IGFBP3 gene promoter methylation analysis and its association with clinicopathological characteristics of colorectal carcinoma.

Promoter methylation mediated silencing of tumor suppressor genes plays an important role in the tumorigenesis of colorectal carcinoma (CRC). Tumor suppressor gene, Insulin-like Growth Factor Binding Protein-3 (IGFBP-3) expression is frequently downregulated in CRC due to promoter methylations. The aim of this study was to analyze the methylation status of IGFBP-3 gene promoter in stage II and III of CRC cases; find its association with clinicopathological characteristics of CRC patients and the methylation patterns as a prognostic biomarker. 58 histopathologically confirmed cases of CRC were included in the study. Methylation status of IGFBP-3 gene promoter was determined by using methylation specific PCR (MS-PCR) and bisulfite sequencing. Kaplan-Meier survival curve and univariate cox regression analysis were used for survival analysis; Chi-square test used for association analysis. IGFBP3 promoter methylation was found in 37 (63.8%) out of 58 CRC cases. This promoter methylation status was significantly associated with lymph-node metastasis (P = 0.013) and the survival period. In stage II CRC cases, unmethylated gene promoter status showed better survival than the methylated. Mean overall survival (OS) of methylated and unmethylated group was 22.23 months, and 49.15 months respectively (P = 0.045), HR = 6.432, 95% CI 0.986-41.943. The IGFBP-3 promoter methylations found in 63.8% CRC cases in this study. The methylations was found to be associated with lymph-node metastasis and overall survival of the patients particularly in stage II CRC patients. However, promoter methylation was not associated with other clinocopathological characteristics such as age, gender, tumor location etc.

Characterization of indoor settled dust and investigation of indoor air quality in different micro-environments.

Present study reports the characterization results of settled dust particles in different indoor micro-environments of an academic institution in India. Field emission scanning electron microscope analysis of indoor dust revealed the presence of mineral particles, fly ash, and soot particles of different morphologies. Energy-dispersive X-ray spectroscopy analysis of indoor dust indicated that crystal particles are comprised of elements such as C, O, Na, Mg, Al, Si, S, Cl, K, Ca, Fe, and Ti. These elements accounted for more than 99% of the samples. The average content of O (42.7%) and C (18.9%) in the dust particles was found to be higher than their natural abundances. The concentrations of PM10, PM2.5, and PM1 were observed in the range of 88-58, 37-33, 23-29 µg/m3, respectively. Except temperature, other parameters such as volatile organic carbon, carbon dioxide concentration, and relative humidity were found to be within comfort limits of American Society of Heating, Refrigerating, and Air-Conditioning Engineers. ABBREVIATIONS: ASHRAE: American Society of Heating, Refrigerating, and Air-Conditioning Engineers; CO2: carbon dioxide; COPD: chronic obstructive pulmonary disease; EDX: energy-dispersive X-ray; FESEM: field emission scanning electron microscope; FTIR: Fourier transform infrared spectroscopy; IAP: indoor air pollution; IAQ: indoor air quality; HAP: household air pollution; OAP: outdoor air pollution; PM: particulate matter; VOC: volatile organic carbon; WHO: World Health Organization.

Surface ozone scenario and air quality in the north-central part of India.

Tropospheric pollutants including surface ozone (O3), nitrogen dioxide (NO2), carbon monoxide (CO) and meteorological parameters were measured at a traffic junction (78°2' E and 27°11' N) in Agra, India from January 2012 to December 2012. Temporal analysis of pollutants suggests that annual average mixing ratios of tropospheric pollutants were: O3 - 22.97±23.36ppbV, NO2 - 19.84±16.71ppbV and CO - 0.91±0.86ppmV, with seasonal variations of O3 having maximum mixing ratio during summer season (32.41±19.31ppbV), whereas lowest was found in post-monsoon season (8.74±3.8ppbV). O3 precursors: NO2 and CO, showed inverse relationship with O3. Seasonal variation and high O3 episodes during summer are associated with meteorological parameters such as high solar radiation, atmospheric temperature and transboundary transport. The interdependence of these variables showed a link between the daytime mixing ratios of O3 with the nighttime level of NO2. The mixing ratios of CO and NO2 showed tight correlations, which confirms the influence of vehicular emissions combined with other anthropogenic activities due to office/working hours, shallowing, and widening of boundary layer. FLEXTRA backward trajectories for the O3 episode days clearly indicate the transport from the NW and W to S/SE and SW direction at Agra in different seasons.

