MMP-2 gene expression at mRNA level in HBV and HCV infected patients.

Matrix metalloproteinases (MMPs) family play a determinative role in the development of liver fibrosis, metastasis, unregulated angiogenesis, and tumor growth. In this study the possible association between the MMP-2 gene expression level and risk of chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections were evaluated in liver transplanted patients. Formalin fixed paraffin embedded (FFPE) liver tissue samples were collected from 225 transplant patients between years 2012 and 2016. The presence of HBV and HCV infections were analyzed in patients studied using molecular and immunologic diagnostic protocols according to the instructions of the manufacturers. Patients were divided to HBV, HCV, and HBV with HCV co-infected groups. A healthy control group was also included. For the quantitative analysis of MMP-2 mRNA gene expression an in-house-SYBR Green Real-Time PCR method was performed. The level of MMP-2 mRNA expression showed a significant increase in all studied viral hepatitis infected patient groups in comparing with healthy controls. The MMP-2 gene expression level increased in HBV infected patients when compared with HCV and HBV with HCV co-infected patients, but not significantly. Results showed a significant increase in MMP-2 expression level in all viral hepatitis single and coinfected liver transplanted patients when compared with the controls and also in HBV infected patients when compered with other viral infected ones, need to confirm in further completed studies.

MMP-2 gene expression at mRNA level in HBV and HCV infected patients

@#Matrix metalloproteinases (MMPs) family play a determinative role in the development of liver fibrosis, metastasis, unregulated angiogenesis, and tumor growth. In this study the possible association between the MMP-2 gene expression level and risk of chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections were evaluated in liver transplanted patients. Formalin fixed paraffin embedded (FFPE) liver tissue samples were collected from 225 transplant patients between years 2012 and 2016. The presence of HBV and HCV infections were analyzed in patients studied using molecular and immunologic diagnostic protocols according to the instructions of the manufacturers. Patients were divided to HBV, HCV, and HBV with HCV co-infected groups. A healthy control group was also included. For the quantitative analysis of MMP-2 mRNA gene expression an in-house-SYBR Green Real-Time PCR method was performed. The level of MMP-2 mRNA expression showed a significant increase in all studied viral hepatitis infected patient groups in comparing with healthy controls. The MMP-2 gene expression level increased in HBV infected patients when compared with HCV and HBV with HCV co-infected patients, but not significantly. Results showed a significant increase in MMP-2 expression level in all viral hepatitis single and coinfected liver transplanted patients when compared with the controls and also in HBV infected patients when compered with other viral infected ones, need to confirm in further completed studies.

Foxo3a transcription factor is a negative regulator of Skp2 and Skp2 SCF complex.

Skp2 (S-phase kinase-associated protein-2) SCF complex displays E3 ligase activity and oncogenic activity by regulating protein ubiquitination and degradation, in turn regulating cell cycle entry, senescence and tumorigenesis. The maintenance of the integrity of Skp2 SCF complex is critical for its E3 ligase activity. The Skp2 F-box protein is a rate-limiting step and key factor in this complex, which binds to its protein substrates and triggers ubiquitination and degradation of its substrates. Skp2 is found to be overexpressed in numerous human cancers, which has an important role in tumorigenesis. The molecular mechanism by which the function of Skp2 and Skp2 SCF complex is regulated remains largely unknown. Here we show that Foxo3a transcription factor is a novel and negative regulator of Skp2 SCF complex. Foxo3a is found to be a transcriptional repressor of Skp2 gene expression by directly binding to the Skp2 promoter, thereby inhibiting Skp2 protein expression. Surprisingly, we found for the first time that Foxo3a also displays a transcription-independent activity by directly interacting with Skp2 and disrupting Skp2 SCF complex formation, in turn inhibiting Skp2 SCF E3 ligase activity and promoting p27 stability. Finally, we show that the oncogenic activity of Skp2 is repressed by Foxo3a overexpression. Our results not only reveal novel insights into how Skp2 SCF complex is regulated, but also establish a new role for Foxo3a in tumor suppression through a transcription-dependent and independent manner.

Longer years of practice and higher education levels promote infection control in Iranian dental practitioners.

BACKGROUNDS: Infection control is one of the primary responsibilities of dental health care personnel. The purpose of this study was to evaluate whether the infection control practices of Iranian dentists and dental nurses working in governmental dental health care centers were influenced by their educational level and years of practice. METHODS: This cross-sectional analytical study was completed in 2009, and it included 63 Iranian dental practitioners. Infection control knowledge was evaluated with a self-administered questionnaire, and infection control practices were evaluated with a checklist of questions by observation with one researcher. RESULTS: The dental practitioners in Mashad had a low level of infection control knowledge. Dental personnel with a higher educational level had significantly greater knowledge than those with less education. Additionally, dental personnel who had more years of practice had a greater knowledge of infection control. CONCLUSION: Since dental practitioners working in Mashad governmental dental health care centers with fewer years of practice and less educational level had a low level of infection control knowledge, we recommend a continuing educational program for this group and dental nurses.

Localization of sense and antisense transcripts of Prdx2 gene in mouse tissues.

