Q plot test were used to verify the normality of the distribution of continuous variables. The Chi‑square/ Fisher’s exact tests were used for categorical variables. Analyses were performed with SPSS Statistics for Windows, Version 20. High overexpression of TROP2 was seen in larger, higher‑grade, deeper‑invasive tumors, tumors with vascular invasion, and pelvic‑lymph‑node metastasis. These results were statistically significant (P ≤ 0.05). Overexpression of TROP2 in endometrioid‑type endometrial carcinoma seems to be a poor prognostic factor; it may be useful in determining the biologically more aggressive tumors before the treatment. This early determination is very important to choose the appropriate surgery, adjuvant‑treatments, and follow‑up.

2 tables, 32 ref

039923   YILDIRIM H T, NERGIZ D, SADULLAHOGLU C, AKGUNDUZ Z, YILDIRIM S, DOGAN S, SEZER C (Pathology Dep, Health Sciences Univ, Antalya- 07050, Turkey, Email: drhulyatosun@gmail.com) : The extent of cyclin D1 expression in endometrial pathologies and relevance of cyclin D1 with the clinicopathological features of endometrioid endometrial carcinoma. Indian J Pathol Microbiol 2020, 63(3), 412-7.

Cyclin D1, a member of the cyclin protein family, is instrumental in the cell cycle due to its influence on the progression from G1 to the S phase. Its overexpression causes reduced doubling time and is also associated with clonogenic growth. The purpose of the present study was to assess cyclin D1 expression in patients with simple hyperplasia (SH), endometrial intraepithelial neoplasia (EIN) and endometrioid endometrial carcinoma, and to evaluate whether there was an association between cyclin D1 expression and the clinicopathological features of endometrioid endometrial carcinoma. Retrospective data were available for 193 patients (30 SH, 40 EIN, and 123 endometrioid endometrial carcinoma cases). To detect cyclin D1 expression, immunohistochemistry staining was performed with tissue microarrays. The percentage of cases with positive cyclin D1 staining were 30 %, 60 % and 78 %, for SH, EIN and endometrioid endometrial carcinoma, respectively (P < 0.001). Carcinomas with higher nuclear grade, histological grade, and FIGO grade displayed higher mean cyclin D1 expression compared to lower grade carcinomas. In addition, patients with lymphovascular invasion (P = 0.006), myometrial invasion (P < 0.001) and lymph node invasion (P < 0.001) had higher mean cyclin D1 expression compared to those without invasion. There was a significant correlation between cyclin D1 expression and clinicopathological features of endometrioid endometrial carcinoma including tumor grade, FIGO grade, lymphovascular invasion, lymph node invasion and myometrial invasion (P < 0.05 for each). Cyclin D1 expression is significantly higher in patients with endometrioid endometrial carcinoma compared to that of the SH and EIN. The extent of cyclin D1 expression is strongly correlated with nuclear and histological grade, myometrial invasion, lymphovascular invasion and lymph node invasion in patients with endometrioid endometrial carcinoma. These findings contribute in several ways to our understanding of cyclin D1 expression and provide a basis for future research on this topic.

2 illus, 2 tables, 25 ref

039922   ABOUHASHEM N S, ABDELBARY E H, ABDALLA M M H, EL-SHAZLY M (Pathology and Urology Dep, Zagazig Univ, Egypt, Email: nehaleldieb@yahoo.com) : Diagnostic utility of amylase α-1A, MOC 31, and CD 82 in renal oncocytoma versus chromophobe renal cell carcinoma. Indian J Pathol Microbiol 2020, 63(3), 405-11.

Renal oncocytoma (RO) and chromophobe renal cell carcinoma (ChRCC) originate from the same cell origin, that is, the intercalated cells of the collecting duct.[1] In most cases, there are clear morphologic differences between RO and ChRCC; however, in some instances, overlapping features may be encountered and the differentiation between the two entities becomes difficult. [2] Several immunohistochemical markers with different expression patterns in ChRCC and RO have been described to rule out this dilemma. About 47 primary renal neoplasms that had been diagnosed as RO or ChRCC were submitted for immunohistochemical staining of amylase α‑1A (AMY1A), MOC 31, and CD 82. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy have been analyzed.  AMY1A positivity was observed in all RO cases in our work with 91.7 % sensitivity and 100 % specificity in the diagnosis of RO. The PPV of its expression was (100 %) and NPV (97.2 %) with a diagnostic accuracy of 97.9 %. A significant high expression of MOC 31 was observed in ChRCC compared to its expression in RO with a statistical significance (P < 0.001). In addition, we obtained 82.9 % sensitivity and 91.7 % specificity of MOC 31 expression in the diagnosis of ChRCC. The positive predictive value (PPV) was (96.7 %), negative predictive value (NPV) (64.7 %) with diagnostic accuracy (85.1 %). In our studied cases, we detected positive immunoexpression of CD 82 in 10 cases (83.3 %) of ChRCC. However, it was lost in all RO cases (100 %). CD 82 sensitivity and specificity in differentiating ChRCC from RO were 100 % and 83.3 %, respectively.  We propose MOC 31 and CD 82 as negative immunostains for RO, as these markers are commonly expressed in ChRCC. In conjunction with AMY1A strong immunopositivity in RO cases, we provide a triple panel of biomarkers (AMY1A, MOC 31, and CD 82) for the distinction between RO and ChRCC.

5 illus, 6 tables, 22 ref

039921   BASU K, KARMAKAR S, SENGUPTA M, ROYCHOWDHURY A, GHOSH A, BANDOPADHYAY M (Pathology Dep, Institute of Post Graduate Medical Education and Research, Kolkata- 700 020, West Bengal, Email: kbasughosh@gmail.com) : Pediatric lupus nephritis -An evil cousin of its adult counterpart: A single-center based experience from a tertiary care hospital of Eastern India. Indian J Pathol Microbiol 2020, 63(3), 397-404.

Systemic lupus erythematosus is an autoimmune multisystem disease with a high predilection for renal involvement. Lupus nephritis develops in 20 % to 75 % within the first two years. Presentation varies from subnephrotic proteinuria to end‑stage renal disease. To study clinical features, biochemical, and serological parameters and correlate with histological activity and chronicity score [modified National Institute of Health (NIH) score]. Retrospective, cross‑sectional, single‑center based study in a tertiary care hospital of Eastern India. We incuded 36 children with lupus nephritis diagnosed from February 2018 to March 2019. Laboratory data included were complete blood count (CBC), blood glucose, urine analysis, serum urea, creatinine, blood urea nitrogen (BUN), albumin, cholesterol, HBsAg, antihepatitis C virus (HCV) antibody, antistreptolysin O (ASO) titer, antinuclear antibody (ANA), myeloperoxidase antineutrophil cytoplasmic antibody (MPO ANCA), proteinase 3 antineutrophil cytoplasmic antibody (PR3 ANCA), double‑stranded DNA (dsDNA), C3, and C4. Clinical parameters were age, sex, blood pressure (BP), skin lesions, arthralgia, edema, obesity. Renal biopsies examined with light microscopy, hematoxylin and eosin (H and E), periodic acid‑Schiff (PAS), silver methanamine, Masson’s trichrome (MT) stains. Immunofluorescence microscopy done with IgG, IgM, IgA, C3c, C1q, kappa, lambda antibodies. Kruskal–Wallis and χ² tests.  Mean age was 15.12 ± 3.49 and 12.5 ± 1.73 years for lupus nephritis (LN) with activity and LN without activity, respectively. Mean dsDNA was higher and mean C3 was lower (52.35 ± 22.21 mg/dl) in active LN. Mean 24‑hour urinary protein was higher in LN without activity. Serum creatinine was raised in active LN. LN class III and IV showed higher activity than chronicity. Pediatric LN is proliferative and more active as compared with adult counterparts. Activity scores are much higher than chronicity scores.

6 illus, 2 tables, 33 ref

039920   WIJESINGHE H D, FERNANDO J, SENARATH U, WIJESINGHE G K, LOKUHETTY M D S (Pathology Dep, Colombo Univ, Colombo 8, Sri Lanka, Email: harshima@path.cmb.ac.lk) : A clinicopathological study of triple-negative breast carcinoma in a patient cohort from a tertiary care center in Sri Lanka. Indian J Pathol Microbiol 2020, 63(3), 388-96.

Triple negative breast carcinoma (TNBC) and basal‑like breast carcinoma (BLBC) are subtypes of breast carcinoma (BCa) that are associated with poor survival. To study the prevalence, clinicopathological profile and survival of TNBC among a Sri Lankan patient cohort and to determine the proportion and predictive histological features of BLBC among TNBCs. A cohort of 221 women undergoing primary surgery for BCa at a tertiary‑care center in Sri Lanka was studied. Clinicopathological and follow‑up information were collected by patient interviews and review of slides and clinical records. Estrogen, progesterone, HER2 receptors, and basal markers (CK5/6, CK14, EGFR, 34βE12) were evaluated immunohistochemically. Data was analyzed with Chi‑square test, multinomial logistic regression, and Cox regression using SPSS20.0. Results: Fifty‑three (24 %) tumors were triple‑negative (95 %CI = 18.37 %–29.63 %). On multivariate analysis, young age (P = 0.002), high Nottingham grade (P = 0.005), moderate to severe tumor necrosis (P = 0.004), absent ductal carcinoma in situ (DCIS) (P = 0.04), reduced vascular density at tumor edge (P = 0.016) and distinct cell margins (P = 0.047) predicted TNBC over luminal subgroups, whereas reduced vascular density (P = 0.004) and low TNM stage (P = 0.011) distinguished TNBC and HER2. BLBC accounted for 45.28 % (95 % CI 32.66 %–58.55 %‑24/53) of TNBC. The presence of extensive necrosis in TNBC correlated significantly with BLBC (P = 0.03). The survival among the TNBC subgroup did not differ significantly from other subgroups. Twenty four percent were TNBCs by immunohistochemical analysis, comparable to studies in the Indian subcontinent, however higher than the West. TNBC status correlated with younger age, high tumor grade, necrosis, absent DCIS, reduced vascular density at tumor edge, and distinct cell margins. The presence of moderate to extensive necrosis in TNBC was predictive of BLBC.