Evaluation of Diagnostic Efficacy of Novel Red Blood Cell Parameters as Potential Screening Test for Detecting Latent Iron Deficiency in Blood Donors.

Iron deficiency anemia (IDA) forms a major share of global burden of anemia. Frequent blood donation is a common iatrogenic cause of iron insufficiency in healthy adults. Serum iron and hemoglobin levels are normal despite low serum ferritin levels, referred to as latent iron deficiency (LID). Aim of the present study was to evaluate the role of novel RBC parameters-percentage of hypochromic RBCs (%HPO), percentage of microcytic RBCs (%MIC), and haemoglobin content of reticulocytes (MCHr) of Abbott Alinity autoanalyzer as indicators of latent iron deficiency in blood donors. 260 consenting and eligible blood donors were included in the study. Complete blood counts including new RBC parameters on Abbott Alinity autoanalyzer and serum iron profile were measured for all donors. Donors were categorized into LID and No LID based on Ferritin and Transferrin saturation (TSAT). Serum transferrin receptors (sTfR) were studied in a subset of samples [LID (n = 46), No LID (n = 18) and IDA (n = 27)]. Statistical analyses was done on IBM SPSS version 22. Among 260 donors, 56 (21.5%) were found to have LID. The difference in mean values for % HPO, % MIC, and MCHr were not found to be statistically significant in LID and No LID groups. sTfR results between LID, No LID and IDA sub-groups revealed significant difference. This study does not support the role of % HPO, % MIC and MCHr measured on Abott Alinity analyzer, as potential screening parameters for LID amongst blood donors. STfr was more informative in this regard. Further research on much larger sample size is required to confirm these findings. Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-023-01683-w.

Primary monophasic synovial sarcoma of the lung: Rare case report.

Primary monophasic synovial sarcoma of the lung is an extremely rare malignant mesenchymal tumor that can develop at any anatomic site. Synovial sarcoma is considered a high grade tumor with a poor prognosis. Metastatic pulmonary sarcoma is much more common. Hence primary lesion elsewhere in the body needs exclusion. No clinical or radiological features are specific for pulmonary sarcoma, often it is confused with bronchogenic carcinoma. Therefore biopsy is needed to establish the diagnosis of this rare tumor. We hereby present two cases of histologically proven primary monophasic synovial sarcoma of lung. The imaging features of this rare disease is reviewed.

A meta-analysis of differentially expressed and regulatory genes with their functional enrichment analysis for brain transcriptome data in autism spectrum disorder.

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition characterized by persistent challenges in social interactions and repetitive behavioral patterns. It is a significant problem emerging worldwide, as one in 100 children is affected by this disorder globally. In this study, a meta-analysis was performed for the identification of differentially expressed genes (DEGs) along with the expression analysis of regulatory genes. Functional enrichment analysis was an integral part of current findings to notify the significant pathways of this complex disorder. The study was conducted with two RNA-Seq datasets, viz., GSE64018 and GSE62098, for ASD patients and control samples from the GEO database. The identification of up-regulatory and down-regulatory genes was performed by the interaction analysis of transcription factors (TF) and DEGs. As an outcome of the meta-analysis, 2543 DEGs were identified as common across both of the datasets in which 1402 DEGs exhibited upregulation and 1130 genes have shown downregulation. In network analysis, upregulatory genes have shown strong interaction while downregulatory genes exhibit weak or null interaction. Further, in the enrichment analysis of screened upregulatory DEGs, three major significant pathways were identified namely the ATP synthesis pathway, FAS signaling pathway, and the Huntington's disease pathway. The common expression of CYC 1 gene in all the identified pathways has indicated that it is an important key regulator gene for the majorly associated pathways. The study concludes that all the potential DEGs were found to show their related high expression in neurobiological regulations specifically with ASD.Communicated by Ramaswamy S. Sarma.

Immunophenotypic characterization of normal and abnormal plasma cells in bone marrow of newly diagnosed multiple myeloma patients.

