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.
the study considers the optical classification of cervical nodal lymph cells and is based on research into the development of a Computer Aid Diagnosis (CAD) to detect the malignancy cases of diseases. We consider 2 sets of features one of them is the statistical features; included Mode, Median, Mean, Standard Deviation and Maximum Probability Density and the second set are the features that consist of Euclidian geometrical features like the Object Perimeter, Area and Infill Coefficient. The segmentation method is based on following up the cell and its background regions as ranges in the minimum-maximum of pixel values. The decision making approach is based on applying of Minimum Dista
This study focused on the expression and regulation of BRCA1 in breast cancer cell lines compared to normal breast. BRCA1 transcript levels were assessed by real time quantitative polymerase chain reaction (RT-qPCR) in the cancer cell lines. Our data show overexpression of BRCA1 mRNA level in all the studied breast cancer cell lines: MCF-7, T47D, MDA-MB-231 and MDA-MB-468 along with Jurkat, leukemia T-lymphocyte, the positive control, relative to normal breast tissue. To investigate whether a positive or negative correlation exists between BRCA1 and the transcription factor E2F6, three different si-RNA specific for E2F6 were used to transfect the normal and cancerous breast cell lines. Interestingly, strong negative relationship was found b
... 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 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 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
... 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 MoreThe inhibitory action of four lactobacilli isolates Lactobacillus bulgaricus, L. acidophilus, L. plantarum and L. fermentum, isolated from four different samples; yoghurt, vinegar, saliva and vagina respectively, on Escherichia coli and Staphylococcus aureus adhesion to uroepithelial cells were investigated. Results showed that all Lactobacillus isolates or their supernatant were able to reduce the number of the uropathogens attached to uroepithelial cells. However, inhibition level of lactobacilli cells was higher than their supernatant. Nevertheless, the human indigenous lactobacilli (L. fermentum and L. plantarum) were more competitive than food lactobacilli (L. acidophilus and L. bulgaricus).
Erythrocytes aggregation is an important physiological phenomenon in the circulation of blood, and is a basic characteristic of normal blood that plays a major role in cardiovascular system especially in the microcirculation. Blood samples have been taken from (30) volunteers (15 male, and 15 female), their ages (20-30) years. The Erythrocytes Sedimentation Rate (ESR) for those subjects was measured at different Packed Cells Volume (PCV) (10%-25%), and also it was measured at different temperature (10oC-25oC). The results show that there was a highly significant decrease (P<0.01) in ESR when the PCV increase and a highly significant increase (P<0.01) in ESR when the temperatures increase. The conclusion from these results is that the ESR va
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