2 illus, 7 tables, 33 ref

039919   RAVAL A P, DESAI U N, JOSHI J S, SHAH F D (Cancer Biology Dep, The Gujarat Cancer and Research Institute, Ahmedabad- 380 016, Gujarat, Email: franky.shah@gcriindia.org) : Role of epithelial- Stromal interaction protein-1 expression in breast cancer. Indian J Pathol Microbiol 2020, 63(3), 382-7.

Epithelial stromal interaction protein‑1 (EPSTI‑1) is originally identified as stromal‑fibroblast – induced gene in breast cancer. It was found to be involved in promotion of EMT, breast cancer invasion, metastasis and anchorage‑independent growth in vitro. Strong expression was observed in various tissues as well as higher expression was observed in invasive breast cancer compared to normal breast. EPSTI‑1 expression was evaluated from 106 pre‑therapeutic breast cancer patients. EPSTI‑1 expression was correlated with known clinico‑pathological parameters of breast cancer to explore its role in breast carcinogenesis. EPSTI‑1 expression was analyzed from the collected synchronous tissues [tumors, Malignant Lymph nodes (LN) and adjacent normal tissues (ANT)] of breast carcinoma patients (N = 106). The statistical correlation was performed using SPSS 16.0. In this study EPSTI‑1 was significantly higher in LN compared to tumors (P < 0.001), and in tumors compared to ANT (P < 0.01) which is also reflected in ROC curve analysis (P < 0.0001). Further the small tumor size, stage I, grade I and tumors without stromal involvement exhibited significant lower expression compared to their counter parts. EPSTI‑1 may have significant role in epithelial stromal interaction and disease extension. Moreover, it may be responsible for aggressive tumor behavior and involved in metastatic process which needs to be validated in larger cohort.

1 illus, 3 tables, 13 ref

039918   VITTHALRAO L P, KHANDEPARKAR S G S, JOSHI A V, GOGATE B P, SOLANKE S G, GORE S H (Pathology Dep, Smt. Kashibai Navale Medical Coll and General Hospital, Pune- 411 057, Maharashtra, Email: siddhigsk@yahoo.co.in) : Immunohistochemical expression of cyclin D1 in invasive breast carcinoma and its correlation with clinicopathological parameters. Indian J Pathol Microbiol 2020, 63(3), 376-81.

Breast cancer (BC) is the most common cancer and leading cause of death in women. This study was conducted to study the cyclin D1 expression in BC and its correlation with other clinicopathological parameters such as tumor size, histological grade, lymph node status, estrogen receptor (ER), progesterone receptor (PR), HER2/neu, and Ki67 status. Fifty cases of BC diagnosed between 2015 and 2018 were included in the study. A technique of manual tissue microarray was employed for the analysis of expression of immunohistochemical (IHC) markers such as cyclin D1, ER, PR, HER2/neu, and Ki67 in all cases. Results were subjected to statistical analysis. Cyclin D1 positivity was seen in 64 % cases of BC cases of which 8 % were triple negative BC (TNBC) molecular subtype. Cyclin D1 expression was statistically significantly associated with ER and PR positivity. Maximum cases showing cyclin D1 expression showed negative HER2/neu expression, Ki67 immunopositivity, absent lymphovascular invasion and were of lower grade and stage. 32 % cases were TNBC. Cyclin D1 was found positive in 25 % TNBC cases. Negative Cyclin D1 expression was seen in TNBC cases of higher grade and higher stage with positive lymph node status, presence of lymphovascular invasion and Ki67 positivity. Cyclin D1 can be potentially used as a prognostic marker and if included in routine IHC workup of BC cases can aid in appropriate patient management with the advent of new targeted therapy that blocks the cyclin D‑CDK4/6 axis.

1 illus, 4 tables, 22 ref

039917   MAHDAVI N, AMINISHAKIB P, NABIYI P, GHANADAN A, GHORBANPOUR M, SOLUK-TEKKESIN M (Tumor Pathology Dep, Istanbul Univ, Istanbul, Turkey, Email: msoluk@istanbul.edu.tr) : Evaluation of the presence of myofibroblasts and matrix metalloproteinase 1 expression in the stroma of oral verrucous hyperplasia and verrucous carcinoma. Indian J Pathol Microbiol 2020, 68(3), 369-75.

Oral verrucous carcinoma is a low‑grade subtype of oral squamous cell carcinoma that should be differentiated from oral verrucous hyperplasia, a premalignant lesion. Stromal activated myofibroblasts known as cancer‑associated fibroblasts have an active role in the initiation and progression of the cancers via secretion of different molecules including matrix metalloproteinases. This study is designed to understand the differences in the presence of myofibroblasts and expression of matrix metalloproteinase‑1 in the adjacent stroma of verrucous carcinoma and oral verrucous hyperplasia (OVH). Cross‑sectional study. Twenty‑seven OVH, 19 oral verrucous carcinoma (OVC), and 8 cutaneous verrucous carcinoma (CVC) specimens were analyzed for immunohistochemical (IHC) expression of α‑smooth muscle actin (αSMA) and MMP‑1. IHC studies for αSMA expression in nonvascular stromal cells of the adjacent stroma revealed mild or no expression in 81.4 %, 73.7 %, and 62.5 % of the cases of OVH, OVC, and CVC groups, respectively. No significant difference was seen in αSMA expression index between OVH and OVC groups (Adj. Sig. = 0.220) and between OVC and CVC groups (Adj. Sig. = 1.00). Pairwise analysis revealed a significant difference in MMP‑1 expression index between the groups. No significant correlation was observed between MMP‑1 expression index and αSMA expression index in OVH (pv = 0.358) and OVC (pv = 0.388) groups. The differences in MMP‑1 expression between OVH and OVC can be used as an adjunctive aid in challenging cases including disoriented or inadequate samples.

3 illus, 3 tables, 30 ref

039916   BAL A, AGRAWAL R, VAIDEESWAR P, ARAVA S, JAIN A (Pathology Dep, Seth GS Medical Coll, Mumbai, Maharashtra, Email: shreeprajai@yahoo.co.in) : COVID-19: An up-to-date review- from morphology to pathogenesis. Indian J Pathol Microbiol 2020, 63(3), 358-66.

The entire world is under a devastating pandemic caused by COVID-19 with a high mortality rate. Knowledge of the viral structure, factors that help in its progression and spread, pathological findings, diagnostic methods and, treatment modalities helps in understanding the viral disease and also in treating the patients in a better way besides preventing the community spread of this deadly infection. The causative agent is a single- stranded RNA virus. The clinical spectrum varies in symptomatic and asymptomatic patients, who later become potential silent carriers, thus unknowingly spreading the virus. The virus constantly undergoes recombination, with reports of cross-species infections. Studies have indicated a strong immunological basis of COVID-19 infection. Not only does it weaken the immune system causing multi-organ involvement but also helps in its progression and spread to others.Multiple organs especially lungs, heart, kidney, gastrointestinal and hepatic system, brain and skin are affected varying in their severity. Similarly, persons with associated co-morbidities are likely to be affected more in terms of the number as well as in the severity. Realtime reverse transcription polymerase chain reaction confirms the presence of COVID-19 infection. Serological diagnosis helps in diagnosing an ongoing outbreak or retrospective infection. Furthermore, it also identifies individuals who have been infected or have recovered from the disease especially the asymptomatic. This helps in the development of an effective vaccine indicating the status of herd immunity in the community. Different treatment modalities are being tried and under trial. This review article thus highlights the global epidemiological status, characteristic of the virus, symptomatology of the patients, role of diagnostic tests available, organs affected including their morphological changes and the latest line of treatment of COVID-19.

1 illus, 2 tables, 59 ref

039915   MISRA V, AGRAWAL R, KUMAR H, KAR A, KINI U, POOJARY A, CHAKRABARTI I, RAI S, SINGHAL A, VIJAY SHANKAR S, IYENGAR J N (Pathology Dep, Rohilkhand Medical Coll and Hospital, Bareilly- 243 006, Uttar Pradesh, Email: drranjan68@gmail.com) : Guidelines for various laboratory sections in view of COVID-19: Recommendations from the Indian association of pathologists and microbiologists. Indian J Pathol Microbiol 2020, 63(3), 350-7.

