Discovery and Validation of Novel microRNA Panel for Non-Invasive Prediction of Prostate Cancer.

BACKGROUND: Early diagnosis remains a challenge for prostate cancer (PCa) due to molecular heterogeneity. The purpose of our study was to explore the diagnostic potential of microRNA (miRNA) in both tissue and serum that may aid in the precise and early clinical diagnosis of PCa. MATERIALS AND METHODS: The miRNA expression pattern analysis was carried out in 250 subjects (discovery and validation cohort). The Discovery Cohort included the control (n = 30) and PCa (n = 35) subjects, while the Validation Cohort included the healthy control (n = 60), benign prostate hyperplasia (BPH) (n = 55), PCa (n = 50), and castration-resistant PCa (CRPC) (n = 20) patients. The expression analysis of tissue (Discovery Cohort) and serum (Validation Cohort) was carried out by quantitative polymerase chain reaction (qPCR). The diagnostic biomarker potential was evaluated using receiver operating characteristics (ROC). Bioinformatic tools were used to explore and analyze miRNA target genes. RESULTS: MiRNA 4510 and miRNA 183 were significantly (p<0.001) upregulated and miRNA 329 was significantly (p<0.0001) downregulated in both PCa tissue and serum. ROC curve analysis showed excellent non-invasive biomarker potential of miRNA 4510 in both PCa (area under the curve (AUC) 0.984; p<0.001) and CRPC (AUC 0.944; p<0.001). The panel of serum miRNAs (miRNA 183 and miRNA 4510) designed for PCa had significant and greater AUC with both 100% sensitivity and specificity. Computational analysis shows that the maximum number of target genes are transcription factors that regulate oncogenes and tumor suppressors. CONCLUSION: Based on ROC curve analysis, miRNAs 4510, 329, and 711 were identified as potential non-invasive diagnostic biomarkers in the early detection of PCa. Our findings imply that a panel of miRNAs 183 and 4510 has high specificity for distinguishing PCa from healthy controls and providing therapeutic targets for better and earlier PCa therapy.

Correlation of Serum Galactose-Deficient IgA1 and Oxford Class in Cases of IgA Nephropathy.

CONTEXT.­: Galactose-deficient immunoglobulin A1 (Gd-IgA1) deposition in the renal mesangium plays a role in the pathogenesis of IgA nephropathy. OBJECTIVE.­: To assess the serum Gd-IgA1 level in biopsy-proven IgA nephropathy cases on diagnosis and 3 months post treatment and its relation with histologic Oxford classification. DESIGN.­: In this hospital-based prospective cohort study, 40 cases and 20 controls were enrolled. Serum samples of biopsy-proven IgA nephropathy cases collected on the day of biopsy and 3 months post treatment were evaluated. Solid-phase ELISA (enzyme-linked immunosorbent assay) was performed for assessment of Gd-IgA1 level. All renal biopsies were scored by using Oxford Classification (C-MEST score). The association of serum Gd-IgA1 levels with other established prognostic parameters was assessed. To estimate the prognostic value of markers, logistic regression analysis and Kruskal-Wallis ANOVA (analysis of variance) were used. RESULTS.­: Significant difference was observed in the serum Gd-IgA1 level values in the IgA nephropathy cases and healthy controls (P = .001) at baseline. However, no significant correlation between serum Gd-IgA1 levels at baseline and 3 months of follow-up (P = .31) or between baseline levels and age, proteinuria, hematuria, or estimated glomerular filtration rate was noted. There was no significant correlation between C-MEST score and serum Gd-IgA1 levels at baseline (P > .05); however, the distribution of Gd-IgA1 at 3 months was found to differ significantly between different grades of S score (P = .008). CONCLUSIONS.­: Serum Gd-IgA1 levels may be of utility in predicting disease progression in IgA nephropathy cases. Measurement of serum Gd-IgA1 levels for the diagnosis and prognosis of IgA nephropathy may preclude the need for invasive renal biopsies.

Evaluation of serum adipokines (omentin-1 and visfatin) in coronary artery disease at a North Indian hospital.

