Biomedical subjects

E Nagy

Publications and source records attributed to E Nagy.

At least 163 records · Page 9Linked to original sources

PubMed1996

Partial transcriptional mapping of the fowlpox virus genome and analysis of the EcoRI L fragment.

Several fowlpox virus (FPV) DNA fragments were selected by differential hybridization using cDNA of transcripts that were strongly transcribed early and/or later after infection of QT-35 cells. The EcoRI L fragment contained three strongly transcribed FPV genes: L1L, a late 1452 bp partial (amino end) ORF; L2R, an early/late 522 bp ORF; and L3R, a late 948 bp ORF. The protein products of L1L, L2R and L3R shared homology with the products of vaccinia virus (VV) genes H4L (RAP94), H5R (Ag35) and H6R (topoisomerase), respectively, suggesting a conservation of gene structure and order between VV and FPV. The 5' upstream non-coding sequences of L1L and L3R were A + T rich and the sequence 5' TAAATG 3' overlapped the predicted translation start codon. Primer extension analysis of the L2R transcript mapped the transcriptional start sites of early and late mRNAs 14 nt downstream of a VV early promoter-like critical region sequence, AAAATTGAA-AAAAAAA. A VV-like TAAAT late transcriptional element was present 20 nt upstream of the L2R ATG translational start codon. A plasmid with the putative early L2R promoter cloned upstream of the Newcastle disease virus haemagglutinin-neuraminidase (HN) cDNA as a reporter gene was at least 6-fold more effective in generating HN MRNa than plasmids containing the P7.5 or P11 VV promoters in transient expression assays in FPV-infected CEF cells treated with cytosine arabinoside. The L2R promoter was also able to express an amount of HN mRNA equal to that expressed by the VV promoters late in infection.

Amino Acid Sequence↗

PubMed1996

Binding of extracellular matrix proteins to the surface of anaerobic bacteria.

The binding of fibronectin, vitronectin, collagen and sialoprotein to 65 anaerobic strains was investigated by means of latex agglutination tests. The binding of fibronectin, collagen and lactoferrin to the same strains was also tested by means of 125I-labelled proteins. The strains were isolated from abdominal infections (55%), from the faeces of healthy subjects (29%) or from the depths of tonsils removed at tonsillectomy (16%). The binding of fibronectin and collagen to Bacteroides fragilis strains, tested by the latex agglutination assay, was stronger than their binding to other species. The vitronectin binding of the strains was less common, but was always accompanied by fibronectin binding. Binding to fibronectin-coated beads was inhibited by pre-incubation of the bacterial cells with soluble fibronectin and by heat or protease treatment of the bacterial suspension. No inhibition of the binding was observed with carbohydrates. None of the 65 strains exhibited any binding to fetuin or asialofetuin; 8% of the strains had a binding site for mucin. The binding to mucin-coated beads was inhibited by pre-incubation of the cells with mucin. The radiolabelling method indicated a low binding to 125I-fibronectin. The binding of 125I-collagen-I and 125I-lactoferrin was higher for the anaerobic strains tested.

Bacteria, Anaerobic↗

PubMed1996

Development and validation of high-performance liquid chromatographic method for the determination of folic acid.

A high-performance liquid chromatographic (HPLC) method with ultraviolet detection at 278 nm is presented for the determination of folic acid with 5-hydroxyindole-3-acetic acid as internal standard. Chromatographic separation was based on ion-pair HPLC with tetrabutylammonium hydroxide as the ion-pairing compound. The method is sensitive, lower limit of detection was 0.1 ng/ml, the limit of quantitation was 0.25 ng/ml, using a 25-microliter sample volume. The systems precision intra- and interassay CVs for folic acid were 1.4% and 2.5%, respectively.

Chromatography, High Pressure Liquid↗

PubMed1995

The effect of redox-related species of nitrogen monoxide on transferrin and iron uptake and cellular proliferation of erythroleukemia (K562) cells.

