045459   KOHLI N, CHAUHAN R S (Veterinary Pathology Dep, GB Pant Univ of Agriculture and Technology, Pantnagar, Uttarakhand, Email: profchauhan58@gmail.com) : Type III hypersensitivity-related disorders in animals. J Immunol Immunopathol 2020, 22(1), 48-55.

Immune complex diseases encompass a variety of disease states in man and animals due to the deposition of circulating antigen–antibody complex in body tissues, leads to the activation of complement cascade pathway that triggers the generation of chemical mediators of inflammation thereby the development of pathological alterations. Normally the immune complexes are formed in the body are taken care by the various phagocytic cells, which are capable of removing foreign antigen from the system. However, presence of persistent infection or repeated exposure to an antigen or in case of self-antigen, the formation of antigen–antibody complexes get enhanced and which may escape from the phagocytic cells and get deposited in tissues that provoke Type III hypersensitivity or immune complex-mediated hypersensitivity reaction. There are two types: systematic/generalised immune complex-mediated reaction, for example, serum sickness and Localised immune complex-mediated reaction, for example, Arthus reaction. There are certain diseases where circulating antigen–antibody complexes are thought to play an important pathogenic role such as glomerulonephritis, rheumatoid arthritis and systemic lupus erythematous.

1 table, 55 ref

045458   RATHOR C, CHAUHAN R S (Veterinary Pathology Dep, GB Pant Univ of Agriculture & Technology, Pantnagar-263 145, Uttarakhand, Email: profchauhan58@gmail.com) : Type II hypersensitivity reaction and related disorders in animals. J Immunol Immunopathol 2020, 22(1), 35-47.

Antibody binding to the cell surface antigen causes destruction of cells through a mechanism called type II hypersensitivity. In the Gell–Coombs formulation, type II hypersensitivity reactions are characterized by antigen– antibody interactions, resulting in the local production of anaphylotoxin (C5a), the recruitment of polymorphonuclear leukocytes (PMNs) and subsequent tissue injury due to the release of hydrolytic neutrophil enzymes after their autolysis. Incompatible blood transfusion and hemolytic diseases are the best examples for the type II hypersensitivity reaction. In transfusion reactions, the antibodies present in the recipient blood destroy the red blood cells of the donor. The antigens found on the surface of erythrocytes are known as blood group antigens. Glycophorin is one of the more common RBC membrane antigens, which is targeted by auto-antibodies that spans over the plasma membrane. The specifications on the erythrocyte, which react with the different reagent or antisera are called as antigenic factors or blood factors. Blood factors are determined by genes at the same locus which belong to the same blood group system. Erythrocytes of man and animals are provided with the different blood group antigens. They vary among different species with regard to their structure, antigenicity and complexity. Some blood group antigen such as ‘B’ system in cattle and ‘A’ antigen in cats are more important from a clinical point of view and are widely distributed in the population. The immune mediated destruction of circulating red blood cells (RBC) occurs by two different mechanisms i.e. by the intravascular destruction of erythrocyte by complement lysis, which is initiated by antibodies that are often of IgM class. The second mechanism is extravascular destruction by immune cells, which identify IgG and complement bound to erythrocytes. Hemolytic syndrome includes immune mediated hemolytic anemia, autoimmune hemolytic anemia, alloimmune (transfusion) hemolytic anemia and drug-induced hemolytic anemia. The Immune-mediated hemolysis occurs due to anti-erythrocyte antibodies, secondary to malignancies, autoimmune disorders, some drugs, and transfusion reactions. A hybrid foetus of sheep and goat dies due to haemolytic anemia when antibodies to their own red blood cells cross the placenta from the maternal circulation. Other transfusion hazards like acute and delayed transfusion reactions, transfusion-related acute lung injury (TRALI), graft-versus-host disease (GVHD), transfusion-associated graft-versus-host disease (TA-GVHD), transfusion-induced immunomodulation and lower nephron nephrosis also occurs in animals. These hazards are infectious and noninfectious in origin.

1 table, 88 ref

041924   SINGLA T, SINGLA G, RANGA S, SINGLA S, ARORA R (Pathology Dep, Vardhman Mahavir Coll and Safdarjung Hospital, New Delhi - 110 029, Email: gauravjoncy1987@gmail.com) : Role of platelet aggregation in metastatic breast cancer patients. Indian J Pathol Microbiol 2020, 63(4), 564-9.

Breast cancer is the most common female cancer in the world. Although early detection and systematic adjuvant therapy has improved survival, distant metastasis remains the leading cause of breast related mortality. The relationship between tumor and the hemostatic system is increasingly recognized as an important regulator of breast cancer progression. Tumors have the ability to induce platelet aggregation which is referred as tumor cell induced platelet aggregation (TCIPA). This study highlights that increased platelet aggregation plays an important role in metastasis of breast cancer. The aim here is to study the role of platelet aggregation in metastatic breast cancer patients using: • ADP • Thrombin. 30 cases (n = 30) of metastatic breast cancer and 30 controls (n = 30) of non‑metastatic breast cancer which were clinically diagnosed and histopathologically confirmed were included in this study. Platelet aggregation studies in vitro using ADP and Thrombin were performed using an optical aggregometer in both cases and controls. Other parameters like platelet count, histological grade and surrogate molecular classification was also correlated with platelet aggregation. In this study, increased aggregation was seen with ADP and thrombin in the metastatic cases and none showed increased aggregation in the non‑metastatic breast cancer patients. Also, high platelet count and higher histological grade correlated with increased aggregation. However, no correlation was seen between platelet aggregation and the surrogate molecular classification. It was concluded from this study that platelet aggregation has an important part to play in the tumor metastasis of breast cancer patients.

4 illus, 6 tables, 18 ref

041923   SINGH V, KAUR N, MANDAL S, MALLYA V, TOMAR R, KHURANA N, BAINS L (Pathology and Pediatric Surgery Dep, Maulana Azad Medical Coll and Associated Hospitals, New Delhi - 110 002, Email: shra_mana@hotmail.com) : Neuroendocrine carcinomas of the breast: Case series with review of literature. Indian J Pathol Microbiol 2020, 63(4), 559-63.