Objective. Adipose tissue is considered to be an endocrine organ that secretes bioactive substances known as adipokines that contribute to the pathophysiology of metabolic and coronary diseases related to obesity. In this study, various novel biomarkers, such as inflammatory markers that are pro-inflammatory (visfatin) and anti-inflammatory (omentin-1), as prognostic indicators for people with coronary artery disease (CAD) were investigated. Methods. In this study, 30 diabetic patients with CAD, 30 diabetic patients without CAD, and 30 healthy control counterparts were included. Serum omentin and visfatin concentrations were evaluated by solid-phase enzyme linked immunosorbent assay (ELISA) kit. Patients with established diagnosis of CAD based on angiography, ECG, and elevated cardiac marker level were included into the study. Patients with cardioembolic stroke, cerebral venous sinus thrombosis, CNS vasculitis, and hemorrhage due to trauma, tumor, vascular malformation, and coagulopathy were excluded. Results. The serum omentin-1 levels were significantly higher in the healthy controls in comparison with the diabetic group (p<0.0001) and serum visfatin levels were significantly higher in the diabetic group in comparison with the healthy controls (p<0.0001). The serum omentin levels were significantly higher in the diabetic group in comparison with the cardio-diabetic group (p<0.0001) and serum visfatin levels were significantly higher in the cardio-diabetic group in comparison with the diabetic group (p<0.0001). The serum omentin-1 showed negative correlation with the serum visfatin in the cardio-diabetic group. Conclusion. The adipokines, such as omentin and visfatin, may be good therapeutic candidates in preventing or ameliorating CAD.

Automated lead toxicity prediction using computational modelling framework.

Background: Lead, an environmental toxicant, accounts for 0.6% of the global burden of disease, with the highest burden in developing countries. Lead poisoning is very much preventable with adequate and timely action. Therefore, it is important to identify factors that contribute to maternal BLL and minimise them to reduce the transfer to the foetus. Literacy and awareness related to its impact are low and the clinical establishment for biological monitoring of blood lead level (BLL) is low, costly, and time-consuming. A significant contribution to an infant's BLL load is caused by maternal lead transfer during pregnancy. This acts as the first pathway to the infant's lead exposure. The social and demographic information that includes lifestyle and environmental factors are key to maternal lead exposure. Results: We propose a novel approach to build a computational model framework that can predict lead toxicity levels in maternal blood using a set of sociodemographic features. To illustrate our proposed approach, maternal data comprising socio-demographic features and blood samples from the pregnant woman is collected, analysed, and modelled. The computational model is built that learns from the maternal data and then predicts lead level in a pregnant woman using a set of questionnaires that relate to the maternal's social and demographic information as the first point of testing. The range of features identified in the built models can estimate the underlying function and provide an understanding of the toxicity level. Following feature selection methods, the 12-feature set obtained from the Boruta algorithm gave better prediction results (kNN = 76.84%, DT = 74.70%, and NN = 73.99%). Conclusion: The built prediction model can be beneficial in improving the point of care and hence reducing the cost and the risk involved. It is envisaged that in future, the proposed methodology will become a part of a screening process to assist healthcare experts at the point of evaluating the lead toxicity level in pregnant women. Women screened positive could be given a range of facilities including preliminary counselling to being referred to the health centre for further diagnosis. Steps could be taken to reduce maternal lead exposure; hence, it could also be possible to mitigate the infant's lead exposure by reducing transfer from the pregnant woman.

A Study of Maternal and Umbilical Cord Blood Lead Levels in Pregnant Women.

