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Biomedical subjects

H L David

Publications and source records attributed to H L David.

At least 73 records · Page 4Linked to original sources

Abortive infection of Mycobacterium leprae by the mycobacteriophage D29.

The interactions of mycobacteriophage D29 and Mycobacterium leprae were examined. It was demonstrated that after adsorption D29 injected its DNA in M. leprae. While the synthesis of host proteins and lipids were inhibited in M. tuberculosis and in M. smegmatis during infection by D29, the results were inconclusive in the case of M. leprae because these bacteria did not incorporate the appropriate substrates.

DNA, Bacterial↗

[Mycobacterium szulgai pulmonary mycobacteriosis, Apropos of a new case].

A case of pulmonary mycobacteriosis due to Mycobacterium szulgai is reported in a 58-year-old man. Clinical and radiological features suggested tuberculosis. The etiologic agent was M. szulgai which was recovered from six sputum samples. The condition has remained unchanged without therapy for four years.

Humans↗

Classification and identification of Mycobacterium africanum by pyrolysis mass spectrometry.

Pyrolysis mass spectrometry was used to classify and identify strains of Mycobacterium africanum and of M. tuberculosis, M. bovis and M. bovis BCG. The multicharacter mass pyrograms were evaluated by computerized data handling procedures that were suited for classification and identification. The results revealed considerable heterogeneity among the African strains, which was shown to be linked to the geographic distribution of the strains. On the basis of a routine mass spectrometric identification key the African strains were identified without exception as belonging to, what is referred to as the 'Tuberculosis complex' (i.e. the clinically relevant group formed by strains of M. Tuberculosis, M. bovis and M. bovis BCG). Classification of the strains by means of discriminant analysis indicated an intermediate clustering for the majority of the African strains and overlap for some African strains with in particular M. bovis. It was concluded that from the mass spectrometric data a species status for the group of African strains was not justifiable.

Africa↗

Spheroplast fusion as a mode of genetic recombination in mycobacteria.

Spheroplasts were prepared from two carotenoid pigment mutants of Mycobacterium aurum named NgR9 and A11, which were obtained by the chemical mutagenesis of the wild type strain A+ with N-methyl-N'-nitro-N-nitrosoguanidine. The carotenoid pigments and the alpha- and beta-mycolic acids were taken as genetic markers and the recombinants were selected on the basis of their colour on Löwenstein-Jensen medium. Spheroplasts of the two mutants were mixed in a 1:1 ratio and were treated with 40% (w/v) polyethylene glycol 6000 for 5 min at 37 degrees C. The frequency of NgR9 X A11 recombination in optimal conditions was about 2.5 X 10(-3). The recombinants selected on the basis of their carotenoid pigment profile were also tested for their alpha- and beta-mycolic acids as a second genetic marker. The results were further confirmed by electron microscopy. The optimal conditions for spheroplast fusion as a mode of genetic recombination in M. aurum are described.

Carotenoids↗

Identification of Mycobacterium fortuitum and Mycobacterium chelonei.

The study of 52 strains of rapidly growing mycobacteria showed that Mycobacterium fortuitum and M. chelonei were clearly distinguished by the aid of seven key tests (nitrate reductase, iron uptake, beta-glucosidase, penicillinase, growth on fructose, resistance to pipemidic acid, and resistance to capreomycin) and by analysis of their respective mycolic acids. However, the subdivision of these species into M. fortuitum var. fortuitum and M. fortuitum var. peregrinum and M. chelonei subsp. chelonei and M. chelonei subsp. abscessus was not satisfactorily accomplished.

Capreomycin↗

[Mycobacterium kansasii: drinking water contaminant of a hospital].

Contamination of tap water by Mycobacterium kansasii in an hospital is described. This contamination was responsible for an increased frequency of M. kansasii isolates from gastric washings. The hygiene measures taken to eliminate the mycobacteria from the water distribution system in the hospital are reported.

Bacteriological Techniques↗

Evaluation of rapid tests for the identification of mycobacteria.

The use of eight rapid tests for the identification of 1307 strains of mycobacteria belonging to 18 species was evaluated. The standard niacin, nitrate-reductase and catalase tests were supplemented by new tests for the detection of beta glucosidase, urease, penicillinase, trehalase and cephalosporinase. This combination of eight rapid tests was not able to replace more conventional procedures but in some cases was of value in discriminating between closely related species.

Cephalosporinase↗

Ultrastructural and chemical studies on wall-deficient forms, spheroplasts and membrane vesicles from Mycobacterium aurum.

Wall-deficient forms of Mycobacterium aurum were prepared by agitating the cells during exponential growth with D-cycloserine, glycine, lysozyme, EDTA and LiCl for approximately the time of three cell divisions (18 h). Wall-deficient forms were then converted to spheroplasts by gentle stirring with lysozyme and EDTA in a Tris/HCl buffer containing sucrose until all the cells appeared spherical by phase contrast microscopy. Subsequent lysis by nucleases followed by osmotic shock produced membrane vesicles. Ultrastructural and chemical properties of the spheroplasts and membrane vesicles are described. The spheroplasts were susceptible to lysis by 0.25% (w/v) sodium dodecyl sulphate and were permeable to certain enzyme substrates.

Anti-Bacterial Agents↗

Growth and cell division of Mycobacterium avium.

