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.
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
... Show MoreCoagulation is the most important process in drinking water treatment. Alum coagulant increases the aluminum residuals, which have been linked in many studies to Alzheimer's disease. Therefore, it is very important to use it with the very optimal dose. In this paper, four sets of experiments were done to determine the relationship between raw water characteristics: turbidity, pH, alkalinity, temperature, and optimum doses of alum [ .14 O] to form a mathematical equation that could replace the need for jar test experiments. The experiments were performed under different conditions and under different seasonal circumstances. The optimal dose in every set was determined, and used to build a gene expression model (GEP). The models were co
... Show MoreDespite extensive investigations, an effective treatment for sepsis remains elusive and a better understanding of the inflammatory response to infection is required to identify potential new targets for therapy. In this study we have used RNAi technology to show, for the first time, that the inducible lysophosphatidylcholine acyltransferase 2 (LPCAT2) plays a key role in macrophage inflammatory gene expression in response to stimulation with bacterial ligands. Using siRNA- or shRNA-mediated knockdown, we demonstrate that, in contrast to the constitutive LPCAT1, LPCAT2 is required for macrophage cytokine gene expression and release in response to TLR4 and TLR2 ligand stimulation but not for TLR-independent stimuli. In addition, cells transfe
... Show MoreAcute myeloid leukemia represents the most prevalent type of acute leukemia in adults. Mutations in the tumor protein (TP53) gene have been found in more than half of all human cancers. This study was done to investigate the relationship between TP53 gene expression and the appearance and progression of acute myeloid leukemia in Iraq. This study included 100 subjects, divided into 60 patients suffering from pre-diagnostic acute myeloid leukemia and 40 healthy individuals. The difference in TP53 gene expression between acute myeloid leukemia patients and healthy individuals has been investigated, and the gene expression of TP53 has been measured after extraction of total RNA at concentrations (15–83 n
... Show MorePseudomonas aeruginosa is an opportunistic pathogen that causes a number of infections in immunocompromised patients. This organism appears to improve resistance to many antimicrobial agents and a high percentage of clinical isolates of P. aeruginosa exhibit multidrug resistance (MDR) phenotype . The purpose of this study is to screen the antibiotic susceptibility patterns and the prevalence of qacE delta1 gene among bacterial isolates. Accordingly, 145 samples were collected from different clinical sources from patients who admitted to different hospitals in Baghdad city in a period ranged 23/8/2018-1/1/2019. The isolates were diagnosed as P. aeruginosa based on routine b
... Show MoreDetecting protein complexes in protein-protein interaction (PPI) networks is a challenging problem in computational biology. To uncover a PPI network into a complex structure, different meta-heuristic algorithms have been proposed in the literature. Unfortunately, many of such methods, including evolutionary algorithms (EAs), are based solely on the topological information of the network rather than on biological information. Despite the effectiveness of EAs over heuristic methods, more inherent biological properties of proteins are rarely investigated and exploited in these approaches. In this paper, we proposed an EA with a new mutation operator for complex detection problems. The proposed mutation operator is formulated und
... Show MoreOut of 120 isolates from different clinical cases, only 75 were found and confirmed that they belong to the Pseudomonas aeruginosa bacteria. The result revealed that the LasB virulent gene was present in 63 isolates with 63% percentage. The gel electrophoresis showed that the molecular weight of LasB gene was 300 bp. DNA sequences of LasB gene was done, and the results showed the presence of some gene mutations like substitution, addition and deletion with 97% identity with the Refseq gene. From the other side, the results of identities of translated nucleotides sequence with the original sequence of amino acids revealed that there are no effects of gene mutations on translation of the product protein.
Abstract: E2F6 is a member of the E2F family of transcription factors involved in regulation of a wide variety of genes through both activation and repression. E2F6 has been reported as overexpressed in breast cancers but whether or not this is important for tumor development is unclear. We first checked E2F6 expression in tumor cDNAs and the protein level in a range of breast cancer cell lines. RNA interference-mediated depletion was then used to assess the importance of E2F6 expression in cell lines with regard to cell cycle profile using fluorescence-activated cell sorting and a cell survival assay using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The overexpression of E2F6 was confirmed in breast tumor cDNA samp
... Show MoreThe Machine learning methods, which are one of the most important branches of promising artificial intelligence, have great importance in all sciences such as engineering, medical, and also recently involved widely in statistical sciences and its various branches, including analysis of survival, as it can be considered a new branch used to estimate the survival and was parallel with parametric, nonparametric and semi-parametric methods that are widely used to estimate survival in statistical research. In this paper, the estimate of survival based on medical images of patients with breast cancer who receive their treatment in Iraqi hospitals was discussed. Three algorithms for feature extraction were explained: The first principal compone
... Show MoreBackground: The study of human leukocytes (HLA) alleles, and haplotype frequencies within populations provide an important source of information for anthropological investigation, organ and hematopoietic stem cell transplantation as well as disease association, certain diseases showed association with specific alleles specially those of known or suspected hereditary origin or immunological basis, whether simple renal cyst is congenital or acquired is still unclear and need to be investigated.Objectives: To study the genetic aspect of simple renal cysts by detecting the gene frequency and the haplotype of HLA class I of patients with simple renal cysts, and to find the presence of these cysts in other family members.Method: Thirty patient
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