Study of gyrA S83R Mutation Rate in Vancomycin Resistant Entrococcus Faecium

Enterococci are among Gram-positive cocci and are common inhabitants of the human gastrointestinal tract and enough potent to cause serious infections such as bacteremia, urinary tract infections (UTIs) and endocarditis. Enterococci are not generally considered as highly infectious bacterium. However, the main reason for treatment failure in enterococcal infections is acquired resistance to glycopeptide antibiotics, specifically vancomycin. Most enterococcal infections in human such as gastroenteritis, intestinal infections, and endocarditis have been caused by E. faecalis and E. facieum. They are holding the second place of most frequent cause of hospital infections since 1990. In present study gyrA S83R polymorphism rate in vancomycin resistant Enterococcus faecium isolated from environment, food industries, and hospitals of Tehran were examined by ARMS-PCR technique. DNA was extracted from the bacterial colonies using standard column method, after separating the samples into two groups of antibiotic resistant and antibiotic susceptible by antibiogram test. A fragment of gyrA gene was amplified using PCR method to investigate point mutation of S83R position. ARMS-PCR technique was applied to detect the presence or absence of mutation using a set of specified primers which can be annealed when the mutation is present. The results were statistically analyzed by chi-square test (p < 0.05) using SPSS 19th version. The results showed that there was a significant correlation between the presences of S83R polymorphism with vancomycin resistance trait in Entrococcus faecium. Therefore, this technique could be used as a diagnostic tool to detect vancomycin resistance cases of E. faecium in patients and environment.HIGHLIGHTS•There is a strong association between the presence of point mutation at the position of Ser83 and vancomycin resistant phenotype.•Resistance in Enterococcus spp. is highly mediated by mutations in gyrA gene which plays a major role in interaction with vancomycin.•Vancomycin is the only drug that can be consistently relied on for the treatment of infections caused by multidrug resistant enterococci.•VRE has presented a serious challenge for the Iranian medical community

Measurement of APRIL serum levels as a tumor marker for diagnosis of pancreatic cancer

Background: Members of the tumor necrosis factor (TNF) superfamily of ligands and their receptors (TNFR) are critical regulators of the adaptive immune system. A proliferation inducing ligand (APRIL) is a member of tumor necrosis factor superfamily. APRIL was identified via database mining in 1998 by Hahne, et al. APRIL allows tumor cells to proliferate at a reasonable rate even in low serum. APRIL is abundantly expressed in many tumor cells and tumor tissues. Increasing level of APRIL expression related to replacement of -Arg-Lys-Arg-Arg- motif by -Ala-Lys-Arg-Ala- between amino acids 101-104 and thus abrogated APRIL processing. Previous studies have shown a correlation between APRIL expression with some autoimmune disease, breast cancer, stomach cancer, esophagus cancer and colorectal cancer. Herein, we explore correlation between serum APRIL with pancreatic cancer. Methods: Our study is performed in digestive disease research institute (DDRI) of the Shariati Hospital in Tehran City and affiliated Hospital of Tehran University of Medical Sciences. In this study, concentrations of serum APRIL in sera (30 pancreatic cancer patients and 30 healthy controls) from November 2011 to November 2013 collected and level of a proliferation inducing ligand measured by ELISA technique. In this study used from SPSS software, version 22 (IBM SPSS, Armonk, NY, USA) to perform statistical data analysis. Results: The case group measurement results compared with control groups results according to some characteristics such as age, smoking and, diabetes. ELISA analysis of APRIL measurements show that mean serum APRIL level of pancreatic cancer patients (7 ng/ml) was significantly higher than control group (5 ng/ml). The p-value of this study was 0.003. Conclusion: Our results indicate that serum APRIL, as a potential biomarker, has a positive diagnosis and prognosis value for pancreatic cancer

Molecular investigation of c-MYC oncogene amplification in diabetic patients of Karaj County, Iran: Investigating c-MYC Amplification in Diabetics

