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&H) which might be pathogenic in high risk subjects including blood transfused patients and hemodialysed patients and low risk subjects as healthy blood donors. Subjects and methods: This study included 75 multitransfused patients, 60 of them were hemodialysed and the remaining were blood diseased including haemophilics, anaemics and leukemics. The study included also 25 healthy blood donors as a control. They were enrolled consecutively at the department of Internal Medicine, Assiut University Hospital. The sera were separated and SENV DNA was detected by polymerase chain reaction. Results: A higher prevalence of SENV infection was detected in patients groups than in blood donors (46.7% versus 20%).No significant relation was found between SENV infection and age, duration of haemodialysis or liver enzymes. However, there was significant difference between SENV positive and negative patients as regards gender and number of blood transfusions. Conclusions: SENV is commonly present in blood transfused and haemodialysed patients attended to Assiut University Hospitals as well as in blood donors at comparable 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.

Extended spectrum β lactamases producing Klebssiella pneumoniae (ESBL-KP) are an important cause of nosocomial infections in Intensive Care Units (ICUs). We conducted a prospective study on 650 patients who were admitted to different adult ICUs at Assiut University Hospital to determine the incidence of ESBL-KP by phenotypic and genotypic methods. Phenotypic tests for ESBL were combined disc method, double disc synergy test (DDST) and E-test. Genotypic detection of ESBL bla TEM and bla SHV genes was carried out by polymerase chain reaction amplification (PCR). The overall nosocomial infection incidence rate was 20% (130 patients). Klebsiella pneumonia was isolated from 44 patients (34%), in which 23 isolates were found to be phenotypically ESBL producers. ESBL-KP was most frequently isolated from chest ICU (47.8%) and blood was the most frequent site of infection (8 isolates, 34%). Based on Clinical and Laboratory Standards Institute (CLSI) screening test for ESBL, the combined disc method was the most sensitive (23/23, 100%) followed by the E-test (95.6%) and lastly the DDST (91.3 %). SHV gene was present in 8 isolates, TEM gene in 2 isolates, both SHV and TEM in 11 isolates and none of TEM or SHV in 2 isolates. Out of 950 environmental samples, Klebsiella pneumoniae was isolated from 48 samples (16.4%) in which 7 isolates (14.5 %) were ESBL and genotyping revealed SHV in 4 strains, and both SHV and TEM in 3 strains. Conclusion: This study revealed the high incidence of ESBL-KP in adult ICUs. SHV genotype was more prevalent than TEM type. Strict implementation of basic infection control measures seems to be the most effective means for controlling the spread of ESBL organisms.

Auto-immune hepatitis (AIH) in children is a rare chronic progressive liver disorder. It is characterised serologically by high aminotransferase levels, elevated immunoglobulin G (IgG) and the presence of autoantibodies. AIH is divided into two types according to the autoantibody profile. This study aims to assess frequency, clinical manifestations, biochemical features and outcome of AIH in children attending Assuit University Hospitals in Upper Egypt with acute icteric hepatitis and seronegative viral markers (anti-hepatitis A virus (HAV) IgM, HbsAg, anti-hepatitis C virus (anti-HCV) Ab). Patients and methods The study includes 34 children with AIH, diagnosed on the basis of the International Scoring Criteria of Auto-immune Hepatitis, recruited from Assuit University Hospitals, during the period from January 2005 to December 2009. All patients received prednisolone 2 mg kg–1 day–1. Follow-up was done for 1 year. Results Among 34 children diagnosed as AIH, 24 were females (70.5%) and 10 were males (29.5%). Jaundice represented the most consistent finding in all patients. According to the autoantibody profile, 25 children were classified as type 1 and nine children were classified as type 2. Corticosteroid therapy was started. Complete remission was observed in 67.6% of patients and partial remission in 17.6%. There was no significant statistical difference in clinical and biochemical features of AIH in patients regarding the response to treatment. Mild side effects of steroid therapy were encountered in 48.2% of patients. After complete withdrawal of corticosteroids, six patients (20.7%) developed relapse. Conclusion AIH type 1 was the main form of AIH in children referred to Assiut University Hospitals. Girls were more affected than boys. AIH type 1 exhibited a more active, ongoing immunologic process. Steroid alone can be used successfully in most cases. Children with AIH type 2 had a higher frequency of relapse after corticosteroid withdrawal. Further studies on a larger number of cases and long-term follow-up are recommended.

