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γH2AX assay has been used for DNA damage assessment at higher doses of radiation exposure. Its expression has not been studied in cases with diagnostic low dose radiation exposure. Concerns have been raised about the after-effects of... more
γH2AX assay has been used for DNA damage assessment at higher doses of radiation exposure. Its expression has not been studied in cases with diagnostic low dose radiation exposure. Concerns have been raised about the after-effects of radiation in diagnostic procedures like Computed Tomography (CT) scan, Angiography etc especially when such scans are repeated within short span of time. The purpose of the present study was to assess immediate DNA damage after exposure to low level of ionizing radiation by the flow cytometric method of gamma-H2AX. Study sample includes total 60, cases and controls with two groups Group I-Normal controls (n = 15); Group II-Low dose, further divided in three groups: Group IIA-single CT scan (n = 15); Group IIB-Multiple CT scans (n = 15); and Group IIC-angiography single exposure (n = 15). For Low dose group blood was collected within 1 h after exposure in EDTA vaccutainers and immediately kept on ice. Lymphocytes were isolated and were fixed in 80% chill...
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Background: Radiation causes oxidative lesions and strand breaks in DNA of exposed cells. Extended length PCR is a reliable method for assessing DNA damage. Longer DNA strands with DNA damage are difficult to amplify compared to smaller... more
Background: Radiation causes oxidative lesions and strand breaks in DNA of exposed cells. Extended length PCR is a reliable method for assessing DNA damage. Longer DNA strands with DNA damage are difficult to amplify compared to smaller DNA strands by PCR. The present study was aimed to evaluate DNA damage caused by ionising radiation exposure in therapeutic and diagnostic medicine. Materials and Methods: The study group comprised 50 cases with low dose single exposure (LDS), low dose multiple exposure (LDM) and low dose angiography (LDA) which were compared with 25 high dose controls (HDC) and 25 controls with no exposure (NEC). Blood samples were collected within 1 hour of radiation exposure. DNA was isolated using a kit based protocol, 50 ng aliquots of DNA were used to amplify a long 13kbp DNA fragment of the β-actin gene by conventional PCR and band intensity was then quantified. Relative amplification was calculated and damage was expressed in terms of lesions per kilobase (kb...
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Research Interests: India, HUMAN PAPILLOMAVIRUS, Humans, Tobacco Use, Female, and 13 moreMale, Coinfection, Risk factors, Middle Aged, Oral Squamous Cell Carcinoma (OSCC), Genotype, Mouth Neoplasms, Adult, Public health systems and services research, Sex Factors, Survival Rate, Risk Factors, and Papillomavirus Infections
Gall bladder Carcinoma (GBC) is the fifth most common cancer of the digestive tract and frequently diagnosed in late stage of disease. Estimation of circulating free DNA (cfDNA) in serum has been applied as a... more
Gall bladder Carcinoma (GBC) is the fifth most common cancer of the digestive tract and frequently diagnosed in late stage of disease. Estimation of circulating free DNA (cfDNA) in serum has been applied as a "liquid biopsy" in several deep seated malignancies. Its value in diagnosis of gall bladder carcinoma has not been studied. The present study was designed to assess the role of cfDNA in the diagnosis of GBC and correlate levels with the TNM stage. Serum was collected from 34 patients with GBC and 39 age and sex matched controls including 22 cholecystitis and 17 healthy individuals. Serum cfDNA levels were measured through quantitative polymerase chain reaction (qPCR) by amplification of β-globin gene. Performance of the assay was calculated through the receiver operating characteristic (ROC) curve. The cfDNA level was significantly lower in healthy controls and cholecystitis (89.32 ± 59.76 ng/ml, 174.21 ± 99.93 ng/ml) compared to GBC (1245.91 ± 892.46 ng/ml, p = <0.001). The cfDNA level was significantly associated with TNM stage, lymph node involvement and jaundice (0.002, 0.027, and 0.041, respectively). Area under curve of ROC analysis for cancer group versus healthy and cholecystitis group was 1.00 and 0.983 with sensitivity of 100 %, 88.24 % and specificity of 100 % respectively. Quantitative analysis of cfDNA may distinguish cholecystitis and gall bladder carcinoma and may serve as new diagnostic, noninvasive marker adjunct to imaging for the diagnosis of GBC.
