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Biomedical subjects

J Lesko

Publications and source records attributed to J Lesko.

At least 19 recordsLinked to original sources
PubMed2001

Isotachophoretic determination of naproxen in the presence of its metabolite in human serum.

An isotachophoretic method with conductivity detection was developed to determine naproxen in the presence of its metabolite 6-O-desmethylnaproxen in human serum. The leading electrolyte contained 10 mM hydrochloric acid, beta-alanine, pH 4.0 and 0.1% methylhydroxypropylcellulose. The terminating electrolyte was 10 mM 2-(N-morpholino)ethanesulfonic acid-tris(hydroxymethyl)aminomethane, pH 6.9, containing 20% (v/v) of ethanol. Naproxen was determined in serum supernatant after simple deproteination of the sample with ethanol. The isotachophoretic results were compared with those obtained by synchronous fluorescence spectrometry.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2000

Leptin responses to glucose infusions in obesity-prone rats.

The secretion of leptin is dually regulated. In fasting animals, plasma leptin concentrations reflect body fat stores, whereas the incremental leptin response to fasting or refeeding most likely reflects insulin-mediated energy flux and metabolism within adipocytes. Impaired secretion of leptin in either pathway could result in obesity. We therefore measured plasma leptin concentrations in fasted animals and plasma leptin concentrations after an intravenous glucose infusion in a rat model of obesity. Young Sprague-Dawley (S-D) and Fischer 344 (F344) rats had similar percent body fat and fasting glucose and fasting leptin concentrations. However, F344 animals had higher insulin concentrations and leptin responses to intravenous glucose than did the S-D animals. The animals were then fed a control or high-fat diet for 6 wk. High-fat fed animals gained more weight and body fat than did the control fed animals. Control and high-fat fed F344 animals gained approximately 40% (P < 0.0001) more weight and >100% (P < 0.01) more body fat than did the S-D animals. Fasting leptin concentrations and leptin concentrations after intravenous glucose infusions and feeding were more than double (P < 0.05) in F344 animals compared with S-D animals. Whether an animal is fed a control or high-fat diet had little effect on the leptin response to intravenous glucose. In conclusion, young, lean F344 animals, before the onset of obesity, demonstrated a greater acute leptin response to intravenous glucose than similarly lean S-D animals. After a 6-wk diet, F344 animals had a greater percent increase in body weight and insulin resistance and exhibited higher fasting leptin concentrations and a greater absolute leptin response to intravenous glucose compared with the S-D animals. The chronic diet (control or high fat) had little impact on the acute leptin response to intravenous glucose. F344 animals exhibit leptin resistance in young, lean animals and after aging and fat accumulation.

Absorptiometry, Photon↗
PubMed1999

Enhanced proliferation and progesterone production by porcine granulosa cells cultured with pseudorabies virus growth factor (PRGF).

The objective of this research was to study possible interactions of pseudorabies virus growth factor (PRGF) with ovarian tissue. Granulosa cells isolated from porcine ovaries were cultured as monolayers for 6 days in a control medium without PRGF and in medium supplemented with different doses of this agent. Increased population density and change towards more fibroblastic-like shape of cells cultured with 10(9) I.U PRGF was observed when compared with control culture. The cells divided significantly faster during 6 days of culture under the influence of 10(3), 10(4), 10(5), 10(6), 10(7), 10(8) and 10(9) I.U./ml of PRGF at a dose dependent manner. PRGF in a dose 10(9) I.U. added to cultured cells isolated from small and medium follicles did not influence progesterone secretion . An increase of progesterone secretion under the influence of PRGF in all investigated days of cultures was observed in cells isolated from large preovulatory follicles. The marked increase in progesterone content in PRGF treated culture in doses of 0.5x10(7), 0.5x10(8), 0.5x10(9) I.U. was observed during 4 and 6 days of culture. The rise of progesterone content was not connected with increased number of secretory cells, but with a stimulation of production per cell. PRGF exerted no visible effect on progesterone secretion by granulosa cells from small and medium follicles cultured for 6 days. The presented in vitro data provide evidence for a local action of PRGF in the follicle depending on the stage of follicular development and duration of exposure. Precise relevance of the interaction of PRGF with follicular development requires further study.