Declared as a pandemic by WHO on March 11, 2020, COVID‑19 has brought about a dramatic change in the working of different laboratories across the country. Diagnostic laboratories testing different types of samples play a vital role in the treatment management. Irrespective of their size, each laboratory has to follow strict biosafety guidelines. Different sections of the laboratory receive samples that are variably infectious. Each sample needs to undergo a proper and well‑designed processing system so that the personnel involved are not infected and also their close contacts. It takes a huge effort so as to limit the risk of exposure of the working staff during the collection, processing, reporting or dispatching of biohazard samples. Guidelines help in preventing the laboratory staff and healthcare workers from contracting the disease which has a known human to human route of transmission and high rate of mortality. A well‑knit approach is the need of the hour to combat this fast spreading disease. We anticipate that the guidelines described in this article will be useful for continuing safe work practices by all the laboratories in the country.

34 ref

039914   BAIG M A (Pathology Dep, BLDE's Shri B.M. Patil Medical Coll Hospital and Research Centre, Bijapur, Karnataka, Email: drasifbaig@yahoo.com) : Comparative analysis of "APTT vs RVVT" based activated protein C resistance assay in the diagnosis of Factor V Leiden mutation. Indian J Pathol Microbiol 2020, 63(2), 247-50.

Thrombophilia is a hypercoagulable state characterized by increased venous thrombosis. The most common cause of heritable thrombophilia is Factor V Leiden (FVR506Q) homozygous state, with a relative risk of 10–80 times as compared to normal individuals and Lupus anticoagulant is the most common cause of acquired thrombophilia. The main objective of this study is to compare the sensitivity of activated partial thromboplastin time (APTT) vs dilute Russell viper venom test (DRVVT) based APCR assays with predilution in Factor V‑deficient plasma for diagnosis of Factor V Leiden mutation. The coagulometer used for APCR test was Sysmex CS‑5100. APTT reagent used is Pathrombin SL supplied by Siemens. All data were expressed as mean ± SD. Statistical analysis was done using unpaired students t‑test and a P value <0.05 was considered as statistical significance. A total of 300 cases of APCR (200 cases of Factor V Leiden mutation was confirmed by PCR and 100 acquired) were studied. The sensitivity of screening APTT‑based APCR for detection of Factor V Leiden mutation is 67 % and for the noncarrier state, it is 62 %. The sensitivity of modified APTT and DRVVT with predilution in FV‑deficient plasma for detection of Factor V Leiden mutation is 82 % and 84 %, respectively and for acquired causes, it is 48 % and 86 %, respectively. Screening APTT test has increased in activated protein C resistance (APCR) due to Factor V Leiden mutation as well as acquired causes such as patients on direct‑acting oral anticoagulants, warfarin, lupus anticoagulants, and oral contraceptive pills which are independent risk factors of venous thrombosis. Modified DRVVT with predilution in FV‑deficient plasma is more sensitive than screening and modified APTT‑based APCR test in the diagnosis of Factor V Leiden mutation and the former test can distinguish homozygous and heterozygous states from normal individuals.

1 illus, 4 tables, 11 ref

039913   DAS S, SHASTRY S, RAI L, BALIGA P B (Immunohematology and Blood Transfusion Dep, Kasturba Medical Coll, Manipal- 576 104, Karnataka, Email: shameeshastry@gmail.com) : Frequency and clinical significance of red cell antibodies in pregnancy- A prospective study from India. Indian J Pathol Microbiol 2020, 63(2), 241-6.

For appropriate management of hemolytic disease of the fetus and newborn (HDFN), it is important to detect irregular red cell antibody in the antenatal period. Though it is a simple one‑step method, it is not part of routine antenatal screening in many developing countries.To reiterate the importance of antenatal antibody screening, we have assessed the frequency and clinical significance of irregular red cell antibodies in our patient population. A prospective study was carried out from October 2013 to May 2015 at a tertiary care center from south India. All antenatal samples received by the laboratory for red cell antibody screening were screened using a commercial three‑cell screening panel. Antibody identification along with further Immunohematological techniques as required were performed for cases with positive screening results. Neonates of the alloimmunized cases were followed up to determine the clinical significance of the antibody. A total of 2336 antenatal mothers were screened for red cell antibodies. The overall rate of alloimmunization in the study group was 2.27 %. Alloimmunization rate among RhD‑negative pregnancies was 6.9 %. Other than anti‑D (49 %), we identified anti‑D + anti‑C (5 %), anti‑G (5 %), anti‑c (5 %), anti‑E (2 %), anti‑e (2 %), anti‑H (Bombay phenotype) (7 %), anti‑M (2 %), anti‑Lea (2 %), anti‑Leb (12 %), and autoantibodies (9 %) in the maternal serum. Anti‑D, anti‑D + anti‑C, anti‑G, anti‑c, and anti‑H were found to be clinically significant in this study. This study showed that 1 in 125 RhD‑positive pregnancies can develop red cell alloantibodies. Hence, implementing routine antenatal antibody screening irrespective of RhD status is essential.

1 illus, 4 tables, 30 ref

039912   AMANULLAH N A R, POOTHIODE U, VILASINIAMMA L (Neuroscience Technology Dep, Imam Abdulrahman Bin Faisal Univ, Saudi Arabia, Email: naamanullah@iau.edu.sa) : Expression of p53 in epithelial ovarian tumors. Indian J Pathol Microbiol 2020, 63(2), 235-40.

Ovarian cancers remain the most lethal of all gynecological malignancies despite major developments in their treatment. To study the rate of expression and staining patterns of p53 in various histological types and grades of epithelial ovarian tumors (EOT). Sixty EOTs received in a tertiary care center were studied for gross, microscopy, and p53 immunohistochemistry (IHC) expression patterns. Parameters such as age, laterality of tumor, ascites, capsule rupture, tumor size, stage at presentation, metastasis, tumor grade, and number of mitosis were correlated. Of the sixty cases studied, 23 (38.3 %) were malignant. Serous carcinomas were the largest group with 17 cases (74 %) followed by mucinous with 4 cases (17 %) and 2 clear cell carcinomas (9 %). All benign and borderline EOT were p53 negative. 65.2 % of the malignancies were p53 positive and all of them were serous malignancies. 15 out of 16 high‑grade serous carcinomas were p53 positive (94 %), while one case was negative (6 %). 10 cases (63 %) showed intense diffuse positivity of more than 60 % of the nucleus, while 5 cases (31 %) showed aberrant null staining <5 % staining of the nucleus. All mucinous, clear cell carcinomas, and the only low‑grade serous carcinoma in the study were p53 negative. P53 staining had positive correlations with variables like capsule rupture, ascites, laterality, and CA 125. The study highlights the different rates of expression and staining patterns of p53 and the need for correct interpretation of p53 IHC for the diagnosis of various EOT.

5 illus, 4 tables, 35 ref

039911   ABDUL-ILAH B W, GAIDAN H A, AL-KAABI M M (Pathology Dep, Mustansiriyah Univ, Baghdad, Iraq, Email: dr.methaq.mueen@gmail.com) : Immunohistochemical expression of interlukin10 (IL10) and heat shock protein-90 (HSP-90) in prostatic carcinoma. Indian J Pathol Microbiol 2020, 63(2), 230-4.

Specific cytokines are related to pathologically changed prostate, propose that the balance in cytokine differs in normal and pathological prostate. Of these cytokines the interleukins 10, due to its “pleiotropic” actions in inflammation and angiogenesis, and HSP‑90 due to its expression in tumor cells at high levels, suggesting that it has an important role for growth and/or survival of tumor cells. Evaluation of HSP‑90 and IL10 immunoreactivity in benign prostatic hyperplasia (BPH) and prostatic carcinoma and to correlate this expression with clinicopathological parameters. A retrospective study in which 83 Paraffin‑embedded tissue specimens including (43) BPH, (40) prostatic carcinoma and (20) normal prostate as control were included between the period of January 2015 and January 2017. Patients, All the cases were evaluated histopathologically and stained immunohistochemically for IL10 and HSP‑90. Only cytoplasmic staining was considered as positive. Immunoreactivity scoring for both markers expression was calculated based on both staining intensity and percentage. Statistical Analysis: Was done using SPSS Version 21 statistical analysis software. P value of <0.05 was considered statistically significant. Statistical analysis of HSP‑90 and IL10 expression revealed a highly significant correlation of expression of these two markers in advanced Gleason grading and tumor, node, and metastasis (TNM) staging cases of prostatic carcinoma. High expression of IL10 and HSP‑90 is associated with high grade and stage of prostatic carcinoma. This provides a base for further studies and researches on the role of these investigated proteins as prognostic markers immunotherapy targets for carcinoma of the prostate.

1 illus, 4 tables, 26 ref

039910   VAVILAPALLI S, MADIREDDY N, UPPIN M S, KALIDINDI K, GUDITHI S, TADURI G, RAJU S B (Pathology Dep, Nizam’s Institute of Medical Sciences, Punjagutta- 500 082, Hyderabad, Email: drmsuppin@gmail.com) : Anti-glomerular basement membrane disease: A clinicomorphological study of 16 cases. Indian J Pathol Microbiol 2020, 63(2), 226-9.

Antiglomerular basement membrane disease manifests as rapidly progressive glomerulonephritis and alveolar hemorrhage. It encompasses 10–15 % of crescentic glomerulonephritis and is associated with poor outcome. In this study, we have elaborated on the clinical details, morphological features, and outcome of anti‑GBM glomerulonephritis. All the consecutive biopsy‑proven cases of anti‑GBM glomerulonephritis over a period of 4½ years were analyzed, retrospectively.  Sixteen cases were diagnosed as anti‑GBM glomerulonephritis during the study period.Twelve patients presented with rapidly progressive renal failure of which four patients required hemodialysis at the time of presentation. Good pasture’s syndrome was noted in two patients. Thirteen cases were positive for circulating anti‑GBM antibodies and two patients showed double positivity for both anti‑GBM antibodies and ANCA. Fifteen biopsies revealed crescentic glomerulonephritis with linear deposition of IgG along the glomerular basement membrane in all the 16 cases. Renal biopsy analysis is important in the diagnosis of Anti GBM nephritis. Morphology is an important predictor of disease progression.