Lead is a highly toxic element which can cross the placental barrier and enter the fetus during pregnancy. Parental lead exposure has adverse effect on infant as well as on maternal health. As part of our program to investigate the lead poisoning in human population we investigated the maternal blood lead levels (MBLL) and umbilical cord blood lead (UBLL) levels in 200 pregnant women and collected their socio-demographic details. In the study we found high lead levels in both maternal and umbilical cord blood samples. The results showed 47.5% maternal blood (n = 95) detected with lead while 38.5% umbilical cord blood (n = 77) samples had lead concentration higher than that of reference range of ≤ 5 µg/dL. We also found that the Spearman's correlation coefficient (rs) revealed a strong positive correlation between the MBLL and UBLL (rs = 0.63). The results from socio-demographic questionnaire demonstrated that the recent home painting (p = 0.002) and residing close proximity to traffic congestion (p = 0.05) were significantly associated with MBLL. Education, mother age, fuel and water sources were not significantly associated with MBLL. Iron and calcium deficiency along with tiredness, lethargy, abdominal pain were also reported in women having high lead level > 5 µg/dL. Concludingly, on the basis of results obtained it may be stated that we found elevated BLLs in both pregnant women as well as in umbilical cord blood. The prevalence of elevated lead levels in mothers will expose the fetus to lead through placental barriers mobilization and it can have long term adverse effects on the developing fetus. Therefore, it is recommended that screening of blood lead levels be carried out in high-risk women based on their social, occupational, environmental, and individual factors. In addition, stringent regulations on lead-based products are also required from government agencies/authorities to reduce environmental lead burden and toxicity. Moreover, public awareness programs should be organized on hazardous effect of lead.

Prevalence of specific micronutrient deficiencies in urban school going children and adolescence of India: A multicenter cross-sectional study.

INTRODUCTION: Childhood and adolescence require adequate amount of micronutrients for normal growth and development. The primary objective of study was to assess the prevalence of deficiencies of Vitamins (Vitamin A, 25 Hydroxy Vitamin D, Vitamin B12 and Folate) and minerals (Calcium, Zinc, Selenium and Iron), among urban school going children aged 6-11 and 12-16 years in ten cities of India. Secondary objective was to find the association between micronutrient deficiencies with sociodemographic and anthropometric indicators. METHODS: A multi-center cross-sectional study was conducted across India. Participants in the age groups of 6 to 11 years (group 1) and 12 to 16 years (group 2) were selected from randomly chosen schools from each center. Data on socio economic status, anthropometric measures was collected. Blood samples were collected for biochemical analysis of micronutrients. Point estimates and 95% confidence intervals was used to assess the prevalence of deficiencies. Associations were observed using chi square, student t test and ANOVA test. RESULTS: From April 2019 to February 2020, 2428 participants (1235 in group 1 and 1193 group 2) were recruited from 60 schools across ten cites. The prevalence of calcium and iron deficiency was 59.9% and 49.4% respectively. 25 Hydroxy Vitamin D deficiency was seen in 39.7% and vitamin B12 in 33.4% of subjects. Folate, Selenium and Zinc were deficient in 22.2%, 10.4% and 6.8% of subjects respectively. Vitamin A deficiency least (1.6%). Anemia was prevalent in 17.6% subjects and was more common among females. CONCLUSION: One or more micronutrient deficiencies are found in almost one half of school going children in urban area. Hence efforts must be made to combat these on priority. TRIAL REGISTRATION NUMBER: CTRI/2019/02/017783.

Chemical Characterization, In-silico Evaluation, and Molecular Docking Analysis of Antiproliferative Compounds Isolated from the Bark of Anthocephalus cadamba Miq.