The breast tumors with neuroendocrine differentiation show features similar to their counterparts in other organs. Neuroendorine carcinomas account for less than 0.1 % of all breast carcinomas. To study the demographics and clinicopathological prameters ten cases showing neuroendocrine carcinoma breast. Material and Methods: Ten cases showing neuroendocrine carcinoma were studied. The data was analysed for demographics and clinicopathological prameters. The Immunohistochemistry for ER, PR, Her2neu, Synaptophysin, Chromogranin, NSE, Ki67 index and EMA were done in these cases. Nine Trucut biopsies were reported as infiltrating duct carcinoma and one case as IDC with neuroendocrine differentiation with focal mucinous areas.The histopathological slides of breast excision specimens revealed clusters of cells arranged in sheets and small nests separated by thin fibrous septae in eight of the cases. Trabeculae were noted in two case and in another rosettes were noted. DCIS component was noted in two cases. Infiltration into fat in five of the cases. One case showed pools of mucin. The tumour cells were positive for synaptophysin in 5/10 cases, chromogranin in 8/10 cases and NSE in 9/10 cases. Estrogen receptor positivity was noted 6 cases (6/10), progesterone receptor positivity in 8 cases (8/10) and Her2neu positivity in 5 cases (5/10). NECB cases are more likely to ER/PR positive with variability of expression of neuroendocrine markers. These tumors are more aggressive with propensity for distant metastasis. Endocrine therapy may be more beneficial than standard chemotherapy. Anti-angiogenic markers are an exciting new approach for these case, which is yet to be explored.

3 illus, 2 tables, 12 ref

041922   ORAL O, UNVERDI H, KUMCU E, TURKBEY D, DOGAN S, HUCUMENOGLU S (Pathology Dep, Manavgat State Hospital, Antalya, Turkey, Email: onur_dr@windowslive.com) : Associations between the expression of mucins (MUC1, MUC2, MUC5AC and MUC6) and clinicopathologic parameters of human breast carcinomas. Indian J Pathol Microbiol 2020, 63(4), 551-8.

The aim of this study is to evaluate the relationships between the expression of mucins in invasive breast carcinomas and clinicopathologic parameters. We examined 150 cases of invasive breast carcinoma, using the 2012 World Health Organization (WHO) classification of the tumors of the breast. We studied the expression of MUC1, MUC2, MUC5AC, and MUC6 by immunohistochemistry. We also evaluated normal breast tissue and ductal carcinoma in situ (DCIS) lesions in nearby invasive tumor areas. In invasive breast carcinomas, MUC1, MUC2, MUC5AC, and MUC6 were expressed in 98.6 %, 11.3 %, 9.9, and 8.5 % of cases, respectively. MUC2, MUC5AC, and MUC6 were overexpressed in invasive tumors and DCIS lesions were compared with normal breast tissue. The apical pattern of MUC1 was correlated with low grade and ER expression. MUC2 was correlated with mucinous carcinoma and an inverse association with invasive ductal carcinoma, not otherwise specified (NOS). MUC6 expression was associated with lymphovascular invasion. Most invasive breast tumors express MUC1 and the apical pattern of MUC1 is correlated with low grade and ER expression. MUC6 expression is associated with indicators of poor prognosis. Further comprehensive studies need to evaluate the role of mucins as a potential biomarker and to be used as a specific therapeutic target against breast cancer.

3 illus, 3 tables, 21 ref

045457   SINGH J, CHAUHAN R S ( Veterinary Pathology Dep, GB Pant Univ of Agriculture & Technology, Pantnagar- 263 145, Uttarakhand, Email: profchauhan58@gmail.com) : Type I hypersensitivity associated disorders in animals. J Immunol Immunopathol 2020, 22(1), 25-34.

Type I hypersensitivity is a sudden inflammatory reaction caused against antigens also known as allergens, due to release of primary mediators and secondary mediators from the mast and basophil cells. Primary mediators include histamines, serotonins and secondary mediators include prostaglandins and leukotrienes. The antibodies produced in this reaction are mainly the IgE, which binds with the surface of mast cells and basophils, and crosslinking of two IgE antibodies by allergen leads to the degranulation of the mast and basophil cells severe form of type I hypersensitivity is known as anaphylactic shock. The IgE antibody produced in type I hypersensitivity is mainly produced inside the lymphoid tissue associated with skin and respiratory tract. These antibodies are seen during the incidences of parasitic infestation, allergic conditions, food allergies, urticaria, anaphylactic shock and atopic dermatitis. The IgE antibodies concentration is also increased in various autoimmune disorders such as systemic lupus erythematosus, rheumatoid arthritis and psoriasis. Type I hypersensitivity is very common and it is short lived while some diseases of type I hypersensitivity are very fatal like anaphylaxis. Anaphylaxis is mainly of two types local and systemic anaphylaxis in which systemic anaphylaxis is very fatal which may result in death. For control of the hypersensitivity reaction exposure of the allergen should be avoided. Prevention is a better approach in case of the hypersensitivity reaction to reduce the chances of any severe acute or chronic disease and to increase the quality of life.

79 ref

041921   LEE S A, CHOI J H, CHO S J, CHANG J W, MAENG Y H (Jeju National Univ School of Medicine, Jeju City, Republic of Korea, Email: yhmaeng@jejunu.ac.kr) : The clinical usefulness of chemokine C X C Motif Ligand 12 as a diagnostic marker for papillary thyroid carcinoma. Indian J Pathol Microbiol 2020, 63(4), 544-50.

Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer worldwide. It is essential to develop methods for the accurate diagnosis of PTC to avoid unnecessary surgery. The chemokine C‑X‑C motif ligand 12 (CXCL12) is associated with various cancers. We aimed to investigate the efficacy of CXCL12 in the diagnosis of PTC in fine‑needle aspiration (FNA) specimens. We prospectively collected samples from 58 patients who were scheduled for surgical treatment of PTC from 2013 to 2015. Tissue samples of 31 people with benign thyroid conditions were used as controls. Immunocytochemical and immunohistochemical staining for CXCL12 was performed on FNAs and corresponding tissue specimens. B‑type Raf kinase (BRAF) V600E mutant protein expression and gene mutation were also analyzed to compare the clinical usefulness. The mean age of the patients was 49.1 ± 1.4 years and 88.1 % were women. Positive CXCL12 staining was observed in 6.5 % of benign and in 98.3 % of PTC samples; positive BRAF V600E mutant protein expression was found in 19.4 % of benign and 93.1 % of PTC samples. For the diagnosis of PTC for CXCL12 staining of FNA specimens, the calculated values were 93.1 % sensitivity, 90.3 % specificity, 94.7 % positive predictive value, 87.5 % negative predictive value, and 89.1 % accuracy. CXCL12 had 100 % sensitivity and specificity for the 12 cases of atypia of undetermined significance (AUS) diagnosed in FNA specimens. CXCL12 may be a useful diagnostic tool for PTC, especially when the FNA specimen is classified as AUS.

2 illus, 6 tables, 33 ref

046731   AL-JOBORAE S F F, ALI I R A A, AL-JOBORAE F F M (Babylon Univ, Hillah, Iraq, Email: qaisajam1981@gmail.com) : Unexpected adverse medical & surgical health conditions in neonates after elective cesarean sections within the context of the influence of local socidemographic factors in Babylon. Med Legal Update 2020, 20(1), 10.37506/v20/i1/2020/mlu/194407.