1 illus, 1 table, 22 ref

039909   LEE J H, LEE J, AHN B K, PAIK S S, KIM H, LEE K H (Surgery Dep, Hanyang Univ, Seoul- 04763, Republic of Korea, Email: leekh@hanyang.ac.kr) : Loss of ASXL1 expression is associated with lymph node metastasis in colorectal cancer. Indian J Pathol Microbiol 2020, 63(2), 221-5.

The function of ASXL1 in colorectal cancer (CRC) has not been investigated yet. The purpose of this study was to investigate the clinicopathological and prognostic impact of ASXL1 expression on CRC. The intensity of expression was scored as 0–3, and the extent of staining was scored as 0–4, based on the percentage of positive cells. The immunoreactivity score (IRS) was calculated by multiplying the two scores. We performed immunohistochemical staining of ASXL1 using tissue microarrays of 408 CRCs, 46 normal colonic mucosae, 48 adenomas, and 92 metastatic lymph nodes. Clinicopathological variables were compared using Fisher’s exact test, χ2‑test, or unpaired Student’s t‑test, depending on the nature of the data. A negative expression of ASXL1 was observed in 10.9 % of normal mucosae, 27.1 % of adenomas, 55.6 % of adenocarcinomas, and 71.7 % of metastatic lymph nodes (P < 0.001). With respect to the IRS cut‑off score, lymph node metastasis and lymphatic invasion were more frequent in the IRS 0–6 group than in the IRS 8–12 group (56.3 % vs. 33.3 %, P = 0.034; 56.0 % vs. 33.3 %, P = 0.035). The 5‑year disease‑free survival rate was significantly lower in patients with IRS 0–6 group than those with IRS 8–12 group (78.7 ± 2.5 vs. 100 %, P = 0.034). ASXL1 might act as a tumor suppressor in CRC. The loss of ASXL1 expression might be associated with lymph node metastasis and lymphatic invasion in CRC.

2 illus, 2 tables, 24 ref

039908   JAIN P, GOYAL S, CHAUHAN G, MAJUMDAR K, ALI S, SAKHUJA P, AGARWAL A K (Pathology Dep, GIPMER, New Delhi, Email: pujasak@gmail.com) : HER-2/neu over expression in gall bladder adenocarcinoma: A quest for potential therapeutic target. Indian J Pathol Microbiol 2020, 63(2), 214-20.

Gall bladder carcinoma (GBC) is an aggressive malignancy with high mortality and aggressive course, with palliation as the only available option. To evaluate frequency of HER‑2/neu overexpression in GBC and to seek its correlation, if any with conventional clinicopathological parameters and survival. Immunohistochemistry (IHC) was performed on 200 cases of GBC, 32 cases of dysplasia, and 100 cases of chronic cholecystitis. Fluorescent in situ hybridization (FISH) was performed on 30 randomly selected cases of GBC to validate IHC. HER‑2/neu overexpression (IHC 3+/FISH amplification ≥2.2) was correlated with clinicopathological parameters by Chi‑square test. P < 0.05 was considered significant. Survival analysis was done by log‑rank test and Kaplan‑Meier analysis. HER‑2/neu overexpression was seen in 14 % (28/200) GBC cases but was not found in dysplasia and chronic cholecystitis. Majority of these cases were ≤grade 2 and in advanced stage, however this was not statistically significant. A lower mean survival in HER‑2/ neu positive group as compared to HER‑2/neu negative group (17.1 ± 2.3 month versus 67.6 ± 8.5 month, respectively) was observed. Concordance between IHC and FISH was seen in 18/19 cases. This study delineates a subset of GBC patients with HER‑2/neu overexpression, in whom targeted therapy can offer a survival benefit.

1 illus, 5 tables, 24 ref

039907   SHARMA S, SHULANIA A, ACHRA A, JERAM H, KANSRA S, DUGGAL N (Microbiology Dep, Dr. RML Hospital, New Delhi, Email: dranuradha.pgirml@gmail.com) : Diagnosis of pulmonary tuberculosis from gastric aspirate samples in nonexpectorating pediatric patients in a tertiary care hospital. Indian J Pathol Microbiol 2020, 63(2), 210-3.

The aim of this study was to assess the utility of Xpert assay, Ziehl–Neelsen (ZN) staining, and Mycobacteria Growth Indicator Tube (MGIT™) culture for diagnosis of pediatric pulmonary tuberculosis from gastric aspirate (GA) samples and to compare Xpert assay and ZN staining with MGIT rapid liquid culture. GA samples from 210 nonexpectorating children, aged between 6 months to 12 years, presenting to the pediatric out‑patient department (OPD) with clinical suspicion of tuberculosis (TB) were collected. The samples were tested by GeneXpert, ZN staining, and MGIT liquid culture. GeneXpert is a more sensitive method for rapid and early diagnosis of pediatric TB when compared with microscopy.

2 tables, 18 ref

039906   SAKRI M S M, RAHMAN W F W A, DIN T A D A T, IDRIS F M, JAAFAR H (Pathology Dep, Sains Malaysia Univ, Kelantan, Malaysia, Email: wfaiziah@usm.my) : Microvessel density and vascular endothelial growth factor receptors in breast carcinoma under the influence of rapamycin and platelet factor 4. Indian J Pathol Microbiol 2020, 63(2), 205-9.

Vascular endothelial growth factor receptors (VEGFRs) are major endothelial growth factor receptors that influence the growth of a tumor. Microvessel density (MVD) is the quantification method of various aspects of tumor vasculature that indicates angiogenic activity. This study aims to analyze the correlation between MVD to the expression of VEGFRs on breast cancer tissue. A total of 60 N‑methyl‑N‑nitrosourea (MNU)‑induced breast carcinomas in rats were suppressed by using antiangiogenic drugs. The rats were then sacrificed, and the tumor was fixed in 10 % formalin, paraffin embedded, and immunohistochemistry stained using VEGFRs and CD34. One-way ANOVA test showed a significant difference in all markers that have been used (P < 0.05) on MNU-breast tumor treated with rapamycin (M = 90.1664, SD = 7.4487), PF4 (M = 93.7946, SD = 7.1303) and rapamycin + PF4 (M = 93.6990, SD = 1.8432). We obtained a significant reduction of MVD count on breast carcinoma for rapamycin group (M = 25.6786, SD = 9.7075) and rapamycin + PF4 group (M = 30.5250, SD = 13.6928) while PF4 group (M=47.7985, SD=4.8892) showed slightly increase compared to control (M = 45.1875, SD = 4.4786). There was a moderately strong, positive correlation between angiogenic markers; Flt-1 (r = 0.544, n=60, P < 0.005) and Flt-4 (r = 0.555, n = 60, P < 0.005) while Flk-1 (r = 0.797, n = 60, P < 0.005) showed a strong, positive correlation with MVD. MVD was strongly correlated to the VEGFRs expression on breast carcinoma.

3 illus, 1 table, 30 ref

039905   PYLA R D, POTEKAR R M, PATIL V S, REDDY A K, SATHYASHREE K V (Pathology Dep, BLDE Deemed to be Univ, Vijayapura- 586 103, Karnataka, Email: ratnakar.potekar@bldeuniversity.ac.in) : Quantitative mast cell analysis and hormone receptor study (ER, PR and HER2/neu) in invasive carcinoma of breast. Indian J Pathol Microbiol 2020, 63(2), 200-4.

Breast cancer constitutes nearly one third of cancers among women. Immune responses caused by neoplastic cells lead to the accumulation of inflammatory cells like mast cells (MCs), macrophages, lymphocytes, and plasma cells around the tumor tissue forming the tumor microenvironment. The study aims at quantifying the role of MCs in different grades of invasive carcinoma of breast with respect to estrogen receptor (ER), progesterone receptor (PR), and Human Epidermal growth factor Receptor 2 (HER2/neu). This study included 60 cases of invasive carcinoma of breast. Toluidine blue staining was used for quantitative MC analysis and correlated with immunohistochemistry analysis for hormonal markers’ positivity—ER, PR and HER2/neu.  The mean age was 52 years (range: 25–75 years). The average number of MCs in Grade I, II, and III were 24.05, 18.4, and 7.9, respectively, with a significant P value. ER, PR, and HER2/neu positivity was found in 60 %, 55 %, and 32 % of the cases, respectively. ER positivity with mean MC count of 23.55 was found in 36 cases, and 33 cases were positive for PR with a mean MC count of 24.18 and a significant P value. HER2 positive cases were 28 with a mean MC count of 20.82. The presence of MCs in breast cancer is inversely proportional to the grade of tumor, i.e., a maximum number of MCs were seen in low grade tumors. In addition, there is a positive correlation between ER and PR receptor positivity with the presence of MCs in the stroma of breast cancer.

2 illus, 6 tables, 15 ref

039904   GAJARIA P K, TAMBE S, PAI T, PATIL A, DESAI S B, SHET T M (Histopathology Dep, Tata Memorial Centre, Mumbai- 400 012, Maharashtra, Email: tanujashet5@gmail.com) : Dual-color dual-hapten in situ hybridization (D-DISH)- Comparison with fluorescence in situ hybridization (FISH) for HER2/neu testing in breast cancer. Indian J Pathol Microbiol 2020, 63(2), 194-9.

