Investigating Post-translational Changes of Histones, DNA Methylation Level, and ERβ Protein Level in the Cumulus Cell Genome of Infertile Women with Endometriosis

Background & aim: Among the important factors affecting female infertility, endometriosis (with a prevalence rate of up to 50% in infertile women) has a special place. Endometriosis, which is known as the presence of endometrial glands and stroma outside the uterine tissue, causes a wide range of functional disorders in the process of follicular development and changes in the follicular microenvironment, which ultimately leads to the creation of an egg that is of suitable quality for the formation of The fetus does not have Therefore, the aim of the present study was to determine and investigate post-translational changes of histones, DNA methylation level and ERβ protein level in the genome of cumulus cells of infertile women with endometriosis. Methods: The present case-control study was conducted at the infertility treatment clinic of Royan Research Institute in 2014. Twenty-four patients were divided into two equal groups. Cumulus cells were obtained from 12 infertile patients with endometriosis and 12 women with male factor infertility (as a control group) under ovulation stimulation protocols for intracytoplasmic sperm injection. Extraction of chromatin from cumulus cells was done after stabilization of DNA binding proteins and then cell lysis and preparation of soluble chromatin. The binding and incorporation levels of MeCP2 protein (as DNA methylation marker), two epigenetic markers related to histone modifications (H3K9me2 and H3K9ac), and ERβ protein to chromatin of cumulus cells were evaluated using the corresponding primary antibodies, secondary antibodies conjugated and Nucleosome-ELISA technique. Data were analyzed using independent t-test t and Levene's test. Results: MeCP2 protein incorporation into DNA was significantly higher in the endometriosis group than in the control group. The level of two epigenetic marks H3K9ac and H3K9me2 in the chromatin of cumulus cells of the patient group showed a significant increase compared to the control group (P<0.05). In addition, an increased level of binding of ERβ protein to the genome was observed compared to the control group. Conclusion: Epigenetic changes, including histone hyperacetylation and hypermethylation, and DNA hypermethylation of the whole genome of cumulus cells of patients with endometriosis had occurred, accompanied by an increase in the level of ERβ protein binding.

MIC susceptility testing of nosocomial infections at PICU in Children's Medical Center, Iran

Background: The aim of this study was to determine the minimal inhibitory concentration (MIC) of microorganisms causing nosocomial infections in the pediatric intensive care unit (PICU) of the Children's Medical Center in Tehran. Methods: All patients with nosocomial infections in the PICU were enrolled in the study. Causative microoraganisms were coagulase positive, and coagulase negative Staphylococci and Pseudomonas aeruginosa . MIC of many antibiotics was determined by microbroth dilution according to NCCLS. Findings: Within a period of 18 months, thirty patients developed nosocomial infection including 17 cases with P.aeruginosa and 9 individuals with Staph aureus infection. The remaining 4 patients were involved with coagulase negative Staphylococci. The most common sources were respiratory tract, blood stream, wound and soft tissue. Multi-antimicrobial resistance (resistance to Amikacin, Ceftazidim, Imipenem and Ciprofloxacin) was common among P. aeruginosa species All strains of Staph aureus were resistant to Methicillin (MRSA). These microorganisms were also resistant to clindamycin and ciprofloxacin in 88% of cases.Conclusion: Resistance to antimicrobial agents was high in our study, therefore routine MIC examination is necessary in PICU

Epigenetic alterations of CYP19A1 gene in Cumulus cells and its relevance to infertility in endometriosis