Introduction: There is growing evidence revealing that genetic factors could be involved in the etiology of insulin resistance and diabetes. Recent studies now suggest that c-MYC oncogene amplification in beta-cells can cause downregulation of insulin gene expression and leading to diabetes. The present study was carried out to examine gene amplification level of c-MYC gene in healthy individuals compared to those with diabetes. Materials and Methods : This case control study consisted of 70 subjects (34 diabetic patients and 36 healthy controls). The genomic DNA was extracted from blood samples using standard phenol-chloroform method, and Differential Quantitative PCR (DQ PCR) was accomplished. Subsequently, the mean Band Intensity Ratio (BIR) of c-MYC to γ-IFN was applied to determine the amplified products' molecular band intensity. The Real-time PCR was used to reconfirm the obtained results. Results: The mean BIR of c-MYC to γ-IFN was 1.29 in diabetics, whereas in healthy individuals, the mean BIR was equal to 1.16.The findings indicate that the mean BIR in diabetics was almost 1.1 times higher than healthy controls; however, it was not statically different (P-value= .168). Conclusions: Since no significant difference was found in terms of BIR of c-MYC to γ-IFN  in case and control groups, it can be concluded that c-MYC may not have a role to trigger the disease in this limited Iranian population. Further studies and larger statistical populations are needed to confirm our findings. This study showed that the DQ PCR technique could be reliable to screen gene amplification in large populations

Association of rs12487066, rs12044852, rs10735781, rs3135388, rs6897932, rs1321172, rs10492972, and rs9657904 Polymorphisms with Multiple Sclerosis in Iranian Population

Abstract Objectives: Multiple sclerosis (MS) is a chronic disease of the central nervous system. The pathogenesis of MS is best described by a multifactorial model incorporating interactions between genetic and environmental factors with the role of genetic factors increasingly taken into account. The main goal of this study was to investigate the associations of rs12487066, rs12044852, rs10735781, rs3135388, rs6897932, rs1321172, rs10492972, and rs9657904 polymorphisms with MS in the Iranian population. Methods: A total of 83 patients with MS (82.0% female and 18.0% male; mean age = 35.2±8.6 years) and 100 physically and mentally healthy subjects (81.0% female and 19.0% male; mean age = 40.4±6.4 years) were selected using convenient sampling. A 5 mL blood sample was taken from each case and control patient. We used the tetra-primer ARMS-PCR method to genotype the desired polymorphisms. The associations between polymorphisms and the disease were studied based on codominant, dominant, recessive, and overdominant models. Results: The rs10735781 polymorphism was codominantly (p = 0.029), overdominantly (p = 0.008), and dominantly (p = 0.009) associated with the disease. The rs6897932 was also found to be codominantly (p = 0.012), dominantly (p = 0.019), and recessively (p = 0.011) associated with the disease. Conclusions: We found an association between the rs10735781 and rs6897932 polymorphisms on the EVI5 and IL7RA genes, respectively, with increased MS in the Iranian population. Therefore, single nucleotide polymorphisms in the EVI5 and IL7RA genes can be considered a prognostic marker of MS

Interleukin-6 and tumor necrosis factor-alpha gene polymorphisms in chronic idiopathic urticaria

Background - This study was performed to evaluate association of gene polymorphisms among proinflammatory cytokines and susceptibility to chronic idiopathic urticaria (CIU). Methods - Ninety patients with prolonged urticaria more than 6 weeks were included as case group. Single nucleotide polymorphisms (SNPs) of IL-6 (G/C −174, G/A nt565) and TNF-α (G/A −308, G/A −238) were evaluated, using polymerase chain reaction (PCR); and the results were compared to the control group. Results - G allele was significantly higher in the patients at locus of −238 of promoter of TNF-α gene (pConclusions - Pro-inflammatory cytokine gene polymorphisms can affect susceptibility to CIU. TNF-α promoter polymorphisms as well as IL-6 gene polymorphisms are associated with CIU