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.
Background: EOS (encoded by the IKZF4 gene) is a member of the zinc finger transcription factor IKaros family, and plays a critical role in Treg suppressor functions, and maintaining Treg stability. IL-6 is a soluble mediator with a pleiotropic effect on inflammation, immune response, and hematopoiesis. Aim: To estimate serum IL-6 level and EOS gene expression in Iraqi patients with psoriasis. Method: Twenty-two patients with psoriasis (8 females, 14 males) with age ranged 18-72 years, were recruited from Baghdad Teaching Hospital, Dermatology Clinic, Baghdad, and 24 healthy donors. The serum levels of IL-6 by ELISA and the gene expression of IKZF4 (EOS gene) by RT-qPCR technique. Results: The results showed a non-significant diffe
... Show MoreFluconazole was used to test the susceptibility of Candida albicans isolated from different clinical samples, and to detect mutations in ERG11 gene, and their relationship to fluconazole resistance. Forty-eight isolates of Candida albicans were tested for susceptibility using the disc diffusion method (M-44). ERG11 genes of six isolates were amplified (four resistant, two susceptible) and sequenced. The sequenced genes were analyzed to detect the mutations. Out of 48 isolates of Candida albicans, 4 (8%) were resistant to fluconazole. Sixteen-point mutations were detected included 13 silent mutations, and three missense mutations. The mutations of A945C (E266D) and G1609A (V488I) were found only in susceptible Candida albicans isolates, whil
... Show MoreThis study is the first investigation in Iraq dealing with genotyping of
Background: Fruits and their by-products are the primary sources of bioactive chemicals in plants. Because of its phytochemical richness, Annona squamosa fruits have gained the alertness of people willing in health-promoting diets. The purpose of this in vitro study was to evaluate the cytocompatibility effect of ethanolic crude extract of Annona squamosa pulp against a human normal cell line as a mouthwash for children. Material and method: The ethanolic extract of Annona squamosa pulp was extracted using the ultrasonic method and then lyophilized to make it powder. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) test was performed to investigate the cytotoxic activity of the pulp extract on a human normal cell l
... Show MoreThe process involved isolating E. faecium from the gut of honeybees, screening the bacterium for bacteriocin-like inhibitory substance (BLIS), evaluating its impact on the expression of the mexA gene in multidrug-resistant (MDR) P. aeruginosa, and determining the role of bacteriocin in treating infected wounds in mice through histopathological examination. After evaluating the best circumstances for producing BLIS, it was discovered that glucose was a superior carbon source and yeast extract was the best source of nitrogen. The pH was found to be 5, the ideal incubation time was 72 hours, and ammonium sulfate salt was used for partial purification at 80% saturation. The identification of MDR P. aeruginosa isolates from pus infection
... Show MoreThis study was conducted during the period 1/9/2014 – 1/2/2015 and aimed to identify the polymorphisms of IGF-1 gene in broiler chickens and their effects on some biochemical traits. A total of 300 one-day-old broiler chicks (Cobb500, n=150; Hubbard F-15, n=150) were evaluated in this study. Blood samples were individually collected from all birds for DNA extraction. PCR-RFLP method being used for determination the genotypes of IGF-1 gene which then correlated with biochemical traits studied. Cobb500 broilers with TT genotype had significantly (p˂0.05) higher serum triglycerides values than those of TC and CC genotypes. Low density lipoprotein (LDL) were significantly (p˂0.05) higher in Hubbard F-15 broilers with TT genotype than those
... Show MoreBackground: There is a significant molecular response to imatinib given at standard doses in individuals with chronic myeloid leukemia (CML) whose ABCB1 polymorphisms are present. Objective: To investigate the impact of the polymorphism in the ABCB1 gene rs1128503 on the effectiveness of nilotinib or imatinib therapy. Methods: From May 2022 until the end of January 2023, the current study was carried out in a single research institution, the National Center of Hematology, Baghdad Teaching Hospital at Medical City, Iraq. 76 people with chronic phase myeloid leukemia (CML-CP), who had previously received a diagnosis using the European Leukemia Net (ELN) criteria, enrolled in the trial. The PCR product was delivered to Macrogen Corpora
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