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Biomedical subjects

H Mayer

Publications and source records attributed to H Mayer.

At least 289 records · Page 16Linked to original sources

Enterobacterial common antigen in mutant strains of Salmonella.

A good correlation was found to exist between the serologically determined presence of enterobacterial common antigen (ECA) and the amount of the rare sugar constituent D-mannosaminuronic acid. Strains classified by serological techniques as ECA+, ECA-, and ECAtrace were found to possess the expected amounts of mannosaminuronic acid in the ECA-enriched phenol-soluble PL-L fractions. This correlation provides strong evidence on the identity of the mannosaminuronic acid-glucosamine polymer with the ECA as defined by Kunin (J. Exp. Med. 118:565-586, 1963).

Acetylglucosamine↗

Restriction endonucleases: general survey procedure and survey of gliding bacteria.

Among 120 strains of gliding bacteria which were screened for restriction endonucleases, 27 were found positive. Additionally, three strains carried enzymes able to release the supercoiled state of closed circular DNA. By using a new rapid method, restriction endonuclease activity was released by stirring about 0.5 g of cells (fresh weight) in a motor-driven glass homogenizer in buffer containing Triton X-101, ethylenediaminetetraacetic acid, and mercaptoethanol. A yield from 60 to 80% of the total activity present in the cells was obtained with minimal destruction of the cells. The enzyme activity in the crude extract was measured semi-quantitatively by digestion of DNA and subsequent separation of the fragments on an agarose slab gel. The method appears to be generally applicable for the extraction of restriction endonucleases from gram-negative bacteria on an analytical scale and in a modified form for large-scale preparation of restriction enzymes.

Coliphages↗

Replication of the mini-R1 plasmid Rsc11 and Rsc11 hybrid plasmids.

Replication of the multicopy mini-R1 plasmid, Rsc11, is dependent on host replication functions dna A, B, C, E and G but independent of polA1. Chloramphenicol immediately stops its replication. A stable relaxation complex is not formed. Composite plasmids were constructed with Rsc11 and other small replicons like pSC101, ColE1 and mini-ColE1. In all combinations the amount of hybrid plasmid DNA in the cell never exceeds the amount of Rsc11 DNA itself. This leads to varying copy numbers of the hybrid plasmids depending on the size of the second plasmid. Replication of the composite plasmids proceeds probably always under the control of the Rsc11 part although the second replicon is still functional. The composite plasmids are incompatible with both the parent replicons.

Chloramphenicol↗

Ribitol-containing lipopolysaccharides from Proteus mirabilis and their serological relationship.

Ribitol phosphate was recently identified as a constituent of lipopolysaccharides obtained from 'proteus mirabilis strain D52 giving 1:4-anhydroribitol during acid hydrolysis (Gmeiner, 1975). Two other Proteus mirabilis strains belonging to serogroups O16 and O33 were shown previously to contain an unknown compoound X as lipopolysaccharide constituent (Kotelko et al., 1975). In this report the identification of compound X as 1:4-anhydroribotol by gas-liquid chromatography, mass spectrometry and mass fragmentography is described. Serological investigations using passive hemagglutination, hemagglutination inhbition and semi-quantitative precipitin reactions indicate strongly that ribitol plays a role in the serological specificity of the respective lipopolysaccharides.

Hemagglutination Inhibition Tests↗

Biological activities of lipopolysaccharides and lipid A from Rhodospirillaceae.

The lipopolysaccharides and free lipid A from several strains of Rhodospirillaceae were assayed comparatively with those of Enterobacteriaceae in a number of biological tests. Free lipid A's from Rhodopseudomonas gelatinosa and Rhodospirillum tenue exhibited strong serological cross-reactions with each other and with free lipid A from Salmonella. Lipid A's from Rhodopseudomonas viridis and Rhodopseudomonas palustris, although cross-reacting with each other, did not do so with either the lipid A of R. gelatinosa or R. tenue or with that of Salmonella. The presence or absence of the above cross-reactions agreed with corresponding similarities or differences in the chemical structure of the lipid A preparations. The lipopolysaccharide of R. gelatinosa was highly toxic for adrenalectomized mice and pyrogenic for rabbits; however, it exhibited no anti-complementary activity. The activity of the R. tenue lipopolysaccharide was very low in both the lethality and pyrogenicity tests. Its corresponding free lipid A also exhibited low pyrogenic activity; however, its lethal toxicity for adrenalectomized mice was considerably higher than that of the intact parent lipopolysaccharide. Both intact lipopolysaccharide and, unexpectedly, the free lipid A exhibited no anti-complementary activity. The lipopolysaccharides of R. viridis and R. palustris were virtually nontoxic for mice and nonpyrogenic for rabbits. Both lipopolysaccharides were highly potent in their interaction with complement. They therefore represent the first example of nontoxic lipopolysaccharides exhibiting high anti-complementary activity.