AIMS: The present study aimed to isolate and characterize chemical compounds from Anthocephalus cadamba Miq. bark and evaluate their anticancer activity by in silico, molecular docking, and in vitro studies. BACKGROUND: Anthocephalus cadamba is a traditionally used Indian medicinal plant. The anticancer and phytochemical properties of this plant remain unexplored except for a few studies. OBJECTIVES: The objective of the study was to evaluate the antiproliferative activity of extract and fractions against breast cancer and prostate cancer cell lines and isolate and characterize active compounds from bio-active guided fractions. Moreover, the anticancer activity of isolated compounds against breast and prostate cancer cell lines was also evaluated, in addition to in silico and molecular docking interactions of isolated compounds with VEGFR2 and PDGFRα target proteins. METHODS: The compounds were isolated and purified with the help of repeated column chromatography, and spectral techniques, such as 1D, 2D NMR, and GC-MS/MS, were used to identify and elucidate the structure of the compounds. Moreover, prediction of activity spectra for substances, physiochemical properties, bioactivity radar prediction, bioactivity score, natural-product likeness, ADME, and toxicity parameters of isolated compounds (AC-1 to AC-4) was performed through various in-silico databases and servers. To evaluate the docking interaction profile and binding energies of compounds, three docking tools were utilized, such as AutoDock, AutoDock Vina, and iGEMDOCK, against two targets VEGFR2 and PDGFRα. MD simulation was performed through ligand and receptor molecular dynamic server (LARMD). RESULTS: It was found that the A. cadamba bark chloroform fraction demonstrated a significant inhibitory effect against MDA-MB-231, MCF-7, and PC-3 cells in a dose-time-dependent manner. The bioassay-guided isolation afforded four molecules AC-1 to AC-4 from chloroform fraction. Moreover, the GC-MS/MS profiling identified fourteen new molecules which were not reported earlier from A. cadamba. The in-silico study showed that the isolated compounds (AC-1 to AC-4) followed Lipinski's rule and had good oral bioavailability. While compound AC-4 had positive bioactivity scores except for kinase inhibitor activity. The ADMET profiling revealed that AC-4 was non-toxic and easily absorbed in the human intestine, and transportable in the blood-brain barrier compared to AC-1, AC-2, AC-3, and standard drug doxorubicin. Molecular docking and MD simulation assessment also signified AC-4 anticancer activity with dual inhibitory action against the target proteins VEGFR2 and PDGFRα amongst the studied compounds. The in vitro cell viability assay of isolated compounds demonstrated that AC-1 showed IC50 (µg/mL) value of 34.96 ±3.91, 47.76±3.80 69.1±4.96, AC-2; 68.26±4.22, 54.03±5.14, >100, AC-3; 35.34±4.14, 51.5±51.5, 70.8±5.25 and AC-4; 44.2±3.57, 24.2±2.67, 51.2±2.54 for MDA-MB-231, MCF-7, and PC-3 cancer cell lines, respectively and compared with standard drug doxorubicin. Moreover, fluorescence microscopy confirmed the apoptogenic property of compounds. We also found that AC-4 exhibited significant intracellular ROS production in breast cancer cells, thereby inducing apoptosis and eventually cell death. CONCLUSION: In conclusion, A. cadamba afforded four pure molecules AC-1 to AC-4 with the identification of fourteen new compounds. The entire in-silico studies concluded that the AC-4 compound had better oral bioavailability, bioactivity score, and ADMET profile among studied molecules. Molecular docking analysis and MD simulation also supported AC-4 dual inhibitory action against both VEGFR2 and PDGFRα receptors. Moreover, the isolated molecules AC-1, AC-2, AC-3, and AC-4 were found to be active against MDA-MB-231, MCF-7, and PC-3 cancer cells. The molecule AC-4 was found to induce ROS-mediated apoptosis in breast cancer cells. It was found that the anticancer inhibitory potentiality of AC-4 is directed to its molecular stereochemistry which specifically binds to the target proteins of breast cancer cells with no toxicological effect. Therefore, AC-4 is suggested to be an effective aspirant for novel drug design and discovery.

Assessment of genotoxicity and oxidative stress in pregnant women contaminated to organochlorine pesticides and its correlation with pregnancy outcome.

The present study was aimed to assess the correlation between transplacental transfer of xenobiotics and resulting biochemical alterations (including genotoxicity and oxidative stress) in non-occupational pregnant women of North India along with the effect on pregnancy outcomes. Maternal and cord blood samples were collected from 221 healthy mother-infant couples and divided according to their gestational age and birth weight. Genotoxic effects in mother and cord blood were examined using comet assay. The quantitative determination of Organo-chlorine pesticides in blood serum of study population was carried out using gas chromatography-mass spectrometry (GC-MS). Notably higher Organo-chlorine pesticides levels were observed in maternal blood of preterm than term subjects for almost all of the compounds detected, with the maximum concentration found for aldrin (3.26 mg/l) in maternal blood and dieldrin (2.69 mg/l) in cord blood. The results showed a significant increment in olive tail moment, tail full length, catalase, super-oxide dismutase, and malondialdehyde levels whereas lower glutathione reductase and peroxidase were found in preterm babies when compared with term group and it varied in the order: maternal blood > cord blood. A clear trend was observed for preterm babies with their lower birth weight and cesarean mode of delivery. Therefore, reduction in birth weight in newborns may be the consequence of increased oxidative damage and genotoxicity brought about by pesticides and these markers could be employed for early detection of pesticides related ailments and toxicities. To the best of our knowledge, this was a pioneering study and it may help to increase our knowledge with regard to xenobiotic exposure in biological system and the need for stringent guidelines for agricultural use of pesticides.

