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Introduction Angiogenesis has been proposed as a promising prognostic marker in a variety of tumours. Microvessel density (MVD) measurement is the most reliable method for the semiquantitative evaluation of angiogenesis. Endoglin (CD105) is a recognised marker of proliferating endothelium and is expressed in intra-tumoral as well as peri-tumoral vessels. It is upregulated in various cancers including prostate cancer, therefore, it is an upcoming prime marker for prognosis, tumour imaging and antiangiogenesis therapy. Aim The study was done to evaluate microvessel density (MVD) by using CD105 (endoglin) as an IHC marker in prostatic adenocarcinoma and to find its relationship with Gleason score and grade groups. Materials and methods This was an observational cross-sectional study under which a total of 31 histopathologically diagnosed cases of prostatic adenocarcinoma on TURP chips were included and were graded according to Gleason score and grades.IHC using endoglin (CD105) was done to detect the highest-density areas of stained vessels (hot spots). The highest value obtained in three hot spot fields was reported as microvessel density (MVD) and its relationship with Gleason score and grade groups was analysed. Results Most common age group was 71-80 years (48.39% cases). Maximum cases (45.16%) had a Gleason score of 9. The maximum mean MVD CD105 score was found in the Gleason grade group 5 (45.32%) while the minimum was in grade group 1 (24%). Comparison of mean MVD CD105 scores in different Gleason grade groups revealed a significant association of MVD CD105 with Gleason grade groups. Conclusion Endoglin measures microvessel density and its expression correlated with Gleason score and grade group. Hence it can be used as a potential prognostic marker in adenocarcinoma prostate.

Background Urothelial carcinoma (UC) of the bladder is a major cause of morbidity and mortality throughout the world. The present study was carried out to see the expression of CD105 and CD34 as marker of angiogenesis in UC and study their relationship with grades and muscle invasion. Methods 50 cases of histopathologically diagnosed UC were included. CD105 and CD34 IHC staining was performed. Microvessel density (MVD) evaluation by CD105 and CD34 was compared. The number of microvessels were counted in high power in three hot spots. The highest value obtained amongst three fields were reported as MVD. Statistical analysis was done by (SPSS 28.00 version for windows) using independent t-test. Results There were 45 high grade cases UC 5 low grade cases of UC. Invasive papillary UC was seen in 25 cases (50%) followed by invasive UC seen in 13 cases (26%). Both Lamina propria (LP) invasion and muscularis propria invasion were noted in 16 cases of high-grade urothelial carcinoma. Only LP invasion was seen in 3 cases of low grade and 25 cases of high-grade UC respectively. Evaluation of MVD CD105 and MVD CD34 did not show any significant association of MVD with grade with a p value of 0.214 & 0.429 respectively. Aso there was no significant association of MVD CD105 (p=0.580) & CD34 (p=0.747) with muscularis propria invasion. Overall mean CD105 and CD34 were similar with no statistical difference between the two markers. Conclusion Both CD105 and CD34 can be used as marker to assess angiogenesis. However, there is no significant association of MVD with grade or muscle invasion in UC.

