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Biomedical subjects

A Laszlo

Publications and source records attributed to A Laszlo.

At least 91 records · Page 5Linked to original sources
PubMed1986

Heat shock proteins within the mammalian cell cycle: relationship to thermal sensitivity, thermal tolerance, and cell cycle progression.

We have measured endogenous and induced rates of 70-kD, 89-kD, and 110-kD heat shock proteins in highly pure G1-, S-, or G2-M phase fractions of Chinese hamster fibroblasts (CHO) separated by fluorescence-activated cell sorting (FACS). Relative rates of synthesis of all three polypeptides as measured by two-dimensional gel electrophoresis were similar throughout the cell cycle, and therefore, endogenous levels were unlikely to explain the thermal sensitivity of S-phase cells. Distinct heterogeneity in induced rates of these polypeptides was noted in all phase fractions. Enhanced rates of 70-kD polypeptide were measured in S and G2-M as compared to G1 following heat shock. Little increase in either the 89-kD or 110k-kD heat shock proteins was observed in heated G1 cells. This heterogeneity in induced rates of synthesis was in contrast to the similarity in thermal tolerance expression kinetics between each phase. Finally, enhanced synthesis of these polypeptides appeared unrelated to regulation of either heat-induced cell cycle delay or to the resumption of phase-specific progression after heat shock as measured by simultaneous flow cytometric measurement of incorporated BrdUrd and DNA content.

Animals↗
PubMed1986

Radiometric diagnosis of mycobacteria.

The aim of this study was to determine the impact of recently developed rapid radiometric techniques on the clinical diagnostic operations of a reference laboratory for mycobacteria. Selective inhibition by rho-nitro-alpha-acetylamino-beta-hydroxypropiophenone was used to rapidly screen submitted cultures for identification of mycobacterial species other than Mycobacterium tuberculosis. The radiometric drug susceptibility test was applied only to those cultures presumptively identified as belonging to the Mycobacterium tuberculosis complex. All referred cultures were tested without additional subculture. The results showed that non-pigmented mycobacteria other than Mycobacterium tuberculosis can be screened with about 99% reliability, most of them within 24 hours. Unnecessary drug susceptibility testing of mycobacteria other than tubercle bacilli can be avoided at an early stage, thus shortening the average reporting time of the Mycobacterium tuberculosis complex to nine days following the receipt of the cultures. Ways of limiting erroneous reporting are discussed.

Antitubercular Agents↗
PubMed1985

Amino acid analogs while inducing heat shock proteins sensitize CHO cells to thermal damage.

Amino acid analogs have been shown to induce heat shock proteins (HSPs). We have examined the effect of these analogs on the thermal sensitivity of Chinese hamster fibroblasts (HA-1) and their stable heat-resistant variants. We found that exposure of HA-1 cells and their heat-resistant variants to canavanine or L-azetidine-2-carboxylic acid cause enhanced synthesis of the three major mammalian HSPs (molecular weight 70,000, 87,000, and 110,000 kd). Although the synthesis of HSPs was increased, the analogs did not induce thermotolerance, a transient ability to protect cells from thermal damage. On the contrary, the analog treatment increased the thermal sensitivity of HA-1 cells, but not of the heat-resistant strains, when these cells were exposed subsequently to elevated temperatures. Our tentative explanation for these findings is that the incorporation of amino acid analogs into HSPs or other cellular proteins sensitizes HA-1 cells to heat. The heat-resistant strains contain higher levels of constitutive HSPs. The additional functional HSPs in the heat-resistant variants may protect these cells from thermal stress. The presence of some newly synthesized analog-substituted, perhaps nonfunctional, HSPs need not affect this thermal protection.

Animals↗
PubMed1985

Heat-resistant variants of Chinese hamster fibroblasts altered in expression of heat shock protein.

Heat-resistant variants of the Chinese hamster HA-1 line have been isolated after repeated heat treatments. The heat-resistant phenotype has been stable for over 70 passages. One of the members of the 70-kDa heat shock protein family was found to be synthesized at greater levels in the heat-resistant variants under normal growth conditions. Mild heat treatment of the variant lines induced a transient thermotolerance that was accompanied by additional increase in the synthesis of the 70-kDa heat shock proteins. Cell-free translation of total cellular RNA revealed greater amounts of 70-kDa heat shock protein mRNA in both control and heated variant cells. The greater levels of 70-kDa heat shock protein synthesized in the variant cells presumably are a reflection of altered levels of its messenger mRNA. In addition, we found that translational control plays a role in the elevated expression of heat shock proteins in heat-shocked HA-1 cells and their heat-resistant variants. The association of the heat-resistant phenotype with increased levels of a 70-kDa heat shock protein suggests strongly that this gene product plays a role in protecting cells from damage inflicted by elevated temperatures.

