A clinical isolate of Bacteroides fragilis from Hungary with high-level resistance to imipenem.
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Adult↗
Biomedical subjects
E Nagy
Publications and source records attributed to E Nagy.
Bacterial phage receptors, versatile tools for display of polypeptides on the cell surface.
Four outer membrane proteins of Escherichia coli were examined for their capabilities and limitations in displaying heterologous peptide inserts on the bacterial cell surface. The T7 tag or multiple copies of the myc epitope were inserted into loops 4 and 5 of the ferrichrome and phage T5 receptor FhuA. Fluorescence-activated cell sorting analysis showed that peptides of up to 250 amino acids were efficiently displayed on the surface of E. coli as inserts within FhuA. Strains expressing FhuA fusion proteins behaved similarly to those expressing wild-type FhuA, as judged by phage infection and colicin sensitivity. The vitamin B(12) and phage BF23 receptor BtuB could display peptide inserts of at least 86 amino acids containing the T7 tag. In contrast, the receptors of the phages K3 and lambda, OmpA and LamB, accepted only insertions in their respective loop 4 of up to 40 amino acids containing the T7 tag. The insertion of larger fragments resulted in inefficient transport and/or assembly of OmpA and LamB fusion proteins into the outer membrane. Cells displaying a foreign peptide fused to any one of these outer membrane proteins were almost completely recovered by magnetic cell sorting from a large pool of cells expressing the relevant wild-type platform protein only. Thus, this approach offers a fast and simple screening procedure for cells displaying heterologous polypeptides. The combination of FhuA, along with with BtuB and LamB, should provide a comprehensive tool for displaying complex peptide libraries of various insert sizes on the surface of E. coli for diverse applications.
Bacterial Outer Membrane Proteins↗
The effect of heat-inactivated Helicobacter pylori on the blastogenic response of peripheral blood mononuclear cells of patients with chronic urticaria.
BACKGROUND: Helicobacter pylori, the most important etiologic factor of gastritis and peptic ulcer, has recently been associated with several extradigestive diseases. Previous studies reported conflicting results on H. pylori eradication in chronic urticaria, in that some studies showed a benefit, while others found no effect. METHODS: Peripheral blood mononuclear cells of 24 chronic urticaria patients (13 seropositive/11 seronegative for H. pylori) and 18 healthy controls (9 seropositive/9 seronegative) were stimulated with whole heat-inactivated H. pylori (8 x 10(5), 8 x 10(6 )and 8 x 10(7) bacteria/well), phytohemagglutinin (2 microg/ml) and pokeweed mitogen (5 microg/ml). The proliferative response was determined by (3)H-thymidine incorporation. Helicobacter-specific IgG antibody response was determined by ELISA. RESULTS: There were significantly higher proliferative responses to various concentrations of whole heat-inactivated H. pylori antigen in 6- to 7-day cultures of peripheral blood mononuclear cells of chronic urticaria patients compared to healthy controls. We found a tendency to exhibit a higher proliferative response to either Helicobacter antigens or mitogens in seropositive compared to seronegative patients. CONCLUSION: Our results support the hypothesis that there is an increased lymphocyte reactivity in chronic urticaria, perhaps further enhanced by the presence of H. pylori which, therefore, may be involved as a trigger in the pathogenesis of chronic urticaria.
Adult↗
Olfactory deficit in Alzheimer's disease?
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Adult↗
Occurrence of metronidazole and imipenem resistance among Bacteroides fragilis group clinical isolates in Hungary.
During the period between 1987 and 1997, various surveillances of the antibiotic resistance of B. fragilis group isolates revealed that practically all the isolates tested were susceptible to imipenem, metronidazole and chloramphenicol; very few isolates (2.5%) exhibited resistance to amoxicillin/clavulanic acid. However, similarly as in some southern European countries, the percentages of the isolates that were resistant to ampicillin, tetracycline and clindamycin were high throughout this period, and the resistance to cefoxitin increased from 6% to 16%. In 2000, isolates with intermediate or high resistance to imipenem and isolates with increased MICs to metronidazole were emerging among the clinical isolates of B. fragilis. The presence of the cfiA gene was demonstrated by PCR in 7 of 242 isolates (2.9%); 2 of them with high MICs to carbapenems harboured the IS942 element immediately upstream of the resistance genes. In the 2 B. fragilis isolates with increased MICs to metronidazole, the nim gene could be detected by PCR. The IS1186 element was found in these isolates upregulating the metronidazole resistance gene.