Introduction: The role of immune mediated injury in pathogenesis of Ankylosing Spondylitis (AS) is well established. HLA B27, a Major Histocompatibility Complex (MHC) class I molecule is one of the major genetic risk factors associated with the disease. Various techniques are used for testing HLA-B27 which includes Polymerase Chain Reaction (PCR) based tests, Enzyme Linked Immunosorbent Assay (ELISA) and flowcytometry. Flowcytometry has gained popularity due to ease of procedure, shortened turnaround time and cost-effectiveness. Aim: The aim of the present study was to assess the sensitivity and specificity of flowcytometry for HLA-B27 detection, taking PCR assay as the gold standard along with its association with demographic, clinicopathological and radiological parameters. Materials and Methods: This was a prospective study conducted in Department at Pathology of Shri Guru Ram Rai Institute of Medical and Health Sciences, Dehradun, India, for a period of 18 months from January 2020 to June 2021. The study included 51 patients for which HLA-B27 typing was done cases by flowcytometry and Sequence Specific Allele (SSA) PCR/Real time PCR on peripheral blood samples. The association of HLA-B27 with clinical features {Inflammatory Back Pain (IBP), arthritis, psoriasis, uveitis, dactylitis, Inflammatory Bowel Disease (IBD), cervicitis, urethritis, diarrhoea) along with MRI findings (sacroiliitis)}, laboratory findings {C-reactive protein and Erythrocyte Sedimentation Rate (ESR)} was evaluated. The performance analysis parameters of flowcytometry were evaluated both by excluding and including the cases in grey zone taking PCR as gold standard. Statistical testing was conducted with SPSS 20.0. Chi-square test or Fisher’s-exact test were used and a p-value of less than 0.05 was taken as significant. Results: A significant association of HLA-B27 was seen only with IBP (p-value= 0.001) and sacroiliitis (p-value= 0.03). Of the 22 (43.1%) patients positive for HLA-B27 by PCR, 18(81.8%) patients were positive while the remaining 4(18.1%) were in grey zone by flowcytometry. Of the 29(56.9%) patients testing negative by PCR, 27(93.1%) patients were negative, one (3.4%) was in grey zone and one (3.4%- false positive) tested positive for HLA-B27 by flowcytometry. Sensitivity and specificity of flowcytometry for detection of HLA-B27 was found to be 100% and 96.4% respectively when grey zone cases were excluded. Conclusion: The study brings to light that flowcytometry is a fairly specific and sensitive method for HLA-B27 detection with a high Negative Predictive Value (NPV) (100%) and Positive Predictive Value (PPV) (94.7%). In the COVID-19 era, it reiterates the importance of flowcytometry for HLA-B27 especially when PCR is overburdened.

Understanding changes in Land Use Land Cover (LULC) is essential for managing and monitoring natural resources and development, particularly where urbanization is expanding. So, this study aimed to assess the level of changes in LULC of Nepalgunj Sub-metropolitan city using temporal Landsat satellite imageries of 1996, 2008, and 2020 AD, and the key drivers of LULC change were observed through a purposive household survey (N=140) with a sampling intensity of 0.5%. LULC maps were generated using initial unsupervised and later supervised classification. LULC changes were computed using the post-change detection classification technique. LULC map of 1996 AD, 2008 AD, and 2020 AD showed accuracy of 84.44 %, 85.45%, and 83.64% with a kappa value of 0.8381, 0.8497, and 0.829 respectively. Bareland, Human buildup, and grassland were found to have increased by 13.34%, 5.07%, and 29.62% respectively while sparse vegetation, dense vegetation, and water bodies were found to have decreased by 44.10%, 17.82%, and 13.34% respectively between 1996 and 2008. Likewise, there was decrease in grassland area (-26%), dense vegetation area (-9.48%), sparse vegetation area (-5%), water bodies (-0.12%), and increase in Bareland (+20%) and Human buildup (+20.6%) in between 2008 to 2020. Eight key drivers of LULC, development of infrastructure, government policy, plans, and land market, forest encroachment, forest, and its products, political condition, economic opportunities, and hotel and tourism activities, were recognized in the study area. Further research is required to determine the specific ramifications of the aforementioned LULC change drivers, as well as the area's long-term viability.