The rates of cell division and of protein, DNA and RNA synthesis upon transition of Mycobacterium avium to and from rich medium were examined. The changes in cell morphology (elongation) were also examined by optical and electron microscopy. Upon transfer from poor to rich medium, the rate of synthesis of RNA increased rapidly, followed by an increase in protein synthesis within 3 h and by an increase in DNA synthesis within 7 h; cell division began after a lag of about 10 h. Upon transfer from rich to poor medium, the preshift rates for protein and DNA synthesis changed to postshift rates after 3 h and 7 h, respectively; RNA synthesis stopped immediately, there was a transient fall in total RNA, and synthesis was resumed at a new rate only after 24 h. After the period of adjustment to new medium, the bacteria entered the postshift growth in which cell size, the increase in cell mass (absorbance at 650 nm) and viable counts, and the rates of synthesis of protein, DNA and RNA were constant. Ultrastructural examination of elongated cells during the adjustment period showed that they had septa at different stages of formation, but no evidence of fragmentation was found. It was concluded that cell division in M. avium was by binary fission, and that the notion of a life-cycle was not supported by present findings.

Bacterial Proteins↗

Multiple drug resistance in Mycobacterium avium: is the wall architecture responsible for exclusion of antimicrobial agents?

Whole cells of Mycobacterium avium, characterized by their negative response in the nine biochemical tests used for mycobacterial identification in our laboratory, turned positive for nitrate reductase, Tween-80 hydrolysis, beta-glucosidase, acid phosphatase, alkaline phosphatase, penicillinase, and trehalase after their wall portion was removed to yield spheroplasts. This suggested that the negative results in most of the biochemical procedures were caused by the exclusion mechanism at the wall level. Preliminary transmission and scanning electron microscopic studies showed differences at wall level between laboratory-maintained opaque, dome-shaped (SmD) and host-recycled smooth, transparent (SmT) colony type variants of M. avium and suggested the presence of an outer regularly structured layer in SmT variants. Comparative ultrastructural studies utilizing different polysaccharide coloration methods confirmed the presence of an outer polysaccharide layer in SmT variants which was probably related to their enhanced pathogenicity for experimental animals and drug resistance as compared to that of SmD variants. These findings are discussed with respect to multiple drug resistance, virulence, and gene expression of M. avium.

Anti-Bacterial Agents↗

Differential identification of Mycobacterium fortuitum and Mycobacterium chelonei.

Mycobacterium fortuitum and Mycobacterium chelonei are distinguished unambiguously by the combined use of five test characters: nitrate reductase, beta-glucosidase, acid production from fructose, penicillinase, and trehalase. Typically, M. fortuitum was nitrate reductase positive, beta-glucosidase positive; M. chelonei was nitrate reductase negative, beta-glucosidase negative, penicillinase positive, and trehalase positive and did not produce acid from fructose.

Fructose↗

Effects of antituberculosis and antileprosy drugs on mycobacteriophage D29 growth.

The minimal inhibitory concentrations of antituberculosis and antileprosy drugs were determined for Mycobacterium aurum. The concentrations that reduced the final yield of bacteriophage D29R1 by 50% and the time during the replication cycle at which the drugs completely inhibited phage production were estimated. THe 50% inhibitory concentration/minimal inhibitory concentration ratios were close to 1.0 for clofazimine, colistin, rifampin, and streptomycin; these ratios were high for dapsone (diaminodiphenylsulfone) and isoniazid. Ethambutol (minimal inhibitory concentration, 1.0 micrograms/ml) was without effect on viral growth.

Antitubercular Agents↗

Evaluation of the chloride and bromide salts of cetylpyridium for the transportation of sputum in tuberculosis bacteriology.

The usefulness of cetylpyridium chloride and cetylpyridium bromide, at a final concentration of 1% in 1% solution of sodium chloride, for the transportation of sputum in tuberculosis bacteriology, was evaluated. It was found that these compounds did not alter the staining properties nor the viability of the tubercle bacilli for at least 14 days.

Bacteriological Techniques↗

Mutations affecting pigment synthesis in Mycobacterium aurum.

Pigmentation mutants of Mycobacterium aurum were isolated after chemical mutagenesis. Examination of the pigments extracted from these mutants indicated that at least 15 carotenoids were formed. beta-Carotene was not detected and the major carotene of M. aurum appeared to be leprotene. The possible biosynthetic pathway is discussed on the basis of these results.

Carotenoids↗

[Evaluation of urease and beta-glucosidase activity for the practical identification of mycobacteria (author's transl)].

The urease and the beta-glucosidase for the identification of the mycobacteria were evaluated using 1,019 strains received for reference purposes in this laboratory in 1978. We found that the urease test was useful in the differential identification of BCG which, contrary to Mycobacterium bovis, gave a strong positive reaction in 3 h. The test was also useful to discriminate M. scrofulaceum from pigmented strains of M. avium. The beta-glucosidase test discriminated between M. tuberculosis and M. bovis and was useful to distinguish the niacine-positive strains of M. africanum which were usually negative in this test. The beta-glucosidase test was particularly useful in the differential identification of M. fortuitum and M. chelonei. In this report, the frequency distribution of the species which were submitted to this laboratory in 1978 was also shown and the results obtained for each of the conventional test procedures was presented.

Glucosidases↗