An observational study to estimate the level of essential trace elements and its implications in type 2 diabetes mellitus patients.

BACKGROUND AND OBJECTIVES: Insulin action of reducing blood glucose has been found to be enhanced by trace elements. MATERIAL AND METHODS: This was a cross sectional study including 150 patients with Type 2 Diabetes Mellitus (T2DM) and 50 controls. Serum concentrations of zinc, copper, chromium, selenium and magnesium was measured by colorimetric kit. Fasting Blood Glucose and Glycated Haemoglobin (HbA1c) were assayed using the standard kit. RESULTS: Out of 150 patients, 85.4% (n = 128) of the cases had uncontrolled blood sugar with HbA1c ≥7 and only 14.6% (n = 22) had good control of blood sugar with HbA1c <7%. Hypertension (42%) and hypothyroidism (14%) were the most commonly associated comorbidities among patients with T2DM. Following percentage of diabetic patients had complications such as peripheral neuropathy (45.3%), diabetic retinopathy (36.7%), coronary artery disease (20.7%), diabetic nephropathy (17.3%), peripheral vascular disease (8.7%), and cerebrovascular accident (6%) respectively. The mean level of zinc, copper, selenium and magnesium was significantly lower in patients with T2DM than the control cases (62.89 vs. 74.95 µg/dL, P < 0.05; 116.30 vs. 150.39 µg/dL, P < 0.001; 8.57 vs. 16.16 µg/dL, P < 0.001; 1.92 vs. 2.31 mg/dL, P < 0.05, respectively). Multivariate analysis showed that there was a significant trend between levels of zinc, copper, selenium, and magnesium and the prevalence of T2DM. CONCLUSIONS: The levels of selenium, zinc, copper, and magnesium were significantly lower in patients with T2DM when compared to healthy counterparts.

Telomerase in saliva: An assistant marker for oral squamous cell carcinoma.

INTRODUCTION: Telomerase is a ribonucleoprotein complex responsible for de novo telomere synthesis and addition of telomeric repeats to existing telomeres. Telomerase activity is generally found to be absent in normal tissues. Telomerase is known to be induced upon malignant transformation of human cells. METHOD: In the present study, we analyzed both telomere length and telomerase activity in saliva samples from oral carcinoma patients. The study was done to investigate the presence of telomerase activity in oral squamous cell carcinoma by TRAP assay. RESULT: Telomerase activity was detectable in 79 of 100 human OSCC and 51 of 100 premalignant cases and 8 of 100 normal patients. CONCLUSION: These results indicate that telomerase is activated frequently during the late stage of oral premalignancy and may play a crucial role in OSCC.

MicroRNA-183-5p: A New Potential Marker for Prostate Cancer.

The microRNA (miR)-183-5p is expressed at high level in the majority of cancer. The purpose of present study was to investigate the role of oncogenic miR-183-5p in prostate cancer (PCa) as biomarker. We carried out our experiment in 50 prostate cancer patients and 40 patients of benign prostatic hyperplasia (BPH) and 40 adjacent controls tissue. The expression of miR-183-5p was evaluated through reverse transcription qualitative polymerase chain reaction. We found that the expression of miR-183-5p in PCa tissue was significantly up regulated as compared to BPH patients and adjacent normal tissues as control. Additionally, miR-183 expression was correlated with higher prostate-specific antigen, higher Gleason Score and metastatic condition. A receiver operating characteristic curve analysis revealed that miR-183-5p distinguished PCa patients from BPH patients and also from control. In conclusion, our data suggest that oncogenic miR-183-5p may be useful as a new tissue specific diagnostic biomarker in prostate cancer.