Background: Gastric carcinoma is a major cause of cancer-related morbidity and mortality worldwide. Gastric neoplasms arise from genetic and epigenetic changes in various genes. Present study evaluates the immunoexpression of PTEN, HER2/neu, and Ki-67 in endoscopic gastric carcinoma biopsies and correlates the expression of these proteins with clinicopathological features. Material and Methods: Adequate endoscopic biopsies of 27 cases of gastric carcinoma were evaluated for World Health Organization (WHO) and Lauren's classification subtypes along with HER2/neu, PTEN, and Ki-67 immunoexpression. HER2/neu immunostaining was scored as proposed in the Trastuzumab for gastric cancer (ToGA) trial while PTEN staining and downregulation were assessed using an immunoreactive score. The cut-off for Ki-67 expression was taken as 90th percentile of the values in adjacent non-neoplastic tissue. All statistical analysis was done at 5% level of significance with SPSS v22 statistical software. Results: Tubular adenocarcinoma was the commonest WHO histological subtype and 56% of cases were of intestinal type as per Lauren's classification. 55.6% of cases showed a complete loss of PTEN expression in neoplastic tissue. 17 of the 19 cases with adjacent non-neoplastic tissue showed PTEN downregulation in neoplastic tissue. 81.5% of cases had a high Ki-67 index and HER2/neu overexpression was noted in 36% of cases. All the four cases who died had high Ki-67 proliferation indices; 3 patients had loss of PTEN expression and HER2/neu overexpression. Conclusion: We conclude that these immunomarkers can play important role in the behavior of gastric carcinomas and can be targeted for new therapies.

Introduction: Prostatic diseases cause significant morbidity and mortality. Although histopathological examination is the gold standard for diagnosing prostatic lesions but diagnosis may be challenging in the presence of benign mimickers or a very small focus of malignancy. Immunohistochemical aid to morphology helps in making a timely and accurate diagnosis. Aim: This study was done to evaluate the role of two antibodies panel High Molecular Weight Cytokeratin (HMWCK) and Alpha-Methylacyl-CoA Racemase (AMACR) in improving the diagnostic accuracy of prostatic lesions. Materials and Methods: This was an observational cross-sectional study conducted in the Department of Pathology, Shri Guru Ram Rai Institute of Medical and Health Sciences (SGRRIM and HS), Dehradun, Uttarakhand, India, from May 2019 to October 2020. Haemotoxylin and Eosin (H&E) stained sections of prostatic biopsies were classified into benign and malignant. Amongst malignant lesions, prostatic adenocarcinomas were graded according to Gleason’s grading system and Gleason’s scores were noted. One section from each was subjected to AMACR and HMWCK antibody tests. HMWCK was interpreted as negative/positive and continuous/discontinuous. For AMACR, both location and intensity of stain was observed. The parameters studied were Gleason’s score, group grade, expression of HMWCK and AMACR. Categorical data was presented in form of frequency and percentage. Independent t-test, Yates Chi-square test were used. Data was entered in Microsoft (MS) Excel sheet and analysis was done using CRAN R 2.1. Results: Total of 80 prostatic biopsies were taken, 24 were malignant and 55 were benign and one was Benign Prostatic Hyperplasia (BPH) with a focus suspicious for malignancy showing atypical small acinar proliferation on histopathological examination. The mean age of non neoplastic cases was 67.68±8.56 years, while that of neoplastic lesions was 75.41±9.34 years. Amongst benign, 56.3% (31/55) cases were BPH, 43.6% (24/55) cases were BPH with associated lesions which included 62.5% (15/24) cases of BPH with non specific prostatitis; 29.2% (7/24) cases of BPH with adenosis and 8.3% (02/24) cases of BPH with basal cell hyperplasia. Of malignant cases, 24 cases were of adenocarcinoma with maximum cases having Gleason’s score 9 (11/24;45.8%) and Group grade V (18/24;75%). The sensitivity, specificity, Positive Predictive Value (PPV), Negative Predictive Value (NPV) of HMWCK and AMACR were calculated using histopathology as the gold standard. Conclusion: Although histopathology is the gold standard in prostatic biopsies but immunohistochemistry is additional diagnostic aid in confirmation of diagnosis. Immunohistochemistry not only confirms the histological diagnosis but is of great help in challenging cases. It has markedly increased the diagnostic accuracy.