Purpose: The purpose of the present study was to investigate the epigenetic mechanisms responsible for the aberrant aromatase expression (CYP19A1) in Cumulus Cells (CCs) of infertile endometriosis patients. Method: Cumulus cells were obtained from 24 infertile patients with and without endometriosis who underwent ovarian stimulation for intracytoplasmic sperm injection. Expression of CYP19A1 gene was quantified using Reverse Transcription Q-PCR. DNA methylation, histone modifications, and binding of Estrogen Receptor, ERβ to regulatory DNA sequences of CYP19A1 gene were evaluated by Chromatin ImmunoPrecipitation (ChIP) assay. Results: CYP19A1 gene expression in CCs of endometriosis patients was significantly lower than the control group (P = 0.04). Higher incorporation of MeCP2 (as a marker of DNA methylation) on PII and PI.4 promoters, and hypoacetylation at H3K9 in PII and hypermethylation at H3K9 in PI.4 were observed in CYP19A1 gene in endometriosis patients (P < 0.05). Moreover, a decreased level of ERβ binding to PII and an increased level of its binding to PI.3 and PI.4 promoters of CYP19A1 were observed in endometriosis patients when compared to control. Conclusion: Significant reduction of CYP19A1 gene expression in CCs of endometriosis patients may be the result of epigenetic alterations in its regulatory regions, either by DNA methylation or histone modifications. These epigenetic changes along with differential binding of ERβ (as a transcription factor) in CYP19A1 promoters may impair follicular steroidogenesis, leading to poor Oocyte and embryo condition in endometriosis patients. © 2016, Springer Science+Business Media New York

Evaluation of Toll-like receptor 3 (TLR3) signaling pathway genes and its genetic polymorphisms in ectopic and eutopic endometrium of women with endometriosis

Objective: Toll-like receptors (TLRs, as members of the innate immune system) are expressed in the human endometrium and their aberrant regulation and expression are involved in the pathogenesis of endometrial diseases. This study is aimed at evaluation of TLR3 signaling pathway genes and its genetic changes in endometriosis patients. Materials and methods: Blood samples were collected from 83 endometriosis patients and 93 healthy fertile women and PCR was performed in blood-derived DNA for detection of SNP of TLR3. Also, ectopic (EC) and eutopic (EU) endometrial biopsies were obtained from endometriosis patients (n = 20), as well as endometrium from healthy women (n = 16, CE). Q-PCR was performed for determination of mRNA expression level of TLR3 signaling pathway genes (TLR3, TICAM, NF-kB1A, CXCL10, IRF3, IFN-B1, IL-6 and IL-8). Also, serum protein levels of TLR3, IFN-β, IL-6 and IL-8 were determined using ELISA. Results: The mRNA expression levels of TLR3, NF-kB1A, IFN-B1, IRF3, TICAM1, IL-6 and IL-8 were significantly higher in EU compared to ectopic ones and also compared to CE. SNPs frequency (rs3775291 and rs3775290) was not significantly different between patients and controls. Serum protein levels of TLR3, IFN-β, IL-6 and IL-8 were significantly increased in endometriosis patients. Conclusion: Significant changes were observed in the expression of IL-6 and IL-8 cytokines and other genes in TLR3 cascade in diseased EU, demonstrating that EU similarly to EC is in an intensive inflammatory state. These fundamental alterations in the concept of immune response in EU may lead to its activation, escapes from apoptosis, and displaced implantation of the endometrium. © 2021 Elsevier Masson SA

Regional Differences in the Efficiency of German Savings Banks

German savings banks play a key role in financing German SMEs. Accordingly, the analysis of their efficiency is of economic importance. We apply data envelopment analysis to explore the influence of the financial crisis (2007 to 2008) as well as the Eurozone crisis (since the end of 2009) on the efficiency of German savings banks. Although the Malmquist index, as a measure of efficiency change and technological change, is on average below one in our sample from 2003 to 2014, we conclude from our efficiency results that German savings banks recovered quickly after the financial crisis as well as the Eurozone crisis. We test for and explain regional efficiency differences between Eastern and Western German savings banks. Since Eastern and Western Germany represent two socio-economically different environments, we control for environmental variables. Our results show that Eastern German savings banks are less inefficient than Western German savings banks since other earning assets, i. e., securities and advances to banks, and refinancing costs are advantageous for their efficiency