Reverse transcription PCR assays for detection of borna disease virus in psychiatric patients

Borna disease virus [BDV] is a virus that naturally infects the central nervous system [CNS] of broad range of warm-blooded animals. BDV is an enveloped virus, non- segmented, negativestranded RNA genome and has an organization characteristic of a member of Bornaviridae. It may persists in the central nervous system [CNS] of infected individuals for their entire life span. In the present work the presence of BDV p24 RNA in peripheral blood cells was detected from 37 psychiatric patients [15 patients diagnosed as schizophrenia, 10 as Major depressive disorder and 12 as mood disorder bipolar I most recent episode mania]. Patients were selected from the outpatient as well as the inpatient units of Psychiatric Department in Mansoura University hospital and 37 healthy volunteers as the control group. All subjects were interviewed by structured diagnostic criteria categorized according to the Diagnostic and Statistical Manual of Mental Disorders DSM-IV. The presence of BDV p24 RNA was investigated by nested reverse transcriptase PCR [RT-PCR] using specific primers to p24 from BDV. The median duration of illness was 6, 4 and 10 years in depressive disorder, mood disorder and schizophrenia respectively. The mean age was 40.7, 30.1 and 36.1 in depressive disorder, mood disorder and schizophrenia respectively. There were no significant differences in age and duration of illness among patients groups with psychotic disorders in the presence or absence of p24 RNA of BDV. Frequency of BDV- RNA on patient's groups was 10.8% [4/37]. The detection of BDV-RNA in the peripheral blood cells of patients but not on control group should help our understanding of the pathogenesis in the disease

Characterization of nosocomial methicillin resistant staphylococcus aureus strains with B-lactam-induced vancomycin resistance, isolated from mansoura university hospitals

Background: beta-lactam antibiotics have often been used together with vancomycin, and this treatment has been a common combination for long time, beta - lactam-induced vancomycin-resistant methicillin-resistant Staphylococcus aureus [MRSA; BIVR] is a subtype of MRSA, which is otherwise vancomycin susceptible that shows vancomycin resistance in the presence of beta -lactam antibiotics

Objective: In our study, we aimed to characterize and evaluate the antimicrobial profile of BIVR isolated from Mansoura University Hospitals [MUH] and prove the association between usage of fi-lactam antibiotics and BIVR development

Patients and Methods: We studied 40 MRSA strains, identified by detection of MeCA gene and they were selected to be vancomycin sensitive with minimum inhibitory concentration [MIC] [< 2microg/ml] by subjecting these strains to simplified population analysis, p-lactamase activity was tested by nitrocefin test. BIVR strains were subjected to time kill study

Results: eight of 40 MRSA strains isolated from different nosocomial infections in MUH were BIVR [20.0%]. Wound infection is the common infection from which BIVR strains were isolated [75.0]. Only one strain [12.5%] of BIVR strains is J3 lactamase producer, while the majority of non-BIVR strains were fi lactamase producer [93.8%] [P<0.05]

Conclusion: BIVR phenomenon is induced only in the presence of fi-lactam antibiotics, proving that BIVR cells seem to have a low level of fi-lactamase activity compared with that ofnon-BIVR MRSA

Prevalence of metallo-beta-lactamase producing acinetobacter species carrying Bla in mansoura university specialized medical hospital

Background: Acinetobacter species are emerging as an important nosocomial pathogen. Multidrug-resistant Acinetobacter spp. has limited the option for effective treatment. Although carbapenems are effective for the treatment of such infections, resistance to this drug has recently been reported. Carbapenems are therapeutic choice against infections caused by gram-negative bacilli including strains of Acinetobacter baumannii. The production of Metallo-p-lactamases [MBLS] is one of the resistance mechanisms of Acinetobacter species

Objective: we aimed to determine the prevalence of MBLS in Acinetobacter and to determine the NDM.j gene as the most recent gene detected in Acinetobacter