Introduction: The increasing incidence of sepsis and its associated mortality has urged the search for rapid and reliable diagnostic test. Flow cytometry offers rapid assessment of change in antigen expression by neutrophils in blood in suspected sepsis cases. An increased expression of CD64, a high affinity opsonic surface receptor, by neutrophils is seen in sepsis. Aim: To critically analyse the role of neutrophilic CD64 expression by different possible flow cytometric parameters in sepsis. Materials and Methods: This case-control study was conducted in Department of Pathology of a Sri Guru Ram Rai Institute of Medical and Health Sciences, Dehradun, Uttarakhand, India from February 2017 to July 2019. It included peripheral blood samples from 43 cases of sepsis and their corresponding age matched controls. Flow cytometric analysis of blood samples for CD64 expression by neutrophils was done. The statistical analysis was performed with Statistical Package for Social Sciences (SPSS) version 18.0, Chicago, IL. Results: Flow cytometric analysis revealed a statistically significant increased expression of CD64 by neutrophils in sepsis patients, in terms of all four parameters analysedincreased expression of CD64 as compared to controls, in terms of percentage of neutrophils expressing CD64 (63.88±34.12 in cases), mean Fluorescent Intensity (FI) (2137.19±2319.71), Median Fluorescent Intensity (MFI), (2011.28±2261.89) as well as Neutrophil:Lymphocyte (N:L) CD64 index (7.29±9.66). Sensitivity and specificity of each parameter evaluated in present study varied from 60.4-67.4% and 81.4-83.7%, respectively. Conclusion: Quantitative expression of neutrophil CD64 (nCD64), by flow cytometry, in terms of percentage expression, MFI, mean fluorescent intensity and Neutrophils:Lymphocytes CD64 index is helpful in diagnosing sepsis patients even after 72 hours of onset. This is a rapid, reliable and cost-effective investigation. Hence, enabling prompt and judicious treatment of sepsis.

Background: CD71 or Transferrin receptor is expressed on the surface of erythroid lineage cells. CD71 expression has been found to be significantly increased in rapidly proliferating cells. Methods: This cross-sectional study included 37 bone marrow samples of acute leukemia cases diagnosed between October 2016 to April 2018. The samples were analysed on BD FACS Canto II. We evaluated the expression of CD71 on leukemic blasts and compared median fluorescent intensities (MFI) of blasts in different types of acute leukemias. Results: The 37 cases comprised of 21 Acute Myeloid Leukemia (AML), 13 B-Acute Lymphoblastic Leukemia (B-ALL), 2 T- Acute Lymphoblastic Leukemia (T-ALL) and 1 mixed phenotypic acute leukemia (MPAL), T/Myeloid. CD 71 expression was noted in 70.3% (n= 26/37) of acute leukemia cases. CD71 expression was most commonly observed in AML (n= 15/21;71.4%), followed by B-ALL (n= 9/13;69.2%) and T-ALL (n= 1/2;50%). Single case of MPAL revealed blasts positive for CD71. MFI of leukemic blasts of single CD71 positive T-ALL was found to be highest, followed by AML, MPAL (T/Myeloid) and least in B ALL. Of the AML cases, the blasts of AML-M6, acute promyelocytic leukemia and AML-M1 showed higher CD71 expression in terms of MFI. Conclusions: Surface CD71 expression is not only found in erythroid lineage cells, but also in proliferating cells. CD71 MFI is highest in T lymphoblasts followed by leukemic erythroblasts, myeloblasts and least in B lymphoblasts.

