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1.
Int J Lepr Other Mycobact Dis ; 50(2): 200-4, 1982 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-6811449

RESUMO

An assay system has been developed based on automated radiometric quantification of 14CO2 produced through oxidation of [1-14C] fatty acids by mycobacteria. Two stains of M. tuberculosis (H37Rv and Erdman) and one of M. bovis (BCG) in 7H9 medium (ADC) with 1.0 microCi of one of the fatty acids (butyric, hexanoic, octanoic, decanoic, lauric, myristic, palmitic, stearic, oleic, linoleic and linolenic) were studied. Results previously published on M. lepraemurium (Hawaiian) were also included for comparison. Both strains of M. tuberculosis had maximum 14CO2 production from hexanoic acid. Oxidation of butyric and avid oxidation of lauric acids were also found with the H37Rv strain but not with Erdman. In contrast, 14CO2 production by M. bovis was greatest from lauric and somewhat less from decanoic acid. M. lepraemurium showed increasing oxidation rates from myristic, decanoic and lauric acids. Assimilation studies of M. tuberculosis H37Rv confirmed that most of the oxidized substrates were converted into by-products with no change in those from which no oxidation was found. These data suggest that the radiometric measurement of differential fatty acid metabolism may provide a basis of strain identification of the genus Mycobacterium.


Assuntos
Ácidos Graxos/metabolismo , Mycobacterium/metabolismo , Dióxido de Carbono/análise , Radioisótopos de Carbono , Meios de Cultura , Mycobacterium bovis/metabolismo , Mycobacterium lepraemurium/metabolismo , Mycobacterium tuberculosis/metabolismo , Oxirredução
2.
Int J Lepr Other Mycobact Dis ; 47(2): 126-32, 1979 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-378870

RESUMO

An assay system has been developed based on automated radiometric quantification of 14CO2 produced through oxidation of (1--14C) fatty acids by mycobacteria. With this system, the Hawaiian strain of M. lepraemurium was studied using the K-36 buffer as a suspending solution for the organisms along with 5.0 muCi of one of the following fatty acids: acetate, butyric, hexanoic, octanoic, decanoic, lauric, myristic, palmitic, stearic, oleic, linoleic, linolenic, and malonic. The 14CO2 production by this organism was greatest with lauric, decanoic, myristic, octanoic, stearic, oleic, linoleic, linolenic, and malonic. The 14CO2 production by this organism was greatest with lauric, decanoic, myristic, octanoic, and stearic acids, in decreasing order. Assimilation studies and radiochromatograms confirmed that most of the oxidized substrates were converted into by-products with no change in those from which no oxidation was found. These data suggest that the radiometric measurement of differential fatty acid metabolism may provide a basis of radiometric identification of M. lepraemurium and assessment of the growth requirements of this organism.


Assuntos
Ácidos Graxos/metabolismo , Mycobacterium lepraemurium/metabolismo , Radiometria/instrumentação , Dióxido de Carbono/metabolismo , Cromatografia em Papel , Cromatografia em Camada Delgada
3.
Int J Lepr Other Mycobact Dis ; 44(3): 294-300, 1976.
Artigo em Inglês | MEDLINE | ID: mdl-789261

RESUMO

The radiometric method has been applied for studying the metabolism of M. lepraemurium and the conditions which might force or inhibit its metabolic activity in vitro. These organisms assimilate and oxidize (U-14C) glycerol, and (U-14C) acetate, but are unable to oxidize (U-14C) glucose, (U-14C) pyruvate, (U-14C) glycine and 14C-formate. When incubated at 30 degrees C M. lepraemurium oxidizes (U-14C) acetate to 14CO2 faster than 37 degrees C. The smae effect was observed with increasing concentrations of polysorbate 80 (Tween 80), or the 14C-substrate. No change in metabolic rate was observed when the organisms were kept at -20 degrees C for 12 days. Although tried several times, it was not possible to demonstrate any "inhibitors" of bacterial metabolism in the reaction system. The radiometric method seems to be an important tool for studying metabolic pathways and the influence of physical and biochemical factors on the metabolism of M. lepraemurium in vitro.