Recently, it has been reported that antisense RNAs are transcribed from a large number of genes in various species including human and mouse. The Prdx2 gene, which is indicated to be involved in signal transduction related to platelet-derived growth factor as well as to protection from oxidizing agents, has been shown to produce sense and antisense transcripts. To obtain clues for possible roles of Prdx2 antisense transcripts, we have performed Northern blot analysis and in situ hybridization on tissues of 8-week-old C57BL/6J mice. The Northern blot analysis revealed that major parts of sense and antisense transcripts were poly(A-)-RNA. The analysis of the fractionated RNA of fibroblasts indicated that the poly(A-)-RNA would be localized in the cytoplasm of cells. The in situ hybridization demonstrated that the sense and antisense transcripts were localized in almost the same limited areas of brain, testis, and spleen. It also revealed that the sense and antisense transcripts coexisted in Purkinje cells. In thymus and stomach, the antisense transcripts were detected, but sense transcripts were not. When tissues of BALB/c mice were examined by in situ hybridization, the observations were essentially the same as those of C57BL/6J except that it appeared that the amounts of sense and antisense transcripts in testis of BALB/c were greater than those in C57BL/6J, and that the amounts of antisense transcripts in stomach of BALB/c were much smaller than those in C57BL/6J.

Genomic organization, expression and evolution of porcine CRSP1, 2, and 3.

Recently we identified and characterized porcine calcitonin receptor-stimulating peptide (CRSP) 1, CRSP2 and CRSP3 as members of the calcitonin/calcitonin gene-related peptide (CT/CGRP) family. In the present study, the genomic sequences and organization of CRSP1, 2, and 3 were determined, and the expression of the genes in the porcine brain was investigated using in situ hybridization. Analysis of 5'-upstream regions of the three CRSPs demonstrated that CRSP1 and CRSP2 have almost identical sequences (>98% similarity) and high sequence similarities including functional transcription binding sites with the corresponding region of human CALCA (CT/alpha CGRP), whereas CRSP3 retains less similarity with the above genes. RH mapping of CRSPs demonstrated that they resided in a region of swine chromosome 2 (SSC2). The arrangement of the genes in the region was found to be conserved in corresponding human and mouse regions. In situ hybridization demonstrated sense transcripts of the three genes in cerebrum, hippocampus, hypothalamus, pons/midbrain, and thalamus of 3-month-old pigs, and CRSP2 sense transcripts additionally in tractus opticus. The sense transcripts of alpha CGRP and CALCB (beta CGRP) were detected in cerebrum, hippocampus, and pons/midbrain of newborn mice, and to a lesser extent in pons/midbrain of 8-week-old mice. These results taken together with the chromosomal conservation and phylogenetic clustering of CT/CGRP family indicate that CRSP1, 2, and 3 may be functionally different from alpha CGRP and beta CGRP, though they are indicated to have a common progenitor gene.

Assignment of 115 genes from HSA9 and HSA14 to SSC1q by RH mapping to generate a dense human-pig comparative map.

A large number of significant QTL for economically important traits including average daily gain have been located on SSC1q, which, as shown by chromosome painting, corresponds to four human chromosomes (HSA9, 14, 15 and 18). To provide a comprehensive comparative map for efficient selection of candidate genes, 81 and 34 genes localized on HSA9 and HSA14 respectively were mapped to SSC1q using a porcine 7000-rad radiation hybrid panel (IMpRH). This study, together with the cytogenetic map (http://www2.toulouse.inra.fr/lgc/pig/cyto/genmar/htm/1GM.HTM), demonstrates that SSC1q2.1-q2.13 corresponds to the region ranging from 44.6 to 63.2 Mb on HSA14q21.1-q23.1, the region from 86.5 to 86.8 Mb on HSA15q24-q25, the region from 0.9 to 27.2 Mb on HSA9p24.3-p21, the region from 35.1 to 38.0 Mb on HSA9p13, the region from 70.3 to 79.3 Mb on HSA9q13-q21 and the region from 96.4 to 140.0 Mb on HSA9q22.3-q34. The conserved synteny between HSA9 and SSC1q is interrupted by at least six sites, and the synteny between HSA14 and SSC1q is interrupted by at least one site.

Analysing two dinucleotide repeats of FVIII gene in Iranian population.

Using dinucleotide repeats for carrier detection and prenatal diagnosis of haemophilia A patients, led us to find different alleles and their frequencies in Iranian population. Polymerase chain reaction (PCR) amplification of two short tandem repeat (STR) loci of factor VIII (FVIII) gene was performed, and the PCR products were resolved on 10% native polyacrylamide gel, and samples were analysed with sequenced DNA markers made of PCR cloning of the dinucleotide FVIII gene fragments. Seven different alleles were observed for intron 13 STR, having 18-24 (CA) repeating units and five alleles for intron 22 STR having 24-28 repeating units of (CACT). Bands produced during dinucleotide study were defined in detail so this could improve the genotyping of heterozygotes and homozygotes. Conformational band produced were characterized to specify the dinucleotide pattern. Our results confirm the Hardy-Weinberg proportions of the heterozygosity rate of the 85 analysed individuals. The observed heterozygosity rate for intron 13 and 22 was 52% and 59% respectively. Our data also indicate that our population is closer to caucasians than to any other populations. Finding different dinucleotide repeat alleles and their frequencies has made it possible to identify carriers and provide prenatal diagnosis with more confidence. This allows antenatal diagnosis to be performed in the vast majority of carriers.