Background: Cytological evaluation of specimens from respiratory tract is the initial investigation in patients suspected to have pulmonary diseases. The various cytological specimens submitted for analysis include bronchoalveolar lavage, bronchial wash, brush smears, transbronchial needle aspiration, guided fine needle aspiration cytology (FNAC) smears and pleural fluid. Present study was undertaken to study the spectrum of lesions diagnosed by cytomorphological analysis of various cytological specimens. Materials and Methods: Centrifuged and direct smears from received samples were stained with MGG and PAP stain. Special stains (ZN and PAS) were used wherever required. Cytohistological correlation was done wherever biopsy was available. Results: This study included 671 samples from 583 patients suspected clinicoradiologically of having a respiratory pathology. A male preponderance (65.87%) was noted with 73.59% of patients in age group 40–80 years. Cytological diagnoses were classified as non-diagnostic (14.90%), negative for malignancy (59.76%), specific benign lesions (5.22%), positive for atypical cells (3.87%) and malignant (16.25%). Tuberculosis (TB) was found in 17 cases. Adenocarcinoma (both primary and metastatic) was the commonest malignancy of the 99 cases positive for malignancy. Incidence of primary lung adenocarcinoma and squamous cell carcinoma were found to be equal. Conclusion: Lung carcinoma is presently the leading cause of cancer deaths while TB is still a common cause of death in developing countries. Cytopathology has a definite role in diagnosis of malignant lesions and also contributes in diagnosis of unsuspected chronic infections. Cytological diagnosis is fairly accurate if the specimen obtained is adequate and representative.

Splenic cysts are rare with an overall incidence of 0.07% in a large autopsy series and 0.5% among all the splenectomies done. The parasitic splenic cysts, usually caused by Echinococcus granulosus, account for 60% of all primary splenic cysts. The primary epithelial cysts account for 10% of all splenic cysts. We report a case of 30-year-old female presenting with left upper abdominal pain and heaviness. Computed tomography revealed a multiloculated cyst in spleen. Hydatid serology was negative. Total splenectomy was done. Histopathological evaluation was done and a diagnosis of primary epithelial splenic cyst was given.

Introduction: Lymphocytic thyroiditis is an autoimmune disorder characterised by lymphocytic infiltration of the thyroid gland leading to follicular destruction in addition to immunological alterations attributed to the antibodies circulating in the serum against thyroid peroxidase, thyroglobulin, and thyroid stimulating hormone receptor antigens. Aim: To study the cytomorphic spectrum of lymphocytic thyroiditis, and correlate the cytological grades with the levels of TSH, FT3, FT4 and Anti-TPO antibodies. Materials and Methods: This study was carried out over a period of 24 months in Pathology department of SGRRIM & HS, Dehradun, India. Out of 58 cases diagnosed as lymphocytic thyroiditis on cytology and graded using Bhatia A et al., grading system, the levels of TSH, FT3, FT4 and Thyroid Peroxidase Antibody were known in 31 cases. The cytological grades of lymphocytic thyroiditis were correlated with these parameters and statistical analysis was done using one way ANOVA. Results: Out of 31 cases, majority of patients (30 cases, 96.7%) were females presenting in third decade of life. Cytologically, in 21 cases (67.7%) the patients had grade 2 thyroiditis, followed by grade 3 thyroiditis in 7 cases (22.6%). Anti-TPO was elevated in 26 cases (83.8%) and elevated level of TSH was seen in 17 cases (54.8%). On correlating the cytological grades with these parameters, p-value was found to be 1.00 which is statistically insignificant. Conclusion: While Lymphocytic infiltration of thyroid follicles is pathognomonic and positivity for antithyroid antibodies is strongly associated with lymphocytic thyroiditis, no correlation was observed between the grades of thyroiditis and the levels of Anti-TPO, FT3, FT4 and TSH.

