THE USE OF NEOTETRAZOLIUM IN THE STUDY OF <i>E. COLI</i> METABOLISM*

Narahara, H. T.; Quittner, H; Goldman, Lester M.; Antopol, William

doi:10.1111/j.2164-0947.1950.tb01890.x

Cited by 22 publications

(4 citation statements)

References 5 publications

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“…Electron pictures of the granules, both within and without phage infected cells of E. coli, have been taken by Hillier, Mudd, and Smith and published in Burrows (1949). The polar granules of E. coli and related gramnegative rods have recently been shown to reduce tetrazolium (Lederberg, 1948; Bielig, Kausche, and Haardick, 1949; Winkler, 1950) and neotetrazolium (Narahara et al, 1950). Figure 1 shows a field from a preparation of E. coli grown on a salt-glucose medium and lysed with T2 coliphage.…”

Section: Resultsmentioning

confidence: 99%

Further Evidence of the Existence of Mitochondria in Bacteria

et al. 1951

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The granules of mycobacteria, originally described by Robert Koch as spores, have been shown in a preceding communication to be sites of oxidative-reductive enzymatic activity. In respect to several enzymatic reactions, as well as in tinctorial and morphological characteristics, these mycobacterial granules were shown to correspond to mitochondria (Mudd, Winterscheid, DeLamater, and Henderson, 1951). In the present communication granules in several other species of bacteria have been examined by similar criteria. These granules also have been found to correspond in essential characteristics to mitochondria. METHODS AND MATERIALS The microorganisms used were: Escherichia coli, strain B; Bacillus megatherium, a strain originally given us by Dr. C. F. Robinow; Micrococcus cryophilus, a large gram-variable coccus isolated in the Department of Agriculture (McLean, Sulzbacher, and Mudd, 1951) and utilized for studies on nuclear mitosis (DeLamater and Woodburn, 1951); and Saccharomyces cerevisiae, a strain from baker's yeast and a "cytochromeless" variant, received through the courtesy of Dr. H. M. Davidson, which had been derived as described by Ephrussi and colleagues (Ephrussi, Hottinguer, and Chimenes, 1949; Slonimski and Ephrussi, 1949). The cells of E. coli and B. megatherium used had been grown for 3 to 4 hours at 37 C on Morton and Engley (1945) agar slants or in M. and E. broth. The micrococcus was incubated 24 hours at 20 C on similar agar slants or broth prior to use. Both strains of S. cerevisiae were incubated 24 hours at 37 C onSabouraud's agar slants. The reactions examined for were the reduction of 2,3,5-triphenyltetrazolium and neotetrazolium, probably indicating the action of flavoprotein enzymes, oxidation of the Nadi reagent, indicating cytochrome oxidase, staining with the mitochondrial stain Janus green B, the Baker acid-hematin stain for phospholipid, and the Harman mitochondrial stain. Experimental procedures were followed as in the preceding communication (Mudd, Winterscheid, DeLamater, and Henderson, 1951) with the following modifications: The reduction of triphenyltetrazolium and neotetrazolium was demonstrated I This research has been aided by a grant from the Damon Runyon Fund through the American Cancer Society as recommended by the Committee on Growth of the National Research Council.

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Section: Resultsmentioning

confidence: 99%

Further Evidence of the Existence of Mitochondria in Bacteria

et al. 1951

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“…The teArazolium aspect of this study was stimulated by the reports indicating that the tetrazolium salts are reduced to pigmented formazans by a variety of living tissues including leukocytes and bacteria (11)(12)(13)(14). It was anticipated that viable leukocytes could be differentiated from non-viable leukocytes, and living from dead intracellular brucella by direct microscopic observation.…”

Section: Discussionmentioning

confidence: 99%

The Protection of Intracellular Brucella Against Therapeutic Agents and the Bactericidal Action of Serum

Shaffer

Kucera

Spink

1953

Journal of Experimental Medicine

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A method for the in vitro study of intracellular brucella has been described. Exudative leukocytes containing intracellular brucella have been maintained in vitro in a synthetic tissue culture medium or in human or animal serum. Intracellular brucella are protected in vitro against the lethal action of therapeutic agents or the bactericidal action of serum. This protection of intracellular brucella is dependent upon the presence of an intact, viable host cell. None of the currently available therapeutic agents, whether used alone or in combinations, were capable of killing all intracellular brucella in vitro in 24 hours. A remarkable protection of intracellular brucella against streptomycin has been demonstrated. The most effective reduction in the number of viable intracellular brucella was accomplished by exposure of the host cells to streptomycin plus aureomycin, terramycin, or chloramphenicol. The available evidence suggests that the ability of brucella to localize and remain viable within the cells of an infected host is an important biologic factor in establishing and perpetuating brucella infections, despite therapeutic measures or the operation of the host's humoral defense mechanisms. Reduction of neotetrazolium by leukocytes and brucella in vitro provides a method for assessing the metabolic status of the host cell, but does not discriminate with any degree of certainty a viable from a non-viable intracellular organism.

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“…The possible existence of mitochondria or analogous structures in bacteria has been indicated by several workers (Graffi and Maas, 1940;King and Beams, 1942;Bielig et al, 1949;Narahara et al, 1950). However, it has been only recently that these cytoplasmic structures in bacteria have received definite recognition as mitochondria and have received adequate cyto-found dense areas at the poles of E. coli cells infected with T-even phages; these areas he inter- 1': il:: Figure 8.…”

Section: Discussionmentioning

confidence: 82%

Light and Electron Microscopic Studies of Escherichia Coli-Coliphage Interactions Iii

et al. 1953

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The first two articles of this series (Beutner et al., 1953; Mudd et al., 1953) contain descriptions of the light microscopic cytology and electron microscopy of T2 bacteriophage infection of Escherichia coli, strain B, with particular emphasis on the changes occurring in the chromatinic material, and with the intracellular appearance of possible phage precursors and apparently mature phage progeny. In this paper we show that the cytoplasmic organelles, the mitochondria, in contrast with the nuclei, maintain their integrity throughout the latent period of infection. MATERIALS AND METHODS Escherichia coli, strain B, and phages T2r+ and T2r were used throughout. The phage suspensions, media, basic techniques, and microscopic equipment have been described elsewhere (Beutner et al., 1953; Mudd et al., 1953). In addition, experiments were performed utilizing 2,3,5-triphenyltetrazolium chlorides as an indicator of reduction (Brodie and Gots, 1951, 1952; Kun, 1951; Shelton and Schneider, 1952). The reduction of 2,3,5-triphenyltetrazolium chloride was followed by mixing 3 ml aliquots of deepgrown (unaerated) log phase broth4 cultures of E. coli and 1 ml of a distilled water solution of I This work has been aided by a grant from the United States Atomic Energy Commission, AEC Contract no. AT(30-1)-1342. Aid was also received from the Theresa F. Felsen Memorial Fund.

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THE USE OF NEOTETRAZOLIUM IN THE STUDY OF E. COLI METABOLISM*

Cited by 22 publications

References 5 publications

Further Evidence of the Existence of Mitochondria in Bacteria

Further Evidence of the Existence of Mitochondria in Bacteria

The Protection of Intracellular Brucella Against Therapeutic Agents and the Bactericidal Action of Serum

Light and Electron Microscopic Studies of Escherichia Coli-Coliphage Interactions Iii

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