Objective: The present work was undertaken to investigate the impact of sub inhibitory concentration of gentamicin on hla gene expression in methicillin resistant Staphylococcus aureus isolates. Methods: The bacterial isolates used in this study represent 33 MRSA strains, previously isolated form patients visiting several hospitals in Baghdad. Gentamicin, vancomycin, and oxacillin MIC were determined using broth dilution method. Microtiter plate method was adopted to investigate the biofilm forming capacity. Alpha hemolysin was detected by culturing MRSA isolates on rabbit blood agar. Furthermore, hla gene was detected in MRSA isolates using conventional PCR technique; while, qRT-PCR method was performed to assay the hla expression in planktonic and biofilm cells in presence and absence of gentamicin. Results: the present results demonstrated that 12 (36.36%) isolates were gentamicin-resistant; whereas, all isolates were resistant to oxacillin and sensitive to vancomycin. Out of 33 MRSA, 3, 23, and 7 isolates formed a weak, moderate, and strong biofilm, respectively. Phenotypically, 30 isolates produced alpha hemolysin on rabbit blood agar plates; nevertheless, hla gene was located in 29 isolates. Of considerable interest, the addition of gentamicin significantly (P < 0.05) reduced the hemolysis activity; while, insignificant fold change (less than two) of hla gene was observed in all tested isolates in the presence of sub MIC of gentamicin (16 µg/ml). Conclusion: gentamicin upregulated the hla gene expression in biofilm cells; hitherto, this increment was isolate specific.
Abstract Background: Multidrug-resistant bacteria (MDR) often contaminate hospital environment and cause serious illnesses. Quorum Sensing (QS) regulates a variety of downstream cellular processes, including antibiotics resistance mechanisms and biofilm formation, and causes harm to the host. This study investigates antibacterial susceptibility and biofilm formation of pathogenic bacteria in hospital environment. Methods: Hundred bacterial isolates were collected from various environments in the Medical City hospital. The antimicrobial susceptibility technique was evaluated through disk diffusion method. Next, biofilms formation was detected by the microliter plate assay. Finally, PCR was used to analyze the frequency of QS system gene
... Show MoreOne hundred and fifteen isolates of Staphylococcus aureus were isolated from human infected wound and burn. The resistance of this organism for eight commonly used antibiotics was determined by disc diffusion methods. The minimum inhibitory concentration MIC was determined for each drug by tube dilution method. Results showed significant variations in the MIC values. It was found that two isolates had high resistance for the eight antibiotics used. Effect of ascorbic acid on bacterial growth was studied singly and in combination with antibiotics. Reductions in MIC value were reported for different antibiotics when tested by growth curve method too.
A field experiment was carried out during winter season of 2019-2020 at Al-Mhanawyah Research Station - Agriculture Research Directorate - Babylon Governorate / Iraqi, to study the gene expression of Sgr gene responsible for controlling the duration of staying green in varieties of wheat under effect of plant growth regulator during the two growth stages (vegetative and reproductive) by using quantitative reverse transcription-PCR (RT-qPCR) technique and achieving the highest grain yield for a number of wheat varieties. Randomized complete block design (RCBD) arranged according to split plots used with three replicates. The experiment included twelve wheat varieties (Saberbic, Al-Rasheed, Iraq, Tamoz-3, Al-Adnaniya, Babel, IPA-99, Al-Latife
... Show MorePvcABCD are cluster of genes found in Pseudomonas aeruginosa. The research was designed to examine the relationship between the pvc genes expression and cupB gene, which plays a crucial role in the development of biofilm, and rhlR, which regulates the expression of biofilm-related genes, and to investigate whether the pvc genes form one or two operons. The aims were achieved by employing qRT-PCR technique to measure the gene expression of genes of interest. It was found that out of 25 clinical isolates, 21 isolates were qualified as P.aeruginosa. Amongst, 18(85.7%) were evaluated as biofilm producers, 10 (47.6%), 5 (23.8%), and 3 (14.2%) were evaluated as strong, moderate and weak producers respectively, while, 3 (14.2%) were considered
... Show MoreIntroduction: Biocides are commonly used for disinfection in a variety of contexts. They are generally used to avoid infection by controlling biofilm on medical equipment. However, the literature lacks information on the effect of biocide on efflux pump gene expression. Objective: To determine the influence of biocide on biofilm development and efflux pump acrA and ramA gene expression. Methodology: The microtiter plate method was used to identify biofilm development in 80 isolates of K. pneumoniae. The minimal inhibitory concentrations (MIC) of three biocides (quaternary ammonium compound (QAC), chlorohexidine digluconate, and chloroxylenol) were estimated. The effect of QAC on the intensity and viability of biofilms was investigated as we
... Show MoreThe inhibitory action of four lactobacilli isolates Lactobacillus bulgaricus, L. acidophilus, L. plantarum and L. fermentum, isolated from four different samples; yoghurt, vinegar, saliva and vagina respectively, on Escherichia coli and Staphylococcus aureus adhesion to uroepithelial cells were investigated. Results showed that all Lactobacillus isolates or their supernatant were able to reduce the number of the uropathogens attached to uroepithelial cells. However, inhibition level of lactobacilli cells was higher than their supernatant. Nevertheless, the human indigenous lactobacilli (L. fermentum and L. plantarum) were more competitive than food lactobacilli (L. acidophilus and L. bulgaricus).
Exploring the antibacterial potential of neem oil (Azadirachta indica) in combination with gentamicin (GEN) against pathogenic molds, especially Pseudomonas aeruginosa, has drawn concern due to the quest for natural treatment options against incurable diseases. Prospective research directions include looking for natural cures for many of the currently incurable diseases available now. microbial identification system, were used to identify the isolates. The research utilized a range of methods, such as the diffusion agar well (AWD) assays, TEM (transmission electron microscopy) analysis, minimum inhibitory concentration (MIC) assays, and real-time PCR (RT-qPCR) to analyze bacterial expression and the antibacterial action of neem oil (Azadira
... Show MoreSTAG proteins, which are part of the cohesin complex and encoded by the STAG genes, are known as Irr1/Scc3 in yeast and as SA/STAG/stromalin in mammals. There are more variants as there are alternate splice sites, maybe three open reading frames (ORFs) code for three main proteins, including: SA1 (STAG1), SA2 (STAG2) and SA3 (STAG3). The cohesin protein complex has various essential roles in eukaryotic cell biology. This study compared the expression of the STAG1 gene in four different breast cancer cell lines, including: MCF-7, T-47D, MDA-MB-468, and MDA-MB-231 and normal breast tissue. RNA was extracted from these cell lines and mRNA was converted to cDNA, and then expression of the STAG1 gene was quantified by three sets of specific prim
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