Introduction: Haemoglobinopathy is a worldwide inherited problem as recognized by WHO and care of affected patients incurs heavy expense on the limited resources of developing countries. There is a need for prevention of births of children with clinically significant haemoglobinopathies by population screening. Cation Exchange-High Performance Liquid Chromatography (CE-HPLC) has emerged to be a simple and precise method to quantify HbA2 , HbF and other variant haemoglobins with certain limitations. Most of the variant haemoglobins can be identified by their retention times, percentages and peak characteristics. Aim: The present study was undertaken to assess the prevalence and spectrum of various haemoglobinopathies in patients reporting to a tertiary health care centre in Uttarakhand, India. Materials and Methods: This was a prospective study conducted on 8144 samples. RBC indices were obtained by sysmex XP 100. CE-HPLC was performed on Biorad D10. The variant haemoglobins were identified on the basis of their percentages, retention times and peak characteristics. Peripheral blood film, reticulocyte count, HbH inclusion and sickling test were done in selected cases. Continuous variables were expressed as mean±SD. Categorical variables were expressed as percentages. Results: Antenatal population formed the bulk of the 8144 cases enrolled in this study. Haemoglobinopathy was seen in 5.9% of the cases with β thalassaemia trait being the commonest abnormality (2.82% of cases). HbD (Punjab) trait was the commonest variant haemoglobin encountered in the study population. There was a significant difference in percentages of variant fractions between compound heterozygotes and variant traits. A presumptive diagnosis of alpha thalassaemia trait was rendered based on RBC indices, iron profile and chromatogram study. Molecular studies were recommended in 81 cases with borderline increase in HbA2 levels to rule out silent mutations. Conclusion: A reasonably high frequency (5.9%) of haemoglobinopathies warrants a routine antenatal screening of total population. An accurate diagnosis can be made in majority of cases by haematological parameters, CE-HPLC chromatograms, cascade screening for haemoglobinopathies and spouses of antenatal cases positive for haemoglobinopathy.

Introduction: Fluid cytology and subsequent utilization of immunocytology on cell block is being used commonly for increasing the diagnostic accuracy in effusion cytology. Aim: To do cytological analysis of fluids and evaluate the role of Epithelial Membrane Antigen (EMA) and Calretinin (CAL) to differentiate between reactive and malignant cell in pleural and peritoneal fluids. Materials and Methods: This was a prospective study carried out in Department of Pathology, Shri Guru Ram Rai Institue of Medical and Health Science, for a period of 18 months from January 2014 to June 2015. A total of 253 samples of pleural and peritoneal fluid were studied by May-Gruenwald-Giemsa (MGG) stain and Papanicolaou (Pap) staining. In 73 cases which were suspicious for malignancy, cell blocks were prepared and IHC was done using two immunomarkers- EMA and CAL. Results: A total of 253 cases of effusion cytology were studied out of which 73 were found positive for malignant cells. Maximum cases of malignant cells or atypical cells were seen in peritoneal fluid. A total of 34 cases were histologically positive for malignancy. All these cases showed strong membranous and cytoplasmic positivity for EMA. Most of cases also showed 2%-5% positivity for CAL. There were 38 cases categorized as atypical or reactive mesothelial cell hyperplasia. These cases showed nuclear and cytoplasmic staining for calretinin and none of these were positive for EMA, although 5 cases showed score 2-3, which was considered as negative. Conclusion: The distinction between reactive and malignant mesothelial cells in cytological specimens can be problematic. A combination of CAL and EMA may help in accurate diagnosis.

Introduction: Cytodiagnosis of CNS Space Occupying Lesions (SOLs) by squash smear preparation is a rapid, inexpensive and fairly accurate diagnostic tool. Most of the studies till date have proved its utility in intraoperative consultation. Aim: This study was undertaken to evaluate accuracy of squash smear cytology and to highlight as well as analyze the diagnostic pitfalls. Materials and Methods: This study was conducted in department of pathology, Shri Guru Ram Rai Institute of Medical and Health Sciences. Intraoperative squash smears of two hundred and twenty two cases of CNS SOLs were studied. Cytological diagnoses were correlated with histopathological evaluation which was taken as gold standard. Discordant smears were reviewed and causes were analyzed. Results: An accuracy of 83.78% was achieved by squash smear cytology. The discrepancies due to interpretational errors were seen in 61.1% cases and sampling errors accounted for 38.9% cases. Cyto-histological discordance was noted more in oligodendrogliomas and grading of gliomas. Conclusion: Squash smear cytology is a fairly reliable diagnostic tool in intraoperative neuropathological consultation. Thorough sampling of tissue submitted, clinicoradiological correlation while reporting and awareness about the diagnostic pitfalls help in achieving reasonable accuracy