Objectives: to determine the prevalence of hypovitaminosis D, its association with bone density and to evaluate related determinants among admitted patients to Trauma Unit in Assiut University Hospitals. Materials and methods: A cross sectional study was carried out on patients with proximal femoral fractures aged 50 years and older admitted to Trauma Unit of Assiut University Hospitals, a level-I trauma center in Upper Egypt, from 1st January to end of December 2014. A randomized sample of 133 participants was included. Patients with polytrauma, major accidents and conditions known to affect bone density were excluded. Two well trained nurses filled a structured questionnaire by personal interview. Weight and height measurement, assessment of 25-hydroxy-vitamin D (25OHD) serum and bone mineral density measurement by DXA were done for all participants. A formal consent was taken from all participants prior to the study. Results: Participants aged 50-99 years with mean 69 ± 11.3 years, 48.1% were males and 51.9% were females. 50.4% and 22.6% of them were housewives and farmers respectively. Osteoporosis (T score of neck fe-mur -2.5) prevalence was 72.2%. Prevalence of (25OHD) deficiency (20 ng/ml) and inadequacy (between 20-30 ng/ml) were 60.9% and 15.8% respectively, and 23.3% were normal (> 30 ng/ml). Vitamin D level was positively correlated with T score of greater trochanter, neck femur, L4, L3 and L2 (r=0.23& p= 0.012, r= 0.28& p=0.001, r= 0.196& p=0.023, r= 0.18&p= 0.036 and r= 0.21 &p=0.02). Vitamin D deficiency was significantly associated with age and increase in BMI (p=0.04&0.012). Lack of milk and yoghurt intake had sig-nificant higher prevalence of vitamin D deficiency than others (p=0.02 and 0.01 respectively). No significant relation was found with sex or edu-cation level. Multiple regression analysis was done. Conclusions: Prevalence of vitamin D deficiency is high among hip fracture patients and is associated with low BMD. Vitamin D supplementation is strongly recommended for prevention of hip fractures in those aged >50 years.

This study explored and compared the mechanisms and selective concentration of resistance between a 3rd (pradofloxacin) and 2nd (ciprofloxacin) generation fluoroquinolone. Pradofloxacin- and ciprofloxacin-resistant mutants were selected by stepwise exposure of Escherichia coli (E. coli) to escalating concentrations of pradofloxacin and ciprofloxacin. The sequence of the quinolone resistance determining region (QRDR) and the transcriptional regulator soxS were analyzed, and efflux pump AcrAB-TolC activity was measured by quantitative real-time reverse transcription-PCR (qRT-PCR). First-step mutants reduced the fluoroquinolone sensitivity and one mutant bore a single substitution in gyrA. Four of six second-step mutants expressed ciprofloxacin resistance, and displayed additional mutations in gyrA and/or parC, while these mutants retained susceptibility to pradofloxacin. All the third-step mutants were fluoroquinolone resistant, and each expressed multidrug resistance (MDR) phenotypes. Further, they displayed resistance to all antibacterials tested except cefotaxime, ceftazidime and meropenem. The number of mutations in QRDR of gyrA and parC correlated with fluoroquinolone MICs. Mutations in parC were not common in pradofloxacin-associated mutants. Moreover, one second- and one third-step ciprofloxacin-associated mutants bore both mutations at position 12 (Ala12Ser) and 78 (Met78Leu) in the soxS gene, yet no mutations in the soxS gene were detected in the pradofloxacin-selected mutants. Altogether, these results demonstrated that resistance emerged relatively more rapidly in 2nd compared to 3rd generation fluoroquinolones. Point mutations in gyrA were a key mechanism of resistance to pradofloxacin, and overexpression of efflux pump gene acrB played a potential role in the emergence of MDR phenotypes identified in this study.

Background: There are no large epidemiological studies of Clostridium difficile-associated disease (CDAD) in hospitalized children. Aim: To describe the frequency, demography, clinical features and outcome of nosocomial CDAD in children admitted to Assiut University Children's Hospital, Egypt. Patients and Methods: In this descriptive cross-sectional study, 72 children developed nosocomial diarrhoea between April 2010 and March 2011. A medical history, clinical assessment and culture for Clostridium difficile and direct toxin detection from stool samples by enzyme immuno-assay were undertaken in all patients. Results: CDAD was diagnosed in 17 (23•6%) patients. Those aged ≤12 months were the most commonly affected (eight, 47%). The main cause of admission was respiratory disorders (eight, 47% of cases), followed by surgical problems (three, 17•7%). Ten patients (58•8%) had severe symptoms. There were no statistically significant differences between any of the demographic or laboratory data for children with CDAD and children with other causes of nosocomial diarrhoea. None of the patients developed complications. Seven children with CDAD (41•2%) had recurrence. Conclusion: CDAD is an important cause of nosocomial diarrhoea in children in Assiut University Children's Hospital. Established guidelines should be followed in all hospitals to minimise exposure to the pathogen. Physicians can do much to reduce the risk of a severe outcome in children by early identification and rapid management. Further research should be undertaken to identify the risk factors for recurrence.

Background: There are no large epidemiological studies of Clostridium difficile-associated disease (CDAD) in hospitalized children. Aim: To describe the frequency, demography, clinical features and outcome of nosocomial CDAD in children admitted to Assiut University Children's Hospital, Egypt. Patients and Methods: In this descriptive cross-sectional study, 72 children developed nosocomial diarrhoea between April 2010 and March 2011. A medical history, clinical assessment and culture for Clostridium difficile and direct toxin detection from stool samples by enzyme immuno-assay were undertaken in all patients. Results: CDAD was diagnosed in 17 (23•6%) patients. Those aged ≤12 months were the most commonly affected (eight, 47%). The main cause of admission was respiratory disorders (eight, 47% of cases), followed by surgical problems (three, 17•7%). Ten patients (58•8%) had severe symptoms. There were no statistically significant differences between any of the demographic or laboratory data for children with CDAD and children with other causes of nosocomial diarrhoea. None of the patients developed complications. Seven children with CDAD (41•2%) had recurrence. Conclusion: CDAD is an important cause of nosocomial diarrhoea in children in Assiut University Children's Hospital. Established guidelines should be followed in all hospitals to minimise exposure to the pathogen. Physicians can do much to reduce the risk of a severe outcome in children by early identification and rapid management. Further research should be undertaken to identify the risk factors for recurrence.

