Evaluation of different methods for the detection of KPC b-lactamase in Klebsiella pneumoniae and escherichia coli

The worldwide increase in the occurrence and dissemination of Klebsiella pneumoniae carbapenemase [KPC] among Gram-negative pathogens makes critical the early detection of these enzymes. The aim of this study is the evaluation of the most appropriate method for the detection and identification of KPC producing Klebsiella pneumonia and Escherichia coli. Modified Hodge test [MHT], Boronic acid lest [BAT] and polymerase chain reaction [PCR] were evaluated for the detection of KPC-possessing isolates. A total of 60 urine samples and 60 sputum samples were collected from 79 patients in Chest Intensive Care Unit and Trauma intensive Care Unit in Assiut University Hospital. Sixty four isolates [26 Klebsiella pneumonia and 38 E. coil] were identified by conventional methods. The antibiotic susceptibility testing of isolates showed that 19% were resistant to meropenem, 17% were resistant to imipenem and 95% were resistant to ertapenem. Thirty seven isolates were MHT positive while twenty three only of them were Boronic acid and PCR positive. The results indicate that the MHT lack sensitivity while Boronic acid test is an excellent inhibitor of KPCs. PCR is the standard method for the detection of the KPCs but it's highly expensive

Prevalence of efflux mediated ciprofloxacin and levofloxacin resistance in pseudomonas aeruginosa urinary isolates

Pseudoinonas aeruginosa [P. aeruginosa] represents a phenomenon of antibiotic resistance, and demonstrates practically all known mechanisms of bacterial resistance. Active efflux is an important mechanism of resistance in P. aeruginosa. It contributes to the development of multiple resistances to all strategic antipseudonional antibiotics. More than five hundred urine samples were collected from patients in Assiut University Hospital. Fifty P. aeruginosa isolates were identified by conventional methods. The antibiotic susceptihity testing of isolates showed that 68% of isolates were resistant to ciprofloxacin and 62% were resistant to levofloxacin. A comparison between the effect of three efflux pump inhibitors [Reserpine, Pantoprazole and Carbonyl cyanide m-chlorophenylhydrazone [CCCP]] on the activity of ciprofloxacin and levofloxacin was done by measuring ability of these agents to potentiate effect of ciprofloxacin and levofloxacin against resistant P. aeruginosa isolates. Outer membrane profile of P. aeruginosa isolates was also done using sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE]. Reserpine was able to potentiate effect of ciprofloxacin in 50% of isolates, and in 5.5% for levofloxacin. Pantoprazole results were 33.3% for ciprofloxacin, 16 7% for levofloxacin. Finally CCCP potentiate. Regarding the SDS-PAGE of P. aeruginosa isolates, all isolates produced proteins with apparent molecular masses in the range of 50-54kDa.Reserpine-ciprofloxacin proved to be the best combination against multidrug resistant P. aeruginosa. Over production of 50-54 KDa outer membrane proteins is responsible for emergence of P. aeruginosa strains highly resistant to fluoroquinolones in clinical settings

Prevalence of sen virus infection in hemodialysis and blood diseased patients in Assiut university hospitals

SENV is a blood- borne, circular ss DNA virus and possessing nine genotypes [A to I]. Among nine genotypes, SENV-D and SENV-H genotypes have the strong link with patients with non [A-E] hepatitis infections .Recently, the identification of SEN virus [SENV] as a possible etiologic agent of parenteral transmission hepatitis let to the study of the prevalence of such agent. This study compared SENV prevalence and its two important genotypes [D and H] which might be pathogenic in high risk subjects including blood diseased patients and hemodialysis patients and low risk subjects as healthy blood donors. This study included 75 multitransfused patients, 60 of them were hemodialysis and the remaining were blood diseased including hemophilics, anemics and leukemics. The study included also 25 healthy blood donors as a control They were received consecutively at Department of Internal Medicine, Assiut University Hospital. The sera were separated and SENV DNA was detected by polymerase chain reaction. The results showed a higher prevalence of SENV infection in patients group than blood donors [46.7% versus 20%].No significant relation was found between SENV infection and age, duration of hemodialysis or liver enzymes. However, there was significant difference between SENV positive and negative patients as regards gender and number of blood transfusions. SENV is commonly present in blood diseased and hemodialysis patients attended to Assiut University Hospitals as well as in blood donors at variable rates. SENV infection has been found in only 20% of blood donors but in 46.7% of patients. The results also indicated that other possible routes of SENV infection other than blood transfusion may be included. Its pathogenic role in causing hepatitis is not documented, so far it can be considered as simple guest till further studies have been done

Endothelial cell dysfunction and inflammatory process in type 1 diabetic patients with-and without micro vascular disease

Endothelial cell dysfunction results in altered production of cell adhesion molecules [CAMs] that may be involved in the pathogenesis of diabetic microvascular disease. Increased circulating cytokines may also be involved in this process. The aim of the present study was to evaluate levels of some CAMs and cytokines in children and adolescents with type 1 diabetes. It was also aimed to assess these parameters in relation to microvascular complications and certain risk factors. The study included 45 cases with type 1 diabetes aged 8-22 years of whom, 30 cases had evidence of microangiopathy [retinopathy or nephropathy] and 15 cases had not. Fifteen apparently healthy matchable subjects were included as controls. Cases were subjected to full history taking and physical examination. Direct ophthalmoscopy and fluorescein angiography were used to diagnose retinopathy, while nephropathy was diagnosed by detection of microalbuminuria. Level of glycated hemoglobin [HbA[1c]] and serum levels of sVCAM-1 and sE-selectin as well as IL-6 and TNF alpha were assessed for all patients and controls. The results showed that diabetic patients as a whole had significantly higher serum levels of sVCAM-lt sE-selectin, IL-6 and TNF alpha than controls. Post pubertal age, long duration of illness, obesity and high HbA[1c] level were significant risk factors for higher levels of CAMs. Significant positive correlations were found between levels of HbA[1c] and each of serum levels of sE-selectin and IL-6. Also significant positive correlations were found between each of serum levels of sVCAM-1 and sE-selectin, and IL-6 and TNF alpha. Patients with evidence of microangiopathy had significantly higher level of sVCAM-1 than cases without, and the latter group had significantly higher level of sE-selectin than controls. It is concluded that young patients with type 1 diabetes had significant markers of endothelial cell dysfunction particularly in those with microvascular disease. Screening of diabetic patients with E-selectin may help early diagnosis of endothelial dysfunction. Strict glycemic control and new therapeutic targets are mandatory to improve diabetic outcome in such cases