Patients and Methods: We studied 18 MBLS-producing Acinetobacter strains, that well identified Susceptibility to various antimicrobial agents was determined by the disc-diffusion method. Minimal inhibitory concentration [MIC] was done by E test technique. MBLS production was detected by using double disc diffusion synergy test [DDST]. DNA templates were prepared from clinical isolates for NDM. i detection

Results: In total, 18 Acinetobacter spp. were isolated from 267 culture-positive samples from hospitalized patients and were identified up to species level as A. baumannii [n=15], A. Iwoffii [n=2] and A. calcoaceticus [n=l]. Acinetobacter spp. represent 6.7% of nosocomial infections while S. aureus represent the most isolated nosocomial pathogen during the period of the current study. Eleven [61.1%] of the Acinetobacter spp. isolates were found to be phenotypically MBLS producers as detected by DDST. MBLS Acinetobacter spp. were isolated from 3 wound swabs, 3 urine samples, 2 blood specimens, 2 sputum specimens, and I endotracheal tube aspirate specimens, during the study period. None of MBLS Acinetobacter spp. isolates was sensitive to neither imipenem nor meropenem, whereas 36.4% were sensitive to amikacin. MBLS production was detected in 11 isolates from total 18 Acinetobacter. MBLS production was found only in A. baumannii [11/15]; also, NDM.j was identified by PCR in only two isolates of MBLS A. baumannii [2/11]

Conclusion: MBLS-producing Acinetobacter species in this hospital is alarming and reflect excessive use of carbapenems and selective antibiotic pressure. Therefore, a strict antibiotic policy should be followed

Transforming growth factor-beta2 and connective tissue growth factor levels in primary adult glaucomas

To elucidate the role of cytokines in pathogenesis of glaucoma, we assessed the transforming growth factor-beta2 [TGF- beta2] and connective tissue growth factor [CTGF] in the aqueous humor of eyes with primary adult glaucoma and the relation to filtering bleb development after trabeculectomy. Thirty patients with primary adult glaucoma and twenty patients with senile cataract of matched age and gender were rolled in the present study. Patients were classified into 3 groups; group I comprised 20 patients with senile cataract, group II comprised 15 patients with primary open-angle glaucoma [POAG], and group III comprised 15 patients with primary angle closure glaucoma [PACG]. At the time of elective surgery, aqueous humor samples were obtained. Aqueous humor samples were kept frozen till time of assay of TGF- beta2 [ELISA] and CTGF [ELISA] and another blood sample for TGF- beta2 gene determination. There were significant increase in aqueous humor levels of total TGF- beta2, and mature TGF- beta2 in POAG eyes compared to the corresponding value of eyes with senile cataract and PACG. Moreover, CTGF level in aqueous humor of eyes with POAG was significantly higher than in eyes with senile cataract and PACG. Following trabeculectomy, POAG eyes with favorable bleb development had normal or low mature TGF- beta2 levels. There is increased expression of TGF- beta2 gene in POAG cases when compared to PACG and control groups. Increased levels of aqueous humor total TGF- beta2 and mature TGF-beta2 and CTGF may play an important role in the pathogenesis of POAG. Moreover, the elevated levels of mature TGF- [32 may be a factor in the development of patent or scarred filtering bleb after trabeculectomy

Bacterial DNA and its consequences in patients with cirrhosis and culture negative, non neutrocytic ascites