Caesarean section was first introduced to reduce the risks of normal delivery, and is recommended only when the lives of the mother and fetus are compromised, as it is associated with certain risks. However, some women’s perceptions may be influenced by local culture. According to international statistics , the rate of C-section is increasing globally, and in Iraq it is 2-3 times higher than other nations. More than 1/3 of women in Babylon choose C-section voluntarily, believing it is safer, healthier, and less painful. Identify the unexpected medical and surgical conditions of newborns by elective cesarean delivery in Al-Hillah city, and assess the association between the actual gestational age at birth and decision by which elective caesarean sections were done as influenced by local sociodemographic factors and perceptions; that resulted in those conditions. This study is a cross sectional study conducted in Al-Hilla General Teaching Hospital from the beginning of March 2018 until the end on August 2018, and included a total of 150 neonates born via elective cesarean section who had unexpected medical and surgical conditions. Mean maternal age was (26.27±9.56) year ranging from (16-43). Seventy percent of them had timing of < 39 weeks of gestation, and 36.7 % had actual gestational age at birth of < 37 weeks. Almost half the them chose C-section for being safe. Respiratory compromise was the commonest medical complication among mothers. There was significant relationship between decision basis and actual gestational age at birth, as well as between maternal age and medical and surgical complications (P-value<0.05). Neonates born after elective cesarean delivery in Al-Hillah city have significantly higher rates of respiratory morbidity and longer length of hospital stay, with increased risk of certain medical and surgical complications. The unexpected medical complications are inversely related to gestational age at which elective cesarean section is timed. 

3 illus, 3 tables, 25 ref

041920   TEJASVI M L A, MARAGATHAVALLI G, PUTCHA U K, RAMAKRISHNA M, VIJAYARAGHAVAN R, AVINASH C K A (Prosthodontics Dep, Kamineni Institute of Dental Sciences, Nalgond,Telangana, Email: anulekhaavinash@gmail.com) : Impact of ERCC1 gene polymorphisms on response to cisplatin based therapy in oral squamous cell carcinoma (OSCC) patients. Indian J Pathol Microbiol 2020, 63(4), 538-43.

Cisplatin is one of the major drugs that used in the treatment of oral cancer.Excision repair cross-complementation group 1 (ERCC1) is a key DNA repair gene in the nucleotide excision repair pathway which is activated in the repair of intra- and interstrand DNA crosslink caused by platinum-based treatment. The aim of this study was to investigate the association between polymorphisms in ERCC1 (C118T & C8092A) genes and the response to cisplatin-based chemotherapy. ERCC1polymorphisms (C118T & C8092A) were studied using PCR-RFLP method from 150 OSCC patients as cases as well as 150 normal tissues from the same patients were collected as controls for this study. Frequencies of ERCC1 C118C, C118T and T118T genotypes were 60 %, 28 % and 12 % in OSCC patients and 78 %, 19 % and 3 % in the controls, respectively. The C118T & T118T genotype had a 1.69 and 4.97 -folds increased risk for OSCC. Frequencies of ERCC1 C8092C, C8092A and A8092A were 78 %, 18 % and 4 % in the OSCC patients and 89 %, 10 %, amd 1 % in the controls, respectively. The C8092A genotype showed a 1.97-fold increased risk for OSCC. In conclusion, this study highlights the DNA repair gene polymorphisms that might play a role in mediating susceptibility to oral squamous cell carcinoma and cisplatin therapy. Our data suggest that the ERCC1 C118T, T118T and ERCC1 C8092A genotypes are genetic risk factors for Oral squamous cell carcinoma and ERCC1 118 C/T and C8092A polymorphisms have significant influence on clinical outcome.

2 illus, 5 tables, 11 ref

041919   PAWAR R B, PALASKAR S J, KALAVANKAR S S (Oral Pathology and Microbiology Dep, Sinhgad Dental Coll and Hospital, Pune ? 411 041, Maharashtra, Email: rasikapawar0710@gmail.com) : Assessment of apoptotic index in various grades of oral epithelial dysplasia: A cross‑sectional study. Indian J Pathol Microbiol 2020, 63(4), 534-7.

Oral cancer is a major health problem worldwide. In cancer, the equilibrium between cell proliferation and apoptosis is disturbed. The defect in the apoptotic pathway allows cells to proliferate with genetic abnormalities. Thus, the apoptotic index (AI) can be used to assess the significance of apoptosis as a proliferative marker in oral epithelial dysplasia. To assess the apoptotic index in various grades of epithelial dysplasia. 1) To calculate the apoptotic index in various grades of oral epithelial dysplasia, 2) To compare the apoptotic index between various grades of oral epithelial dysplasia, 3) To predict the biologic behavior of oral epithelial dysplasia based on an apoptotic index. Cross‑sectional tissue analyzing study. This study constituted 30 cases, previously diagnosed with various grades of oral epithelial dysplasia (OED). AI was calculated as the number of apoptotic bodies/cells expressed as a percentage of the total number of cells counted in each case. Statistical analysis was carried out using ANOVA test. A statistically significant difference was observed between mild dysplasia and severe dysplasia where P = 0.002. The mean AI was increased progressively with increasing grades of OED. This study demonstrated the clinical significance of apoptosis in assessing disease progression in Oral Potentially Malignant Disorder (OPMD) which may be used as a prognostic indicator in OED. This would, in turn, help in knowing the prognosis of the disease and to develop targeted drug therapy.

4 illus, 1 table, 21 ref

041918   PAREKH D, KUKREJA P, MALLICK I, ROY P (Oncopathology Dep, Tata Medical Centre, Kolkata - 700 156, West Bengal, Email: paromita.roy@tmckolkata.com) : Worst pattern of invasion – type 4 (WPOI‑4) and Lymphocyte host response should be mandatory reporting criteria for oral cavity squamous cell carcinoma: A re‑look at the American Joint Committee of Cancer (AJCC) minimum dataset. Indian J Pathol Microbiol 2020, 63(4), 527-33.