Anti-Bacterial Agents↗
Post-antibiotic and synergic effects of fluoroquinolones and ceftazidime in combination against Pseudomonas strains.
Infectious complications are still the leading cause of morbidity and mortality in seriously ill patients [1-3]. To combat the resistance of (mainly Gram-negative, non-fermentative) bacteria, e.g. Pseudomonas aeruginosa, to a wide spectrum of antibiotics, drug combination therapy has been widely adopted as standard clinical practice since the late 1990s. beta-Lactam combinations are not optimal and the potential of nephrotoxicity and ototoxicity from aminoglycosides has caused clinicians to evaluate new possibilities, such as combinations of fluoroquinolones and beta-lactams. We examine here the synergic and post-antibiotic effects (PAEs) of ciprofloxacin, ofloxacin and pefloxacin-ceftazidime combinations against 6 clinical Pseudomonas isolates. The fluoroquinolone-ceftazidime combinations were not only synergic against Comamonas (P.) testosteroni, but had double the PAEs of the two drugs alone. The ciprofloxacin-ceftazidime combination had a longer PAE against P. aeruginosa isolate 1 than ciprofloxacin alone. The combinations, however, did not have longer PAEs than those of the single drugs against the other 5 P. aeruginosa isolates.
Anti-Infective Agents↗
Molecular genetic and traditional methods for detection of Mycobacterium tuberculosis complex (discrepancy analysis).
In the past six and half years, 862 different clinical samples [sputum, bronchoalveolar lavage, thorax puncture, cerebrospinal fluid and skin samples] were tested by Gen-probe amplified Mycobacterium tuberculosis direct test (MTD) or ligase chain reaction (LCR) or polymerase chain reaction (PCR). 239 parallel clinical samples were cultivated, and some samples were stained with Ziehl-Neelsen staining. 1-4 samples were tested per patient. 29 (12.13%) samples were positive and 177 (74.05%) samples were negative with both cultivation and molecular genetic methods. 2 (0.83%) samples were positive only on cultivation, and 31 (12.97%) samples were positive only with the molecular diagnostic methods. The differences are undoubtedly explained by the sensitivity of the molecular diagnostic methods.
Bacteriological Techniques↗
Gender-related physiologic differences in human neonates and the greater vulnerability of males to developmental brain disorders.
OBJECTIVE: To examine whether gender-specific physiologic differences are present at birth and can be a basis for gender-specific vulnerability to developmental disorders in males. We report on three studies of male-female physiologic and structural differences in neonates and their relevance to observed differences in the incidence of developmental disorders in males. METHODS: Study I: 56 neonates were examined for cardiac reactivity to the Moro reflex. Study II: 863 neonates' basic anthropometric data were examined to demonstrate gender-specific differences in body proportions as a possible basis for psychophysiologic differences. Study III: Developmental data on 1000 one- to 26-week-old infants were analyzed for gender-specific developmental differences in rhythmic patterns of sleeping and eating. RESULTS: Study I: There were gender-related differences in heart rate reactivity (male > female). Study II: Male newborns had significantly larger head/chest proportions, suggesting that they may have a greater metabolic demand, related to brain size. Study III: Mothers reported that infant males' sleeping rhythm developed significantly later than females', and that they slept for shorter periods at night. CONCLUSIONS: Gender-related vulnerability in brain development is proposed, based on physiologic differences during a specific early sensitive period in development. This hypothesis may help to explain the overrepresentation of males reported for most developmental disorders.
Analysis of Variance↗
The relationship between the occurrence of undifferentiated bovine respiratory disease and titer changes to bovine coronavirus and bovine viral diarrhea virus in 3 Ontario feedlots.
Serological evidence of previous viral exposure (titer at arrival) and current viral exposure (titer increase) during a 28-day study period, was used to determine if bovine coronavirus (BCV) or bovine viral diarrhea virus (BVDV) was associated with the occurrence of undifferentiated bovine respiratory disease (UBRD) in feedlot calves. Neutralizing antibody titers to BCV and BVDV were determined for 852 animals from 3 Ontario feedlots. Calves at 2 of the 3 feedlots (n = 753) received a modified live 4-way viral vaccine containing BVDV. On arrival at the feedlots, 90% of animals were seropositive for BCV, while 39% of animals were seropositive for BVDV. This evidence of previous exposure to both viruses was associated with reduced subsequent UBRD risk. Evidence of exposure to BCV during the study period was common, as 50% of animals showed a 16-fold or greater titer increase; however, treatment for UBRD was not associated with titer change. Although the majority of animals were vaccinated for BVDV at arrival, within a feedlot, animals treated for UBRD had larger titer increases to BVDV than non-treated animals. Based on our findings we infer that BCV was not causally related to UBRD occurrence, however consistent with other literature, BVDV may be causally related to UBRD occurrence.