Promoter variants of TNF-α rs1800629 and IL-10 rs1800871 are independently associated with the susceptibility of coronary artery disease in north Indian.

BACKGROUND: Tumor necrosis factor-alpha (TNF-α) are considered as a pro inflammatory and interleukin-10 (IL-10) anti inflammatory have been shown to predict the risk of incident of coronary artery disease (CAD). The polymorphism at promoter of TNF-α and IL-10 has been shown to increase transcriptional activity of the gene and play a important role in patho physiology of CAD. Aim of present study is to examine the impact of the TNF-α and IL-10 variant allele on various markers of the CAD and to study its relation with circulating TNF-α and IL-10 levels. METHODS: The -308 G/A & -238 G/A of TNF-α and -1082 G/A & -819 C/T of IL-10 gene polymorphism has been studied in 301 diagnosed CAD subjects (Age 51.50 ±â€¯9.28; BMI 25.30 ±â€¯3.58) and 305 healthy controls (Age 51.57 ±â€¯9.50; BMI 24.06 ±â€¯7.26). These polymorphism of TNF-α and IL-10 were detected by real time PCR by using Taqman SNP genotyping assay. Furthermore serum TNF-alpha and IL-10 levels were also measured by ELISA. RESULTS: Allelic and genotypic frequencies did not deviate from Hardy-Weinberg equilibrium in the controls (p > 0.05). On allele contrast, significant association with susceptibility to CAD was detected with polymorphisms in TNF-α -308 G/A, that variant genotype GA + AA (dominant model) (p = 0.030: OR = 1.61: 95% CI = 1.06-2.44) and variant allele (A) (p = 0.006: OR = 1.71: 95% CI = 1.17-2.51) of TNF-α 308 G/A gene was significant highly observed in the cases as compared to control group. Furthermore, variant genotype CT + TT (dominant model) (p = 0.004: OR = 1.62: 95% CI = 1.17-2.24) and variant allele (T) (p < 0.001: OR = 1.49: 95% CI = 1.17-1.89) of IL-10 -819 C/T gene was significant highly observed in the cases as compared to control group. CONCLUSION: Our results suggest that the TNF-α G-308A polymorphism independently associated with DBP, cholesterol, triglyceride, LDL, TNF-α and IL-10 levels which may be leads to the development of coronary artery disease of North Indians.

Lycopene amends LPS induced oxidative stress and hypertriglyceridemia via modulating PCSK-9 expression and Apo-CIII mediated lipoprotein lipase activity.

This study was undertaken to uncover the regulatory role of lycopene in targeting lipopolysaccharide (LPS) induced oxidative stress and inflammatory cascades and subsequent regulation of proprotein convertase subtilisin/kexin type-9 (PCSK-9) expression via sterol regulatory element binding protein-2 (SREBP-2) and hepatocyte nuclear factor-1α (HNF-1α). Further, protein-protein interaction (PPI) studies for Lycopene-Apo-CIII complex against lipoprotein lipase (LPL) were also performed to assess its regulatory role behind the enhanced circulatory TG/TRLs clearance. Lycopene treatment down-regulated hepatocyte PCSK-9 expression via down-regulation of HNF-1α, whereas, LDL-receptor (LDL-R) was up-regulated by subsequent up-regulation of SREBP-2. PPI studies showed that lycopene diminishes the affinity of Apo-CIII to complex with LPL (ΔG: -917.1 Kcal/mol) resulting in increased LPL functionality and TRLs clearance. Moreover, lycopene also ameliorated LPS stimulated oxidative-stress via enhanced total antioxidant and HDL associated PON-1 activity in addition to down-regulate the expression and plasma level of inflammatory mediators. Based on above findings, we concluded that lycopene exhibits dual role in targeting LPS induced oxidative stress and hypertriglyceridemia via down-regulation of PCSK-9, making greater no. of surface LDL-R available for LPS processing and clearance, as well as increased LPL activity through inhibition of Apo-CIII.