A proportion of early‑stage node‑negative oral squamous carcinoma patients fail despite complete surgical resection. Adjuvant treatment in early oral cancer is controversial and is often individualized based on stage, depth, and margin status. We reviewed various histological markers in pT1/T2N0 cases, resected upfront with elective nodal dissection, with an emphasis on tumor‑tissue interface characteristics of the worst pattern of invasion (WPOI), tumor cell nest size (sCNS), budding and lymphocytic host response (LHR), to assess their prognostic significance. Archived blocks of 95 cases were reviewed. Tumor stage, grade, size, depth of invasion, lymphovascular, and perineural invasion, WPOI, LHR, sCNS, and tumor bud (single cells or <5 cell clusters) score were recorded. Prognostic significance was statistically analyzed using SPSS software version 20. Depth of invasion (P = 0.008), WPOI‑ 4 and 5 (P = 0.033), sCNS (<5 cells) at tumor interface (P = 0.010), high bud count (≥3 buds/40 × hpf) (P = 0.021) and poor LHR (P = 0.019) correlated significantly with poor disease‑free survival on univariate analysis. However, on multivariate analysis only LHR and WPOI‑4 (that is presence of small cell nests or buds) were significant, with high hazard ratio of 4.351 (95 % CI 1.290–14.676, P = 0.018) and 5.019 (95 % CI 1.212–20.789, P = 0.026), respectively. We propose mandatory reporting of WPOI‑4 at the tumor interface and absence of LHR, as significant markers of poor prognosis in early‑stage oral cavity squamous carcinoma.

3 illus, 2 tables, 32 ref

041917   VERMA T, KUMARI S, MISHRA S, RASTOGI M, TIWARI V, AGARWAL G R, ANAND N, HUSAIN N (Pathology Dep, Dr. Ram Manohar Lohia Institute of Medical Sciences, Lucknow - 226 010, Uttar Pradesh, Email: drnuzhathusain@hotmail.com) : Circulating free DNA as a marker of response to chemoradiation in locally advanced head and neck squamous cell carcinoma. Indian J Pathol Microbiol 2020, 63(4), 521-6.

Liquid biopsy has moved from bench to bedside as a non‑invasive biomarker for early diagnosis and monitoring treatment response. This study investigated the role of circulating free DNA (cfDNA) as a diagnostic marker in locally advanced head and neck squamous cell carcinoma (HNSCC) and in monitoring response to chemoradiation therapy. Serum was collected from treatment naïve, histopathologically diagnosed tumors in 24 HNSCC cases and 16 normal controls. CfDNA levels were quantified using β globin gene amplification. The cfDNA level was significantly elevated in HNSCC (992.67 ± 657.43 ng/mL) as compared to healthy controls (60.65 ± 30.42 ng/mL, P = <0.001). The levels of cfDNA did not significantly correlate with TNM stage, lymph node involvement and grade. In responders, percentage decrease in cfDNA levels was 9.57% and 29.66%, whereas in nonresponders percentage increase was 13.28% and 24.52% at the end of three months of follow‑up. Our study adds to the evidence that cfDNA levels are significantly higher in HNSCC cases and provides some evidence that levels increase with tumor progression. CfDNA may be a promising prospective non‑invasive marker to predict response in patients undergoing chemo‑radiotherapy.

3 illus, 3 tables, 27 ref

041916   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

041915   REKHI B, KARMARKAR S, GUPTA C, DEODHAR K K, MENON S, PATHUTHARA S, MAHESHWARI A, SHYLASREE T S, GUPTA S (Surgical Pathology Dep, HBNI Univ, 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

041913   CELIK S Y, CELIK O I (Pathology Dep, Dr. Özgür Ilhan Çelik, Sitki Kocman Univ, Kotekli, Mugla, 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

041912   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

041910   BASU K, KARMAKAR S, SENGUPTA M, ROYCHOWDHURY A, GHOSH A, BANDOPADHYAY M (Pathology Dep, nstitute 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, 3 tables, 33 ref

041909   WIJESINGHE H D, FERNANDO J, SENARATH U, WIJESINGHE G K, LOKUHETTY M D S (Pathology Dep, Colombo Univ, Colombo, 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. 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

041908   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

041907   LENGARE P V, KHANDEPARKAR S G S, JOSHI A R, 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

041906   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, 63(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

041905   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

041904   MISRA V, AGRAWAL R, KUMAR H, KAR A, KINI U, POOJARY A, CHAKRABARTI I, RAI S, SINGHAL A, SHANKAR S V ET AL (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.

1 table, 34 ref

041903   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

041902   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

041901   AMANULLAH N A R, POOTHIODE U, VILASINIAMMA L (Neuroscience Technology Dep, Imam Abdulrahman Bin Faisal Univ, Eastern Province, 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

041900   BAKIR W A-L, 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. 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. 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

041899   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, Telangana, 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. Goodpasture’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

041898   LEE J H, LEE J-H, 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

041897   JAIN P, GOYAL S, CHAUHAN G, MAJUMDAR K, ALI S, SAKHUJA P, AGARWAL A K (Pathology Dep, GIPMER, New Delhi, Delhi - 110 002, 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

043798   TAUHEED A, HAMIDUDDIN , KHANAM S , SOOFI G , ALI M A (Ilmul Saidla (Unani Pharmacy) Dep, National Institute of Unani Medicine, Bengaluru - 560 091, Karnataka) : Formulation, physicochemical, and mating behavior evaluation of tablet modified from Safoofe kharekhasak: A Unani Pharmacopoeia aphrodisiac powder. Int J Green Pharm 2020, 14(1), 23-37.

Safoofe kharekhasak (SK), a powder used as an aphrodisiac in Unani Medicine. Its ingredients are Anacyclus pyrethrum DC. root, Zingiber officinale Rosc. rhizome, and Tribulusterrestris Linn fruit and sugar. The present study is attempted to reduce its bulkiness by converting it into extract tablets, to make it more palatable, portable, sugar-free, and to improve patient compliance. Physiochemical and aphrodisiac activity evaluation of the formulated tablet and SK was performed to validate its action. Tablet batch was selected prepared from the authenticated ingredient and excipients on pre- and post-compression evaluation. Physicochemical evaluation with high-performance thin-layer chromatography (HPTLC) fingerprinting with quantitative estimation of diosgenin and screening of mating behavior in rats with parameters mounting frequency (MF), intromission frequency (IF), mounting latency (ML), intromission latency (IL), ejaculatory latency in first series, and post-ejaculatory interval was carried out for the optimised batch of the tablet and SK. Selected batch of tablet containing extract 400 mg (50 %), microcrystalline cellulose, starch and lactose 124 mg (15.5 %) each, SSG 8 mg (1 %), Aerosil-200 16 mg (2 %), and magnesium stearate 4 mg (0.5 %). Hardness in Kg, friability (%), and disintegration time (in min) were 11.0 ± 0.00, 0.279 ± 0.02, and 7.906 ± 0.169, respectively. Standards for loss of weight on drying, pH, ash value, extractive values, qualitative test for various functional groups, and HPTLC fingerprinting were set in. Diosgenin content in tablet was estimated to be 63.85 μg/gm. Mating behavior study revealed significant aphrodisiac effect of the formulated tablet in the dose (140 mg/kg body weight) as per MF and EL with respect to control. Extract tablet was formulated with excipients in compliance with the analytical specification; its physicochemical standards were established. Mating behavior in rats showed potential aphrodisiac effect in rats at a specified dose.