HER2/neu testing in breast cancer is a mandate due to availability of trastuzumab, a monoclonal antibody targeted against this biomarker. Dual‑color dual‑hapten in situ hybridization (D‑DISH) is a new test for assessment of HER2/neu gene overexpression on light microscopy. This was a validation study for D‑DISH in our laboratory and was conducted to study the concordance between fluorescence in situ hybridization (FISH) and D‑DISH for HER2/neu testing in breast cancer. In all, 150 cases of invasive breast carcinoma requested for FISH analysis were selected. Immunohistochemistry by Ventana PATHWAY anti‑HER2/neu (4B5) antibody, FISH by ZytoLight SPEC ERBB2/CEN17 Dual Color Probe, and D‑DISH using the Ventana INFORM HER2 Dual ISH DNA Probe Cocktail Assay was carried out. Cohen’s kappa coefficient was used to calculate concordance between FISH and D‑DISH assays. The ratios and average number of signals were compared with Lin’s concordance correlation coefficient. About 93.1 % of the cases showed concordance between FISH and D‑DISH results. Cohen’s kappa correlation coefficient was 0.836, indicating almost perfect level of agreement. Lin’s concordance correlation coefficient (ρc) showed moderate strength of agreement for HER2/chromosome 17 ratios between FISH and D‑DISH assays (ρc 0.9452). As per the American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) 2018 updated guidelines, four of the cases that were nonamplified on FISH showed low‑level amplification on D‑DISH due to counting errors caused by faint signals or background dust. Genomic heterogeneity and larger red chromosome 17 signals on D‑DISH led to discordance of the six cases amplified by FISH. D‑DISH failure rate was 3.33 %.  Overall, D‑DISH showed good concordance with FISH but needs expertise for reporting.

1 illus, 4 tables, 16 ref

039903   CAKIR E, SAYGIN I, KISIOGLU S (Pathology Dep, Karadeniz Technical Univ, Ortahisar-61080, Trabzon, Email: emelnur7@gmail.com) : A comparison between unifocal papillary thyroid microcarcinoma with noninvasive follicular thyroid neoplasm with papillary like nuclear features and other patterns: A retrospective clinicopathological study. Indian J Pathol Microbiol 2020, 63(2), 188-93.

Papillary thyroid microcarcinoma (mPTC) is defined as a tumor with low malignancy potential. Different treatment protocols have been used at different centers for analyzing this tumor which has common recurrence and metastasis rates. Consequently, in 2016, the definition of noninvasive follicular thyroid neoplasm with papillary‑like nuclear features (NIFTP) was accepted which included the lesions >1 cm. It is important to explain the clinical course and appropriate treatment options for mPTC and its subtypes. In this study, we aimed to describe the clinical course of mPTC with and without NIFTP and to determine different risk groups among these subtypes. We performed microscopic reexamination of about 280 unifocal mPTCs retrieved from our archives between 2007–2018 and analyzed the results of morphological and clinical comparison among these cases that had 0–11‑years of clinical follow‑up. Among 280 unifocal mPTCs, 127 cases (45.4 %) had classical morphology, 58 (20.7 %) had NIFTP, 53 (18.9 %) had infiltrative pattern, 27 (9.6 %) had oncocytic pattern, 12 (4.3 %) showed capsular invasion, and 3 (1.1 %) showed other morphologies. Seven patients were detected with lymph node metastasis and one with distant metastasis at diagnosis. Lymph node metastasis (recurrence) was postoperatively detected in five patients. All patients with recurrence were women. Moreover, three of these patients were in their 30s and two in 70s. The median diameter of the tumor was 3 mm. Both invasive and noninvasive cases have recurred. Contrary to the results of the previous studies, the results of our study did not confirm the indolent course of mPTC with NIFTP. However, metastasis was detected both at the time of diagnosis and during the postoperative period. The malignancy potential of these tumors may not be low. Therefore, more clinicopathological and molecular studies are needed to determine the biological behavior of mPTC cases with different histology.

2 illus, 2 tables, 23 ref

039902   KAR A, PATTNAIK K, KAR T, BISWAL P, MISHRA C, GURU L (Pathology Dep, S.C.B. Medical Coll, Cuttack, Odisha, Email: asarantikar@yahoo.co.in) : Clear cell lesions in pathology: Histomorphologic approach to diagnosis. Indian J Pathol Microbiol 2020, 63(2), 177-87.

There has been remarkable progress in the field of surgical pathology; however, histomorphology has remained the most important and essential tool of the surgical pathologist in everyday practice till now. It is surprising that the hematoxylin–eosin (H and E) stain, introduced more than a century ago, has still remained the gold standard stain for histological examination and diagnosis of human diseases. Besides different findings or clues observed in histopathology sections like inclusions, granules, grooving, globules, halo, or clearing, which would enable the pathologist to provide a precise and accurate diagnosis; observation of clear cells is one of the important findings and clue for reporting. It may also sometimes lead to difficulties and delays in establishing the diagnosis. It can be focal or extensive and primary or rarely it may be secondary. Clear cell changes may be observed in many non‑neoplastic, benign, or malignant tumors of diverse origin. Clear cell tumors contain a preponderance of clear cells. It can be seen in almost all the organs of human body and can be classified according to location or biological behavior. Commonly seen clear‑cell tumors are usually malignant and common organs involved are female genital tract, urogenital tract, head and neck areas, central nervous system, skin, and rarely in bone and soft tissues. For approach to clear cell lesions, one has to decide if the change is artifactual, a mimic of clear cell tumors, or a clear cell tumor in reality. Once the mimics and artifactual/degenerative changes have been ruled out, a tumor either primarily of clear cell origin or showing secondary change has to be decided. The tumor next is to be diagnosed as benign/malignant and epithelial/mesenchymal based on morphology.

7 illus, 1 table, 49 ref

039901   CHOUDHURY M, PAHWA A R (Raheja Atlantis, Gurugram- 122 001, Haryana, Email: drarchnarautela@gmail.com) : Comparison of liquid-based cytology with conventional cytology in the evaluation of abdominal masses. Indian J Pathol Microbiol 2020, 63(1), 73-7.

Liquid‑based cytology. Utility of liquid‑based cytology (LBC) was compared to conventional smear cytology in ultrasound‑guided fine‑needle aspirates of abdominal masses.This was a prospective comparative study conducted in collaboration with surgery and pediatrics surgery departments of our institute. Thirty patients presenting with evidence of abdominal mass were enrolled for the study and underwent fine‑needle aspiration cytology. The material was processed for the preparation of conventional smears and residual material was rinsed into cytolyt for LBC by Thin Prep method and into cell block fluid. The smears prepared from both the methods were compared by two independent and experienced pathologists for adequacy, cellularity, architectural pattern, cytoplasmic preservation, nuclear preservation, and background. Cellularity was frequently higher in the conventional smears than on Thin Prep slides (P value = 0.025). Recognition of architecture was better on the conventional smears (P value = 0.001). Cytoplasm was better preserved on the conventional smears (P value = 0.001) but difference in the preservation of nuclear details was not statistically significant on slides prepared from both the techniques. The background of Thin Prep slides is significantly cleaner than direct smears (P value = 0.001). Non epithelial elements such as mucin and neurofibrillary tangles were better preserved on direct smears (P value = 0.001) but diagnostic accuracy of both the methodologies showed no statistically significant difference (P value = 0.226).The Thin Prep technique utilizes expensive equipment and reagents. It also generates certain morphological artefacts in slides with which a cytologist needs to get familiar.When used in isolation, it may not consistently provide any added advantage in the diagnosis of such lesions and should be used as an adjunct to conventional smears. It may be preferred in situations where material has to be transported for processing or is required for ancillary tests.

4 illus, 3 tables, 16 ref

039900   SHETTY O, PAI T, GURAV M, REKHI B (Surgical Pathology Dep, Tata Memorial Hospital, Mumbai- 400 012, Maharashtra, Email: rekhi.bharat@gmail.com) : Comparison between fluorescence in-situ hybridization (FISH), reverse Transcriptase PCR (RT-PCR) and fragment analysis, for detection of t (X; 18) (p11; q11) translocation in synovial sarcomas. Indian J Pathol Microbiol 2020, 63(1), 64-72.