Animals↗
PubMed1999

Dual effect of pseudorabies virus growth factor (PRGF) displayed on actin cytoskeleton.

Pseudorabies virus growth factor (PRGF) was shown to possess transforming activity as well as transformation repressing activity in in vitro systems. In order to better understand these phenomena we studied actin cytoskeleton and its alterations induced by PRGF using normal human fibroblasts VH-10 and transformed cell line HeLa. For specific detection of filamentous actin cells were stained with phalloidin conjugated with fluorescein isothiocyanate (FITC)-phalloidin. PRGF was applied to VH-10 cells for various length of time from 10 min up to 48 h. The effect was very fast and changes in actin filament composition could be detected already after 10 min. In comparison to untreated cells the staining of treated cells was more diffuse and a number of actin microfilaments in individual stress fibers became reduced. After 30 min thick short actin bundles appeared in the perinuclear region. A 24-h exposure resulted in a large reduction of actin bundles. After additional 24 h a partial restoration of actin cytoskeleton in cells was observed. In transformed HeLa cells PRGF induced opposite process than in normal cells: the number of filamentous actin structures increased. We hypothesise that PRGF may act as a transcription-like factor and may initiate changes in gene expression which consequently result in actin cytoskeleton alterations.

Actins↗
PubMed1997

Production of IL-5 and granulocyte-macrophage colony-stimulating factor by naive human mast cells activated by high-affinity IgE receptor ligation.

BACKGROUND: The late-phase allergic reaction is an eosinophilic inflammatory response that begins several hours after allergen exposure, may persist for 24 hours, and is an important pathogenic mechanism in allergic disease. OBJECTIVE: Cultured naive human mast cells were used to investigate whether mast cells are a direct source of the eosinophil-promoting cytokines IL-5, IL-3, and granulocyte-macrophage colony-stimulating factor (GM-CSF). METHODS: Naive human mast cells were derived from bone marrow mononuclear cells cultured in the presence of stem-cell factor. Cytokine message and protein production in response to high-affinity IgE receptor ligation of cultured mast cells were measured by semiquantitative polymerase chain reaction and ELISA, respectively. RESULTS: IL-5, IL-3, and GM-CSF messenger RNA increased within 2 hours of mast cell activation, with IL-5 and GM-CSF message remaining elevated for 24 hours, whereas IL-3 mRNA rapidly declined. IL-5 and GM-CSF protein were measurable 4 to 6 hours after stimulation and peaked by 24 and 12 hours, respectively. IL-3 protein was not detectable. CONCLUSION: These findings demonstrate that naive mast cells do not constitutively produce IL-5 or GM-CSF protein but are a major source of these eosinophilotropic cytokines on high-affinity IgE receptor ligation.

Base Sequence↗
PubMed1997

Oliguria.

Oliguria is a common finding in adult patients in hospitals and carries a significant morbidity if its origin is not corrected promptly. The differential diagnosis of oliguria is broad; thus a solid knowledge of its causes is required for advanced practice nurses to diagnose oliguria correctly.

Adult↗
PubMed1996

Application of porous carbon for solid-phase extraction of dicarboxyimide fungicide residues from wines in combination with high-resolution capillary gas chromatography and gas chromatography-mass spectrometry.

Solid-phase extraction with a novel porous carbon sorbent CARB GR was used for the clean-up step of dicarboxyimide fungicides residues from variety of Slovak grape wines with subsequent capillary gas chromatography-flame ionization detection, -electron-capture detection (ECD) and -mass spectrometry-ion-trap detection (MS-ITD) analysis. Recovery was tested at various concentration levels of vinclozolin and iprodione in standard solutions (R = 80-97%, R.S.D. < or = 5). The value of recovery in spiked wines is dependent on concentration level (studied in the range of 5.9 micrograms/1-1.96 mg/l) and on the variety of wine (R = 80-96%; R.S.D. = 3-5%). Limits of quantitation (for sample volume 50 ml) were determined to be with GC-ECD for both fungicides in ppt range and with GC-MS-ITD in the multiple ion detection mode monitoring in ptt range for vinclozolin and ppb range for iprodione. Concentration levels of vinclozolin residues were determined in treated wines (with 0.1% Ronilan 50 WP) as well as iprodione residues (with 0.15% Rovral 50 WP) and a strong dependence on the protective term before the harvest is shown.