11 illus, 4 tables, 54 ref

043796   LAOUFI R , YAHIAOUI K , BOUCHENAK O , BENHABILES N , LEFKIR S , ARAB K (M’Hamed Bougara of Boumerdes Univ, Algeria) : Beneficial effect of the Urtica dioica aqueous extract on the crystallization of calcium oxalate in urine. Int J Green Pharm 2020, 14(1), 6-12.

Urtica dioica, belongs to the family of Urticaceae, is widely spread in the world. It is known as one of the most famous medical plants and the more useful. The aim of this research is to quantify the polyphenolic aqueous extract of parts of the U. dioica in the region of Boumerdes and evaluate the antioxidant activity and antilithiasis activity. The antioxidant activity of the leaves extract was evaluated by 1,1-diphenyl-2-picrylhydrazyl ’s method. The antilithiasis activity followed by an observation on electronic microscope to be scanned. Phytochemical screening revealed worth parts in secondary metabolites (polyphenols). The extraction yield of polyphenols is from 2.8 % for 20 g of vegetal powder, with a concentration of 126.28 mg EAG/g powders for all the parts. By coincidence, the obtained results show that the aqueous polyphenolic extract has an important antioxidant power, and of an antilithiasic effect, in particular, the crystallization of calcium oxalate in the urine. In fact, the observation in scanning electron microscope found that the augmentation of the extract concentration favored the reduction in the size of the crystals from 17 μm to 5 μm. Conclusion: These results indicate that aqueous extract from the leaves of U. dioica L. possesses significant antioxidant and antilithiasic potential, deserves to be valued by its integration into therapeutic applications.

13 illus, 1 table, 28 ref

043795   HAZARIKA S , DAS S , GOSWAMI A K , SHARMA H K (Pharmaceutical Sciences Dep, Dibrugarh Univ, Dibrugarh - 786 004, Assam) : Green extraction using goat urine as menstruum and evaluation for in vitro antimycobacterial activity of Curcuma zedoaria and Curcuma caesia rhizomes collected from Assam. Int J Green Pharm 2020, 14(1), 1-5.

In Indian traditional system of medicine, goat urine is believed to have therapeutic value and is also reported its use in the treatment of tuberculosis (TB). On the basis of reported traditional uses for the treatment of TB and/or leprosy, Curcuma caesia and Curcuma zedoaria rhizomes were selected. Aim: It was aimed to study the antimycobacterial activity of goat urine and extracts of the rhizome of the two plants obtained using goat urine as menstruum. The rhizomes were amassed from in and around Dibrugarh. The clean sliced rhizomes were dried at room temperature. The dried rhizomes of both the plant species were extracted using raw and photoactivated goat urine as menstruum by maceration process. In vitro antimycobacterial activity of the rhizome extracts was carried out by disc diffusion method. Crude photoactivated goat urine extracts of both the plants C. caesia (paGuCc) and C. zedoaria were found to have higher antimycobacterial activity against Mycobacterium smegmatis than that of raw goat urine extracts of both the plants C. caesia and C.zedoaria. Among all paGUCc extracts were found to exhibit highest antimycobacterial activity. The extracts obtained using photoactivated goat urine showed higher activity than the extracts obtained using raw goat urine. Goat urine also exhibited antimycobacterial activity, but not as much as the extracts. Thus, it is proved that the extracts and goat urine have antimycobacterial activity and extracting with goat urine and thus have improved activity.

1 illus, 1 table, 18 ref

041896   SHARMA S, SHULANIA A, ACHRA A, JERAM H, KANSRA S, DUGGAL N (Microbiology Dep, Dr. RML Hospital, New Delhi, Delhi - 110 001, 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

041895   SAKRI M S M, RAHMAN W F W A, TENGKU DIN T A D A-A, 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

045440   KHANAL H, KHANAL U , KOIRALA J (PG Kayachikitsa Dep, National Institute of Ayurveda, Jaipur, Rajasthan) : Medicinal plant vasaka could be a therapeutic option for the management of COVID-19 symptoms. J Med Plants Stud 2020, 8(5), 44-8.

Corona virus disease 19 (COVID-19) outbreak is a current pandemic that causes acute respiratory syndrome with huge mortality worldwide. The chief clinical features of COVID-19 is the presence of respiratory symptoms, with severe cardiovascular and renal complications in some patients. This study explores the potentiality of Vasaka (Adhatoda vasica Nees) in the prevention and management of symptoms associated with COVID-19. Vasaka is a well-known natural shrub in Ayurveda system of medicine which has a beneficial effects, particularly in respiratory disorders. First, we reviewed various researches on Vasaka, and its use in the management of respiratory system related diseases, viral diseases and other diseases in various journals and Ayurveda classical texts. Second, a survey based study was conducted to assess the effectiveness of Vasaka in the prevention and management of respiratory related problems. The literature review revealed that Vasaka has been widely studied for its pharmacological activities. It has various action like antiviral, hepatoprotective, antibacterial, anti-inflammatory, cardioprotective, antitubercular and antioxidant. The survey results of the study indicated that only 15 % of the sampled respondents used Vasaka in the treatment of respiratory related diseases. Nevertheless, among the respondents who used the plants, 97 % stated that the use of Vasaka was useful in the treatment of diseases. Interestingly, 36.4 % of the sampled respondents reported that they were completely recovered from the respiratory related diseases. Due to Vasaka multi-modal therapeutic effects and the findings from the survey-based research indicate the potentiality of its use in the management of COVID-19 symptoms.

1 illus, 2 tables, 26 ref

041894   PYLA R D, POTEKAR R M, PATIL V S, REDDY A K, SATHYASHREE K V (Pathology Dep, Shri B M Patil Medical Coll, 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

041893   GAJARIA P K, TAMBE S, PAI T, PATIL A, DESAI S B, SHET T M (Pathology Dep, Tata Memorial Hospital, 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

045435   RUDRASWAMY S, SHANKAR H J, MRUTHUNJAYA K, ARADHYA M (Public Health Dentistry Dep, JSS Dental Coll and Hospital, Mysuru, Karnataka) : Economically viable and ecologically sustainable tree of heaven “Simarouba glauca”. J Med Plants Stud 2020, 8(5), 1-4.

Simarouba glauca commonly known as paradise tree is a versatile fast growing multipurpose oilseed tree belonging to the Simaroubaceae family. It is considered as a potential source of edible biodiesel and also known for various medicinal properties having ability to check the soil erosion and improvement in the ground water balance.