Evaluation of miR-711 as Novel Biomarker in Prostate Cancer Progression

Objective: MicroRNAs (miRs) are class of small non-coding regulatory RNA aberrantly expressed in various types of malignancies including prostate cancer and serves as potential targets to develop new diagnostic and therapeutic strategies. In this quiet we investigated miRNAs expression profile in benign prostatic hyperplasia (BPH) and prostate cancer (PCa) tissue samples and correlated their expression with clinicopathological parameters. Methodology: The miRNAs expression profile as well as their validation has been done by using Microarray and RT-PCR, respectively. Additionally, we also tried to speculate microRNA-mRNA regulatory module through computational target predictions by using Targetscan, Miranda and MirWalk and obtained results were analysed through DAVID software. Result: We observed that miR-711 is significantly deregulated in BPH and PCa, compared to controls. The lower expression of miR-711 was found to be significantly associated with high Gleason score and metastatic disease. Furthermore, the computational target prediction analysis explored miR-711 association to various cancer cells signalling cascade key molecules associated with cancer cell survival.Conclusion: From our observations we suggest that miR-711 may play a critical role in PCa progression, regulation of various cancer cell survival signalling cascades and that it may be a valuable biomarker for prediction of metastatic disease and poor prognosis in PCa.

Identification of the cysteine-rich 61 (CYR61) gene variations in osteosarcoma patients.

BACKGROUND/AIM: Osteosarcoma requires an angiogenesis process. CYR61 is one of the extracellular signaling molecules that promote angiogenesis, tumor growth, and the malignancy of osteosarcoma. In the present study, we investigate the CYR61 gene variations in osteosarcoma and their correlations with clinicopathological findings. MATERIALS AND METHODS: We performed variation analysis of the CYR61 gene in 58 patients with osteosarcoma. With an aim to ascertain the variety of variations in exons 2, 3, 4, and 5 of the CYR61 gene in osteosarcoma, we did a PCR-SSCP followed by DNA sequencing. RESULTS: In osteosarcoma the CYR61 gene variations found were 18.96% (11/58) in exon 2, 3.44% (2/58) in exon 3, 8.62% (5/58) in exon 4, and 15.51% (9/58) in exon 5. In our variation analysis, we detected one missense variation in exon 2 (Arg47Trp), one silent variation in exon 3 (Lys152Lys), one missense variation in exon 4 (Phe213Leu), and two missense variations in exon 5 (Gly315Arg and Asp339Asn). The overall CYR61 variation frequency in exons 2, 3, 4, and 5 was determined to be 46.55% (27/58). CONCLUSION: Our study specifies the role of CYR61 gene variation in osteosarcoma. The result signifies that CYR61 might be used as a prognostic/diagnostic marker in osteosarcoma patients.

Genetic polymorphism of interleukin-10 (-A592C) among oral cancer with squamous cell carcinoma.

OBJECTIVE: Interleukin-10 (IL-10) is a pleiotropic cytokine with either immunosuppressive or immunostimulative activities. It has been reported that in cancer, the promoter region polymorphism of IL-10 (-A592C) alters both the expression and serum levels of this cytokine. In the present study, we have addressed the question as to whether the single nucleotide polymorphisms (SNPs) at positions -592 A/C in the IL-10 gene promoter, could predispose an individual to oral squamous cell carcinoma (OSCC). DESIGN: We analyzed the genotype of the IL-10 (-A592C) gene, in 250 histopathologically confirmed OSCC patients and similar number of healthy volunteers taken as controls, in an Indian population by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Allele and genotype frequencies were analyzed by the Student's t-test and the chi-squared test, and strength of associations by the odds ratio (OR) with 95% confidence intervals. RESULTS: The genotype and allele distribution of IL-10 (-A592C) gene polymorphism was significantly different between OSCC cases and controls (genotype AA vs AC: OR 2.87; 95 % CI 1.50-5.48; p=0.0016 and AA vs CC: OR 4.08; 95 % CI 1.98-8.41; p=0.0002). The -592 C alleles were found to be significantly different among OSCC cases and controls (OR: 1.44, 95% CI: 1.12-1.85, p<0.0051). CONCLUSIONS: The IL-10 gene promoter region (-592) A/C polymorphism is significantly associated with reduced risk of OSCC. The OSCC group had a significantly greater frequency of genotype AA as compared to control group.

