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.
Ankylosing spondylitis (AS) is a common, highly heritable inflammatory arthritis affecting primarily the spine and pelvis. This study was aimed to investigate the relationship between the rs27044 polymorphism in Endoplasmic reticulum aminopeptidase-1 (ERAP-1) with the susceptibility and severity of AS correlated with some biochemical markers such as hematological parameter (Erythrocytes sedimentation rate (ESR)) and immunological parameters (C-reactive protein (CRP), Human leukocyte antigen-B27 (HLA-B27), Interlukin-6 (IL-6) and Interlukin-23 (IL-23)), and oxidative stress parameters (Glutathione (GSH) and Malondialdehyde (MDA)) in a sample of Iraqi population. A total of 60 blood samples were collected from AS patients requited Rhe
... Show More98 samples were collected from various clinical sources included (Burns, wounds, urines, sputums, blood) From the city of Baghdad, After performing the biochemical and microscopic examination, 52 isolates were obtained for Pseudomonas aeruginosa, 17 (32.7%) isolates from burn infection, 12 (23%) isolates from Wound infection 11 (21.2%) isolates from urine infection, 7 (13.5%) isolates of sputum and 5 (9.6%) isolates from blood. Bacteria susceptibility to form biofilm has been detectedby microtiter plate method, The results showed that 80% of the bacterial isolates were produced the biofilm with different proportions, alg D gene (alginate production) has been detected by polymerase chain reaction (PCR) Which plays an essential role in the fo
... Show MoreDrug resistance is a hot topic issue in cancer research and therapy. Although cancer therapy including radiotherapy and anti‐cancer drugs can kill malignant cells within the tumor, cancer cells can develop a wide range of mechanisms to resist the toxic effects of anti‐cancer agents. Cancer cells may provide some mechanisms to resist oxidative stress and escape from apoptosis and attack by the immune system. Furthermore, cancer cells may resist senescence, pyroptosis, ferroptosis, necroptosis, and autophagic cell death by modulating several critical genes. The development of these mechanisms leads to resistance to anti‐cancer drugs and also radiotherapy. Resistance to therapy can increase mortal
In this study, gold nanoparticles were synthesized in a single step biosynthetic method using aqueous leaves extract of thymus vulgaris L. It acts as a reducing and capping agent. The characterizations of nanoparticles were carried out using UV-Visible spectra, X-ray diffraction (XRD) and FTIR. The surface plasmon resonance of the as-prepared gold nanoparticles (GNPs) showed the surface plasmon resonance centered at 550[Formula: see text]nm. The XRD pattern showed that the strong four intense peaks indicated the crystalline nature and the face centered cubic structure of the gold nanoparticles. The average crystallite size of the AuNPs was 14.93[Formula: see text]nm. Field emission scanning electron microscope (FESEM) was used to s
... Show MoreTuberculosis (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
... Show MoreSwarming is one of the most important virulence factors used by bacteria to invade new sites. This study aimed to test the effects of gentamicin on swarming motility of Pseudomonas aeruginosa, both phenotypically and molecularly. The present results revealed that 11/25 isolates had gentamicin MIC of 1024 µg/ml. However, gentamicin at sub-minimal inhibitory concentration significantly (P< 0.05) reduced the diameter of swarming in all P. aeruginosa isolates. Noticeably the mean and median swarming diameter before treatment with gentamicin 5.557 and 5.816 cm respectively had significantly (P < 0.001) reduced to 0.871 and 0.766 cm respectively. At the molecular level, amrZ (a global regulator of multiple genes) and
... Show MoreIntroduction and Aim: Forkhead box P3 (FOXP3) and interleukin-10 (IL-10) are the key regulators controlling the activity of Treg cells, which are crucial for maintaining immune tolerance and reducing autoimmune reactions. The objective of this study was to investigate the potential utility of elevated levels of FOXP3 and IL-10 gene expression as a diagnostic indicator in patients with rheumatoid arthritis (RA). Materials and Methods: The study used quantitative polymerase chain reaction (qPCR) to examine the expression levels of FOXP3 and IL-10 transcripts in whole blood samples from Iraqi patients with rheumatoid arthritis. A group of healthy control subjects were also included in the study. Results: In blood samples taken fr
... Show MoreOsmotin and osmotin-like proteins belong to the PR-5 pathogenesis-related group of proteins and are induced in response to various types of biotic and abiotic stresses in several plant species. Carrot was transformed with a tobacco osmotin gene that encodes a protein lacking the vacuolar-sorting motif that is composed of a 20-amino-acid sequence at the C-terminal end, under the control of the cauliflower mosaic virus 35S promoter, using Agrobacterium-mediated transformation. Transgene integration and expression were confirmed by Southern and western blot analyses, and three selected transgenic lines were evaluated for their ability to tolerate drought stress. Under drought stress conditions, all transformants exhibited slower rates of wilti
... Show More