Background: Identification of plasma cells into abnormal (APC) and normal (NPC) compartments is of utmost importance in flow cytometric (FC) analysis of multiple myeloma (MM) and related plasma cell dyscrasias for diagnosis, prognosis, and follow-up. No single phenotypic marker is sufficient to distinguish NPC from APC. Materials and Methods: 43 newly diagnosed cases of MM and 13 controls were included in the study. Bone marrow (BM) samples from the 2nd pass were processed on the same day with antibodies against CD38, CD138, CD19, CD81, CD45, CD117, CD200, CD56, cytoKappa, and cytoLambda in a 4-color experiment with CD38 and CD138 as gating antibodies. Results: Mean APC% in cases was 96.5%. The expected Immunophenotype (IP) of APC which is CD19-/56+/45-/81-/117+/200+ was found in only 13/43 MM cases. In 30/43 cases, APC revealed deviation from expected IP either for single or a combination of markers. Sensitivity for APC detection was highest for CD19 (95.2%) followed by CD56 (90.4%) and CD81 (83.7%). Specificity was highest for CD19 (100%), CD56 (100%), and CD81 (100%) followed by CD117 (92.3%). Combination of markers with maximum sensitivity to detect APC (97.6%) was CD81- or CD19- and CD200+ or CD56+ (two markers); and for NPC (92.3%) was CD81+ and CD19+ and CD56- (three markers). Conclusion: Plasma cell IP can be highly variable with multiple minor subpopulations in both cases and normal controls. CD 19 and CD56 are highly informative markers for a 4-color experiment. Assessment of multiple markers in an 8-10 color experiment is more informative but the lack of advanced flow cytometers should not limit the use of FC in a 4-color approach. Our results emphasize that even basic equipment with limited fluorochrome can provide meaningful information if used appropriately.

Diagnostic accuracy of flow cytometry in detecting malignant epithelial cells in serous effusions.

INTRODUCTION: This study aims to evaluate diagnostic accuracy of flow cytometry (FCM) in detecting malignant epithelial cells in serous effusions. MATERIALS AND METHODS: Flow cytometric assessment of 96 serous fluids (86 ascitic, 10 pleural) was performed by using epithelial cell adhesion molecule (EpCAM) (in all 96 fluids) and MUC-1 (in a subgroup of 40 fluids) as epithelial markers and CD45 and CD14 as leucocyte markers. The percentage of EpCAM positivity and MUC-1 positivity was calculated in the CD14 and CD45 dual negative population by selective gating. The findings were then correlated with the defined gold standard criteria. RESULTS: The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy for EpCAM was found to be 92.06%, 96.96%, 98.31%, 86.48%, and 93.75%, respectively, while that for MUC-1 was 79.16%, 93.75%, 95%, 71.4%, and 85%, respectively. The sensitivity, specificity, PPV, NPV, and diagnostic accuracy for dual positivity for EpCAM and MUC-1 was found to be 83.3%, 100%, 100%, 80%, and 90% respectively. On combining FCM with cytomorphology the sensitivity, specificity, PPV, NPV, and diagnostic accuracy all increased greatly to 95.3%, 100%, 100%, 91.4%, and 96.8%, respectively. CONCLUSIONS: This study highlights the importance of multicolored flow cytometric analysis in detecting epithelial malignancies in effusions specially in cases belonging to the atypia of undetermined significance and suspicious for malignancy categories and in cases with strong clinical suspicion of malignancy with negative fluid cytology. We recommend the combined use of FCM and cytology for this specific subgroup of patients in routine clinical practice for fast and accurate reporting.

Application of international system for reporting serous fluid cytology (ISRSFC) in effusion samples-a prospective study in an oncology setting.