16 ref

041890   SARMA P, BHATTACHARYYA A, KAUR H, PRAJAPAT M, PRAKASH A, KUMAR S, BANSAL S, KIRUBAKARAN R, REDDY D H, MEDHI B ET AL (Pharmacology Dep, PGIMER, Chandigarh - 160 012, Email: drbikashus@yahoo.com) : Efficacy and safety of steroid therapy in COVID‑19: A rapid systematic review and Meta‑analysis. Indian J Pharmacol 2020, 52(6), 535-50.

Although the use of steroids in the management of COVID‑19 has been addressed by a few systematic review and meta‑analysis, however, they also used data from “SARS‑CoV” and “MERS‑CoV.” Again, most of these studies addressed only one severity category of patients or addressed only one efficacy endpoint (mortality). In this context, we conducted this meta‑analysis to evaluate the efficacy and safety of steroid therapy among all severity categories of patients with COVID‑19 (mild to moderate and severe to critical category) in terms of “mortality,” “requirement of mechanical ventilation,” “requirement of ICU” and clinical cure parameters. 11 databases were screened. Only randomized controlled trials (RCTs) or high quality (on the basis of risk of bias analysis) comparative‑observational studies were included in the analysis. RevMan5.3 was used for the meta‑analysis. A total of 15 studies (3 RCT and 12 comparative‑observational studies) were included. In the mechanically‑ventilated COVID‑19 population, treatment with dexamethasone showed significant protection against mortality (single study). Among severe and critically ill combined population, steroid administration was significantly associated with lowered mortality (risk ratio [RR] 0.83 [0.76–0.910]), lowered requirement of mechanical ventilation (RR 0.59 [0.51–0.69]), decreased requirement of intensive care unit (ICU) (RR 0.62 [0.45–0.86]), lowered length of ICU stay (single‑study) and decreased duration of mechanical ventilation (two‑studies). In mild to moderate population, steroid treatment was associated with a higher “duration of hospital stay,” while no difference was seen in other domains. In patients at risk of progression to “acute respiratory distress syndrome,” steroid administration was associated with “reduced requirement of mechanical ventilation” (single‑study). This study guides the use of steroid across patients with different severity categories of COVID‑19. Among mechanically ventilated patients, steroid therapy may be beneficial in terms of reduced mortality. Among “severe and critical” patients; steroid therapy was associated with lowered mortality, decreased requirement of mechanical ventilation, and ICU. However, no benefit was observed in “mild to moderate” population. To conclude, among properly selected patient populations (based‑upon clinical severity and biomarker status), steroid administration may prove beneficial in patients with COVID‑19.

5 illus, 2 tables, 60 ref

041888   MISHRA K, NATH M, HALDER N, VELPANDIAN T (All India Institute of Medical Sciences, New Delhi - 110 029, Email: tvelpandian@hotmail.com) : Evaluation of the possibility of selective modulation of retinal glucose transporters in diabetic complications: An experimental study. Indian J Pharmacol 2020, 52(6), 495-504.

To identify the possibility of modulating retinal glucose transporters in diabetic conditions to prevent retinal complications of diabetic retinopathy. In silico and in vitro binding assays were performed to assess the effect of genistein and positive controls (pioglitazone and estradiol) on nuclear receptor estrogen receptor beta and peroxisome proliferator‑activated receptor gamma (PPARγ). In vivo effects of compounds were tested on diabetic rats. Structural and functional analysis of retina was performed at 28^th day followed by gene expression analysis of glucose transporters and nuclear receptors. Pioglitazone and genistein levels were analyzed by liquid chromatography with tandem mass spectrometry. Genistein showed equi‑affinity toward PPARγ in in silico experiments contrary to in vitro findings. In multidose study, their therapeutic effects were observed by analyzing the retinal function. Retinal gene expression studies revealed that both test agents significantly up regulated PPARγ, GLUT4, and down regulated GLUT1. Genistein showed significant up regulation of GLUT4 and down regulation of GLUT1 as compared to PGZ which has been well correlated with the Electroretinography (ERG) outcome. This study showed the possibility of selective upregulation of GLUT4 (independent of PPARγ activation) in the retina of diabetic rats using genistein. Selective modulation of retinal glucose transporters as therapeutic target in ocular diabetic complications can be possibly explored.

5 illus, 30 ref

041886   CHADALAVADA V, BABU S M, BALAMURUGAN K (Pharmacy Dep, Annamalai Univ, Chidambaram - 608 002, Tamil Nadu, Email: vineelach99@gmail.com) : Nonprescription sale of schedule H1 antibiotics in a city of South India. Indian J Pharmacol 2020, 52(6), 482-7.

The acquisition of antibiotic without a prescription by the general population is a typical practice found in community pharmacies across India, which is a notable contributor of antimicrobial resistance. According to the present regulation in India, sale of certain antimicrobials included in schedule H1 without prescription is unlawful. In this contest, a program was organized by the Drug Control Administration, Government of Andhra Pradesh, to educate pharmacists regarding schedule H1. The aim of our study is to assess the impact of the program on the rate of antibiotics dispensed at community pharmacies. A cross‑sectional study was designed to investigate the nonprescription sale of antibiotics, from September to December 2018 through 200 community pharmacies located in and around Guntur city located in the state of Andhra Pradesh in India. A simulated client methodology was used in this study. A total of 3 female actors including an author of this present study are prior trained to present a standardized simulation of clinical conditions (sore throat, urinary tract infection, cold, and fever) to the pharmacist at the community pharmacies. Microsoft excel sheet was used for data analysis. The simulated patients successfully obtained antibiotic from 78 % pharmacies with the highest rate of urinary tract infection when compared to other conditions. Pharmacists who objected to dispense antibiotics (22 %) are found in developed locations in the city and appeared well qualified. The present study revealed that the antibiotics are continued to be sold without prescription even after the education program on schedule H1. The deregulation of the act is definitely the problem to be addressed by the government.

3 illus, 3 tables, 19 ref

041885   SRUTHI V, REDDY R N, SOWMINI K, GRACE N S (Pharmacology Dep, Osmania Medical Coll, Hyderabad - 500 095, Telangana, Email: resunehareddy@gmail.com) : To evaluate the efficacy and safety of olopatadine 0.1% ophthalmic solution and bepotastine 1.5% ophthalmic solution in patients with vernal keratoconjunctivitis in a tertiary care hospital. Indian J Pharmacol 2020, 52(6), 476-81.