The iron-responsive element-binding protein (IRE-BP) modulates both ferritin mRNA translation and transferrin receptor (TfR) mRNA stability by binding to specific mRNA sequences called iron-responsive elements (IREs). The regulation of IRE-BP in situ could possibly occur either through its Fe-S cluster and/or via free cysteine sulphydryl groups such as cysteine 437 (Philpott et al, J Biol Chem 268:17655, 1993; and Hirling et al, EMBO J 13:453, 1994). Recently, nitrogen monoxide (NO) has been shown to have markedly different biologic effects depending on its redox state (Lipton et al, Nature 364:626, 1993). Considering this fact, it is conceivable that the NO group, as either the nitrosonium ion (NO+) or nitric oxide (NO+), may regulate IRE-BP activity by S-nitrosylation of key sulphydryl groups or via ligation of NO. to the Fe-S cluster, respectively. This hypothesis has been examined using the NO+ generator, sodium nitroprusside (SNP); the NO. generator, S-nitroso-N-acetylpenicillamine (SNAP); and the NO./peroxynitrite (ONOO-) generator, 3-morpholinosydnonimine hydrochloride (SIN-1). Treatment of K562 cells for 18 hours with SNP (1 mmol/L) resulted in a pronounced decrease in both the RNA-binding activity of IRE-BP and the level of TfR mRNA. In addition, Scatchard analysis showed a marked decrease in the number of specific Tf-binding sites, from 590,000/cell (control) to 170,000/cell (test), and there was also a distinct decrease in Fe uptake. Furthermore, SNP did not decrease cellular viability or proliferation. In contrast, the NO. generator, SNAP (1 mmol/L), increased RNA-binding activity of IRE-BP, the level of TfR mRNA, and the number of TfRs in K562 cells. Moreover, both SNAP (1 mmol/L) and SIN-1 (0.5 mmol/L) reduced cellular proliferation. The results are discussed in context of the possible physiologic role of redox-related species of NO in regulating iron metabolism.

Antioxidants↗

PubMed1995

[The effect of prolonged acenocoumarol therapy on bone density].

The effect of chronic cumarin treatment on bone mineral content was investigated. Bone mineral density was determined by double photon densitometry (Lunar DPXL). The density data (mean +/- SE) of 45 cardiac patients (age: 57.0 = +/- 6.3 y, body mass index: 26.7 +/- 3.8 kp/m2, cardiac stadium score, according to New York Heart Association: 2-3), had been treated by acenocumarol at least for 2 years (duration of treatment: 75.0 +/- 52 months), were compared to the values of 45 age, body mass index, cardiac status matched patients not treated by anticoagulant. The density values of L2-L4 lumbar regions were lower in the treated group (1.041 +/- 0.17 vs. controlls: 1.13 +/- 0.15 g/cm2, p < 0.05), while no differences in ultradistal ulnar and radial regions were detected. No correlation between bone mineral density and the length, or the dose of the cumarin treatment were observed. This observation suggests the importance of the regular bone densitometry control of cumarin treated patient.

Acenocoumarol↗

PubMed1995

Comparison of the inverted terminal repetition sequences from five porcine adenovirus serotypes.

The nucleotide sequences of the region of inverted terminal repetition from representative strains of all five porcine adenovirus (PAV) serotypes were determined and analyzed. The first 17 nucleotides of this region were identical in PAV-1 to 3 and PAV-5, and 10 bp of identical sequence was found in all the PAVs. The closest relationships were among PAV-1 to 3, which shared more common sequences than the other serotypes. PAV-4 had the longest inverted terminal repeat reported for any adenovirus. The proximal 54-bp AT-rich region was partially conserved and the distal GC-rich region was less well conserved among all five serotypes.

Animals↗

PubMed1995

Glyceraldehyde-3-phosphate dehydrogenase selectively binds AU-rich RNA in the NAD(+)-binding region (Rossmann fold).

A 36-kDa protein that binds AU-rich RNA was purified from human spleen and identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). GAPDH has been previously demonstrated to bind tRNA with high affinity. Competition studies suggested that cytoplasmic GAPDH binds the AU-rich elements (AREs) of lymphokine mRNA 3'-untranslated regions with higher affinity than tRNA. The AUUUA-specific RNA binding activity of GAPDH was inhibited by NAD+, NADH, and ATP in a concentration-dependent manner, suggesting that RNA binding of GAPDH might involve the NAD(+)-binding region, or dinucleotide-binding (Rossmann) fold. This hypothesis was supported by experiments that localized RNA binding to the predicted N-terminal 6.8-kDa peptide, known to be involved in the formation of the NAD(+)-binding domain. The direct demonstration of ARE-specific binding protein activity localized to the NAD(+)-binding region of GAPDH supports the general concept that enzymes containing this domain may exhibit specific RNA binding activity and play additional roles in nucleic acid metabolism. Finally, cytoplasmic GAPDH was found in the polysomal fraction of T lymphocytes. Thus, the RNA binding specificity of GAPDH as well as its localization within the cell merit its strong consideration as a protein important in the regulation of ARE-dependent mRNA turnover and translation in addition to its well described role in glycolysis.