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Genomic DNA methylation is one of the most important epigenetic modifications in eukaryotes play vital role in development of severe disease like cancer. Many techniques used for assessment of DNA methylation, bisulfite treatment followed... more
Genomic DNA methylation is one of the most important epigenetic modifications in eukaryotes play vital role in development of severe disease like cancer. Many techniques used for assessment of DNA methylation, bisulfite treatment followed by methylation specific polymerase reaction (MSP) are one of them, which introduce conversion of unmethylated cytosine into uracil. The significant level of bisulfite treated DNA degradation results in the failure of methylation detection. Therefore, this step is to be properly controlled to avoid the degradation of DNA. In the present study, an attempt has been made to access the incubation time of DNA with bisulfate treatment at three time points i.e. 2.5, 4 and 16 hrs to get complete conversion of cytosine to uracil. Currently, the experiments were undertaken using oral cancer tissue, with varying incubation time of bisulfite treatment and 2 representative genes viz MGMT and p16 were selected for the quantitative assessment of methylation by rea...
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Research Interests: Oxidative Stress, Mitochondria, DNA damage, Transcription Factors, Gene expression, and 15 moreMitochondrial DNA, Glucose, Endothelial Cells, Reactive Oxygen Species, Diabetic Retinopathy, Animals, Male, Retina, Electron Transport, Gene Dosage, Superoxide Dismutase, Cattle, Rats, Experimental Diabetes Mellitus Type 1 and 2, and Biochemistry and cell biology
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Research Interests: Biology, DNA replication, DNA damage, Medicine, Gene expression, and 15 moreTransgenic Mice, Mitochondrial DNA, Glucose, Mice, Diabetic Retinopathy, Animals, Retina, Superoxide Dismutase, Experimental Diabetes Mellitus Type 1 and 2, Protein Binding, Superoxides, Protein Transport, Gene Expression Regulation, Mitochondrial Proteins, and Medical biochemistry and metabolomics
Research Interests: Algorithms, Biomedical Engineering, Adolescent, Comparative Study, Anterior Cruciate Ligament, and 15 moreBone graft, Cohort Study, Arthroplasty, Clinical Sciences, Aged, Adult, Achilles tendon, Clinical Assessment, Bone and Bones, Clinical Diagnosis, Clinical evaluation, Case Control Studies, Cohort Studies, Biomechanical Phenomena, and Anterior cruciate ligament reconstruction
Promoter methylation and relative gene expression of O6-methyguanine-DNA-methyltransferase (MGMT) andp16genes were examined in tissue and blood samples of patients with premalignant oral lesions (PMOLs) and oral squamous cell carcinoma... more
Promoter methylation and relative gene expression of O6-methyguanine-DNA-methyltransferase (MGMT) andp16genes were examined in tissue and blood samples of patients with premalignant oral lesions (PMOLs) and oral squamous cell carcinoma (OSCC). Methylation-specific PCR and reverse transcriptase PCR were performed in 146 tissue and blood samples from controls and patients with PMOLs and OSCC. In PMOL group, significant promoter methylation ofMGMTandp16genes was observed in 59% (P=0.0010) and 57% (P=0.0016) of tissue samples, respectively, and 39% (P=0.0135) and 33% (P=0.0074) of blood samples, respectively. Promoter methylation of both genes was more frequent in patients with OSCC, that is, 76% (P=0.0001) and 82% (P=0.0001) in tissue and 57% (P=0.0002) and 70% (P=0.0001) in blood, respectively. Significant downregulation ofMGMTandp16mRNA expression was observed in both tissue and blood samples from patients with PMOLs and OSCC. Hypermethylation-induced transcriptional silencing ofMGMT...