Animals↗
Temporary cerebral histic hypoxia in simulated microgravity.
The authors determined in their previous examinations that the vertical component of diagonal optokinetic nystagmus (DOKN) decreases considerably more in the conditions of hypoxia and antiorthostatic posture than does the horizontal component. Relying on these findings, they reasoned that the tectal and pretectal gaze centers, which organize vertical eye movement, are more sensitive to both hypoxia and antiorthostatic posture than is the pontine gaze center, which directs horizontal eye movement. In their present examinations, the authors sought to discover how cranial shifting of body fluids alters the DOKN and cardiopulmonary dynamics. They established that, at-30-degree antiorthostatic posture during 3 hours' loading, the diminution of pulse rate and systolic blood pressure is accompanied by continuous enhancement of diastolic blood pressure, diminishment of the carbonometric value, and augmentation of respiratory rate. The disturbances of DOKN, which mimicked previous similar findings under conditions of hypoxia and antiorthostatic posture, may be caused by the hypocapnia that is induced by antiorthostatic posture and that produces histic hypoxia. However, hypocapnia does not exclude the possibility of cerebrovascular injury, which was detected during animal experiments in microgravity or in simulated microgravity. To resolve this issue ultimately, further investigations are necessary.
Adolescent↗
[Coronary revascularization on beating heart].
At authors clinic 563 coronary artery bypass graft procedures were done from 1. January 1998 to 15. September 1999. 134 coronary revascularizations were performed from the same surgical team on beating heart using the same surgical technic at normothermia. Out of them, 98 operations were done with and 36 without ECC support respectively. Patients were operated on with this sophisticated technic exclusively during this period. Conversion to cardiac arrest was not necessary in any case. Male/female ratio was 76/58, 84% of the patients suffered from one or more preoperative myocardial infarction. The average bypass number was 2.7/patient. Arterial revascularization was performed in 94%. Postoperative mortality was 2.2% (3 pts). The postoperative inotropic demand, myocardial infarction rate (CK-MB release) and the amount of bleeding was very low. Mean postoperative stay in the ICU decreased to the average of 19 hrs. Patients normally left hospital on the 7th postoperative day. This operative technique--performed by an experienced surgeon--is less invasive for the heart and other organ systems, the incidence of complications can be decreased and the principle of complete revascularization is not compromised.
Coronary Artery Bypass↗
Identification of the NAD(+)-binding fold of glyceraldehyde-3-phosphate dehydrogenase as a novel RNA-binding domain.
There is growing evidence that metabolic enzymes may act as multifunctional proteins performing diverse roles in cellular metabolism. Among these functions are the RNA-binding activities of NAD(+)-dependent dehydrogenases. Previously, we have characterized the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an RNA-binding protein with preference to adenine-uracil-rich sequences. In this study, we used GST-GAPDH fusion proteins generated by deletion mutagenesis to search for the RNA binding domain. We established that the N-terminal 43 amino acid residues of GAPDH, which correspond to the first mononucleotide-binding domain of the NAD(+)-binding fold is sufficient to confer RNA-binding. We also provide evidence that this single domain, although it retains most of the RNA-binding activity, loses sequence specificity. Our results suggest a molecular basis for RNA-recognition by NAD(+)-dependent dehydrogenases and (di)nucleotide-binding metabolic enzymes that had been reported to have RNA-binding activity with different specificity. To support this prediction we also identified other members of the family of NAD(+)-dependent dehydrogenases with no previous history of nucleic acid binding as RNA binding proteins in vitro. Based on our findings we propose the addition of the NAD(+)-binding domain to the list of RNA binding domains/motifs.
Amino Acid Sequence↗
[Potentials of screening for Chlamydia trachomatis in Hungary: cost-benefit analysis].