INTRODUCTION: Serous fluid cytology is a cost-effective procedure that can help in the diagnosis, staging, and origin of the malignancy. Recently introduced International System for Reporting Serous Fluid Cytology (ISRSFC) standardizes the reporting of serous fluid cytology in the 5 categories: Category 1: Nondiagnostic (ND), Category 2: negative for malignancy (NFM), Category 3: atypia of undetermined significance (AUS), Category 4: suspicious for malignancy (SFM), and Category 5: malignant (MAL). Here, we present our experience adopting the ISRSFC. MATERIALS AND METHODS: We implemented ISRSFC in December of 2019 at our institute and included a cohort of 555 prospective effusion samples. The pertinent surgical pathology, radiology, and clinical follow-up were also extracted to assess the risk of malignancy (ROM) and performance parameters. RESULTS: The assessment of interobserver reliability indicated substantial concordance (κ = 0.717) between the 2 investigators for serous fluid categorization. A total of 555 effusion samples were classified as follows: ND, 14 (2.5%); NFM, 394 (71%); AUS, 12 (2.2%); SFM, 13 (2.3%); and MAL, 122 (22%). The ROM for the ND, NFM, AUS, SFM, and MAL categories was 57.1%, 9.9%, 66.7%, 66.7%, and 97.2%, respectively, in peritoneal effusions and 57.1%, 7.1%, 66.7%, 100%, 100%, respectively, in pleural effusions. The ROM for NFM and MAL was 0% and 100%, respectively, in pericardial effusion. CONCLUSIONS: Application of the proposed ISRSFC can help in achieving uniformity and reproducibility in diagnoses and also help in risk stratification in cytology. ISRSFC was successfully adopted by our cytology laboratory and clinicians, with overall diagnostic performance similar to previous studies.

Screening of the c-kit gene missense mutation in invasive ductal carcinoma of breast among north Indian population.

Breast cancer is one of the most frequently diagnosed cancers and the leading cause of cancer deaths among females across the world, accounting for 23 % (1.38 million) of total new cancer cases and 14 % (0.45 million) of the total cancer deaths in 2008. c-kit is expressed in mast cell growth factor, cellular migration, proliferation, melanoblasts, haematopoietic progenitors and germ cells. We have designed our study with aim to explore the c-kit gene mutations in invasive ductal carcinoma (IDC) breast. To ascertain the range of mutations in exon 11, 13 and 17 of c-kit gene in 53 cases of IDC breast, we carried out PCR-SSCP followed by DNA sequencing. The mutation frequency of c-kit gene in exon 11, 13 and 17 were 9.43 % (5/53), 1.88 % (1/53) and 3.77 % (2/53), respectively. During our mutational analysis, we have detected five missense mutations in exon 11 (Pro551Leu, Glu562Val, Leu576Phe, His580Tyr and Phe584Leu), one missense mutation in exon 13 (Ser639Pro) and two missense mutations in exon 17 (Arg796Gly and Asn822Ser). It seems that c-kit mutations might participate in breast cancer pathogenesis and may be utilized as predictive marker, since the loss of c-kit positivity is generally linked with different types of breast cancer. Further molecular studies are necessary to validate the association of c-kit gene mutation in IDC breast pathogenesis.

Cutaneous cholesterol embolization syndrome: A case report.

An 81-year-old woman with chronic kidney disease, systemic hypertension, and a large infra-renal abdominal aortic aneurysm, developed bilateral calf muscle pain, altered sensorium, and deterioration of renal function following endovascular aneurysmal repair. On the third post-operative day she developed symmetrical purpuric macules with erythematous margins on the gluteal region and bluish reticulated patches on the soles and tips of toes. This was followed by melena development on the seventh post-operative day. Histology of the skin confirmed the diagnosis of cutaneous cholesterol embolization syndrome (CES). She was treated with hemodialysis and supportive management and she recovered.

The Frequency of Mutations in Exon 11 of the c-kit Gene in Patients With Leukemia.

OBJECTIVE: To determine the frequency of mutations in exon 11 of the c-kit gene in patients with leukemia. MATERIAL AND METHODS: The study included 50 leukemia patients (31 with acute myeloid leukemia, 5 with acutelymphoblastic leukemia, 9 with chronic myeloid leukemia, and 5 with chronic lymphocytic leukemia) that underwentPCR-SSCP, followed by direct DNA sequencing. RESULTS: In all, 28 of the leukemia patients were male and 22 were female, with a mean age of 31.88 years (range: 2-65years). In total, 20 mutations in 19 patients were identified, including Lys550Asn, Tyr568Ser, Ile571Thr, Thr574Pro,Gln575His, Tyr578Pro, Asp579His, His580Gln, Arg586Thr, Asn587Asp, and Arg588Met, as well as novel point mutationsat codons Ile563Lys, Val569Leu, Tyr570Ser, and Pro577Ser. Ile571Leu substitution was observed in 2 patients andTrp582Ser substitution was observed in 3 patients. CONCLUSION: The results suggest that mutations in exon 11 of the c-kit gene might be useful as molecular geneticmarkers for leukemia.

