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Detection of the effect of synthetic siRNA on efflux pump MexA gene expression and antibiotic resistance in clinical isolates of Pseudomonas aeruginosa
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Introduction and Aim: Pseudomonas aeruginosa is a nosocomial infection with an ability to develop high levels of antibiotic resistance. The efflux pump system is one of the mechanisms that is linked to multidrug resistance in P. aeruginosa. In this study, we employed siRNA loaded on gold nanoparticles against the MexA efflux pump gene to decrease the MexA gene expression in P. aeruginosa and estimated antibiotic resistance after gene silencing.   Materials and Methods: This study examined four strains of P. aeruginosa isolated from patients in various hospitals in Baghdad. Bacteria isolated were identified by biochemical tests and Vitek compact 2 system.  Single-stranded siRNA (33bp) designed in this study was loaded onto gold nanoparticles (AuNPls). Detection of the MexA gene was carried out by conventional PCR technique. The expression of MexA gene was examined by qRT-PCR in order to determine if the siRNA have impacted on MexA gene expression and on the antibiotic resistance in aeruginosa   Results: This study showed that the mRNA expression level of the MexA gene exhibited a decrease in fold change CT -2   in P. aeruginosa (isolates numbers 67, 66,49, and PDR(5p)) when examined in vitro. The specific fold change values observed were (0.202, 0.040, 0.063, and 0.163) respectively. The resistance percentages of antibiotics tested was observed to increase after MexA gene silencing.   Conclusion: Targeting the MexA gene with synthetic siRNA may be a unique approach to diminish P. aeruginosa resistance to antibiotics. However, many unexpected consequences may occur when utilizing any genetic manipulation in bacteria.

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Publication Date
Mon Apr 10 2017
Journal Name
Ibn Al-haitham Journal For Pure And Applied Sciences
Role of Extracted Genomic DNA on Biofilm Formation by Pseudomonas aeruginosa and Klebsiella pneumoniae in vitro
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   Bacteria form complex and highly elaborate surface adherent communities known as biofilms.Biofilm have been shown to be associated with several human diseases ,and to colonize a wide variety of medical devices . The current study focuses on contribution of extracted genomic   DNA in  biofilm formation by   P. aeruginosa and  K. pneumoniae isolates   .The percentages of Pseudomonas aeruginosa recovery from drinking water  in this study were  10%(20 positive P. aeruginosa  samples ) and K. pneumonia.,  7%(14 positive K. pneumonia samples).The results showed that all P.aeruginosa and K. pneumoniae isolates (100%) were slime producer but in different degrees by forming  of black

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Publication Date
Sun Sep 22 2019
Journal Name
Baghdad Science Journal
Detection of CTX-M-type ESBLs from Escherichia coli Clinical Isolates from a Tertiary Hospital, Malaysia
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The present study aims to detect CTX-M-type ESBL from Escherichia coli clinical isolates and to analyze their antibotic susceptibility patterns. One hundred of E. coli isolates were collected from different clinical samples from a tertiary hospital. ESBL positivity was determined by the disk diffusion method. PCR used for amplification of CTX-M-type ESBL produced by E. coli. Out of 100 E. coli isolates, twenty-four isolates (24%) were ESBL-producers. E. coli isolated from pus was the most frequent clinical specimen that produced ESBL (41.66%) followed by urine (34.21%), respiratory (22.23%), and blood (19.05%).  After PCR amplification of these 24 isolates, 10 (41.66%) isolates were found to possess CTX-M genes. The CTX-M type ESBL

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Publication Date
Fri May 09 2025
Journal Name
Iraqi Journal Of Biotechnology
DETECTION OF GENE EXPRESSION OF SERINE PALMITOYLTRANSFERASE (SPT2) IN MOUSE CELL LINE RAW264.7 INFECTED WITH LEISHMANIA MEXICANA AMASTIGOTES
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Leishmania species are the causative agent of a tropical disease known as leishmaniasis. Previous studies on the old world species Leishmania major, showed that the amastigotes form which resides inside the macrophage of the vertebrate host, utilize host’s sphingolipids for survival and proliferation. In this study, gene expression of serine palmitoyltransferase (SPT) subunit two (MmLCB2) of the mouse macrophage cell line (RAW264.7), which is the first enzyme in the de novo sphingolipid biosynthesis, was detected in both infected and non-infected macrophages. This was detected under condition where available sphingolipid was reduced, with the new world species Leishmania mexicana. Results of qPCR analysis showed that there was no differen