Aminoimidazole Carboxamide↗
PubMed1996

Clinical experience with preimplantation genetic diagnosis of cystic fibrosis (delta F508).

Preimplantation genetic diagnosis (PGD) was attempted in 12 couples in whom both parents carry the common delta F508 deletion causing cystic fibrosis (CF). In vitro fertilization (IVF) was followed by cleavage stage biopsy on days 2 and 3 and removal of one or two cells for genetic analysis by nested polymerase chain reaction (PCR) and heteroduplex formation. A total of 18 cycles resulted in 137 normally fertilized embryos, of which 115 developed to cleavage stages and 114 were successfully biopsied. Genetic analysis was successful in 83 embryos (73 per cent). With the remaining embryos, either results from two or more cells were discordant or amplification failed. In 15 cycles, one or two either normal or carrier embryos were transferred and five (33 per cent) clinical pregnancies were established. Five singletons have been born and at birth all five babies have been confirmed as homozygous for the normal allele. Our experience demonstrates that IVF and cleavage stage biopsy consistently provides sufficient embryos, diagnosed as unaffected, for transfer in this autosomal recessive disease and that pregnancy rates are comparable to those following IVF.

Adult↗
PubMed1996

Analysis of metalaxyl residues in wines by SPE in combination with HRCGC and GC/MS.

Solid phase extraction (SPE) with the porous carbon sorbent CARB GR was used for the preconcentration of metalaxyl residues from a variety of Slovak grape wines with subsequent capillary GC and GC/MS analysis. Recovery was tested at various concentrations of metalaxyl in standard solutions (recovery, R > or = 92%, relative standard deviation RSD < or = 4.3%) and in wines. The value of recovery in spiked wines was dependent on the concentration (studied in the range 0.02-1.96 mg/l) and on the variety of wine (R = 80-99%; RSD = 2-7%). Limits of quantitation (for a sample volume of 50 ml) were determined to be 0.75 microgram/l with GC-FID and 0.50 microgram/l with GC/MSITD. Concentration levels of metalaxyl residues were determined in treated wines (with 0.25% Ridomil plus 48 WP) and a strong dependence on the protective term before the harvest was shown.

Alanine↗
PubMed1994

Morphological transformation of Syrian hamster embryo cells by pseudorabies virus related growth factor.

Syrian hamster embryo (SHE) clonal morphological transformation assay has been used for the in vitro testing of transforming activity of Pseudorabies virus (PRV) related growth factor (PRGF). It was found that PRGF induces morphologically transformed colonies of SHE cells in the wide titer scale (in the range 1 x 10(7) to 1 x 10(13) U/ml). The concentrations of PRGF which induced the transforming phenotype of SHE cells in the colonies did not cause significant cytotoxic effect.

Animals↗
PubMed1994

Determination of the antigenic variability of pseudorabies virus field isolates with monoclonal antibodies.

Five stable hybridoma cell lines secreting monoclonal antibodies (MAbs) were obtained by fusing spleen cells of pseudorabies virus (strain BUK) immunized BALB/c mice with mouse myeloma cell line SP2/0. Plate-trapped and antibody-trapped antigen ELISAs were done to compare the interaction of MAbs with TOP and "K" strains and 7 Slovak field isolates. Only 2 Slovak isolates reacted with all the MAbs, 3 MAbs reacted with TOP and "K" strains. One Slovak isolate gave no reaction with any MAbs.

Animals↗
PubMed1994

Pseudorabies virus growth factor can be resolved into two active components.

Pseudorabies virus (PRV) growth factor (PRGF) which induces a transformed phenotype in normal MK-2 cells and represses the transformed phenotype of Hela cells was partially purified and resolved into two components (M(r) < 300 and < 180). Each of the PRGF components retained the transforming activity of the original factor in MK-2 cells but lost its transformation-repressing activity in Hela cells. The latter activity of PRGF could be reconstituted by simultaneous application of its two components. Two monoclonal antibodies against gII glycoprotein of PRV were able to neutralize both PRGF activities, thus supporting the previously suggested hypothesis that the PRV gene for glycoprotein gII might be involved in PRGF synthesis.