Synovial sarcoma (SS) is an aggressive, but a relatively chemosensitive soft tissue sarcoma, characterized by a specific, t (X;18) (p11;q11) translocation, leading to formation of SS18–SSX chimeric transcript.This translocation can be detected by various techniques, such as fluorescencein‑situ hybridization (FISH), reverse transcriptase PCR (RT‑PCR) and fragment analysis. To compare the results of detection of t (X;18)(p11;q11) translocation, across three different platforms, in order to determine the most optimal and sensitive technique.  Formalin‑fixed paraffin embedded (FFPE) tissue sections of 45 soft tissue sarcomas were analyzed, including 16 cases of SS confirmed by histopathology, immunohistochemistry and molecular technique (s) (Group 1); 13 cases, wherein SS was one of the differential diagnosis, preceding molecular testing (Group 2) and 16 cases of various other sarcomas (Group 3). Various immunohistochemical (IHC) markers studied, including INI1/SMARCB1. All cases were tested for t (X;18) translocation, by fragment Analysis, FISH and RT‑PCR.  There were 23 cases of SS, including 16 of group 1 and 7 of group 2. By fragment analysis, t (X;18)(p11;q11) translocation was detected in 22/23 cases (95.6 %). By FISH, SS18 gene rearrangement was detected in 18/22 cases (78.2 %), whereas by RT‑PCR, SS18‑SSX transcripts were detected in 15/23 cases (65.2 %).Immunohistochemically, a unique “weak to absent”/reduced INI1 immunostaining pattern was exclusively observed in 12/13 cases of SS (92.3 %). Fragment analysis and FISH were relatively more sensitive techniques. Unique “weak to absent”INI1 immuno expression significantly correlated with positive t (X;18)translocation results (P = 0.0001).  The present study constitutes first such study from our subcontinent. Fragment analysis is a promising technique for detection of t (X;18)(p11;q11) translocation. FISH and INI1 immunostaining pattern were also relatively more sensitive, over RT‑PCR.

5 illus, 4 tables, 26 ref

039899   NAIR I R, BALAN S, PHALAK P, DANIEL M (Pathology Dep, Amrita Institute of Medical Sciences, Kochi - 682 041, Kerala, Email: indurn@aims.amrita.edu) : Clinicopathologic spectrum of necrotizing lymphadenitis. Indian J Pathol Microbiol 2020, 63(1), 60-3.

Necrotizing lymphadenitis represents a group of diseases characterized by non-granulomatous inflammation and necrosis of the lymphnode, caused by a variety of infective and inflammatory diseases, most common being Kikuchi-Fujimoto disease, acute Epstein Barr viral infection and systemiclupus erythematosis (1). To study the morphological features in lymph nodes in cases of necrotizing lymphadenitis, to correlate them with specific etiological conditions.Materials and methods-58 cases of necrotizinglymphadenitis were reviewed and categorized into different etiological subtypes, i.e. acute EBV lymphadenitis, lupus lymphadenitis and the rest asKikuchis lymphadenitis. Morphological features studied were presence of vascular proliferation, periadenitis, foamy macrophage, neutrophil and plasmacell infiltrate. Clinical follow up was done.  62.2 % of cases were Kikuchis lymphadenitis. Both lupus and Kikuchis had a female preponderance(78 % and 62 % respectively). Among the morphological parameters, plasmacell infiltration and vascular proliferation showed significant association with lupus lymphadenitis. Kikuchis and EBV lymphadenitis showed self-limiting course, with only 2 cases of Kikuchis developing recurrence .4 cases developed complications. All cases of lupus lymphadenitis needed long term therapy. Kikuchis lymphadenitis is the most common cause of necrotizinglymphadenitis, followed by lupus and acute EBV lyphadenitis.Young females were commonly affected in the first 2 groups. It is worthwhile to classify the cases of necrotizing lymphadenitis into etiological sub groups as the prognosis and treatment differ (2). Among the morphological features studied, plasmacell infiltrate and vascular proliferation were significantly associated with lupuslymphadenitis, hence can be used to predict etiology.

3 illus, 1 table, 18 ref

039898   GULERIA P, KUMAR L, KUMAR S, BHATLA N, RAY R, SINGHAL S, MEENA J, MATHUR S R (Pathology Dep, All India Institute of Medical Sciences, New Delhi - 110 029, Email: mathuraiims@gmail.com) : A clinicopathological study of granulosa cell tumors of the ovary: Can morphology predict prognosis?. Indian J Pathol Microbiol 2020, 63(1), 53-9.

Granulosa cell tumors (GCT) are low‑grade malignant sex cord‑stromal tumors (SCST) with late metastasis/recurrences and long disease‑free periods. We performed a clinicopathological evaluation of GCT to ascertain features having prognostic impact.  All cases of GCT of ovary from January 2006 to December 2018 were assessed for architectural patterns, nuclear grooves, and Call‑Exner bodies. Each featurewas graded on frequency of occurrence: not present (0)–very frequent (3). Anisonucleosis, necrosis, and inflammation were noted. Cases were grouped on mitotic count; <10 mitosis/10 High power field (HPF) or >=11 mitoses/10HPF and Ki‑67 index; <10 % Ki‑67 and >=11 % Ki‑67. GCT formed60.1% of SCST. Sixty cases’ ages were in the range of 15–78 years (median 45). Clinical details were available in 37. Commonest presentation was abnormaluterine bleeding. Serum CA125 was raised in 16.1 % and Inhibin in 58.8 %.Seventy percent were in stage I. Disease recurrence was associated with higher stage (P = 0.007). The most frequent pattern was diffuse sheets (47 %).Call‑Exner bodies were absent in 22.2 %. Grooves with score 1, 2, and 3 were seen in 35.8 %, 23.5 %, and 13.6 %, respectively. Anisonucleosis was present in 26.7 %, necrosis in 11.1 %, and lympho‑plasmacytic infiltrate in 43 %. Out of total, 93.3 % had <10 mitosis/10 HPF and 43.2 % had recurrence, most with high Ki‑67 (P = 0.064). Our study outlines histomorphological spectrum of GCT and emphasizes its frequent occurrence in lower stages with late recurrences. The presence of grooves may indicate granulosa‑cell origin. Call‑Exner bodies are not a necessity. Histomorphological features are not prognostically important. However, prognostic value of Ki‑67 cannot be excluded. Limitation of the study was a small number of cases with follow‑up.

4 illus, 2 tables, 34 ref

039897   HU J, LU Y, TANG N, LI L, GUO P, ZHANG Y (Pathology Dep, Maternal and Child Health Hospital of Hubei Province, Wuluo Road - 430 070, Wuhan, Email: cqjbhu@163.com) : Expression of alpha-methylacyl-coa racemase in vaginal gastric-type adenocarcinoma and uterine clear cell carcinoma. Indian J Pathol Microbiol 2020, 63(1), 49-52.

Alpha‑methylacyl‑coenzyme A racemase (AMACR, P504S) is a commonly used marker in immunohistochemical diagnosis of prostate cancer. Recent studies identified P504S markers of the clear cell histotype in the ovary and/or endometrium. Gastric‑type adenocarcinoma (GAS) is difficult to diagnose histologically, particularly when there is crossover with clear cell carcinoma (CCC). However, the significance of P504S for differentially diagnosing GAS and CCC is unclear. To evaluate P504S as a potential diagnostic marker of GAS and CCC. We analyzed P504S expression in 48 cervical carcinomas (32 GAS and 16 CCC), as well as the expression of other markers including hepatocyte nuclear factor‑1 beta (HNF‑1β) and NapsinA.  The expression differences of HNF‑1β,NapsinA, and P504S in GAS and CCC were detected by immunohistochemistry. Immunohistochemical histoscores based on the intensity and extent of staining were calculated.The positive rates of HNF‑1β in GAS and CCC were 90.32 % and 75 %, respectively. (χ2 = 2.251, P = 0.663). The positive rates of NapsinA in GAS and CCC were 19.36 % and 81.25 %, respectively. (χ2 = 47.332,P < 0.01). The positive rates of P504S in GAS and CCC were 16.13 % and 81.25 %, respectively. (χ2 = 41.420, P < 0.01). HNF‑1β was frequently expressed in GAS and CCC, while NapsinA and P504S were frequently expressed in CCC, and reduced or lost in GAS. NapsinA and P504S can be used to differentiate between GAS and CCC.

2 illus, 1 table, 19 ref

039896   ALJERIAN K (Pathology Dep, King Saud Univ, Riyadh- 12372, Saudi Arabia, Email: kaljerian@ksu.edu.sa) : Chorioamnionitis: Establishing a correlation between clinical and histological diagnosis. Indian J Pathol Microbiol 2020, 63(1), 44-8.

Chorioamnionitis that is associated with high rates of morbidity and mortality needs an early diagnosis for effective treatment.However, views are conflicting on the effectiveness of a clinical versus a histological diagnosis of the disease. The accuracy of clinical diagnoses should be evaluated by determining their correlation with histopathological data.  A total of 696 placental records from single and multiple pregnancies between January 2011 and February 2018 were collected and reviewed to determine if chorioamnionitis was present. Of the 696 records, 255 had histological data available, and of these, histological evidence for chorioamnionitis was recorded in 135 (52.9 %). Clinical chorioamnionitis diagnosis was insensitive (26.7 %; 95 % confidence interval 19.43 %–34.96 %)and inaccurate (61.1 %; 95 % confidence interval 54.90 %–67.19 %). As well,73.3 % of histologically positive chorioamnionitis cases were missed us ingclinical indicators. Clinical diagnosis for chorioamnionitis is inaccurate; in our study, most of the positive cases were not diagnosed using clinical indicators. However, of the clinical indicators examined, maternal and fetal tachycardia were the most reliable.

2 illus, 2 tables, 20 ref

039895   CAROLI-BOTTINO A, MAURICIO A, PANNAIN V N (Pathology Dep, Clementino Fraga Filho Univ, Rio de Janeiro-21941-590, Brazil, Email: caroli_bottino@yahoo.com) : CD57 as a routine neuroendocrine marker for liver metastasis. Indian J Pathol Microbiol 2020, 63(1), 38-43.