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Publication Date
Mon May 22 2017
Journal Name
Ibn Al-haitham Journal For Pure And Applied Sciences
Role of Omeprazole in Enhancement of Antibiotic Resistance in E. coli
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E. coli was isolated, and it was Gram-negative rod bacteria that was colony circular, regular edged, thick somewhat glitter and  viscous(less). It was lactose fermenter bacteria and belongs the family of Enterobacteriaceae. E. coli showed sensitivity to all used antibiotics except Erythromycin (E), Cloxacellin (CX), Rifampin (RA), Cephalothin (KF), Ampicillin (AM), and Penicillin (P). The experimental results of antibiotic sensitivity of E. coli in media containing different concentrations of omeprazole, a proton pump inhibitor, showed an enhancement of resistance by decreasing the sensitivity of E. coli inversely with drug concentration against the antibiotics that E. coli was sensitive to. It seems that omeprazole changed cell mem

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Publication Date
Sat Jul 03 2021
Journal Name
Indian Journal Of Forensic Medicine & Toxicology,
The Effect of Aqueous Extract of Quercus infectoria Plant on Pap E gene Expression of Uropathogenic Escherichia coli
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One hundred twelve urine samples were collected from Baghdad hospitals and examined by different identification techniques. Seventy isolates (62.5%) were diagnosed as Escherichia coli after microscopic and cultural identifications. The result of PCR product electrophoresis on the isolates showed that thirteen isolates (18.57%) have Pap E gene which are uropathogenic E. coli. Antibiotic susceptibility test was done, and four high resistant strains were mixed with aqueous extract of Quercus infectoria plant in 96 well ELISA plate and incubated for different times. After 0, 6, and 12 hr. of incubation, the effect of the plant extract on the bacterial growth was determined by ELISA reader, and the effect on the expression of P

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Publication Date
Tue Sep 08 2020
Journal Name
Baghdad Science Journal
Silver Nanoparticles and their Role in Gene Expression of Motility Gene motB and Repression of AI-2-Controlled Gene
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Antibiotic resistance is the capability of the strains to resist or protect themselves from the effects of an antibiotic. Such a resistance towards the current antimicrobials leads to the search of novel antimicrobials. Nanotechnology has been promising in different field of science and among it is the use of nanoparticles as antibacterial agents. The gastrointestinal tract seems to be the primary reservoir of uropathogenic E.coli (UPEC) in humans. UPEC strains harbour the urinary tract and cause urinary tract infection. They cause serious ailments in terms of humans. They develop resistance and increase their virulence by forming biofilms. They also show a remarkable locomotory movement with the aid of autoinducer controlled ge

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Publication Date
Mon Jan 01 2024
Journal Name
Pakistan Journal Of Life And Social Sciences (pjlss)
Antibacterial and Antibiofilm Activity of Phenolic Compounds Extracted From Camellia Sinensis And Evaluate The Effect On The Gene Expression (Clfa) In Staphylococcus Aureus
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Publication Date
Sun Oct 06 2024
Journal Name
Ukrainian Journal Of Nephrology And Dialysis
Effect of Lactobacillus supernatant on swarming-related gene expression in Proteus mirabilis isolated from urinary tract infections
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Proteus mirabilis isolates have been intensively researched for their capacity to cause urinary tract infections (UTIs) and their swarming motility, although little is known about this phenomenon. Probiotic Lactobacillus species, which are beneficial bacteria, are being studied worldwide as therapeutic and preventative agents against bacterial infections. This study investigated Lactobacillus supernatants as a potential new treatment against Proteus mirabilis. In addition to testing their antimicrobial and anti-swarming activities, the research also aimed to understand the genetic mechanisms behind the observed phenotypic changes. Methods. A total of 150 urine specimens were collected from UTI patients at various hospitals in Baghdad. Dire

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Publication Date
Mon Apr 29 2024
Journal Name
Journal Of The College Of Basic Education
Detection Of Biofilm Formation By Beta- Lactam Resistance Klebsiella Pneumoniae Isolated From Clinical Specimens And Aquatic Samples
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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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