Heterogeneity and Biological Activity of Endotoxic Glycolipid from Salmonella minnesota R595

Chen, C. H.; Johnson, A. G.; Kasai, Nobuhiko; Key, B. A.; Levin, Jack; Nowotny, Alois

doi:10.1093/infdis/128.supplement_1.s43

Cited by 54 publications

(25 citation statements)

References 18 publications

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“…Nowotny et al (2,17) reported that endotoxins were extremely heterogeneous. Intramolecular heterogeneity of the glycolipid seems to result from the difference in fatty acid contents and the occurrence of phosphate or 4-amino-4-deoxyarabinose.…”

Section: Discussionmentioning

confidence: 99%

Chemical and Ultrastructural Differences in Endotoxic Glycolipids from Salmonella Minnesota Re Mutant Extracted with Various Solvent Systems

Amano

Fukushi

1984

Microbiology and Immunology

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Endotoxic glycolipids (ReGl) extracted from the whole cells (WC) and cell walls of heptose‐less Re mutant of Salmonella minnesota with hot phenol‐water (PW), phenol‐chloroform‐petroleum ether (PCP), and chloroform‐methanol (CM) were analyzed chemically and examined with an electron microscope. ReGl‐PW‐(WC) contained mannose and proteins as contaminants, ReGl‐PCP(WC) consisted of an excess amount of amino compound (cadaverine), and ReGl‐CM(WC) consisted of proteins and cadaverine, in addition to the ReGl constituents. The ultrastructure of ReGl‐PW(WC) resembled onions when stained with uranyl formate, and was spherical when stained with uranyl acetate, sodium phosphotungstate and ammonium molybdate, whereas those of ReGl‐PCP(WC) and ReGl‐CM(WC) were shaped like ribbons. However, the shadowed ultrastructures of ReGl‐PCP(WC) and ReGl‐CM(WC) showed small pieces of flat and wide sheets, respectively. ReGl‐PCP(WC) re‐extracted by the PW method was found to be converted into the onion‐like structure which was similar to that of ReGl‐PW(WC), while an intermediate form (fingerprint‐like) was observed after re‐extraction of ReGl‐PW(WC) with PCP. It was strongly suggested that the ultrastructural arrangement of ReGl was dependent on the solvent systems used for extraction.

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Section: Discussionmentioning

confidence: 99%

Chemical and Ultrastructural Differences in Endotoxic Glycolipids from Salmonella Minnesota Re Mutant Extracted with Various Solvent Systems

Amano

Fukushi

1984

Microbiology and Immunology

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“…The smooth to rough conversions (R mutation) of Salmonella minnesota LPS have been charac terized by partial loss of the polysaccharide moiety. The endotoxins including LPS and R mutant glycolipids (RGl) were extracted with hot phenol-water (PW) (25), phenol-chloroform-petroleum ether (PCP) (8), or chloroform-methanol (CM) (4). Generally, the PW method is used for extraction of all kinds of endotoxins, while the PCP and CM methods are used for extraction of RGl.…”

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Chemical and Ultrastructural Comparison of Endotoxins Extracted from Salmonella minnesota Wild Type and R Mutants

Amano

Fukushi

1984

Microbiology and Immunology

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Endotoxic lipopolysaccharide and glycolipids (RGl) extracted from Salmonella minnesota wild type and R mutant cells (chemotypes Ra, Rb, Re, Rd1, and Rd2), respectively, with hot phenol‐water (PW) and phenol‐chloroform‐petroleum ether (PCP) were analyzed chemically and electron microscopically. All RGl extracted with PW (RGl‐PW) contained excess amounts of phosphate, O‐ester linked fatty acids and neutral sugars, while all RGl extracted with PCP (RGl‐PCP) contained excess amounts of free amino groups and fatty acids, in addition to the RGl constituents. Polyamine (cadaverine), phosphoethanolamine, and an unidentified amino compound were contained in RGl‐PCP as free amino groups. When stained with uranyl formate, the ultrastructure of RGl‐PW showed a spherical form (onion‐like form), whereas the micrographs of RGl‐PCP showed a filamentous structure, regardless of strain differences. On the other hand, the micrographs of RGl‐PW represented spherical and doughnut‐shaped forms, and the micrographs of RGl‐PCP showed filamentous or stick forms, when stained with uranyl acetate. Thus, it is suggested that the ultrastructures of RGl were dominated by the solvent systems used for extraction, and not by the strains used here.

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“…The limulus gelation test, developed by Levin and others (13,14), has provided a useful in vitro means to assess an endotoxin activity which corresponds with in vivo indices of toxicity (3,15,24). We have previously found this assay to be useful in evaluating endotoxin inactivation processes (4).…”

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Inactivation of Endotoxin by Polymyxin B

Cooperstock

1974

Antimicrob Agents Chemother

124

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The limulus gelation assay was utilized to investigate endotoxin inactivation by a number of antibiotics in vitro. Endotoxin activity was sharply reduced by polymyxin B and sodium colistimethate. The effect of the polymyxin was not significantly inhibited by 0.001 M calcium or 90% serum. Crude endotoxins from a variety of aerobic gram-negative bacteria, including several not previously studied, could be inactivated 1 or more logs by as little as 1 Og of polymyxin B per ml, whereas Bacteroides fragilis endotoxin was poorly detoxified. A 10,000-fold range in the relative susceptibility of different endotoxins to inactivation by polymyxin B was found. The endotoxin most susceptible to polymyxin B was derived from an organism resistant to polymyxin B by disk sensitivity testing, suggesting that the bacteriocidal and endotoxin detoxifying properties of polymyxin need not be directly related.Recent studies have demonstrated the capacity of polymyxin B to protect animals from a variety of toxic effects of endotoxin, including the generalized Schwartzman reaction (5, 23), diffuse intravascular coagulation (6, 7), renal cortical necrosis (6, 7), leukopenia (6, 7), and death (6,12,21,23). The limulus gelation test, developed by Levin and others (13,14), has provided a useful in vitro means to assess an endotoxin activity which corresponds with in vivo indices of toxicity (3,15,24). We have previously found this assay to be useful in evaluating endotoxin inactivation processes (4). The present study was designed to evaluate the effect of polymyxin B and other antibiotics upon endotoxin, the conditions of the reaction, and the relative polymyxin susceptibility of endotoxins from a variety of gram-negative bacteria. MATERIALS AND METHODS

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Heterogeneity and Biological Activity of Endotoxic Glycolipid from Salmonella minnesota R595

Cited by 54 publications

References 18 publications

Chemical and Ultrastructural Differences in Endotoxic Glycolipids from Salmonella Minnesota Re Mutant Extracted with Various Solvent Systems

Chemical and Ultrastructural Differences in Endotoxic Glycolipids from Salmonella Minnesota Re Mutant Extracted with Various Solvent Systems

Chemical and Ultrastructural Comparison of Endotoxins Extracted from Salmonella minnesota Wild Type and R Mutants

Inactivation of Endotoxin by Polymyxin B

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