Background: There are no large epidemiological studies of Clostridium difficile-associated disease (CDAD) in hospitalized children. Aim: To describe the frequency, demography, clinical features and outcome of nosocomial CDAD in children admitted to Assiut University Children's Hospital, Egypt. Patients and Methods: In this descriptive cross-sectional study, 72 children developed nosocomial diarrhoea between April 2010 and March 2011. A medical history, clinical assessment and culture for Clostridium difficile and direct toxin detection from stool samples by enzyme immuno-assay were undertaken in all patients. Results: CDAD was diagnosed in 17 (23•6%) patients. Those aged ≤12 months were the most commonly affected (eight, 47%). The main cause of admission was respiratory disorders (eight, 47% of cases), followed by surgical problems (three, 17•7%). Ten patients (58•8%) had severe symptoms. There were no statistically significant differences between any of the demographic or laboratory data for children with CDAD and children with other causes of nosocomial diarrhoea. None of the patients developed complications. Seven children with CDAD (41•2%) had recurrence. Conclusion: CDAD is an important cause of nosocomial diarrhoea in children in Assiut University Children's Hospital. Established guidelines should be followed in all hospitals to minimise exposure to the pathogen. Physicians can do much to reduce the risk of a severe outcome in children by early identification and rapid management. Further research should be undertaken to identify the risk factors for recurrence.

Multidrug resistance (MDR) is associated with fluoroquinolone (FQ) resistance in companion animal Escherichia coli (E. coli). In this study, gyrA, gyrB, parC, and parE quinolone resistance determining regions (QRDR) were sequenced among uropathogenic E. coli isolates with different resistant phenotypes. Also determined were porin, efflux pump and regulatory gene expression based on quantitative real-time reverse transcriptase PCR (qRT-PCR), the impact of efflux pump inhibition (Phe-Arg-β-naphthylamide) and the presence of plasmid-mediated quinolone resistance (PMQR). Using enrofloxacin as the prototypic FQ, we found that (i) the number of mutations in target genes correlate well with minimum inhibitory concentrations (MICs). A single mutation (Ser83Leu) in gyrA increases FQ MIC in susceptible isolates; subsequent mutations result in resistance that increases from low (enrofloxacin MICs 4-16 μg/ml) to high level (enrofloxacin MICs≥128 μg/ml) with each progressive mutation. (ii) as MIC increase, acrB activity and the number of drug classes contributing to the MDR phenotype increases; (iii) a consistent relationship between regulatory gene expression and MIC could not be identified; and (iv) qnrS and aac(6')-Ib-cr gene were detected in 14 and 5 ENR(R)-MDR isolates containing the target mutation, respectively. Of 13 isolates expressing PDR isolates, 10 (77%) were positive for qnrS gene, and 4 (40%) carried both qnrS and aac(6')-Ib-cr gene. These findings demonstrated that MDR-associated FQ resistance in canine and feline uropathogenic E. coli reflects a combination of point mutations, enhanced efflux pump activities, and PMQR mechanisms. Point mutations in DNA gyrase, however, are necessary to achieve a clinical level of FQ resistance.

Escherichia coli respond to selective pressure of antimicrobial therapy by developing resistance through a variety of mechanisms. The purpose of this study was to characterize the genetic mechanisms of antimicrobial resistance in fecal E. coli after the routine use of 2 popular antimicrobials. Fourteen resistant E. coli isolates, representing predominant clones that emerged in healthy dogs' feces after treatment with either amoxicillin (11 E. coli isolates) or enrofloxacin (3 E. coli isolates), were tested for mutations in DNA gyrase (gyrA and gyrB) and in topoisomerase IV (parC) and for the presence of β-lactamases (bla(TEM), bla(SHV), bla(PSE-1) and bla(CTX-M)) and plasmid-mediated quinolone resistance (qnrA, qnrB, qnrS, aac(6')-Ib, and qepA), by polymerase chain reaction. Escherichia coli isolates cultured following amoxicillin therapy only expressed single-drug resistance to β-lactams, while the isolates cultured from dogs receiving enrofloxacin therapy expressed multidrug resistance (MDR). The use of RND efflux pump inhibitors increased the susceptibility of the 3 MDR E. coli isolates to doxycycline, chloramphenicol, enrofloxacin, and ciprofloxacin, which indicates a role of the efflux pump in the acquisition of the MDR phenotype. Amplification and sequencing of AcrAB efflux pump regulators (soxR, soxS, marR, and acrR) revealed only the presence of a single mutation in soxS in the 3 MDR isolates.