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.
miRNAs regulate protein abundance and control diverse aspects of cellular processes and biological functions in metabolic diseases, such as obesity and diabetes. Lethal-7(Let-7) miRNAs specifically target genes associated with diabetes and have a role in the regulation of peripheral glucose metabolism. The present study aimed to describe the gene expressions of the let-7a gene with the development of diabetes in Iraq and the difference in the expression of this gene in patients with diabetes and healthy individuals. The association between age and gender with the development of diabetes was studied in this study and the results were compared with those of healthy individuals in the group of control. Based on the obtained results, there was
... Show MoreBackground: Chronic myelogenous leukemia is a malignant hematological disease of hematopoietic stem cells. It is difficult to adapt treatment to each patient's risk level because there are currently few clinical tests and no molecular diagnostics that may predict a patient's clock for the advancement of CML at the time of chronic phase diagnosis. Biomarkers that can differentiate people based on the outcome at diagnosis are needed for blast crisis prevention and response improvement. Objective: This study is an effort to exploit the SLC25A3 gene as a potential biomarker for CML. Methods: RT-qPCR was applied to assess the expression levels of the SLC25A3 gene. Results: In comparison to the mean ΔCt of the control group, which was found to b
... Show MoreTo study the genetic effect of gestational diabetes mellitus by study IRS1gene expression in female with Gestational diabetes mellitus. It is characterized high level of blood glucose, especially during first trimester then increased during the 2nd and 3rd trimester of the pregnancy period. The blood samples taken from one hundred twenty healthy women and female with gestational diabetes mellitus in 3rd trimester period of pregnancy, level of fasting blood glucose (FBG) also HbA1c% measured to diagnose GDM, in addition to lipid profile (cholesterol, triglyceride, HDL, LDL, and VLDL), molecular study consist of RNA extraction and qRT- PCR for IRS1gene expression determination. The fasting blood glucose mg/
... Show MoreFor the period from February 2014 till May 2014, one hundred and nine lactose fermenter clinical isolates from different samples (urine, stool, wound swab, blood, and sputum) were collected from Alyarmok, Alkadimiya, and Baghdad teaching hospitals at Baghdad governorate. Identification of all Klebsiella pneumoniae isolates were carried out depending on macroscopic, microscopic characterizations, conventional biochemical tests, and Api 20E system. Fifty-three (48.62%) isolates represented K. pneumoniae; however, 51.73% represented other bacteria. Susceptibility test was achieved to all fifty-three K. pneumoniae isolates using five antibiotic disks (Ceftazidime, Ceftriaxone, Cefotaxime, Imipenem, and Meropenem). Most of tested isolates (90
... Show MoreIn this paper an atmometer apparatus were used in the greenhouses for estimating reference evapotranspiration values. Experimental work was conducted in the agriculture research center in the College of Agriculture-University of Baghdad west of the city of Baghdad. One atmometer was used in eggplant greenhouse and in cucumber greenhouse through the winter growing season 2013-2014. FAO Penman-Monteith equation was applied outside the greenhouse and used only 65% from the value of ETo in the greenhouses for estimating the reference evapotranspiration in the greenhouse. Moreover, Penman-Monteith equation was applied in greenhouses for the evaluating the performance of the atmometer. The results show that the erro
... Show MoreBackground: Klebsiella pneumoniae were considered as normal flora of skin, and intestine. It can cause damage to human lungs; the danger of this bacterium is related to exposure to the hospital surroundings. materials and methods: the detection of Klebsiella pneumoniae on morphological and biochemical tests and then assured with VITEK 2 system. Resistance to antibiotics was determined by Kirby-Baeur method. And genotyping of IMP-1 in isolates was done by PCR technique, then biofilm formation was identified by Micro titer plate method. Results: The present study included a collecting of 50 specimens from different clinical specimens, (blood 40%, urine 30%, sputum 20%, wound infection 10%); 10 isolates were identified as K
... Show MoreThe present work aimed to investigate the neuraminidase (nan1) gene expression in 32 different clinical isolates of Pseudomonas aeruginosa to explore the role of the enzyme in different types of infection and might give a better understanding of host cell-pathogens interaction. In addition, the effect of monosaccharide D-mannose on neuraminidase gene expression in eight isolates was studied by utilizing a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results demonstrated that the highest expression of nan1 gene was in otitis samples (208,913.81) which were significantly higher than that from other infections (P < 0.01). While, the concentrations of gene copies obtained from urin
... Show MoreThis study was aimed to detect weather copA gene(copper resistance gene ) presence in A.baumannii A92 genome(AbaR genomic islands). The full genomic sequence of A.baumannii A92 not published in NCBI genome similarity was detected between two strains so the sequence of A.baumanniiIS-116(Genebank-AMGF0100000.1 ) was used to design the primers that were used for amplify of copA gene of A.baumannii A92.
Two primers contain two sites for restriction enzymes (KpnI,XohI) and PWSK29 vector were used in the cloning, double digestion has been performed for vector and gene. Then the re-ligation was completed to form recombinant molecule,after that, transformation have been performed for the recombinant molecule by using chemical competent E.coli
due to the presence of chemoresistance and the risk of tumor recurrence and metastasis. There is a pressing necessity to develop efficient treatments to improve response for treatment and increase prolong survival of breast cancer patients. Photodynamic therapy (PDT) has attracted interest for its features as a noninvasive and relatively selective cancer treatment. This method relies on light-activated photosensitizers that, upon absorbing light, generate reactive oxygen species (ROS) with powerful cell-killing outcomes. Nuclear factor kappa B (NF-κB), a transcription factor, plays a key role in cancer development by regulating cell proliferation, differentiation, and survival. Inhibiting NF-κB can sensitize tumor cells to chemotherapeuti
... Show MoreEvolutionary algorithms (EAs), as global search methods, are proved to be more robust than their counterpart local heuristics for detecting protein complexes in protein-protein interaction (PPI) networks. Typically, the source of robustness of these EAs comes from their components and parameters. These components are solution representation, selection, crossover, and mutation. Unfortunately, almost all EA based complex detection methods suggested in the literature were designed with only canonical or traditional components. Further, topological structure of the protein network is the main information that is used in the design of almost all such components. The main contribution of this paper is to formulate a more robust E
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