Adenine↗

PubMed1995

Modulation of natural killer cell-mediated cytotoxicity by tamoxifen and estradiol.

BACKGROUND: The nonsteroidal antiestrogenic drug, tamoxifen, inhibits the growth of estrogen receptor-positive tumors by interfering with the growth-stimulatory effect of estradiol. However, there is compelling evidence that tamoxifen treatment also is beneficial for patients with estrogen receptor-negative tumors. The hypothesis that tamoxifen is capable of enhancing the immunologic defense of tumor-bearing hosts was been investigated as a possible method for targeting receptor-negative neoplasms. METHODS: Natural killer (NK) cells in the spleen of female Fisher and Wistar-Furth rats were used against the YAC-1 murine lymphoma target in 51Cr-release assays. The effect of various concentrations of estradiol and tamoxifen (1 nM-1 microM) and of the metabolic inhibitors actinomycin D and cyclohexamide on target-cell killing was investigated. RESULTS: Tamoxifen enhanced and estradiol inhibited killing if applied for the entire 5 hours of the cytotoxic reaction. When applied jointly in this experimental setup, estradiol interfered with the enhancing effect of tamoxifen. Pretreatment of target cells with tamoxifen led to highly significant enhancement of cytotoxicity; estradiol also enhanced target cell killing, but to a lesser extent. After joint treatment, the level of cytotoxicity was comparable with that obtained with tamoxifen alone. Both pharmacologic (100 nM and 1 microM) and physiologic (1 or 10 nM) concentrations of estradiol and equimolar tamoxifen enhanced target cell lysis. However, pharmacological levels of estradiol inhibited effector cells when applied alone or in combination with tamoxifen. Highly significant enhancement of target-cell destruction occurred if both target and effector cells were pretreated with tamoxifen, whereas estradiol treatment of both cell types led to slight enhancement or no effect on cytotoxicity. Treatment of the target cells with actinomycin D or cycloheximide inhibited the lysis of untreated and tamoxifen- or estradiol-exposed cells. Treatment of YAC-1 target cells with tamoxifen or estradiol also enhanced the NK cell-mediated release of the nuclear label, 3H-thymidine, indicating DNA degradation. Similarly treated P815 cells resisted lysis by NK cells, but showed sensitization when the NK cells were stimulated by interleukin-2 for 48 hours before the lytic reaction. Estradiol and tamoxifen changed the kinetics of 3H-thymidine incorporation by YAC-1 cells, but the cells were capable of growing with the highest drug concentrations (1 microM) used in the cytotoxicity experiments. YAC-1 cells have no cytosolic estradiol receptors and are weakly positive for cytosolic progesterone receptors. CONCLUSIONS: These experiments indicate that NK cell-mediated target-cell destruction can be enhanced by tamoxifen primarily through sensitizing the target for lysis. Estradiol also sensitizes the target but inhibits the effector cells simultaneously so that little or no change results in cytolysis. Target-cell sensitization is not mediated by classical estrogen receptors and requires the active metabolic participation of the cells treated. A likely mechanism of this phenomenon is priming the target cell for apoptosis.

Animals↗

PubMed1995

Antibiotic susceptibility of Bacteroides fragilis group strains in Hungary.

Resistance rates to different antibiotics of 495 Bacteroides fragilis group strains were followed between 1987 and 1994 in Hungary. In 1992 the strains were collected in three different laboratories, whereas during the other periods strains were isolated in one centre. Metronidazole, chloramphenicol, imipenem and amoxicillin/clavulanic acid were the most active drugs. A high level of resistance was observed in 1987 for ampicillin (88% at > 4 mg/L), erythromycin (51% at > 4 mg/L), tetracyclin (53% at > 8 mg/L) and clindamycin (27% at > 4 mg/L). The same level of resistance was seen during the further years for clindamycin and ampicillin. Resistance to cefoxitin increased from 6% to 11% between 1987 and 1993/1994. No differences in resistance rates were observed between the strains collected in the three centers. For 100 strains, the results of the E test were compared with those of the micro-broth dilution test, both being used routinely for testing the antibiotic susceptibility of Bacteroides fragilis group strains in this period.

Journal Article↗

PubMed1995

Combined application of in vivo UV-crosslinking and in vitro label transfer in the examination of AU-rich sequence binding protein-RNA interactions.