Association of -330 interleukin-2 gene polymorphism with oral cancer.

BACKGROUND & OBJECTIVES: Cytokines play an important role in the development of cancer. Several single-nucleotide polymorphisms (SNPs) of cytokine genes have been reported to be associated with the development and severity of inflammatory diseases and cancer predisposition. This study was undertaken to evaluate a possible association of interleukin 2 (IL-2) (- 330A>C) gene polymorphisms with the susceptibility to oral cancer. METHODS: The SNP in IL-2 (-330A>C) gene was genotyped in 300 oral cancer patients and in similar number of healthy volunteers by polymerase chain reaction (PCR)-restriction fragment length polymorphism and the association of the gene with the disease was evaluated. RESULTS: IL-2 (-330A>C) gene polymorphism was significantly associated with oral cancer whereas it was neither associated with clinicopathological status nor with cancer pain. The AC heterozygous genotype was significantly associated with oral cancer patients as compared to controls [odds ratio (OR): 3.0; confidence interval (CI): 2.14-4.20; P<0.001]. The C allele frequency was also significantly associated with oral cancer (OR: 1.80; CI: 1.39-2.33; P<0.001). IL-2 (-330A>C) gene polymorphism was also associated with oral cancer in tobacco smokers and chewers. INTERPRETATION & CONCLUSIONS: Our results showed that oral cancer patients had significantly higher frequency of AA genotype but significantly lower frequency of AC genotype and C allele compared to controls. The IL-2 AC genotype and C allele of IL-2 (-330A>C) gene polymorphisms could be potential protective factors and might reduce the risk of oral cancer in Indian population.

ROS mediated pro-apoptotic effects of Tinospora cordifolia on breast cancer cells.

The inevitable development of chemoresistance and unmanageable side effects are the major therapeutic challenges in management of breast cancer imposing an urgent need for identification of novel therapeutic agents. In the present investigation, we report anti-proliferative activity of chloroform fraction of Tinospora cordifolia (TcCF), an Ayurvedic medicinal plant, on breast cancer cells. We found that TcCF inhibited growth of breast cancer cells, MDA-MB-231 and MCF-7. More interestingly, we observed TcCF treatment increased intra-cellular ROS levels, altered expression of pro and anti-apoptotic genes, decreased colony formation ability and induced apoptosis in breast cancer cells. We also found that inhibition of ROS abrogated TcCF induced apoptosis in breast cancer cells, emphasizing the role ROS in TcCF induced breast cancer cell death. Furthermore, we identified the presence of pharmacologically active compounds like rutin and quercetin which account for the anti-cancer property of TcCF against breast cancer cells. These data show TcCF is a promising anti-cancer agent against breast cancer cells.

Retrospective case-control study of correlation between MTHFR gene and OSCC risk in North India.

BACKGROUND: Oral squamous cell carcinoma (OSCC) occurrence appears to be the number one among all cancers in India. Folate is a methyl donor during DNA methylation, as it provides substrate for methylenetetrahydrofolate reductase (MTHFR) to convert 5,10-MTHF to 5-MTHF and subsequently metabolizes it to methionine. The purpose of this study was to identify MTHFR C677T gene polymorphism in patients with OSCC. MATERIALS AND METHODS: A total of 350 OSCC cases and 350 healthy controls participated in this study. MTHFR C677T single-nucleotide polymorphism was evaluated by PCR-RFLP. RESULTS: In the present study, MTHFR gene 677CC, CT, and TT genotype frequencies of the total OSCC cases were 74.8; 19.4 and 5.71; and 88.5, 9.42, and 2.0 % in controls. The average frequency of the MTHFR 677T allele was 15.4 % in OSCC cases compared to 6.71 % in the controls. The CT genotype occurrence prevailed more in patients than controls in contrast to TT genotype, although both the genotypes were statistically significant for OSCC. Moreover, we found that T allele was significant in cases of smoking and tobacco chewing. CONCLUSIONS: In this study, we found that the homozygous mutant T allele appeared to have significantly higher risk of OSCC especially in late stages and therefore supporting in OSCC susceptibility and its progression.