Pluripotent stem cells (PSC) possess unlimited proliferation, self-renewal, and a differentiation capacity spanning all germ layers. Appropriate culture conditions are important for the maintenance of self-renewal, pluripotency, proliferation, differentiation, and epigenetic states. Oxygen concentrations vary across different human tissues depending on precise cell location and proximity to vascularisation. The bulk of PSC culture-based research is performed in a physiologically hyperoxic, air oxygen (21% O2) environment, with numerous reports now detailing the impact of a physiologic normoxia (physoxia), low oxygen culture in the maintenance of stemness, survival, morphology, proliferation, differentiation potential, and epigenetic profiles. Epigenetic mechanisms affect multiple cellular characteristics including gene expression during development and cell-fate determination in differentiated cells. We hypothesized that epigenetic marks are responsive to a reduced oxygen microenvironment in PSCs and their differentiation progeny. Here, we evaluated the role of physoxia in PSC culture, the regulation of DNA methylation (5mC (5-methylcytosine) and 5hmC (5-hydroxymethylcytosine)), and the expression of regulatory enzyme DNMTs and TETs. Physoxia enhanced the functional profile of PSC including proliferation, metabolic activity, and stemness attributes. PSCs cultured in physoxia revealed the significant downregulation of DNMT3B, DNMT3L, TET1, and TET3 vs. air oxygen, accompanied by significantly reduced 5mC and 5hmC levels. The downregulation of DNMT3B was associated with an increase in its promoter methylation. Coupled with the above, we also noted decreased HIF1A but increased HIF2A expression in physoxia-cultured PSCs versus air oxygen. In conclusion, PSCs display oxygen-sensitive methylation patterns that correlate with the transcriptional and translational regulation of the de novo methylase DNMT3B.
Abstract: E2F6 is a member of the E2F family of transcription factors involved in regulation of a wide variety of genes through both activation and repression. E2F6 has been reported as overexpressed in breast cancers but whether or not this is important for tumor development is unclear. We first checked E2F6 expression in tumor cDNAs and the protein level in a range of breast cancer cell lines. RNA interference-mediated depletion was then used to assess the importance of E2F6 expression in cell lines with regard to cell cycle profile using fluorescence-activated cell sorting and a cell survival assay using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The overexpression of E2F6 was confirmed in breast tumor cDNA samp
... Show MoreBackground BK polyomavirus is one of the common post-transplant viral infections, affecting ∼15% of renal transplantation recipients (RTR), leading to graft loss in more than half of cases. Objectives Study the rate of detection of BK virus (BKV) in RTRs in Pap-stained urine cytology specimens. Methods A single center study, urine samples were collected from 99 RTR patients, with 15 Living Donors (LD) and 15 patients with chronic kidney disease (CKD) were taken as controls. And urine cytology smears were Pap stained for detection of decoy cells (DCs). Results Out of the 99 RTRs, 27 (27.3%) patients were decoy positive, 8 out of these 27 patients had uncommon DCs, and 5 out of these 27 cytology positive patients (18.5%) had biopsy proven B
... Show MoreE-learning applications according to the levels of enlightenment (STEM Literacy) for physics teachers in the secondary stage. The sample consists of (400) teachers, at a rate of (200) males (50%), and (200)females (50%), distributed over (6) directorates of education in Baghdad governorate on both sides of Rusafa and Karkh. To verify the research goals, the researcher built a scale of e-learning applications according to the levels of STEM Literacy, which consists of (50) items distributed over (5) levels. The face validity of the scale and its stability were verified by extracting the stability coefficient through the internal consistency method “Alf-Cronbach”. The following statistical means were used: Pearson correlation coefficient,
... Show MoreCarnosol, a phenolic diterpene, is one of the effective anticancer agents naturally occurring in rosemary, sage, parsley, and oregano. The chemoresistance problem increased with the routinely used chemotherapy. Therefore, the efforts to find a substitute with safe and low cost have become crucial worldwide. The current study attempts to inspect the anticancer molecular mechanisms of Carnosol on modulating up- and down- regulation of multiple genetic carcinogenesis pathways. The cytotoxicity of Carnosol on Hela cells was evaluated by MTS assay. Flow cytometry was used to assess apoptosis and cell cycle arrest. The apoptotic morphological changes were obvious by dual apoptosis assay. The differential gene expression after treatment wi
... Show MoreAccording to the prevalence of multidrug resistance bacteria, especially Pseudomonas aeruginosa, in which the essential mechanism of drug resistance is the ability to possess an efflux pump by which extrusion of antimicrobial agents usually occurs, this study aims to detect the presence of mexB multidrug efflux gene in some local isolates of this bacteria that show resistance towards three antibiotics, out of five. Sensitivity test to antibiotics was performed on all isolates by using meropenem (10μg/disc), imipenem (10μg/disc), amikacin (30 μg/disc), ciprofloxacin (5μg/disc) and ceftazidime (30 μg/disc). Conventional PCR results showed the presence of mexB gene (244bp) in four isolates out of ten (40%). In addition,25, 50μg/ml of cur
... Show MoreHepatitis B infection is a prominent infectious disease caused by hepatitis B virus (HBV), which infect liver and is considered as the main cause of liver cirrhosis, fibrosis and liver cancer worldwide. A pro-inflammatory cytokine Interleukin32 is believed to have a role in chronic HBV infections. Since its role in CHB infections is remain unclear, this study was done to detect IL-32 gene expression in CHB patients in order to identify its exact role. A total number of 110 blood samples were collected from Gastroenterology and Hepatology Teaching Hospital in Baghdad Medical City from CHB patients for both males and females with different age groups according to the research ethics form then sent to Central Public Health Laboratory (CPHL),
... Show MoreObjective: The present work was undertaken to investigate the impact of sub inhibitory concentration of gentamicin on hla gene expression in methicillin resistant Staphylococcus aureus isolates. Methods: The bacterial isolates used in this study represent 33 MRSA strains, previously isolated form patients visiting several hospitals in Baghdad. Gentamicin, vancomycin, and oxacillin MIC were determined using broth dilution method. Microtiter plate method was adopted to investigate the biofilm forming capacity. Alpha hemolysin was detected by culturing MRSA isolates on rabbit blood agar. Furthermore, hla gene was detected in MRSA isolates using conventional PCR technique; while, qRT-PCR method was performed to assay the hla expression in plank
... Show MoreIntroduction and Aim: Beta-thalassemia is a serious inherited genetic disorder and an increasing health burden globally. Beta -thalassemia is caused by genetic globin abnormalities within the hemoglobin beta (HBB) gene. This study aimed to characterize the HBB gene mutations in beta -thalassemia among southern Iraqi patients. Materials and Methods: The study included 30 beta -thalassemia patients referred to the Thi-Qar Center for Genetic Diseases, Iraq and 15 control samples from a random group of apparently healthy individuals. Genomic DNA was isolated from blood sample collected from each individual. The DNA was amplified for specific regions of the HBB gene and the amplified products sequenced. The sequences generated were analysed for
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