Untreated chlamydial infections of the genital tract may cause a wide range of serious complications, which could be prevented by screening. Cost-effectiveness analysis based on previously reported epidemiological study among pregnant women was carried out to develop and evaluate cost-effectiveness of age-based screening (women younger than 20 years old) for Chlamydia trachomatis in Hungary. Three kind of screening strategies were used: a) no screening, b) screening by using ELISA test and c) screening by amplified Gen-Probe diagnostic test. Young age and unmarried status were significant predictors of the chlamydial infection. Prevalence of chlamydial infection was 11.4% in the group of age younger then 20 years. Strategy b) was more cost-effective than neither testing nor treating. Strategy c) was less cost-effective than non screening, unless certain conditions were satisfied. If the cost of the diagnostic test were less or equal than 2,000 HUF or the prevalence of infection in women were greater than 17%, screening strategy c) would be more cost-effective than no screening. Although screening by using ELISA test was more cost-effective than screening by amplified Gen-Probe test, additionally 10,000 infected cases could be prevented using amplified Gen-Probe method for screening Chlamydia trachomatis.
Adolescent↗
Immunogenicity of porcine transmissible gastroenteritis virus spike protein expressed in plants.
Transgenic plants expressing recombinant proteins from pathogenic microorganisms provide an inexpensive edible vaccine for induction of local immunity. Three transgenic plant lines were generated expressing the spike (S) protein of transmissible gastroenteritis virus (TGEV), a protein crucial for establishing mucosal immunity. All three of them were driven by a strong plant promoter. One construct contained the 3.7 kb 5' end of the native S gene sequence. In the second construct part of the S gene, from nucleotide 49 to 1785, was modified for optimal plant recognition and was fused to a plant signal peptide coding sequence. The third construct contained the D epitope-coding region of the S gene, from nucleotide 1201 to 1591, which was fused to the alfalfa beta-amylase gene. The S gene products were detected by enzyme-linked immunosorbent assay (ELISA) and Western blotting. Antigens from all three transgenic plant lines induced TGEV-specific immune responses in pigs as determined by virus neutralization and ELISA, and the resultant antibody titers for all three constructs were similar.
Animals↗
The role of anaerobic bacteria in prostatitis.
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Adult↗
The role of Chlamydia trachomatis in asymptomatic and symptomatic urogenital infections.
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Chlamydia Infections↗
Apoptosis inhibitory effect of the isothiourea compound, tri-(2-thioureido-S-ethyl)-amine.
Sulfhydril compounds, some of them with immunomodulatory activity have also been shown to modulate the induction of apoptosis. This study was performed to assess the possible apoptosis inhibitory effect of the immunomodulatory compound tri-(2-thioureido-S-ethyl)-amine (K-1) in U-937 and HL-60 leukaemia cell lines as model systems. Treatment of U-937 and HL-60 cells with K-1 inhibited etoposide (ETO)-induced apoptosis in both cell lines in a dose-dependent manner, IC(50)=200 microg/ml. The results indicate that inhibition of ETO-induced apoptosis occurs downstream of ETO-mediated cleavable complex formation but early in, or at the level of the mitochondria events of the apoptotic pathway. Further, like other isothioureas, K-1 proved to be a potent inhibitor of nitric oxide synthase. This raises the possibility that where the activation of nitric oxide synthase is involved in apoptosis induction, K-1 might also be effective. These findings suggest that K-1 may serve as a potent inhibitor of apoptosis initiated by ETO or nitric oxide.
Animals↗
Sequence analysis and deletion of porcine adenovirus serotype 5 E3 region.
A 3000 basepair (bp) region corresponding to the E3 region, the flanking pVIII and part of the fiber protein genes, of the two prototype strains (HNF-61 and HNF-70) of porcine adenovirus serotype five (PAdV-5) was sequenced. A potential E3 promoter and poly-A signals were identified. The size of the E3 region was 2039 (strain HNF-61) and 2020 bp (strain HNF-70) the largest E3 so far reported among PAdVs. Three open reading frames (ORF2-4) were identified within the E3 region. Based on the predicted amino acid (aa) sequences ORF2 was similar to other adenovirus E3 ORFs, ORF3 showed some similarity to a bovine adenovirus (BAdV-1) ORF. ORF4 was unique to PAdV-5. E3 mRNA transcripts were detected early in infection by Northern blot analysis. Genomic clones of HNF-70 with a 1505 or 1237 bp deletions in the E3 region were constructed to map non-essential regions. After transfection of the DNA into swine testicle cells, virions were recovered for only the shorter 1237 bp deletion. At least 60% of the E3 region was not essential for virus replication, bringing the theoretical vector capacity of a helper independent PAdV-5 to 2.9 kb.
Adenovirus E3 Proteins↗