Animals↗
PubMed1985

Interlaboratory drug susceptibility testing of Mycobacterium tuberculosis by a radiometric procedure and two conventional methods.

A total of 224 recent isolates of Mycobacterium tuberculosis from 163 patients selected to have multidrug resistance were tested against streptomycin (SM), isoniazid, rifampin, and ethambutol (EMB) by the rapid radiometric BACTEC method and two conventional proportion methods: the World Health Organization (WHO) method, using Lowenstein-Jensen medium; and the Veterans Administration reference laboratory for mycobacteria (VA) method, using Middlebrook 7H10 agar medium. The results were compared, focusing on the concentrations of the drugs in all three methods. Among the four drugs tested, most of the discrepancies in measured activity were observed with SM and EMB, generally because of differences in the drug concentrations used by the three methods. A 4-micrograms amount of SM in the BACTEC method was found to be slightly less active than 10 micrograms in the VA method and significantly more active than 4 micrograms of dihydrostreptomycin in the WHO method. With EMB, 2.5 micrograms in BACTEC was similar to 5 micrograms in the VA method and 2 micrograms in the WHO method, while 10 micrograms in the BACTEC method was found to be more active than 10 and 2 micrograms in the VA and WHO methods, respectively. To attain close agreement, drug concentrations used in the BACTEC method should be carefully selected when a comparison is to be made with any conventional method employed in a laboratory. Standardization of in vitro susceptibility testing is greatly needed to achieve uniformity among the test methods used to evaluate tuberculosis therapeutics.

Drug Resistance, Microbial↗
PubMed1985

Intralaboratory comparison of drug susceptibility testing of Mycobacterium tuberculosis by BACTEC and conventional methodology.

The BACTEC radiometric method of drug susceptibility testing of Mycobacterium tuberculosis is a reliable and rapid diagnostic tool in clinical mycobacteriology. However, large scale comparative studies have also shown that the level of agreement with standard methodology was less satisfactory with strains resistant to ethambutol and streptomycin than with strains resistant to rifampin and to isoniazid. Since disagreement with drug resistance strains is far more frequent than with drug susceptible strains, it was felt that only the comparison of a large number of resistant strains would be needed to further refine this new technique. The analysis of BACTEC-derived data for isoniazid and rifampin shows that the level of agreement with conventional methodology falls well within accepted limits. Statistical analysis of the radiometric versus conventional comparisons shows no significant differences between the two methods in the case of isoniazid, rifampin, and ethambutol (3 mg/L). Streptomycin and two other ethambutol concentrations tested showed lower levels of agreement and significant statistical differences with conventional methodology.

Anti-Bacterial Agents↗
PubMed1984

Evaluation of a rapid radiometric differentiation test for the Mycobacterium tuberculosis complex by selective inhibition with p-nitro-alpha-acetylamino-beta-hydroxypropiophenone.

This study is an evaluation of a rapid technique for the differentiation of the Mycobacterium tuberculosis complex from other mycobacteria, using p-nitro-alpha-acetylamino-beta- hydroxypropiophenone (NAP) as a selective inhibitory agent. A total of 416 coded cultures, 234 cultures belonging to the M. tuberculosis complex and 182 cultures belonging to 35 other mycobacterial species, were tested in two laboratories for p-nitro-alpha-acetylamino-beta- hydroxypropiophenone inhibition to concentrations of 5 and 10 micrograms of NAP per ml in Middlebrook 7H12 liquid medium. Two testing modes were compared: the indirect, in which a large bacterial inoculum was used from an isolated culture on a solid medium, and the direct, which used a small inoculum from 7H12 medium. A decrease or no increase in daily 14CO2 output as measured by a BACTEC system was considered evidence of inhibition. The data presented show that a concentration of 5 micrograms of NAP per ml can effectively separate the M. tuberculosis complex from other mycobacterial species in 4 to 6 days. The direct test data show that, unlike other conventional biochemical tests, it does not require a heavy inoculum of mycobacteria and can therefore be performed soon after growth is detected by the radiometric method.

Bacteriological Techniques↗
PubMed1983

TPA induces simultaneous alterations in the synthesis and organization of vimentin.