Microsatellite Instability in Sporadic Colorectal Malignancy: A Pilot Study from Northern India.

BACKGROUND: Three molecular pathways are described as the genetic basis of colorectal tumorigenesis. Among these, microsatellite instability (MSI) has shown greatest promise in serving as a biomarker to determine disease aggression by tumour biology, recurrence, and response to chemotherapy. METHODOLOGY: This prospective observational pilot study included patients of colorectal cancers, in a population subset coming to a tertiary care hospital in northern India, who were operated with curative or palliative intent over a period of one year and followed up for a maximum of 55 months. The post-operative pathological assessment was done for MSI status using PCR technique, and an attempt was made to evaluate its correlation with conventional clinical and histological parameters, early recurrences, disease-free survival and overall survival in comparison to MSS type tumours in sporadic cases of colorectal malignancies. RESULTS: Out of 38 patients of colorectal cancer, 26 were included in the study. Male to female ratio was 7:6 (n=14:12). Mean age of presentation was 48±14.2 years. Incidence of MSI was n=4 (15.4%). On subgroup analysis, age of presentation (p=0.044) and evidence of perineural invasion (p=0.017) was found to have significant statistical association with MSI tumour biology. Recurrence was seen in seven of the seventeen patients who previously had no synchronous or metastatic disease (41.2%). The mean disease-free survival for MSS was 21.32 months and was 25.25 months for MSI group which was statistically insignificant (p = 0.277). Out of four MSI tumour biology patients one was alive and without recurrence at 47 months. While the other two were alive and without recurrence till 27 months of follow-up.   Conclusion: Age and perineural invasion showed statistically significant association with MSI tumour biology. Due to the small sample size statistical significance was not established with site, recurrence rate, DFS and OS.

The Utility of Ancillary Techniques in the Cyto-Diagnosis of Malignant Scalp Lesions.

BACKGROUND: Fine needle aspiration cytology (FNAC) is rapid, inexpensive, and easy technique to establish the diagnosis of scalp lesions. The use of ancillary techniques such as immunocytochemistry (ICC), immunohistochemistry (IHC), and flow cytometry on aspiration material aids in accurate diagnosis which is additionally beneficial for management and prognosis. AIMS: The objective of this prospective case series was to evaluate the utility of ancillary techniques in the accurate cyto-diagnosis of malignant scalp lesions. MATERIALS AND METHODS: This study was a prospective case series that included 64 cases of scalp lesions in which FNAC had been performed for diagnosis. The lesions were categorized as Non-diagnostic/Inadequate, Inflammatory, Benign and Malignant. In all the cases that were categorized as malignant additional material was collected for ancillary testing that included ICC, cell block preparation followed by IHC and flow cytometry. RESULTS: Non-diagnostic/inadequate aspirates were 17.19% (n = 11/64), 25% (n = 16/64) aspirates were inflammatory, 35.93% (n = 23/64) aspirates were benign and 21.87% (n = 14/64) aspirates were categorized as malignant. With the aid of ancillary techniques, 57.14% malignant scalp aspirates were accurately categorized as epithelial origin. Lesions of bone and soft tissue constituted 28.57% (n = 4/14) of cases and lesions of hematolymphoid origin constituted 14.29% (n = 2/14) of all cases. CONCLUSION: This is a novel study where accurate categorization of malignant scalp tumors has been done with the use of ancillary techniques. This is useful as it may help in defining the tumor type, may aid in patient management. The material obtained can also be triaged for molecular testing.

Grossing and reporting of bone tumor specimens in surgical oncology: Rationale with current evidence and recent updates.

Primary bone tumors, including sarcomas, are rare tumors and require a multidisciplinary approach, including inputs from a radiologist, pathologist, medical oncologist, and surgical and radiation oncologist, for optimal management. Over the years, there has been a paradigm shift toward the treatment of bone sarcomas, from radical resections to conservative surgical procedures, to achieve improved clinical and functional outcomes. This has led to receiving and processing various types of specimens in orthopedic oncopathology. Grossing and reporting of bone tumors require expertise. This review focuses upon the types of biopsies, grossing techniques of various specimens in orthopedic oncology and reporting, with rationale and recommendations from pathologists, actively involved in reporting and pursuing a special interest in bone tumors, based on current evidence. Furthermore, there is a section on some of the updates in the diagnosis of bone tumors, based on the recent fifth edition of the World Health Organization classification of tumors of soft tissues and bone.