Animals↗

Demonstration of enterobacterial common antigen by bacterial agglutination.

Potent antisera against the enterobacterial common antigen (ECA) agglutinate R bacteria of the Enterobacteriaceae family that possess unimpaired R-core structures of the Escherichia coli R1 or E. coli R4 core type. In these strains, known to be ECA immunogenic, ECA is most probably linked to the lipopolysaccharide R core. R mutants of other core types (e.g., Salmonella Ra, E. coli R2 or R3) or R mutants with incomplete core structures of the E. coli R1 type, as well as an rfaL mutant deficient in the O-translocase system, agglutinate to a much lesser extent or not at all. All the later mutants are nonimmunogenic; they possess the ECA in a free form, not linked to the R core. None of the S forms tested from many different enterobacterial genera was found to be agglutinable with the ECA antiserum. The dynamics of the ECA agglutinin formation in rabbits parallels the ECA hemagglutinin formation, indicating that the same antibody class might be involved in bacterial agglutination and hemagglutination.

Agglutination↗

Lipophilic O-antigens in Rhodospirillum tenue.

Lipopolysaccharides of eight wild-type strains of the phototrophic bacterium Rhodospirillum tenue have been analyzed. All of the lipopolysaccharides are highly lipophilic. The compositions of preparations obtained by the phenol-water or by the phenol-chloroform-petroleum ether procedure are very similar. The polysaccharide moiety, obtained by mild acid hydrolysis of lipopolysaccharide, consists mainly of aldoheptoses: L-glycero-D-mannoheptose is present in all strains, whereas D-glycero-D-mannoheptose is an additional constituent in some strains. Galactosaminuronic acid and two unknown ninhydrin-positive components were detected in the lipopolysaccharides of six strains. Spermidine and putrescine are present in large amounts in a salt-like linkage in the lipopolysaccharides from three strains. 2-Keto-3-deoxyoctonate forms the linkage between the polysaccharide moiety and lipid A. The lipid A fraction contains all the glucosamine and all the D-arabinose present in the lipopolysaccharide. D-Arabinose is an invariable constituent of the lipid A from the Rhodopseudomonas tenue lipopolysaccharides investigated. The principal fatty acids are beta-hydroxycapric, myristic, and palmitic acids. The isolated R. tenue lipopolysaccharides (O-antigens) react with rabbit antisera prepared against homologous cells. The titers in passive hemagglutination are low, similar to those found with enterobacterial R-lipopolysaccharides. R. tenue O-antigens containing only L-glycero-D-mannoheptose and those containing both the L- and D-epimers of glycero-D-mannoheptose could not be differentiated by serological means.

Amines↗

Cloning of calf thymus satellite I DNA in Escherichia coli.

The 1400 base pair repeat produced by digestion of calf satellite I DNA (phi = 1.714 g/cm3) with EcoRI, was cloned in E. coli. The hybrid plasmid (pGM 214) which contains the ColE1-Ap vector (pSF 2124) and the 1400 base pair fragment replicates stably in E. coli and can be amplified by chloramphenicol treatment. No clone was found in which more than one "repeat unit" of the satellite I DNA was present in the chimaera plasmid. Digestion of the original satellite I and the plasmid pGM 214 with R-SmaI shows that the satellite DNA replicated in E. coli is cleaved by the restriction endonuclease SmaI whereas the original satellite I DNA from calf thymus is not, suggesting that the satellite I contains a large amount of modified cytosine or guanosine, probably 5-methyl-cytosine. R-EcoRI* produces a number of fragments with the satellite I in the range of 300 base pairs to 1400 base pairs. A physical map of pGM 214 (and pSF 2124) with R-EcoRI, R-HincII, R-HindIII, R-SmaI, R-BamI and R-EclI was constructed. The 1400 base pair "repeat unit" in the pGM 214 is efficiently transcribed in vitro by purified RNA polymerase, starting from a pSF 2124 promoter. The restriction enzyme EclI produces a 350 base pair repeat with calf satellite II (phi = 1,722 g/cm3), whereas the satellite I is not cut by this enzyme.