We have found an increased rate of vimentin synthesis in TPA-treated chicken embryonic fibroblasts, as shown by two-dimensional electrophoretic separation of newly synthesized polypeptides. The capacity of TPA to elicit this effect correlated with its activity as a tumor promoter. Treatment with TPA also altered the organization of the vimentin-containing intermediate filaments, as revealed by immunofluorescence. Treatments which inhibited the TPA-induced rearrangements of vimentin filaments did not prevent the increase in the rate of vimentin synthesis, indicating that gross alterations in cytoskeletal organization were not the immediate cause of the altered vimentin synthesis. On the other hand, alterations in the rate of vimentin synthesis may be a necessary, but not sufficient condition for alterations in vimentin filament organization. TPA as a positive modulator of vimentin synthesis may serve as a useful probe for an eventual understanding of the dynamics of the mechanisms that control the assembly and organization of vimentin filaments.

Animals↗
PubMed1983

Conventional and radiometric drug susceptibility testing of Mycobacterium tuberculosis complex.

A recently developed method of drug susceptibility testing of Mycobacterium tuberculosis which measures the evolution of labeled CO2 from [1-14C]palmitic acid (BACTEC 460 system) was compared to three conventional methods. The proportion method of drug susceptibility testing was the standard against which all test results were compared. Indirect drug susceptibility to isoniazid, streptomycin, rifampin, and ethambutol of 245 isolates belonging to the M. tuberculosis complex was determined. In 95% of the cases, results obtained by the radiometric method were available within 1 week, as opposed to 3 to 6 weeks needed in conventional methodology. Overall agreement was 96.4%. Specificity values ranged from 0.98 to 1.0; sensitivity values of 1.0 for rifampin, 0.96 for streptomycin, 0.91 for isoniazid, and 0.18 for ethambutol were obtained. The specificity of the absolute concentration and resistance ratio drug susceptibility testing methods were 0.99 and 1.0, respectively. The sensitivity of the former was higher than that of the radiometric method (0.99 verus 0.92), whereas that of the latter was lower (0.88 verus 0.96). Further testing indicated that the low sensitivity determined for ethambutol may be due to the choice of the critical concentration used, rather than to a shortcoming of the procedure. The radiometric method thus does not significantly differ in reliability from conventional methods of drug susceptibility testing of M. tuberculosis.

Antitubercular Agents↗
PubMed1981

Tumor promoters alter gene expression and protein phosphorylation in avian cells in culture.

We have investigated the effect of 12-O-tetradecanoylphorbol 13-acetate (TPA) on the synthesis and modification of polypeptides in normal avian cells and cells infected by wild-type and temperature-sensitive Rous sarcoma virus (RSV). Using two-dimensional gel electrophoresis, we have detected alterations in both the abundance of cellular polypeptides and in their phosphorylation that seem unique to TPA treatment. However, the state of phosphorylation of the major putative substrate for the action of the src gene-associated protein kinase, the 34- to 36-kilodalton protein, was not altered. Moreover, examination of the phosphorylated amino acid content of total cellular phosphoproteins revealed that the response to TPA was not associated with detectable increases in their phosphotyrosine content. These results make it unlikely that TPA acts by the activation of the phosphorylating activity of the cellular proto-src gene or by the activation of other cellular phosphotyrosine-specific kinases. We have shown previously that temperature-sensitive RSV-infected cells at nonpermissive temperature demonstrate an increased sensitivity to TPA treatment [Bissell, M. J., Hatie, C. & Calvin, M. (1979) Proc. Natl. Acad. Sci. USA 76, 348-352]. Our present results indicate that this is not due to reactivation of the phosphorylating activity of the defective src gene product or to its leakiness, and they lend support to the notion of multistep viral carcinogenesis.

Animals↗
PubMed1981

Hyperuricaemia associated with 18q deletion. Atypical Lesch-Nyhan syndrome?

The existence of a clinically typical Lesch-Nyhan syndrome was observed in a male infant with 18q deletion syndrome. Indirect hypoxanthine-guanine-phosphoribosyl transferase activity determination demonstrated a normal value, and thus the possibility of Lesch-Nyhan syndrome linked to the X-chromosome may be excluded. It is assumed that the uric acid metabolism must be under the primary or secondary effect of one or other of the gene loci on the long arm of chromosome 18, since the existence off a hyperuricaemic syndrome was observed in this 18q deletion patient.

Chromosome Deletion↗
PubMed1979

Differentiation of rapidly growing mycobacteria with trimethoprim (Tmp).

Inhibition of Mycobacterium smegmatis, M. vaccae, and M. diernhoferi by Trimethoprim (Tmp) in both liquid and solid media is described. Other mycobacteria were not inhibited by the same concentrations. The selective inhibition of the above strains, particularly M. smegmatis, by Tmp could be used for differentiation of these species from other fast-growing acid-fast bacteria.

Culture Media↗