The characterization of hepatic metastases as having neuroendocrine origins is essential and the main markers currently used are chromogranin A (CgA) and synaptophysin (Syn). However, these markers may exhibit certain limitations, and the use of CD56 and CD57 can also be considered, although, due to low specificity, their use is discouraged. This study sought to compare the immunohistochemicalexpression of these markers in hepatic metastases of neuroendocrineneoplasms (NEN). Eighteen samples, were used for immunohistochemical staining with CgA, Syn, CD56, and CD57 antibodies.The immunostaining reactions were compared according to its intensity (I), the percentage of labeled cells (P), and a final score (I × P). Statistical agreement between the markers was also evaluated. CD57 was expressed in the highest number of cases and also showed the most intense expression. CgA showed the highest number of cases with more than 80 % positively stained area (72.2 %), followed by CD57 (61.1 %). The highest average score (I × P) was obtained for CD57 (9.1 ± 4.1). The best indices of agreement were between CgA and CD57 with respect to positivity (P = 0.021) and score (P = 0.014). According to the primary site, stomach/duodenum, lungs, and undetermined subgroups showed the highest average scores for CD57, followed by CgA. For the small bowel sub group, the highest average score was obtained for CgA, followed by CD57. Our results highlight the importance of CD57 in the evaluation of hepatic metastases of NEN and indicate that this marker should be included with CgA and Syn in routine diagnostic panels.

2 illus, 3 tables, 36 ref

039894   SADEK S A, REHIM D M A, FATIMA S (Pathology Dep, King Khalid Univ, Kingdom of Saudi Arabia, Email: sohailafatima@gmail.com) : The role of tumor budding in colorectal adenocarcinoma: Possible involvement of the intestinal cancer stem cell marker Lgr5. Indian J Pathol Microbiol 2020, 63(1), 32-7.

Tumor budding (TB) is a promising prognostic factor in colorectal cancer (CRC) that is independent of tumor‑node‑metastasis (TNM) staging.Leucine‑rich repeat‑containing G‑protein‑coupled receptor 5 (Lgr5) is a stem cell marker and a member of the canonical Wnt‑signaling cascade. It is involved incolorectal carcinogenesis. However, its role in CRC progression and TB needs tobe clarified. TB was assessed in both H and E and CK immunostained sections of 92 CRC cases. Associations between TB grade and different clinicopathological parameters were evaluated. Lgr5 expression in CRC cases and its association with TB grade and other clinicopathological features was also evaluated. H and E stained sections revealed low‑ and high‑grade budding in 55 (59.8 %) and 37 (40.2 %) tumors, respectively, whereas Cytokeratin Immunohistochemistry (CK‑IHC) showed low‑ and high‑gradebudding in 31 (33.7 %) and 61 (66.3 %) tumors, respectively. TB grade (in H and Eand CK stained sections) was significantly associated with adverse pathologicalprognostic variables including vascular invasion (P = 0.03 and 0.001), lymphnode metastasis (P = 0.001 and 0,001), advanced Dukes (P = 0.000 and 0.000),and TNM (P = 0.001 and 0.000) stages and inversely associated with Tumor infiltrating lymphocytes (TILS) (P = 0.02 and 0.0001) which is known to be agood prognostic indicator. Lgr5 protein was positively expressed in 52.2 % (48/92) of the CRCs. Immunoreactivity of Lgr5 was significantly associated with histological grade (P = 0.01), lymph node metastasis (P = 0.002), vascularinvasion (P = 0.02), TNM stage (P = 0.000), Dukes stage (P = 0.000), and TILS (P = 0.03). Furthermore, Lgr5 was found to be significantly associated with TB estimated in both H and E and CK stained tumors (P = 0.003 and 0.001respectively). This study supported the relevance of TB in the assessment of CRC aggressiveness. It also revealed that Lgr5 expression is related to morphologic features in the invasive front of CRC. Lgr5 could have an important role in forming a morphologic feature at the invasive front associated with the aggressiveness of the tumor.

2 illus, 4 tables, 27 ref

039893   OGUT B, EKINCI O, CELIK B, UNAL E R, DURSUN A (Pathology Dep, Gazi Univ, Ankara- 06560, Turkey, Email: betulcimer@gmail.com) : Comparison of the efficiency of transgelin, smooth muscle myosin, and CD31 antibodies for the assessment of vascular tumor invasion and free tumor deposits in gastric, pancreatic, and colorectal adenocarcinomas. Indian J Pathol Microbiol 2020, 63(1), 25-31.

This study aimed to compare CD31, smooth muscle myosin (SMM), and transgelin antibodies for their efficiency in detecting venous invasion (VI) and the nature of free tumor deposits (TDs) in gastric, pancreatic, and colorectal adenocarcinomas. Eleven Whipple, 5 gastrectomy, and 3 colectomy specimens and 1 low anterior resection specimen were reviewed and examined, revealing 254 probable foci. Foci were reviewed and divided into 3 types: Type A, the “orphan artery” pattern; Type F, free TDs in the periorganadipose and connective tissue without an unaccompanied artery; and Type X,a focus that could be detected only with the immunohistochemical procedures mentioned. No foci were positive for CD31. Transgelin staining was more sensitive than SMM staining in all focus types, Type A only and Type Fonly (P < 0.001, P = 0.001, and P = 0.10, respectively). In free TDs (Type F),35.7 % of the samples were negative for all four stains, and 64.2 % of the samples were positive for SMM and transgelin. We did not make the distinction between a metastatic lymph node and VI in positive foci. We conclude that hematoxylin and eosin (H and E) staining is inadequate and that smooth muscle markers, such as transgelin and/or SMM, are more effective than endothelial markers, such as CD31, in revealing VI and lymph node/large extramural invasion.

5 illus, 2 tables, 29 ref

039892   VARMA K, CHAUHAN A, BHARGAVA M, MISRA V, SRIVASTAVA S (Pathology Dep, M.L.N. Medical Coll, Allahabad, Uttar Pradesh, Email: kachnar12@gmail.com) : Association of different patterns of expression of beta-catenin and cyclin D1 with pathogenesis of breast carcinoma. Indian J Pathol Microbiol 2020, 63(1), 13-8.

Beta‑catenin and cyclin D1 have attracted considerable attention in recent studies as potential proto‑oncogenes in many human cancers especially colonic cancer. Beta‑catenin plays multiple roles within the cell such as canonical Wnt signaling where cyclin D1 has been identified as one of its target genes.The role of beta‑catenin and cyclin D1 in breast cancer has been evaluated inmany studies but not established yet. The expression ofbeta‑catenin and cyclin D1 was evaluated in 82 cases of breast carcinoma (BCa) and 32 cases of ductal carcinoma in situ (DCIS) by immunohistochemistry (IHC).Their relationship with clinicopathological features was also investigated. Statistical analysis was done to establish an association. Abnormal expression of beta‑catenin (ABE) was seen in 80.2 % cases of invasive ductalcarcinoma (IDC) and 47 % cases of DCIS, while the cyclin D1 positive expressionrate was 60.9 % and 50 %, respectively. In the cases showing ABE, cyclin D1 positivity was 88.1 %. ABE showed significant association with high‑grade BCa.The most common pattern of ABE was loss of membrane with nuclear positivity which is associated with worst prognosis. In addition, ABE in cases of BCa and DCIS showed concordant patterns. Therefore, an association exists between ABE and cyclin D1 in BCa and its precursor lesions implying that Wnt/beta‑catenin oncogenic pathway may have a definite role in breast carcinogenesis and can be used for targeted therapy. Also, different patterns of beta‑catenin expression may have prognostic and predictive value. 

2 illus, 4 tables, 21 ref

039891   AGARWAL A, GARG C, GANESH M S, REDDY S (Pathology Dep, Rohilkhand Medical Coll and Hospital, Uttar Pradesh- 243 001, Email: cheena.garg@gmail.com) : Molecular mechanisms of tobacco induced oral and oropharyngeal cancer: Results of a tissue microarray and immunohistochemistry-based study from a tertiary cancer center in India. Indian J Pathol Microbiol 2020, 63(1), 7-12.

It is well established that chronic exposure to tobacco induces head and neck cancers but the exact etiopathogenesis is not known. Though studies have shown expression of TIMP1, EPS8 and AXL in cancers, their rolein tobacco‑induced cancers is not known. We aimed this study to evaluate the role of these molecules in oral and oropharyngeal squamous cell cancers (SCC). In this single institutional study, 31 patients of oral and oropharyngeal SCC with history of chewing tobacco were included.Smokers were excluded from the study. After informed consent biopsies were taken from affected and contralateral normal mucosa. Paraffin blocks were made and tissue microarray (TMA) were constructed using these blocks.Immunohistochemistry (IHC) for TIMP1, EPS8, AXL kinase was carried out on these tissue microarrays. The intensity of staining was scored from 0 to 3+, related to expression of each of the three molecules.  The expression of TIMP1,EPS8 and AXL kinase was significantly more in the cancerous mucosa versusnon‑cancerous mucosa (P = 0.000 in all three) in oral and oropharyngeal SCC exposed to chewing tobacco. Immunohistochemical expression of these molecular markers in oral and oropharyngeal SCC correlated with their molecular based studies. Significant IHC expression of TIMP1, EPS8 and AXL establishes their role in the pathogenesis of oral and oropharyngeal squamous cell carcinomas. Novel targeted therapies may be researched that can detect and target these molecules at an earlier stage of pathogenesis of these tumors.

