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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
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
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
Fri Jun 07 2024
Journal Name
Asian Pacific Journal Of Cancer Prevention
The Effect of NF-κB Deactivation on Cancer Cell Response to ALA Mediated Photodynamic Therapy
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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

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Publication Date
Thu Feb 16 2023
Journal Name
Iraqi Journal Of Pharmaceutical Sciences ( P-issn 1683 - 3597 E-issn 2521 - 3512)
A Novel Single Nucleotide Polymorphism of Interleukin-10 Gene is Linked to Type 2 Diabetes Mellitus in Iraqi Patients with Toxoplasmosis(Conference Paper )#
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Type 2 diabetes mellitus (T2DM) is a chronic disorder that is a serious health concern all over the globe, it is linked to Interleukin-10 (IL-10) single nucleotide polymorphisms (SNPs) at the promoter region. On the other hand, diabetes influences the cellular and humoral immunity predisposing the patient to a variety of opportunistic parasites one of them is Toxoplasma gondii (T. gondii), which may infect any nucleated cell, including pancreatic cells. The purpose of this research was to explore the association of IL-10 genetic polymorphisms with T2DM and latent toxoplasmosis among Iraqi patients with T2DM. Fifty-five and fifty-eight venous blood samples were taken from T2DM patients and age-matched non-diabetic person

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Publication Date
Fri Mar 29 2024
Journal Name
Iraqi Journal Of Science
Involvement of Total Antioxidant Activity and eNOS Gene rs1799983/ rs2070744 Polymorphisms in Breast Carcinogenesis
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Globally, breast cancer is the common malignancy affecting women and understanding its associated molecular events could help in disease prevention and management strategies. The present study was set to investigate an association between total antioxidant capacity (TAC) and endothelial nitric oxide synthase (eNOS) polymorphisms with breast cancer. For this purpose, 100 subjects were participated in this work, including 50 female patients diagnosed with breast cancer recruited from Oncology hospital, Baghdad - Iraq and 50 healthy women as a control group. The concentration of antioxidants was measured in the serums collected from blood samples of breast cancer patients and healthy controls. While eNOS SNPs (rs1799983, G894T and rs2070744, T

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Publication Date
Fri Jun 16 2023
Journal Name
Iraqi Journal Of Pharmaceutical Sciences ( P-issn 1683 - 3597 E-issn 2521 - 3512)
Role of Fasting Mimicking Diet in Farnesoid x Receptor for Suppressing Epithelial-to-Mesenchymal Transition, Cell Cycle Progression, and Viability of Prostate Cancer Cells
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The systemic and resistant nature of metastatic castration-resistant prostate cancers (mCRPC) renders it largely incurable even after intensive multimodal therapy. Proliferation, survival, and epithelial-mesenchymal transition (EMT) are three fundamental events that are deeply linked to carcinogenesis.  Hence, it is necessary to find a new combination of several therapies, targeting those vital mechanisms without causing side effects. Significant research works have shown differential low expression of the metabolic Farnesoid X receptor (FXR) in primary and metastatic prostate cancer suggesting their importance in prostate pathogenesis. Obticholic acid (INT 747), a potent FXR agonist is widely used in primary biliary chola

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Publication Date
Mon Aug 22 2022
Journal Name
Biochemical And Cellular Archives
Deregulation of autophagy flux and gene expression induced by tobacco smoke among Iraqi smokers.
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: Cigarette smoking is a lifestyle behavior that causes significant adverse health effects. Cigarette smoke contains chemicals, many of which are lead to the production of reactive oxygen species (ROS), which can lead to apoptosis and autophagy. To estimate the association of Cigarette smoking with the autophagy and immunity, technology of real time polymerase chain reaction (RTPCR) for gene expression of (LC3A, LC3B, LC3C, myd88) was used. Enzyme-linked immunosorbent assay (ELISA) technique was utilized to measurement the amount of TNF-α protein. The ratios of LC3A/LC3B and LC3B/LC3C were calculated to estimate the autophagy flux. The results indicate the expression of LC3B, LC3C and Myd88 genes in smokers is increased significantly (p

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Publication Date
Sun Oct 01 2023
Journal Name
Baghdad Science Journal
Evaluation of the cytotoxic effects of the colchicine compound isolated from the leaves of Calotropis procera (Ait) against MCF-7 and SK-GT-4 cancer cell lines.
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Alkaloids are regarded as important nitrogen-containing chemical compounds that serve as a rich source for discovering and developing new drugs where most plant-origin alkaloids have antiproliferation effects on different kinds of cancers. Alkaloids’ continence of Calotropis procera leaves are detected by two biochemical alkaloid reagents. Also GC-MS analysis for leaf alkaloid extract was done that showed the existence of one type of alkaloid compound at retention time12.8min detected as colchicine (C22H25N06( by comparing it with colchicine standard reference (Sigma Aldrich) with M.wt 399g/mol and percentage area 7.1%. Furthermore, identification, separation, and purification

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Publication Date
Tue May 05 2020
Journal Name
Book
Study the inhibitory effect of miswak extract on biofilm gene.
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This book presents the problem of tooth decay due to bacteria Streptococcus mutans one of methods of treatment using 3 extracts of S. persica (miswak) (aqueous, acetone and methanol) and prove its effectiveness and its impact on the gtf (B, C, and D) genes that code the glucosyltransferase (Gtf) enzymes that cause decay membrane compared to the usual means used for the prevention of tooth decay

Publication Date
Wed Oct 26 2022
Journal Name
Iraqi Journal Of Science
Gene Expression Analysis via Spatial Clustering and Evaluation Indexing
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The density-based spatial clustering for applications with noise (DBSCAN) is one of the most popular applications of clustering in data mining, and it is used to identify useful patterns and interesting distributions in the underlying data. Aggregation methods for classifying nonlinear aggregated data. In particular, DNA methylations, gene expression. That show the differentially skewed by distance sites and grouped nonlinearly by cancer daisies and the change Situations for gene excretion on it. Under these conditions, DBSCAN is expected to have a desirable clustering feature i that can be used to show the results of the changes. This research reviews the DBSCAN and compares its performance with other algorithms, such as the tradit

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