The detection of bacterial DNA [BactDNA] in serum and ascitic fluid [AF] from patients with liver cirrhosis and ascites is interpreted as molecular evidence of intestinal bacterial translocation [BT] and considered sufficient to activate the cellular immune response leading to greater cytokine synthesis. In the present work we study whether BactDNA and Tumor necrosis factor-alpha [TNF- alpha] in cirrhotic patients with culture negative, non neutrocytic ascites have been implicated in various complications of cirrhosis such as hepatorenal syndrome [HRS], spontaneous bacterial peritonitis [SBP] and mortality. We studied 34 patients with liver cirrhosis and culture negative, non neutrocytic ascites [22 patients without BactDNA [group I] aged [48.3 +/- 7.85y] and 12 patients with BactDNA [group II], aged [49.7 +/- 6.5y]]. Full history and complete clinical examination were done with the following investigations in the first admission and subsequent admissions during follow up for 24 weeks: complete blood picture, S. creatinine, S. bilirubin, S. albumin, S. transaminases [AIT and AST], AF and plasma TNF-alpha, AF protein and polymorphnuclear leucocytes [PMNL], both blood culture and AF aerobic and anaerobic cultivation, and detection of blood and ascitic fluid BactDNA using PCR. Plasma and ascitic TNF-alpha were significantly higher in cirrhotic patients with compared to those without BactDNA during first admission [54.5 +/- 22.56 vs 35.2 +/- 17.97; 123.2 +/- 49.32 vs 82.6 +/- 29.58 pg/ml respectively, P<0.05]. These changes became highly significant at the end of follow up of both groups [119.3 +/- 27.19 vs 40.2 +/- 16.08; 518.8 +/- 91.11 vs 97.6 +/- 17.81 pg/ml respectively, P<0.001]. There is non significant change of plasma and ascitic TNF-alpha in group I at first admission compared to those at the end of follow up [P>0.05]. However, in group II, there is highly significant increase in both plasma and ascitic TNF-alpha at the end of follow up compared to those at the first admission [P<0.001]. The relative risk of deaths, HRS and SBP were higher in patients with compared to those without BactDNA after follow up for 24 weeks [2.73, 27.37 and 18.18 respectively]. There were significant positive correlation between both plasma and ascitic TNF-alpha and each of serum creatinine and PMNL in the studied patients at the end of follow up. [r= 0.590, p= 0.002 ; r= 0.535, p= 0.005 ; r=0.499, p=0.009 ; r= 0.589, p= 0.002, respectively]. -Patients with BactDNA had more advanced liver disease after 24 weeks follow up compared to patients without BactDNA. We conclude that cirrhotic patients with culture negative, non neutrocytic ascites and BactDNA have significant higher level of AF and plasma TNF-alpha and higher risk of HRS, SBP and morality compared to those without BactDNA during follow up for 24 weeks which could suggest that both BactDNA and TNF-alpha have been implicated in these complications of liver cirrhosis

Nosocomial infections caused by klebsiellae species with plasmids carrying bla shv-5 extended-spectrum beta-lactamase gene in Mansoura university hospitals

SHV-5 is a variant of SHV-1 and it is considered now one of the most important ESBL enzymes produced by Klebsiellae species. In this study we aimed to detect the prevalence of plasmid encoded SHV-5 among Klebsiellae strains causing nosocomial infections in Mansoura University Hospitals [MUH]. One hundred and seventy four Klebsiellae strains were isolated from overall 680 cases of nosocomial infections [25.59%] acquired within MUH over 4 months period from July to November 2004. One hundred and thirty six isolates [78.16%] of them were K. pneumoniae and 38 isolates [21.84%] were K. oxytoca. MICs [micro g/ml] of the isolated strains was done for augmentin, cefoperazone and ceftazidime using E test. One hundred and forty one [81.03%] of them were beta- lactamase producer as detected by nitrocefin discs, where 77 isolates from ?-lactamase producing strains were ESBL producers constituting 44.25% of Klebsiellae isolates. Sixty six ESBL producing strains of total 77 were isolated from cases of blood stream infections [85.71%]. Sixty five ESBL producing strains were isolated from neonatal intensive care unit [NICU] [84.42%]. All ESBL producing strains [n=77] posses at least one large plasmid > 23 kbp. SHV-5 gene was amplified by PCR after plasmid isolation from ESBL producing Klebsiellae isolates, reveal that 68 isolates [88.31%] were harbored SHV-5 gene on their large plasmids. In conclusion we found that the SHV-5-producing K. pneumoniae isolates were recovered from different wards of Mansoura University Hospital during the studied period. Thus, it was hypothesized that one clone may have persisted in that hospital. We recommend that infection control measure of endemic ESBL producers should include : the consumption of the broad-spectrum cephalosporins needs to be restricted to reduce the selection pressure which enables the proliferation of ESBL producers in hospital, continuous application of infection control program as; surveillance, hand washing and contact isolation procedure