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.
Various assays are used to determine the toxic effects of drugs at cellular levels in vitro. One of these methods is the dye exclusion assay, which measures membrane integrity in the presence of Trypan blue. Trypan blue the dye which was used in this study to investigate cytotoxic effect of a new Cis –dichloroplatinum (II) complex [(Qu)2PtCl2] on the viability of polymorphonuclear cells (PMNs). Three concentrations of platinum complex were prepared (70, 35and 17.5 µg/ ml) and the results revealed that the percentage of cell viability decreased as the platinum complex concentration increased in comparison with control.
... Show MoreThe aim of the study was extraction of arial part of Euphorbia cyathophora constituents with methanol and evaluate its effect on mitotic index and total chromosomal aberration bone marrow cell and spleen cell in mice 200 gm of E. cyathophora fine powder was defatted then extracted by cold maceration 80% ethanol for seven days. The extract was filtered and dried in a rotary evaporator then the dried extract was suspended with water and consecutively extracted using chloroform, ethyl acetate for each. The aqueous layer was then mixed with 100ml methanol. These fractions are dried under reduced pressure to obtain the dry extract. Twenty-four Albino mice were used for the experiment. The animals were divided into four groups: Gr
... Show MoreBackground: Dental implants provide a unique treatment modality for the replacement of a lost dentition .This is accomplished by the insertion of relatively an inert material (a biomaterial) into the soft and hard tissue of the jaws, there by providing support and retention for dental prostheses. Low level laser therapy (LLLT) is an effective tool used to prompt bone repair and remodeling, this has referred to the biostimulation effect of LLLT. The Aim of this study was to evaluate the effects of inflammatory cells on osseointegration of CpTi implant irradiated by low level laser. Materials and Methods: thirty two adult New Zealand white rabbits, received titanium implants were inserted in the tibia. The right side is considered as experime
... Show MoreObjective: The present study investigates whether the exposure to low-power diode laser induces denaturation in red blood cell (RBC) membrane protein composition, and determines the irradiation time for when denaturation of membrane protein process begins. Background: A low-energy laser has been used extensively in medical applications. Several studies indicated significant positive effects of laser therapy on biological systems. In contrast, other studies reported that laser induced unwanted changes in cell structure and biological systems. The present work studied the effect of irradiation time of low-power diode laser on the structure of membrane proteins of human RBCs. Materials and methods: The RBC suspension was divided into five equa
... Show MoreBiotreatment using immobilized cells (IC) technology has proved to be the most promising and most economical approach for the removal of many toxic organic pollutants found in petroleum-refinery wastewater (PRW) such as phenol. This study was undertaken to evaluate the degradation of phenol by Pseudomonas cells individually immobilized in two different bio-carrier matrices including polyvinyl alcohol-guar gum (PVA-GG) and polyvinyl alcohol-agar agar (PVA-AA). Results of batch experiments revealed that complete removal of phenol was attained in the first cycle after 150 min using immobilized cells (IC) in both PVA-GG and PVA-AA. Additional cycles were confirmed to evaluate the validity of recycling beads of immob
... Show MoreA study has been performed to compare the beddings in which ductile iron pipes are buried. In water transmission systems, bends are usually used in the pipes. According to the prescribed layout, at these bends, unbalanced thrust forces are generated that must be confronted to prevent the separation of the bend from the pipe. The bed condition is a critical and important factor in providing the opposite force to the thrust forces in the restraint joint system. Due to the interaction between the native soil and the bedding layers in which the pipe is buried and the different characteristics between them. Also, the interaction with the pipe material makes it difficult to calculate the real forces opposite to the thrust forces and the way they
... Show MorePeriodontitis is one of the most prevalent bacterial diseases affecting man with up to 90% of the global population affected. Its severe form can lead to the tooth loss in 10-15% of the population worldwide. The disease is caused by a dysbiosis of the local microbiota and one organism that contributes to this alteration in the bacterial population is Prophyromonas gingivalis. This organism possesses a range of virulence factors that appear to contribute to its growth and survival at a periodontal site amongst which is its ability to invade oral epithelial cells. Such an invasion strategy provides a means of evasion of host defence mechanisms, persistence at a site and the opportunity for dissemination to other sites in the mouth. However, p
... Show MoreThis study was aimed to determine the mutations and single nucleotide polymorphisms (SNPs) in exon 3 and 7 of estrogen receptor beta (ESR2) gene in women with breast cancer from Iraq. Different samples (blood, fresh tissue with blood from same patient, and formalin fixed paraffin embedded, FFPE) were collected from women with breast cancer. Molecular analysis exon 3 and 7 in ESR2 has been studied by using PCR. It was found exon 3 and 7 in ESR2 were revealed as a single band with size 151 and 157 bp, respectively. There was no SNP in exon 3 has been identified. While three novel polymorphisms (ACT, AGG and GCA) were detected in exon 7, the type of those polymorphisms deletion for ACT and AGG while substitution polymorphism for GCA. From this
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