Grossing and reporting of a soft tissue tumor specimen in surgical pathology: Rationale, current evidence, and recommendations.

Soft tissue tumors, including sarcomas are complex and diagnostically challenging tumors. This is as a result of their heterogeneity and overlapping clinicopathological, immunohistochemical and also molecular features, the latter to some extent. More than 80 types of sarcoma have been described. Current management, which is best offered at centers with active multidisciplinary care, is based on balancing oncologic and functional outcomes in such cases. This has transcended into the types of specimens received for grossing these rather uncommon tumors. Over the years, diagnostic specimens have reduced in their sizes from, open biopsies to core needle biopsies. These specimens need to be adequately and judiciously triaged for ancillary techniques, such as molecular testing. Conservative surgeries have led to resected specimens for marginal assessment. Lately, post neoadjuvant (chemotherapy or radiation therapy)-treated resection specimens of soft tissue sarcomas are being submitted for surgical pathology reporting. This article focuses on the grossing of soft tissue tumors, including sarcomas, in terms of types of specimens, grossing techniques including rationale, tissue triage, reporting, and recommendations from the surgical pathologists actively engaged in reporting musculoskeletal tumors, based on current evidence.

Hypolipidemic activity of Hibiscus rosa sinensis root in rats.

The hypolipidemic activity of Hibiscus rosa sinensis (family Malvaceae) root extract was studied on triton and cholesterol-rich high fat diet (HFD) induced models of hyperlipidemia in rats. In triton WR-1339-induced hyperlipidemia, feeding with root extract (500 mg/kg body wt/day p.o.) exerted lipid-lowering effect, as assessed by reversal of plasma levels of total cholesterol (TC), phospholipids (PL) and triglycerides (TG) and reactivation of post-heparin lipolytic activity (PHLA) of plasma. The other model was fed with cholesterol-rich HFD and root extract (500 mg/kg body wt/ day p.o.) simultaneously for 30 days. This also caused lowering of lipid levels in plasma and liver homogenate and reactivation of plasma PHLA and hepatic total lipoprotein lipase activity. The hypolipidemic activity of Hibiscus rosa sinensis root was compared with a standard drug guggulipid (200 mg/kg body wt/day p.o.), a known lipid- lowering agent in both models. Histopathological findings in rat liver supported the protective role of H. rosa sinensis root extract in preventing cholesterol-rich HFD-induced hepatic steatosis.

Prognostic Relevance of SFRP1 Gene Promoter Methylation in Colorectal Carcinoma

Background: The development of colorectal carcinoma (CRC) involves many genetic and epigenetic alterations and methylation being an important epigenetic event has been described as a diagnostic and prognostic biomarker. Secreted Frizzled- Related Protein 1 (SFRP1) gene regulates diverse physiological processes via the Wnt signaling. Promoter hypermethylation of SFRP1 gene is an epigenetic regulation mechanism that downregulates SFRP1 protein level in the tumor, and happens to be one of the significant events in colorectal carcinogenesis. We studied the clinicopathological relationship of CRC including survival outcomes with SFRP1 gene promoter methylation. Methods: We evaluated promoter methylation status of SFRP1 gene by methylation-specific PCR (MS-PCR) in the tumor tissue in 54 cases of stage II-III CRC patients in north India. The MS-PCR result was further validated by bisulfite sequencing. Results: SFRP1 gene was methylated in 72.2% cases and un-methylated in 27.8%. We found, that SFRP1 gene methylation in tumor was associated with lymph node invasion (p=0.05). The mean overall survival was 22.318 months and 45.173 months respectively for patients with methylated and unmethylated SFRP1 gene (p= 0.010, log rank test), (HR = 17.313, 95% CI: 2.021-148.290 P=0.009). Conclusion: Study indicates that promoter methylation of SFRP1 gene is associated with lymph-node metastasis and poor mean overall survival and it can be a prognostic marker in CRC.