Vernal conjunctivitis comprises 0.5 % of allergic eye diseases. The study is intended to collate the effectiveness of drugs by observing the reduction in signs and symptoms. The objective of the study is to evaluate the effectiveness and safety of olopatadine 0.1 % ophthalmic drops with bepotastine besilate 1.5 % ophthalmic drops in patients with vernal keratoconjunctivitis (VKC). A randomized, open‑label, comparative study conducted in Sarojini Devi Eye Hospital, Telangana. The study included 50 patients diagnosed with VKC, of which Group A and Group B were given olopatadine 0.1 % ophthalmic drops and bepotastine besilate 1.5 % ophthalmic drops, respectively, twice a day for 8 weeks. The reduction in signs and symptoms in both groups was compared. The observations and results were tabulated accordingly, and data were analyzed using the SPSS. The unpaired t‑test is used as the test of significance in between two groups. P value is statistically significant when it is less than 0.05. Overall, 50 cases were included in the study, 72 % of total patients were in the age group of 5–10 years, and 28 % were in the age group of 11–15 years. There were 39 males and 11 females. After 8 weeks of follow‑up, the mean reduction in the scoring of symptoms and signs provided better and quicker relief of watering, ocular discomfort, and conjunctival hyperemia with bepotastine 1.5 % eye drops. Olopatadine 0.1 % eye drops provided faster improvement in papillary hypertrophy. Both drugs were equally effective in reducing itching. Laboratory findings of absolute eosinophil count had no statistical significance in between the two groups. In this study, based on the evaluation of therapeutic performance, bepotastine eye drops proved quicker relief of symptoms and signs compared to olopatadine eye drops but was not statistically significant which would prove beneficial for the patients.

2 illus, 5 tables, 29 ref

041884   BIST A, SAVITHA A, GUMMA K M (Pharmacology Dep, ACS Medical Coll, Chennai - 600 012, Tamil Nadu, Email: ankitabist9@gmail.com) : Efficacy of valacyclovir and famciclovir in herpes zoster: A comparative study. Indian J Pharmacol 2020, 52(6), 472-5.

The objective was to evaluate the efficacy of antiviral agent valacyclovir compared with famciclovir in the treatment of herpes zoster. A comparative study was conducted over a period of 1 year. Data relevant to the study were collected from 60 patients, with active herpes zoster presenting to the outpatient department within 72 hr of the first occurrence of zoster rash. They were divided in to two groups of 30 patients each. The first group of patients received valacyclovir tablet 1000 mg thrice daily, whereas those in the second group were given famciclovir tablet 500 mg thrice daily. Both the drugs were given for 7 days. Periodic follow‑up till 29^th day was done for assessment of the effects of given drugs. Significant decrease was observed on comparison of pain scores between the two groups using the visual analog scale, with the drug valacyclovir, than in the famciclovir group at day 29. Furthermore, valacyclovir treatment accelerated the resolution of zoster associated pain in more number of patients compared with famciclovir. Oral valacyclovir administered during acute zoster infection for a period of 7 days offers significant benefit compared to famciclovir by providing a well tolerated and greater resolution of pain while maintaining the favorable safety profile, making valacyclovir more efficacious and a better drug in management of Herpes Zoster in comparison to famciclovir.

2 illus, 2 tables, 15 ref

041883   MISHRA R K, CHAKRAVARTY R, SIDDIQUE N, PANDEY K R (Obstetrics and Gynaecology Dep, Armed Forces Medical Coll, Pune - 411 040, Maharashtra, Email: rcpublication20@gmail.com) : Pregnancy outcomes following exposure to efavirenz based antiretroviral therapy in indian women. Indian J Pharmacol 2020, 52(6), 467-71.

Mother‑to‑child transmission of HIV has witnessed a significant reduction due to effective antiretroviral therapy (ART). Efavirenz has been introduced as a part of ART since last few years in the national  Prevention of Parent to Child Transmission (PPTCT) program for pregnant women living with HIV. However, data related to adverse pregnancy outcomes associated with efavirenz‑based ART are limited in the Indian scenario. The present study evaluated pregnancy outcomes in HIV‑infected pregnant women who were given efavirenz‑based ART during pregnancy. It is a retrospective, observational, analytic study carried out at a referral hospital in Western India. Collection of data was done for a period of 5 years, and various adverse outcomes were studied which included preterm delivery, low birth weight (LBW), stillbirths, congenital anomaly, and neonatal death. This study showed a preterm birth rate of 19 % and LBW in 36 % of cases. There was no significant association with congenital anomaly, stillbirth, or neonatal death. There was an association of exposure to efavirenz with an increased incidence of adverse pregnancy outcomes, especially LBW infants. This study emphasizes the requirement of large prospective studies to investigate fetomaternal outcomes in pregnant women exposed to efavirenz.

3 tables, 21 ref

041882   SARANGI S C, SOPORY P, PATTNAIK S S, REETA K H (Pharmacology Dep, All India Institute of Medical Sciences, New Delhi - 110 029, Email: reetakh@gmail.com) : Antibody–drug conjugates, cancer immunotherapy, and metronomic chemotherapy as novel approaches in cancer management. Indian J Pharmacol 2020, 52(5), 402-13.

Treatment of cancer is a major challenge even though the pathophysiology is becoming clearer with time. A number of new chemical entities are developed to target cancer growth inhibition, but the targeted delivery of these products still needs novel research. This is of utmost importance not only for higher efficacy but also for a reduction in systemic toxicity and cost of treatment. Although multiple novel targets and molecules are being researched, most of them could not pass the regulatory approval process, due to low benefit–risk ratio and lack of target specificity. Failure of a majority of these drugs was in part due to their superiority claimed via surrogate markers. Despite these, currently, more than 100 chemotherapeutic agents are in practice. This review paper discusses in detail the molecular basis, drug discovery, and pros and cons over conventional treatment approaches of three novel approaches in cancer therapy, i.e., (i) antibody–drug conjugates, (ii) cancer immunotherapy, and (iii) metronomic chemotherapy. All the drugs developed using these three novel approaches were compared against the established treatment regimens in clinical trials with clinical end points, such as overall survival, progression‑free survival, and quality of life.

5 illus, 3 tables, 88 ref

041881   EFTEKHARZADEH M, SIMORGH S, DOSHMANZIARI M, HASSANZADEH L, SHARIATPANAHI M (Toxicology and Pharmacology Dep, Iran Univ of Medical Sciences, Tehran 1449614535, Iran, Email: shariatpanahi.m@iums.ac.ir) : Human adipose‑derived stem cells reduce receptor‑interacting protein 1, receptor‑interacting protein 3, and mixed lineage kinase domain‑like pseudokinase as necroptotic markers in rat model of Alzheimer’s disease. Indian J Pharmacol 2020, 52(5), 392-401.