Animals↗

Isolation and characterization of the lipopolysaccharide of Chromatium vinosum.

Lipolysaccharide was isolated from Chromatium vinosum by phenol/water extraction. The lipopolysaccharide is found exclusively in the phenol phase and can be cleaved into a sugar moiety and a lipid A fraction by hydrolysis in 10% acetic acid at 100 degrees C for 3-4 h. The sugar moiety contains the neutral sugars 3-O-methyl-D-ribose, D-ribose, L-arabinose, mannosamine and glucose, and smaller quantities of D-rhamnose, D-glycero-D-manno-heptose (tentatively identified), quinovosamine and 2-keto-3-deoxyoctonate. L-glycero-D-manno-heptose was not detected. The 2-keto-3-deoxyoctonate linkage in C. vinosum lipopolysaccharide is more resistant to acid hydrolysis than that of Escherichia coli. The lipid A fraction contains glucosamine, mannose and the fatty acids of the lipopolysaccharide. The major fatty acid is beta-hydroxymyristic acid, with smaller amounts of lauric and palmitic acids as well as 14-carbon mono-unsaturated fatty acid, also being present. The phosphorus content of the C. vinosum lipopolysaccharide was found to be approximately 0.1%. Erythrocytes sensitized with alkali-treated C. vinosum lipopolysaccharide were agglutinated by antisera prepared against heat-killed cells. Untreated or heat-treated lipopolysaccharide did not sensitize erythrocytes. The lethal toxicity to mice of the C. vinosum lipopolysaccharide is about one-tenth as that from Salmonella abortus equi.

Animals↗

Immunochemical studies on lipopolysaccharides from wild-type and mutants of Escherichia coli K-12.

Lipopolysaccharides from a number of mutants of Escherichia coli K-12 were investigated by means of chemical and serological methods. Inhibition of passive hemagglutination and inhibition of precipitation show that L-rhamnose is the immunodominant sugar in the lipopolysaccharide from wild-type E. coli K-12. The disaccharide rhamnosyl-KDO (where KDO is 3-deoxy-D-manno-octulosonic acid) was isolated and characterized after mild acid hydrolysis of the lipopolysaccharide. It is concluded that rhamnose is present in the innermost part of the core as a side-chain substituent on KDO. From crosses between an E. coli K-12 donor and E. coli O8, hybrids were obtained which contained either one or both of the donor rfa and rfb clusters. Serum absorption studies with lipopolysaccharides from these hybrids indicated that the histidine-linked rfb cluster is responsible for the presence of rhamnose in the K-12 core oligosaccharide. Using paper chromatography of 32P-labelled lipopolysaccharides we have found heterogeneous lipopolysaccharide in two strains as well as some differences between two wild-type strains. The latter difference is believed to be due to varying contents of KDO-linked ethanolamine phosphate. The overall results presented together with those described in the companion paper clearly show that the core oligosaccharide in E. coli K-12 has a structure different from the types previously described for other strains of E. coli (designed coli R1 to coli R4).

Alleles↗

Antigenic determinants of murein lipoprotein and its exposure at the surface of Enterobacteriaceae.

Murein lipoprotein from the outer membrane of Escherichia coli could be fixed to erythrocytes without pretreatment of the erythrocytes. Passive hemagglutination or immune hemolysis could thus be used as sensitive assays to determine antibodies against lipoprotein. In rabbit antisera prepared against whole cells of E. coli, Salmonella, Arizona, and Shigella antibodies against lipoprotein were present. The respective titers were lowest in encapsulated smooth strains and highest in rough mutants. Antisera against deep rough mutants showed even higher anti-lipoprotein titers than anti-R-lipopolysaccharide titers. Correspondingly,absorption of lipoprotein antibodies with enterobacterial strains was most pronounced with deep rough mutants and lowest with smooth strains. Lipoprotein becomes increasingly an immunogen as well as an antigen the more sugar residues are missing in the lipolysaccharide on the cell surface. In wild-type cells lipoprotein is buried in the outer membrane; its exposure in mutant cells is related to defects at the cell surface.

Animals↗