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Reliable Reference Gene for Normalization of RT- qPCR Data in Human Cancer Cell Lines
Subjected to Gene Knockdown
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Quantitative real-time Polymerase Chain Reaction (RT-qPCR) has become a valuable molecular technique in biomedical research. The selection of suitable endogenous reference genes is necessary for normalization of target gene expression in RT-qPCR experiments. The aim of this study was to determine the suitability of each 18S rRNA and ACTB as internal control genes for normalization of RT-qPCR data in some human cell lines transfected with small interfering RNA (siRNA). Four cancer cell lines including MCF-7, T47D, MDA-MB-231 and Hela cells along with HEK293 representing an embryonic cell line were depleted of E2F6 using siRNA specific for E2F6 compared to negative control cells, which were transfected with siRNA not specific for any gene. Using RT-qPCR, Ct (threshold cycle) values of 18S and ACTB were determined in transfected cells and compared with control cells. In the selection of the above cell lines, 18S was identified as the most stably expressed reference gene than ACTB in gene knockdown experiments.

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Publication Date
Mon May 23 2022
Journal Name
International Journal Of Molecular Sciences
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

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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
Mon May 09 2022
Journal Name
مجلة كلية التربية الاساسية الجامعة المستنصرية
Detection of sul1 resistance gene in Acinetobacter baumannii from different Clinical cases
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Publication Date
Thu Feb 27 2020
Journal Name
Iraqi Journal Of Science
Gene Expression of pelA and pslA in Pseudomonas Aeruginosa under Gentamicin Stress
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     Pseudomonas aeruginosa produces an extracellular biofilm matrix that consists of nucleic acids, exopolysaccharides, lipid vesicles, and proteins. Alginate, Psl and Pel are three exopolysaccharides that constitute the main components in biofilm matrix, with many biological functions attributed to them, especially concerning the protection of the bacterial cell from antimicrobial agents and immune responses. A total of 25 gentamicin-resistant P. aeruginosa selected isolates were enrolled in this study. Biofilm development was observed in 96% of the isolates. In addition, the present results clarified the presence of pelA and pslA in all the studied isolates. The expression of these genes was very low. Even though all biof

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Publication Date
Tue Jan 30 2024
Journal Name
Iraqi Journal Of Science
Differential Diagnosis of Entamoeba spp. Using the 18SrRNA Gene in Gastroenteritis Patients
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In order to accurately diagnose Entamoeba spp., this study's major goal was to develop a proof-of-concept method for simultaneously detecting pathogenic and non-pathogenic amoebae using DNA. During amoebiasis, two diagnostic techniques (microscopic inspection and PCR techniques with particular primers) were evaluated. About 100 feces samples from Fallujah individuals who had clinical symptoms were taken. The outcome reveals that only 20 samples have Entamoeba spp. infections. According to this study, the two species had distinct infection percentages. Entamoeba histolytica was the most prevalent infection, at 85%, followed by Entamoeba dispar, which was 15% of all the Entamoeba-positive sampl

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Publication Date
Tue Jan 04 2022
Journal Name
Iraqi Journal Of Science
New Mutations in GyrA Gene of Escherichia Coli Isolated form Iraqi Patients
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The present study included a collecting of 165 specimens form different sources,
93 isolates were identified as Escherichia coli depending on morphological and
biochemical tests in addition to automated systems such as VITEK 2 and api 20E.
All isolates under study developed high resistance toward cefotaxime, ceftazidime,
ceftriaxone, and ciprofloxacin estimated by minimum inhibitory concentration. Stool
and wound specimens characterized by harbouring the highest resistant isolates in a
percentage reached 100% against antibiotics under study. Insignificant differences
were found between isolates collected from males and females. Upon using disk
displacement method to detect extended spectrum beta lactamases (ESBL),

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Publication Date
Tue Feb 28 2023
Journal Name
Iraqi Journal Of Science
Assessment of miR-146a Gene Polymorphisms in Patients with Systemic Lupus Erythematosus
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      Systemic lupus erythematosus (SLE) is a multifactorial chronic autoimmune disease, with a wide spectrum of effect. The main feature of the disease is the production of a wide variety of autoantibodies as a result of immune tolerance loss. The work aims to evaluate the miRNA-146a gene polymorphism potential association with disease activity and chronicity changes in SLE patients. The study included 100 SLE patients and 50 matched controls. The systemic lupus erythematosus disease activity index (SLEDAI) was assessed. The single nucleotide polymorphism (SNP) of miR-146a gene (rs2910164) polymorphism was assayed by polymerase chain reaction (PCR) and sequencing technique in patients and control. 100 SLE pati

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Publication Date
Tue Feb 28 2023
Journal Name
Iraqi Journal Of Science
Gentamicin Upregulates the Gene Expression of hla and nuc in Staphylococcus aureus
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The current study aimed to detect the effect of gentamicin stress on the expression of hla (encodes hemolysin) and nuc (encodes nuclease) genes of Staphylococcus aureus. Fifty-eight isolates identified as S. aureus were isolated locally from different clinical specimens. Disk diffusion method was used to detect the resistance to S. aureus. The minimum inhibitory concentration (MIC) of gentamicin was estimated by broth microdilution method. hla and nuc genes were determined by polymerase chain reaction technique. The biofilm was evaluated using the microtiter plate method in the presence and absence of gentamicin at sub-MIC. The results showed that 18 (31%) and 40 (69%) S. aureus isolates were sensitive and resistant to gentamicin, respectiv

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Publication Date
Mon Jan 01 2018
Journal Name
Current Research In Microbiology And Biotechnology
Serodiagnosis of Human Cytomegalovirus in Iraqi Breast cancer and fibroadenoma patients
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Human cytomegalovirus (HCMV) has a worldwide distribution and extremely common infections. The presence of HCMV genome and antigens has been detected in many kinds of human cancers especially breast cancer. In Iraq, the incidence of breast cancer generally exceeds any other type of malignancies among Iraqi population. The study was performed in the period between October 2016 and June 2017 in Central public health laboratory/Baghdad. It involve samples from 90 women including 60 breast cancer patients, 20 benign tumor patients, and 10 normal breast tissues. A blood sample was obtained from each woman included in this study. Anti-HCMV IgG antibody was presented in 9/10 (90%) of normal women, benign breast tumor patients 19/20 (95%) and malig

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Publication Date
Tue Mar 15 2022
Journal Name
Gene Reports
Genotyping of Human Cytomegalovirus Glycoprotein N in Iraqi Breast cancer Patients
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Human Cytomegalovirus (HCMV) is an enveloped ubiquitous ds-DNA virus that has been implicated in several types of malignancies. The current work was conducted in the period extending from (November 2018 to the end of October 2019) and aimed to assess the frequency of glycoprotein N (gN) genotypes of HCMV. A total number of 91serum and plasma specimens were collected to fulfill this purpose from females (71 breast cancer patients, and a control group of 20 females) attending Al-Amal hospital for cancer management and Baghdad teaching hospital. The molecular part of this data was achieved through both PCR and Multiplex PCR for detection of HCMV gN (UL73) entire gene as well as for genotyping. gN was detected in 36/71 (50.7%) of breast cancer

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