3 illus, 3 tables, 19 ref

039890   SADHU S, MAJUMDER P D, BISWAS J (Uvea and Ocular Pathology Dep, Sankara Nethralaya, Chennai - 600 006, Tamil Nadu, Email: drjb@snmail.org) : Biological therapy in refractory cases of uveitis and scleritis: An analysis of 18 cases from a tertiary eye care center from South India. Indian J Ophthalmol 2020, 68(9), 1929-33.

To evaluate the effectiveness of biologic therapy in a cohort of patients with various types of refractory non‑infectious uveitis and scleritis. A retrospective observational study on patients with non‑infectious uveitis and scleritis who were not responding or had a high recurrence rate with the conventional treatment and had received biologic therapy. We studied 18 patients (33 eyes) who received biological therapy between January 2017 and November 2019. The mean age was 30 ± 17 years and mean duration of uveitis was 36.8 months (range 1–120 months). Anterior uveitis (27.7 %) was most commonly observed followed by scleritis, panuveitis, posterior, and intermediate uveitis. The most common etiology was Behçet’s disease (4 patients, 22.2 %) followed by juvenile idiopathic arthritis (3 patients, 16.6 %), granulamotosis polyangitis, and idiopathic (2 patients each, 11.1 %). Majority had trialled one or more immuno suppressive and were refractory in nature. Maximum patients had received adalimumab (61 %) followed by infliximab (22 %), rituximab (12 %), and golimumab (6 %). The median prednisolone dose was reduced from 30 mg (range 7.5–60 mg) to 5 mg (range 0–10 mg) after biological therapy (P = 0.002). Significant visual improvement was observed post biologic therapy (mean log mar VA 0.41 ± 0.62 improved to 0.23 ± 0.48 at the final visit, P = 0.008). Maximum number of patients (16 patients, 89 %) responded well with biological therapy. Three patients developed recurrence and systemic complications were observed in two patients. Biologic therapy is effective in non‑infectious refractory uveitis who were resistant to conventional therapy and may prolong disease recurrence.

2 illus, 2 tables, 12 ref

039889   PATNAIK G, SUDHARSHAN S, GEORGE A E, GANESH S K, BISWAS J, MAJUMDER P D (Uvea Dep, Sankara Nethralaya, Chennai- 600 006, Tamil Nadu, Email: drparthopratim@gmail.com) : Clinical profile of patients with anterior nodular scleritis in India. Indian J Ophthalmol 2020, 68(9), 1925-8.

To report the clinical profile of a series of anterior nodular scleritis in Indian population. We conducted a retrospective review of medical records of 140 eyes of 123 consecutive patients with nodular scleritis who presented to a tertiary eye care institute between 2007 and 2018. The mean age at presentation was 46.8 ± 13.1 years and 70.7 % of the patients were female. Bilateral involvement was observed in 14 % patients. The most common presenting symptom was redness (92.6 %) and ocular pain (69.1 %). Twenty‑seven patients (22 %) had some systemic association and rheumatoid arthritis (5 %) was the most common autoimmune disease. Presumed ocular tuberculosis was diagnosed in 13 % patients. Methotrexate was the most common immunosuppressive used in these patients and an additional immunosuppressive was required in 6.5 % patients. Recurrence of inflammation was observed in 74.8 % patients. Deterioration of vision noted in 2.8 % eyes. Tuberculosis remains an important cause of nodular scleritis in India. Recurrence of scleritis is common in nodular scleritis and cases with non infectious nodular scleritis often require treatment with immune suppressives.

2 illus, 1 table, 18 ref

039888   BALBABA M, DAL A, ÇOLAKOGLU N, BULMUS O, ULAS F, YILDIRIM H, AYDEMIR O, ERÖKSÜZ Y (Ophthalmology Dep, F-rat Univ, Turkey, Email: mbalbaba@yahoo.co.uk) : Anti-inflammatory effect of cortistatin in rat endotoxin-induced uveitis model. Indian J Ophthalmol 2020, 68(9), 1920-4.

To evaluate the anti‑inflammatory effect of cortistatin (CST) in endotoxin‑induced uveitis (EIU) model and to compare the results with corticosteroid treatment. A total of 35 healthy Wistar albino rats were randomly divided into five groups. EIU was induced by a single subcutaneous injection of lipopolysaccharide (LPS). Group I received intraperitoneal (ip) normal saline (NS), Group II received ip 150 μg LPS plus NS, Group III received ip 150 μg LPS plus 250 μg/kg CST, Group IV received ip 150 μgLPS plus 1mg/kg dexamethasone, and Group V received ip 250 μg/kg CST only. The aqueous humor was collected 24 h after injection and the infiltrating cells were determined. Moreover, histopathological and immunohistochemical examinations were also performed. The clinical score and infiltrated cell count were reduced in Groups III and IV compared with Group II (P < 0.001). The pathological findings of Groups III and IV were significantly reduced compared with Group II (P < 0.001). These findings were similar between Groups III and IV (P = 1.000). Tumor necrosis factor‑alpha (TNF‑α) and interleukin 1 beta (IL‑1β) immunoreactivity in the ciliary body of Group III and Group IV were significantly reduced compared with Group II (P < 0.001). TNF‑α and IL‑1β immunoreactivity in the ciliary body of Group III and Group IV were similar compared with Group I and Group V (range of P values was 0.539–0.958). CST administration as a therapeutic agent might ameliorate the severity of intraocular inflammation in uveitis patients. In conclusion, effect of CST and dexamethasone in EIU model was comparable.

4 illus, 28 ref

039887   KAWALI A, SRINIVASAN S, MAHENDRADAS P, SHETTY B (Uveitis and Ocular Immunology Dep, Narayana Nethralaya, Bangalore, Karnataka, Email: akawali332@gmail.com) : Epidemic retinitis and community outbreaks. Indian J Ophthalmol 2020, 68(9), 1916-9.

The objective of this study was to correlate seasonal variation of epidemic retinitis (ER) with concurrent community outbreaks. This is a retrospective, observational, comparative study conducted in a tertiary care eye hospital in south India. Monthly variation in number of ER cases in comparison with reported community outbreaks by Integrated Disease Surveillance Program (IDSP) from 2009 to 2020 in the same region were studied. Month‑wise graphs against number of patients were plotted for ER and for each community outbreak. ER was diagnosed in 163 patients. Diagnosis of presumed rickettsial ER was made in 48 cases (29.44 %), chikungunya in 5, dengue in 3 and typhoid in 6 cases, while in other cases the etiological diagnosis remained uncertain (n = 101). Multiple positives erological tests were seen in 6 patients (Weil Felix Test (WFT) with WIDAL in 4 and chikungunya IgM with WFT in 2 patients). Relevant reported outbreaks by IDSP were: Pyrexia of unknown origin (PUO) (n = 5148),Chikungunya (n = 6577), Dengue (n = 7350), Measles (n = 1422), Mumps (n = 881), Rubella (n = 288),Malaria (n = 2262), Chicken Pox (n = 2385), Typhoid (n = 597), Kyasanur Forest Disease (n = 381), ScrubTyphus (n = 13), Typhus fever (n = 4), Japanese Encephalitis (n = 15). None of the outbreak graphs pattern was identical or similar to the graph of ER. Inverse relation of the graph of dengue outbreak with ER was observed. Inverse correlation between dengue and ER should be further studied for causation,which we believe may prove dengue as least common cause. Reporting of rickettsial outbreaks should be enhanced by undertaking statewide awareness and procurement of gold standard tests.

2 illus, 1 table, 22 ref

039886   KAWALI A A, MOHAN A, MEHTA R, MAHENDRADAS P, SRINIVASAN S, SHETTY B (Uveitis and Ocular Immunology Dep, Narayana Nethralaya, Bengaluru- 560 010, Karnataka, Email: akawali332@gmail.com) : Anti-vascular endothelial growth factor in the treatment of macular edema in epidemic retinitis. Indian J Ophthalmol 2020, 68(9), 1912-5.

To study efficacy of anti‑vascular endothelial growth factor (anti‑VEGF) in resolution of macularedema in epidemic retinitis (ER).  In this retrospective, comparative study, patients diagnosed as ER with central macular thickness (CMT) ≥ 600 μm on SD‑OCT at presentation were studied. Eyes which did not receive intravitreal anti‑VEGF formed group A and eyes receiving additional anti‑VEGF formed group B. Eyes receiving anti‑VEGF monotherapy were studied separately. Cases with subsequent OCTscans with interval of more than 20 days and cases without OCT scan at the resolution were excluded.Treatment details, visual outcome, and days to resolution of macular edema were studied. MeanCMT in group A (n = 8) was 820.1 μm (range 607‑1004 μm) and in Group B (n = 4) was 756.0 μm (range 603‑1000 μm). Macular edema resolved in 34.8 days (range: 16‑65) and 39.0 days (range: 21–45) in group Aand B, respectively. Two eyes with anti‑VEGF monotherapy recovered in 45 and 18 days, respectively. Mean corrected distance visual acuity (CDVA) at presentation in group A was 19.1 (range: 0–61) ETDRS letters and in group B was 14.3 (range: 0–35) ETDRS letters. Mean CDVA improved to 65.7 (range: 0–85) and 50.8 (range:20–76) ETDRS letters in group A and B, respectively. Anti‑VEGF monotherapy eyes improved from 35 and 46 ETDRS letters to 70 and 85 ETDRS letters, respectively.  Additional anti‑VEGF therapy hasno added advantage in speed of resolution of macular edema due to ER. A randomized controlled trial with steroids sparing “anti‑VEGF monotherapy” may verify our observations.

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