Alzheimer’s disease (AD) is a constant, developing brain impairment that is described as the aggregation of misfolded amyloid‑beta‑peptide (Aβ) and abnormal tau protein in the brain. Stem cell therapy became a favorable candidate for the regeneration of neurodegenerative disorders like AD, but there is still shortage of knowledge about the underlying mechanisms. The goal of this survey was the determination of the necroptotic pathway as the possible mechanism for the effect of human adipose‑derived stem cells (hADSCs) in the rat model of AD. Twelve rats were consumed, dividing into four groups: Control, sham, AD model and AD + stem cell groups. We utilized Nissl and Thioflavin S staining for determining histological changes and immunofluorescent techniques for evaluating necroptotic markers in different regions of the hippocampus. The confirmation of AD model was approved with histological examination. The findings indicated more distinct Thio‑S stain and an increased number of dead cells in AD rats comparing to other groups. Alternatively, the dead cells number in the CA3 area significantly lessened in AD + stem cell group comparing to AD group. Data showed that hADSCs significantly decreased the expression of necroptotic markers (receptor‑interacting protein 1, receptor‑interacting protein 3 and mixed lineage kinase domain‑like pseudokinase (MLKL)) in AD + stem cell group compared to AD group in different regions of the hippocampus. Our findings revealed that the intravenous injection of hADSCs reduced necroptosis and consequently declined the death of neuronal cells in the hippocampus of AD rats.

6 illus, 30 ref

041880   BAGALI S, NERUNE S M, REDDY R C, YENDIGERI S M, PATIL B S, NAIKWADI A A, KULKARNI R V, DAS K K (Physiology Dep, Shri B. M. Patil Medical Coll, Vijayapura - 586 103, Karnataka, Email: kusaldas@bldedu.ac.in) : Low oxygen microenvironment and cardiovascular remodeling: Role of dual L/N‑type Ca²⁺ channel blocker. Indian J Pharmacol 2020, 52(5), 383-91.

Patients exposed to chronic sustained hypoxia frequently develop cardiovascular disease risk factors to ultimately succumb to adverse cardiovascular events. In this context, the present study intends to assess the role of cilnidipine (Cil), a unique calcium channel blocker that blocks both L‑type and N‑type calcium channels, on cardiovascular pathophysiology in face of chronic sustained hypoxia exposure. The study involved Wistar strain albino rats. The group‑wise allocation of the experimental animals is as follows ‑ Group 1, control (21 % O₂); Group 2, chronic hypoxia (CH) (10 % O₂ , 90 % N); Group 3, Cil + 21 % O₂ ; and Group 4, CH (10 % O₂ , 90 % N) + Cil (CH + Cil). Cardiovascular hemodynamics, heart rate variability, and endothelial functions (serum nitric oxide [NO], serum endothelial nitric oxide synthase [NOS3], and serum vascular endothelial growth factor [VEGF]) were assessed. Cardiovascular remodeling was studied by histopathological examination of the ventricular tissues, coronary artery (intramyocardial), and elastic and muscular arteries. Normalized wall index of the coronary artery was also calculated. The results demonstrated altered cardiovascular hemodynamics, disturbed cardiovascular autonomic balance, increased levels of VEGF and NOS3, and decreased bioavailability of NO on exposure to chronic sustained hypoxia. The histopathological examination further pointed toward cardiovascular remodeling. Treatment with Cil ameliorated the cardiovascular remodeling and endothelial dysfunction induced by CH exposure, which may be due to its blocking actions on L/N‑type of calcium channels, indicating the possible therapeutic role of Cil against CH‑induced cardiovascular pathophysiology.

4 illus, 4 tables, 31 ref

041879   GARG V K, GOYAL M K, KHULLAR M, SAIKIA B, MEDHI B, PRAKASH A, PRABHAT N, TANDYALA N, MAHESH K V, SACHDEVA J ET AL (Neurology Dep, Postgraduate Institute of Medical Education and Research, Chandigarh - 160 012, Email: juliesachdeva188@gmail.com) : Presence of allele CYP3A4*16 does not have any bearing on carbamazepine‑induced adverse drug reactions in North Indian people with epilepsy. Indian J Pharmacol 2020, 52(5), 378-82.

The objectives of this study were to determine the relationship between genetic polymorphisms in gene encodings for CYP3A4 and carbamazepine (CBZ)‑induced dose‑related side effects in North Indian people with epilepsy. The current prospective study included 37 patients with CBZ‑induced dose‑related side effects and 102 patients who did not experience side effects while on CBZ. The genotyping for CYP3A4 allele (CYP3A4*16) was done using real‑time polymerase chain reaction (RT‑PCR) in Applied Biosystems 7500 RT‑PCR System (USA). CBZ was administered in all patients at a dose varying from 15 to 20 mg/kg daily. Various demographic variables were comparable between the groups except that control of seizures was far better in controls. After testing, it was found that none of our patients had the presence of CYP3A4*16 allele. CYP3A4*16 allele is not represented significantly in North Indian people with CBZ‑induced dose‑related side effects.

2 tables, 13 ref

041878   KADLA S A, DAR M A, SHAH N A, KHAN B A, SHAH A I, PATHANIA R, PARVEEN S (Gastroenterology Dep, Govt. Medical Coll, Srinagar, Jammu and Kashmir, Email: nisarshah19@gmail.com) : Outcome of hepatitis C patients in a community with predominant genotype 3 with standard‑of‑care treatment before and after advent of direct‑acting antivirals: A retrospective‑cum‑prospective study. Indian J Pharmacol 2020, 52(5), 372-7.

Chronic hepatitis, cirrhosis, and hepatocellular carcinoma are mainly caused by hepatitis C infection. It is a worldwide predominant pathogen and is one of the main causes of healthcare problem in Asia. In the last few decades, there has been a considerable change in the treatment regimen for hepatitis C virus. The objective of this research was to relate the treatment response with sustained viral response in various therapies which have been the standard of care from time to time. This hospital‑based, retrospective‑cum‑prospective research span over a period of 2 years; we enrolled hepatitis C patients who attended the Department of Gastroenterology and Hepatology, Government Medical College, Srinagar, since June 2015 till May 2017. Subsequently, the database was prepared, containing all the relevant information about these patients. (i) In retrospective group: The overall efficacy (sustained viral response at 24 weeks [SVR‑24]) of pegylated interferon α2a and ribavirin regimen was 90.96 %. (ii) In prospective group: The efficacy (SVR) of different regimens was found to be as: sofosbuvir + ribavirin + daclatasvir (SVR‑24, 83.33 %); sofosbuvir + ribavirin (SVR‑12, 94.57 %); and sofosbuvir + daclatasvir (SVR‑12, 98.00 %).

3 illus, 3 tables, 27 ref