A combination of in vivo UV light-induced crosslinking of nucleic acids to proteins and in vitro label transfer assay was applied to investigate specific interactions between AU-rich sequences (ARE) in the 3' UTR of lymphokine mRNAs and cytoplasmic AU-rich sequence element binding proteins (AUBP) in normal human lymphoblasts and MLA 144 gibbon lymphoid tumor cells. We demonstrate that a pool of cytoplasmic AUBP can be effectively crosslinked to RNA in vivo, suggesting a close association of these proteins with ARE sequences in the cytoplasm. We also show that the UV-crosslinked AUBP pool is markedly reduced in malignantly transformed MLA 144 cells compared with normal lymphoblasts, indicating weaker interaction between lymphokine ARE and AUBP in these tumor cells. Similar differences in AUBP-RNA associations were found between the membrane-bound polysomal subfractions of the two cell types where most of the AUBP activity was localized. We suggest that the decreased AUBP-mRNA association in MLA 144 cells might reflect a process concerned with disturbances of mRNA metabolism in the neoplastic phenotype.

Animals↗

PubMed1995

Molecular cloning and physical mapping of porcine adenovirus types 1 and 2.

The 25R strain of porcine adenovirus type 1 (PAV-1) and the A47 strain of PAV-2 were propagated in ST cells, and DNA was extracted from the infected cells by a modified Hirt method. The DNA of each virus was digested by each of nine restriction endonucleases, and restriction enzyme fragments representing the entire genome were cloned. The genomic size of each virus was approximately 33 kb. Physical maps for the nine restriction endonucleases were constructed from the results of double digestion and Southern blot hybridization experiments, and oriented with respect to the PAV-3 genome. PAV-1 and PAV-2 were found to be related genetically to PAV-3, and there was a closer relationship between PAV-1 and PAV-3 than between PAV-1 and PAV-2 or between PAV-2 and PAV-3.

Animals↗

PubMed1995

Sequence analysis of putative pVIII, E3 and fibre regions of porcine adenovirus type 3.

Sequence analysis of a region of the genome of porcine adenovirus type 3 from map unit 79.5 to map unit 92 was performed. Homology studies revealed genes coding for the hexon-associated protein pVIII on the left and for the fibre protein on the right of the sequenced region. By analogy with the genomic organization of other adenoviruses, the 1179 bp sequence between the pVIII and fibre open reading frames, extending from map unit 81.3 to map unit 84.7, was identified as the equivalent of the E3 region of human adenoviruses. The deduced amino acid sequence of one of the three open reading frames of the putative E3 region showed homology with the 13.3K E3 protein of canine adenovirus type 2. The primary structure of the putative fibre protein was similar to that described for human adenovirus types 2 and 5, with a 14 pseudorepeat motif in the shaft region of the fibre. A 742 bp tandem repeat starting in the middle of the fibre gene and extending beyond the termination codon of this gene was observed.

Adenoviridae↗

PubMed1995

Restriction endonuclease analysis and physical mapping of the genome of porcine adenovirus type 5.

The HNF61 and HNF70 isolates of porcine adenovirus type 5 (PAV-5) were cultivated in PK-15 cells, and viral DNA was extracted from the infected cells by a modified Hirt procedure. The DNAs were digested by each of 9 restriction endonucleases, and fragments representing the entire genomes were cloned. Based on the sizes of the restriction enzyme fragments, the genome of each isolate was estimated to be 33.2 kb. Physical maps for the 9 restriction endonucleases were constructed. The physical maps of the two isolates were identical for 5 of the restriction endonucleases, but 4 enzymes revealed differences in restriction sites occurring mainly between map units 78 and 83, which may include the E3 region of the genome. There were no similarities between the physical maps of PAV-5 and those described for the other 4 serotypes of PAV.

Animals↗

PubMed1995

Nitroglycerin-induced direct protection of the ischaemic myocardium in isolated working hearts of rats with vascular tolerance to nitroglycerin.

We investigated whether nitroglycerin (NTG) was able to produce an anti-ischaemic effect in isolated working hearts of rats with vascular tolerance to NTG. Hearts isolated from tolerant and non-tolerant rats were subjected to 10 min coronary occlusion in the presence of 10(-7) M NTG and/or its solvent. NTG alleviated ischaemia-induced deterioration of cardiac function and decreased lactate dehydrogenase release whilst having no effect on coronary flow nor the area of the ischaemic zone both in hearts isolated from NTG-tolerant and non-tolerant rats. The magnitude of the effect was similar in the two groups. These results suggest that the anti-ischaemic effect of NTG involves direct myocardial mechanisms independent of its vascular action and that vascular tolerance to NTG does not affect this direct protective action.

Animals↗

PubMed1995

NaCl injections in brain induce natriuresis and blood pressure responses sensitive to ANG II AT1 receptors.