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Promoter methylation and relative gene expression of O 6-methyguanine-DNA-methyltransferase (MGMT) and p16 genes were examined in tissue and blood samples of patients with premalignant oral lesions (PMOLs) and oral squamous cell carcinoma... more
Promoter methylation and relative gene expression of O 6-methyguanine-DNA-methyltransferase (MGMT) and p16 genes were examined in tissue and blood samples of patients with premalignant oral lesions (PMOLs) and oral squamous cell carcinoma (OSCC). Methylation-specific PCR and reverse transcriptase PCR were performed in 146 tissue and blood samples from controls and patients with PMOLs and OSCC. In PMOL group, significant promoter methylation of MGMT and p16 genes was observed in 59% (í µí± = 0.0010) and 57% (í µí± = 0.0016) of tissue samples, respectively, and 39% (í µí± = 0.0135) and 33% (í µí± = 0.0074) of blood samples, respectively. Promoter methylation of both genes was more frequent in patients with OSCC, that is, 76% (í µí± = 0.0001) and 82% (í µí± = 0.0001) in tissue and 57% (í µí± = 0.0002) and 70% (í µí± = 0.0001) in blood, respectively. Significant downregulation of MGMT and p16 mRNA expression was observed in both tissue and blood samples from patients with PMOLs and OSCC. Hypermethylation-induced transcriptional silencing of MGMT and p16 genes in both precancer and cancer suggests important role of these changes in progression of premalignant state to malignancy. Results support use of blood as potential surrogate to tissue samples for screening or diagnosing PMOLs and early OSCC.
Changes in eating habits and sedentary lifestyle are main contributors to type 2 diabetes (T2D) development, and studies suggest that epigenetic modifications are involved with the growing incidence of this disease. Regular exercise... more
Changes in eating habits and sedentary lifestyle are main contributors to type 2 diabetes (T2D) development, and studies suggest that epigenetic modifications are involved with the growing incidence of this disease. Regular exercise modulates many intracellular pathways improving insulin resistance and glucose uptake in skeletal muscle, both early abnormalities of T2D. Mitochondria dysfunction and decreased expression of glucose transporter (GLUT4) were identified as main factors of insulin resistance. Moreover, it has been suggested that skeletal muscle of T2D subjects have a different pattern of epigenetic marks on the promoter of GLUT4 and PGC1, main regulator of mitochondrial function, compared with nondiabetic individuals. Recent studies have proposed that regular exercise could improve glucose uptake by the attenuation of such epigenetic modification induced at GLUT4, PGC1 and its downstream regulators; however, the exact mechanism is still to be understood. Herein we review the known epigenetic modifications on GLUT4 and mitochondrial proteins that lead to impairment of skeletal muscle glucose uptake and T2D development, and the effect of physical exercise at these modifications.
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Trichloroethylene (TCE) is major industrial pollutant that contaminate environment. Its exposure may lead to hepato-renal toxicity along with the cancer progression. Although extensive research is done on its toxicity still not much is... more
Trichloroethylene (TCE) is major industrial pollutant that contaminate environment. Its exposure may lead to hepato-renal toxicity along with the cancer progression. Although extensive research is done on its toxicity still not much is known about its genotoxic potential on humans in relation to genetic polymorphism. Cytochrome P450 (CYP P-450) and glutathione-S-transferases (GSTs) are important in cellular detoxification of TCE. Variations in gene sequences result in population specific regional genetic variations (polymorphism). Genotyping of CYP1A1, GSTM1, GSTT1 and GSTP1 polymorphism was performed in 220 normal and 97 solvent-exposed individuals from northern part of India using real time PCR, PCR and restriction digestion techniques. The parameters examined to study genotoxicity were chromosomal aberration (CA) and cytokinesis block micronucleus assay (CBMN) in lymphocyte culture in vitro. The observed average frequencies for GSTM1 (null) and GSTT1 (null) were 41, 22 and 12.7%,...
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ABSTRACT Real Time PCR is a sensitive technique with multidisciplinary applications such as detectionand quantification of genetically modified organisms(GMO’s), pathogens (bacteria, viruses), highthrough put screening of Single... more
ABSTRACT Real Time PCR is a sensitive technique with multidisciplinary applications such as detectionand quantification of genetically modified organisms(GMO’s), pathogens (bacteria, viruses), highthrough put screening of Single Nucleotide Polymorphism (SNP) within a population and expressionstudies. SYBR Green I an intercalating dye, used to discriminate genotypes/strains by meltingcurve analysis. Present paper discusses the discrepancy in melting temperature (Tm) for thesimilar PCR product using SYBR Green dye master mixes procured from different suppliers.Genes used in this study are ?-globin, Lectin, SPD a surfactant protein, Salmonella hinH2, Nosterminator and Virus (35S promoter). The article highlights the variation in melting temperaturethat may not be overlooked while performing SYBR green analysis. (On the basis of our results,one can conclude that proper recording of different kits used in specific experiments, isimportant for replicating the experiment in different laboratory setting. Tm parameter seemsto be a good tool to discriminate between PCR products of various GMO’s/strains provided onemake sure to use same kit.