Animals↗
PubMed1993

Canine distemper virus replication in cells on microcarriers.

Chick embryo and Vero cells were grown on Gelaspher M gelatin microcarriers in suspension culture. The microcarriers had no adverse effects on cell morphology and growth. Microcarrier cell cultures were used for large-scale production of canine distemper virus. Virus yields (TCID50 per ml) were more than 10-times higher as compared to stationary cell culture.

Animals↗
PubMed1992

Gas chromatographic-mass spectrometric detection of trace amounts of organic compounds in the intravenous solution Infusio Darrowi.

A gas chromatographic-mass spectrometric procedure was employed to confirm the presence of trace amounts of organic compounds in the intravenous solution Infusio Darrowi. Organic contaminants in the solution analysed were concentrated by microextraction with n-pentane. The main compounds detected were 2,6-di-tert.-butyl-4-methylphenol, 2,6-di-tert.-butyl-4-ethylphenol, 2,6-di-tert.-butyl-4-methoxyphenol, benzothiazole, isomeric C9 alkyl phenols and di(n-butyl) phthalate. These impurities were leached from rubber stoppers during their sterilization into the intravenous solution at levels ranging ca. from 5 x 10(-6) to 5 x 10(-8) g/l.

Benzothiazoles↗
PubMed1992

Herpes simplex virus type 2 and pseudorabies virus associated growth factors and their role in the latency in vitro.

A putative herpes simplex virus type 2 (HSV-2) growth factor (HSGF-2) was detected in a crude extract from virus infected mouse embryo cells. This factor, similar to previously described pseudorabies virus (PRV) associated growth factor (PRGF) was shown to have ability to morphologically transform non-transformed cells and to repress the transformed phenotype of transformed cells. Both activities could be neutralized with two, out of seven monoclonal antibodies directed against glycoprotein B of HSV-2. Both PRGF and HSGF-2 were detected in human embryo lung cells latently infected with PRV or HSV-2 either at 41 degrees C, or in the presence of phosphonoacetic acid. Human alpha-2 interferon, when present in medium of latently infected cells enhanced the production of both HSGF and PRGF. On the contrary, when latently infected cells were treated with 5-azacytidine the synthesis of both PRGF and HSGF-2 was completely blocked and the virus reactivated from latency replicated to higher titers than in non-treated cells. The role of PRGF and HSGF-2 in the establishment, maintenance and reactivation of latency, as well as in cellular transformation is discussed.

Animals↗
PubMed1992

The glycoprotein B gene and its syn3 locus of herpes simplex virus type 1 are involved in the synthesis of virus-associated growth factor (HSGF-1).

A putative growth factor (HSGF-1) associated with herpes simplex virus type 1 (HSV-1), which is similar to PRGF associated with pseudorabies virus, and/or HSGF-2 associated with HSV-2, was described. Experiments with four syncytial (syn) and four nonsyncytial (syn+) HSV-1 strains showed that the ability of this virus to produce HSGF-1 in infected cells is associated with the syn+ phenotype. Double infection of cells with syn+ and syn strain resulted either in enhancement or complete inhibition of HSGF-1 production, depending on the chosen pair of syn+ and syn strains. The studies with the recombinants between the syn+ strain KOS and syn strain ANGpath in the gene for glycoprotein B (gB) and syn3 locus revealed that the gB gene and its syn3 locus play a role in the HSGF-1 synthesis.

Animals↗
PubMed1990

Biohazard in microbiology, virology and in work with cell cultures.

Possible biohazards are referred to in this report resulting from person's manipulation with microbes, viruses and cell cultures as well as risky contamination of examined biological material with the worker's microflora. Eventual intercontaminations are described throughout the hypo- and cryothermic conservation including the freeze-drying of biological material. Working security regulations are listed to conclude the report.

Cells, Cultured↗