RESUMO
Background: Polycystic ovary syndrome (PCOS) is an endocrine disorder and the criteria are specified by common complex genetic hyperandrogenism, oligomenorrhea or amenorrhea and polycystic ovary morphology. It is a leading cause of female infertility. The prevelance of PCOS among reproductive age women has been estimated to be 4-12%. The association between PCOS and FSH receptor (FSHR) polymorphism attracts wide attention. The aim of the present study was to evaluate whether polymorphism of FSHR at Ala307Thr codon is associated with PCOS and with clinical features of PCOS patients in Egypt. Results: PCOS patients (n=64) and control subjects (n=65) in the reproductive age were recruited from the outpatient clinic of Obstetrics and Gynecology Department, Mansoura University, Egypt. The Ala307Thr polymorphism in FSHR, and the frequency of respective genotypes was studied and statistical analysis was performed. We found that the heterozygote Ala/Thr genotype was associated with PCOS (64.1%, OR=2.68, CI=0.97, P= 0.033) compared with controls. Conclusion: The variant of Ala307Thr polymorphism of FSHR was associated with PCOS but it may be related to high total testosterone levels. In addition the FSHR polymorphism was not associated with either luteinizing hormone or follicular stimulating hormone. The present study suggests that the variant of the FSHR gene may act as a causative factor of PCOS in Egyptian women.
RESUMO
Aim: the present study aims to estimate the phenotypic and genotypic characteristics of efflux pump systems in Enterobacteriacae clinical isolates
Methods: a total of 60 isolates of Escherichia coli and Klebsiella pneumoniae were studied for antibacterial susceptibility pattern, Effect of efflux pump inhibitors [reserpine, carbonyl cyanide m-chloropheylhydorazone [CCCP] and omperazole] on the MIC of antibacterial agents and ethidium bromide [EtBr] efflux assay, preparing outer membrane proteins, polymerase chain reaction for the detection of efflux genes. Sequencing of some efflux genes, constructing the phylogenetic tree and determining the copy number of the resistance genes in both chromosomal and plasmid DNA
Results: nearly all the clinical isolates were multiple resistant as they were resistant to most antimicrobial classes used in this study. CCCP was found to have the highest effect on decreasing the MIC of different antibiotics comparable with reserpine and omperazole. It was found that all tested E. coli and K. pneumoniae isolates extrude EtBr resulting in decrease in fluorescence over the time of assay. In E. coli isolate No.25 and K. pneumoniae No. 12, only the control cells extrude EtBv resulting in significant loss in fluorescence. In presence of each of reserpine, CCCP and omperazole, an insignificant loss in fluorescence was observed, reflecting blockage of EtBr by these compounds at different levels. All E. coli isolates produced high amount of outer membrane proteirns with apparent molecular mass of 37 and 39 and 50 KDa. These proteins may be designated as OmpC, OmpF and TOIC respectively. Regarding K. pneumonia, all the isolates produced high amount of outer membrane proteins with apparent molecular mass of 37 and 50 KDa and only six isolates produced an outer membrane protein of apparent molecular mass of 39 KDa these proteins may be designated as ompK35 and TOIC and ompK 36 respectively. AcrA and AcrB and TOIC genes was detected and amplified in most of the tested isolates of E. coli and K. pneumoniae genomic and plasmid DNA. AcrA amplicons were visualized at 500 bp for E. coli and 200 bp for K. pneumoniae, AcrB amplicon was detected at 500 bp and TOIC amplicon was detected at 700 bp for both E. coli and K. pneumonia isolates. Moving to the sequencing results, the phylogeny was used to determine the relatedness between different isolates with the observation that different genes like AcrA or AcrB were very closely related to each other on chromosomal DNA and that was the same observation for their gene sequence on plasmid DNA that differs from the chromosomal sequence. It was observed that the chromosomal copy number of AcrA and AcrB genes of the E. coli and K. pneumoniae isolates was nearly the same and after the acquisition of the plasmid, the copy number of' these genes was significantly increased
RESUMO
Aim: the present study aims to determine the efflux pump resistance by phenotypic and genotypic Methods
Methods: a total of 40 isolates of Pseudomonas aeruginosa were studied for antibiotic susceptibility pattern, , Effect of efflux pump inhibitors [reserpine, carbonyl cyanide mchloropheylhydorazone [CCCP] and omperazole] on the MIC of antibacterial agents, ethidium bromide [EtBr] efflux assay, outer membrane proteins, polymerase chain reaction for the amplification of resistance genes
Results: nearly most of the isolates were multiple resistant as they were resistant to most antimicrobial classes used in this study. CCCP was found to have the highest effect on decreasing the MIC of different antibiotics comparable with reserpine and omperazole. It was found that all tested Pseudomonas aeruginosa isolates extrude EtBr resulting in decrease in fluorescence over the time of assay. In Pseudomonas aeruginosa isolate No.3 and 14 only the control cells extrude EtBr resulting in significant loss in fluorescence. In presence of each of reserpine, CCCP and omperazole/e, an insignificant loss in fluorescence was observed, reflecting blockage of EtBr by these compounds at different levels. All the isolates produced high amount of outer membrane proteins with apparent molecular mass of 54 KDa and 50 KDa. These proteins may be designated as OprJ and OprM or OprN. MexA and MexB genes was detected and amplified in most of the tested isolates of Pseudomonas aeruginosa genomic and plasmid DNA. The amp/icons were visualized at 500 bp and 1000 hp respectively. While OprM gene was amplified successfully in the twenty tested isolates and the amp/icons were visualized at size of 900 bp