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Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells
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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.

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Publication Date
Fri Dec 01 2023
Journal Name
Baghdad Science Journal
Gene Expression and Methylation Levels of PCSK9 Gene in Iraqi Patients with Coronary Artery Disease
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The expression of the Proprotein Convertase Subtilisin/Kexin Type 9 gene (PCSK9) is inextricably related to lipid levels and a risk of atherosclerotic coronary artery disease (ASCAD). The present study aims to measure the quantity of PCSK9 gene expression and the effect of methylation on its expression level taking part in the pathogenesis of acute coronary artery disorder.

A current study included 150 subjects from the Iraqi population, 100 ASCAD patients and 50 healthy controls. The concentration of PCSK9 in each serum sample was determined by the ELISA technique, the expression levels of the PCSK9 gene in whole blood were estimated by RT-qPCR – Quantitative Reverse Transcription PCR method, and DNA

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Publication Date
Thu Jun 01 2023
Journal Name
Baghdad Science Journal
In vitro isolation and expansion of neural stem cells NSCs
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   Neural stem cells (NSCs) are progenitor cells which have the ability to self‑renewal and potential for differentiating into neurons, oligodendrocytes, and astrocytes. The in vitro isolation, culturing, identification, cryopreservation were investigated to produce neural stem cells in culture as successful sources for further studies before using it for clinical trials. In this study, mouse bone marrow was the source of neural stem cells. The results of morphological study and immunocytochemistry of isolated cells showed that NSCs can be produced successfully and maintaining their self‑renewal and successfully forming neurosphere for multiple passages. The spheres preserved their morphology in culture and cryopreserved t

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Publication Date
Tue Feb 28 2023
Journal Name
Iraqi Journal Of Science
Global DNA Methylation Levels in Epstein-Barr-Virus-Positive Iraqi Patients with Acute Lymphoblastic Leukaemia
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       Acute lymphoblastic leukemia (ALL) is one of the commonest hematological malignancies affecting children and adults. Recent evidence suggests an involvement of Epstein-Barr virus (EBV) in ALL pathogenicity. Epigenetic aberration, especially altered DNA methylation marks, is a key event of cancer development. The present study aims to investigate how the ALL epimethylome reacts to viral infection through the assessment of the total 5-methylcytosine (5mC) levels in ALL patients, according to EBV infection. The 5mC global DNA methylation levels in 50 diagnosed ALL patients (age mean 26.23 yrs; age range 10-60 yrs) and 25 age-matched healthy controls were assessed using MethylFlash™ Methylated DNA Quantification Kit. Acute pri

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Publication Date
Sat Dec 21 2024
Journal Name
Gastroenterology
Evaluation of global DNA methylation, homocysteine and vitamin B12 levels among patients with celiac disease
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Celiac disease (CD) is an immune-mediated disorder caused by gluten in genetically susceptible individuals characterized by chronic inflammation that essentially affects the small intestine. Objective: this study was designed to measure the potential role of some serological biomarkers including vitamin B12 and homocysteine (HCY) in the progression of CD as well as their relations to global DNA methylation (5mC). Materials and methods. Forty CD patients were enrolled in the study with an average age of (36.60 ± 2.03) years (range between 15 and 60). The diagnosis of the disease was confirmed by serological examinations and intestinal endoscopy in Gastroenterology and Liver Teaching Hospital in the Medical City Hospital in Baghdad

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Publication Date
Wed Mar 15 2023
Journal Name
Journal Of Survey In Fisheries Sciences
The Correlation of DAZ1 Gene Methylation with Azoospermia in Iraqi Infertile Men
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After about twelve months or maybe more, some people can’t achieve pregnancy. This might be a sign of infertility as a reproductive system disease. The following study was carried out to investigate the DAZ 1 gene methylation level and its association with azoospermia in Iraqi patients. One hundred and fifty human blood samples were collected from from different regions in Baghdad governorate, including (private medicals Labs and the high institute for infertility diagnosis assisted reproductive techniques and Kamal Al- Samara'ay IVF Hospital) from both fertile and infertile men. The control group consists of 50 samples ranging from 22 to 51 years old, while the patient (infertile group) consists of 100 samples ranging between 25 and 51 y

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Publication Date
Thu Mar 30 2023
Journal Name
Iraqi Journal Of Science
Human Bone Marrow Mesenchymal Stem Cells Isolation, Expansion and Identification
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The present study aimed to isolate and diagnose mesenchymal stem cells derived from human bone that is the source generating cells that are the best types of treatment for tissue diseases.
Cells were isolated from the back bone of the human pelvis, separated using density gradual sedimentation method and then the cells were grown on the culture media RPMI-1640 \ 20% FBS.
To detect the purity of cells that have been isolated and have been transplanted immune use the method using CD44 (mesenchymal stem cells marker) CD43, a specific marker for hematopoietic cells Nestin, (the neurons private marker).
The present study has shown that mesenchymal cells that have been isolated and expanded in this experiment has reached up 99.7% for

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Publication Date
Fri Jan 03 2025
Journal Name
World Academy Of Sciences Journal
Effects of global epigenetic methylation changes and interleukins‑15 and ‑29 on the progression of rheumatoid arthritis
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Publication Date
Wed Jun 26 2024
Journal Name
Opera Medica Et Physiologica
The Impact of Global DNA Methylation and Hypoxia-Inducible Factor 1 Alpha Levels in the Progression of Breast Cancer
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Publication Date
Thu Jan 16 2025
Journal Name
Journal Of Baghdad College Of Dentistry
Immunohistochemical study of CD34 in tooth eruption by using amniotic stem cells
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Background: Tooth eruption is a more general process, however, which includes certain posteruptive tooth movements. There are two fundamental requirements for both tooth eruption to occur: (1) Require soft tissue, intervening between tooth structure and alveolar bone, which plays an important role in regulating the remodeling of adjacent tissues. (2) Require bone turnover that is temporally and spatially regulated to facilitate specific translocations of teeth through alveolar bone These amniotic stem cells are multipotent and able to differentiate into various tissues, which may be useful for human application and recently it used in many medical branches. CD34 is an endothelial marker that is extensively used in immunohistochemistry a

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Publication Date
Sun Mar 17 2019
Journal Name
Baghdad Science Journal
DNA Methylation Patterns of Interferon Gamma Gene Promoter and Serum Level in Pulmonary Tuberculosis: Their Role in Prognosis
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Tuberculosis (TB) still remains an important medical problem due to high levels of morbidity and mortality worldwide. A series of innate immune mechanisms that create a cytokine network control the pathogenesis of tuberculosis and this response has the capacity to modify the host genomic DNA structure through epigenetic mechanisms such as DNA methylation which could constantly alter the local gene expression pattern that can modulate the metabolism of the tissues and the immune-response. Interferon-gamma (IFN-γ) is an important pro-inflammatory cytokine regulator of the innate immune response to TB. This study aims to determine DNA methylation patterns of INF-γ gene promoter and measure serum IFN- γ level in newly diagnosed TB patient

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