Assuntos
Mycobacterium lepraemurium/metabolismo , Acetatos/metabolismo , Animais , Dióxido de Carbono/biossíntese , Radioisótopos de Carbono , Feminino , Congelamento , Glucose/metabolismo , Glicerol/metabolismo , Glicina/metabolismo , Fígado/microbiologia , Camundongos , Camundongos Endogâmicos , Ácidos Oleicos/farmacologia , Polissorbatos/farmacologia , Piruvatos/metabolismo
4.
Int J Lepr Other Mycobact Dis ; 43(3): 234-8, 1975.
Artigo em Inglês | MEDLINE | ID: mdl-176120

RESUMO

A simple radiometric method has been developed for evaluating the effect of drugs on the metabolism of M. lepraemurium. The method is based on the measurement of the 14CO2 produced through bacterial metabolism of acetate-U-14C. Seventeen drugs were tested: bacitracin, cephaloridine, chloramphenicol, cycloserine, dactinomycin, DDS, ethionamide, INH, kanamycin, methenamine mandelate, nitrofurantoin, oxacillin, polymyxin B, rifampicin, streptomycin, sulfadimethoxine and vancomycin. The drugs which caused most marked inhibition were chloramphenicol, INH, ethionamide and nitrofurantoin in order of increasing effectiveness. The radiometric study which is completed in 15 days permits direct study of the drug effect on the metabolism of M. lepraemurium and a more rapid screening of antileprosy drugs than has previously been possible. Currently, these observations are being extended to studies of the structure-activity relationships of antileprosy drugs and the metabolism and drug susceptibility of M. leprae in vitro.


Assuntos
Antibacterianos/farmacologia , Infecções por Mycobacterium/tratamento farmacológico , Mycobacterium lepraemurium/efeitos dos fármacos , Bacitracina/farmacologia , Cefaloridina/farmacologia , Cloranfenicol/farmacologia , Dactinomicina/farmacologia , Dapsona/farmacologia , Etionamida/farmacologia , Isoniazida/farmacologia , Canamicina/farmacologia , Metenamina/farmacologia , Testes de Sensibilidade Microbiana , Nitrofurantoína/farmacologia , Oxacilina/farmacologia , Polimixinas/farmacologia , Radiometria , Rifampina/farmacologia , Estreptomicina/farmacologia , Sulfadimetoxina/farmacologia , Vancomicina/farmacologia
5.
Appl Microbiol ; 28(3): 452-5, 1974 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-4608779

RESUMO

A sensitive and nondestructive radiometric method has been applied to the detection of metabolism of Mycobacterium lepraemurium, as a model for the study of the metabolism and substrate requirements of M. leprae. The method is based on the measurement of the (14)CO(2) produced through the bacterial conversion of [U-(14)C]acetate or [U-(14)C]glycerol by 7 x 10(9) bacteria suspended in 10 ml of either a simple buffer system (K-36) or a complex medium (NC-5). Metabolism of the bacilli was easily detected within 3 days after inoculation and was measured daily. NC-5 medium supported metabolism of M. lepraemurium for several weeks longer than the simple K-36 buffer. The radiometric technique shows promise as a rapid and efficient system for evaluating the metabolism of mycobacteria without introducing any changes in the physiologic state of the organisms, studying their metabolic pathways, determining conditions potentially favorable for multiplication of these organisms in vitro, and studying their susceptibility to inhibition by drugs.


Assuntos
Mycobacterium lepraemurium/metabolismo , Radiometria/métodos , Acetatos/metabolismo , Dióxido de Carbono/biossíntese , Radioisótopos de Carbono , Estudos de Avaliação como Assunto , Glicerol/metabolismo
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