In the present study we tested the hypothesis that the natriuretic and pressor effects of intracerebroventricularly (icv) injected hypertonic saline involve a central angiotensinergic pathway. All experiments were performed in conscious Wistar rats. Bolus injections of hypertonic saline (0.19, 0.23, 0.30, and 0.60 M icv; injection volume 5 microliters) induced a concentration-dependent increase of renal sodium excretion without affecting urinary flow. The increase in renal sodium excretion after the two highest saline concentrations was accompanied by significant increases in mean arterial blood pressure (MAP). Pretreatment with the angiotensin (ANG) AT1 receptor antagonist, losartan (5 micrograms icv), reduced the natriuretic effect of 0.23 and 0.30 M saline but did not affect the natriuresis induced by 0.60 M saline. The increase in MAP after 0.30 and 0.60 M saline icv was markedly attenuated by intracerebroventricular pretreatment with losartan. Our results demonstrate the involvement of a central angiotensinergic mechanism in the natriuretic and pressor responses to hypertonic saline. In addition to the ANG II-mediated natriuresis, an additional natriuretic mechanism, independent of ANG II and associated with the saline-induced pressor effect, seems to be recruited with increasing concentrations of saline in the cerebrospinal fluid.

Angiotensin II↗

PubMed1995

Passive protection of piglets by recombinant baculovirus induced transmissible gastroenteritis virus specific antibodies.

Sera of pigs immunized with parts of the transmissible gastroenteritis virus (TGEV) spike (S) protein expressed by recombinant baculoviruses were tested, together with a TGEV hyperimmune antiserum, for their abilities to protect three-day-old piglets against TGEV infection. The piglets were infected with virulent TGEV and the sera were given orally 3 h before infection, together with the virus, and every 6 h postinfection during the 30 h of the experiment. Virus shedding was monitored by TGEV isolation from rectal swab samples. The sera containing antibodies induced by the complete S protein or the amino terminal half of the S protein showed protective properties, indicated by delayed onset of clinical signs and virus shedding, similar to the TGEV hyperimmune serum. Those immune sera containing antibodies induced by shorter recombinant proteins were not protective.

Animals↗

PubMed1995

Resistance to beta-lactam antibiotics and beta-lactamase production of Bacteroides, Porphyromonas and Prevotella strains.

Resistance to beta-lactam antibiotics of 183 clinical isolates belonging to Bacteroides, Porphyromonas and Prevotella was tested by a micro-broth dilution MIC method. Beta-lactamase production was screened by using nitrocefin sticks. Prevalence of resistance to different beta-lactam antibiotics was higher among the Bacteroides strains other than B. fragilis parallel with a beta-lactamase production (88% and 96%, respectively). Resistance was observed less frequently among Porphyromonas and Prevotella strains. Cefoxitin resistance was 11.5% and amoxicillin/clavulanic acid 3.5% among Bacteroides isolates, whereas no resistance was found to these antibiotics among the Porphyromonas and Prevotella strains. All strains tested were susceptible for imipenem. Beta-lactamase production of selected isolates was tested quantitatively. Beta-lactamase of B. fragilis 1 and B. levii differed in their isoelectric points, substrate profiles and inhibition by clavulanic acid, sulbactam and tazobactam.

Anti-Bacterial Agents↗

PubMed1994

[Experience in the management of patients with artificial heart valves (analysis of the material of the Békés County Cardiology Clinic)].

Authors report of 152 patients, 146 of whom underwent single mitral (n = 70), single aortic (n = 64), double aortimitral (n = 17) and aortic-mitral-tricuspid (n = 1) valve replacement with mechanical prostheses. 6 patients received bioprostheses. The mean age was 55 years. Follow-up ranged from 1 to 23 years (mean 5.1 years). The methodology of follow-up consisted patients examination in our office (at least twice a year) and questionnaire. Valve related complications were defined as paravalvular leak (6 cases), valve thrombosis (3 cases), thromboembolism (13 cases), anticoagulant-related hemorrhage (21 cases), infectious endocarditis (6 cases). During care 14 patients died. The most common causes of death were left-ventricle insufficiency (n = 6), infectious endocarditis (n = 3), prosthetic valve thrombosis (n = 1), cerebral hemorrhage (n = 2) and sudden death (n = 2). 138 patients (56 percent) are retired or invalid. Authors emphasize the importance of regular control and the significance of anticoagulant policy (in the target international normalized ratio to 2.5-3.5 and the provision of the antibiotic prophylaxis to avoid the late complications.

Anti-Bacterial Agents↗