Emergence of multiple drug resistance in Vibrio cholerae O1 El Tor from Ecuador.
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Anti-Bacterial Agents↗
Biomedical subjects
B Said
Publications and source records attributed to B Said.
Vibrio cholerae serogroup 0139 in England and Wales.
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Cholera↗
Heterogeneity in expression of lipopolysaccharides by strains of Escherichia coli O157.
A total of 47 strains of Escherichia coli belonging to serogroup O157 were examined for the expression of long-chain lipopolysaccharide (LPS) by means of SDS-PAGE and silver staining. Strains belonged to 10 different flagellar (H) types or did not express flagella. Nine strains carried genes encoding Vero cytotoxin (VT). Strains of E. coli O157 expressed one of three LPS SDS-PAGE profiles designated A, B and C. Strains expressing profile A belonged to H-types 6 and 19, while those expressing LPS profile B belonged to H-types 2, 16, 20, 39, 42 and 45. Strains of E. coli expressing profile C belonged to H-types 7 and 8; strains producing VT expressed LPS profile C only, although not all strains with H-types 7 and 8 carried genes for VT. Regardless of H-type or the LPS profile expressed, serum antibodies produced by patients infected with strains of E. coli belonging to serogroup O157 would be detected by serological tests.
Antigens, Bacterial↗
Virulence properties of Escherichia coli strains belonging to serogroups O26, O55, O111 and O128 isolated in the United Kingdom in 1991 from patients with diarrhoea.
Some strains of Escherichia coli belonging to serogroups O26, O55, O111 or O128 produce Vero cytotoxin (VT). These serogroups are included in the range of enteropathogenic E. coli (EPEC) serogroups for which commercial antisera are available. In an attempt to obtain information on VT-producing strains other than those of serogroup O157, 122 strains belonging to these four serogroups and isolated in 1991 from patients with diarrhoea in the United Kingdom were tested for hybridization with VT probes. Only 18 of the 122 strains were VT-positive and these were O26 or O128. However 90 strains hybridized with the E. coli attaching and effacing (eae) probe (including 14 VT-positive strains) and 17 with the enteroaggregative E. coli (EAggEC) probe. For 78 eae-positive and 9 EAggEC-positive strains, tissue culture tests correlated with the probe results as the strains gave, respectively, either localized adhesion and a positive fluorescent-actin staining test or a characteristic aggregative attachment. A total of 111 of the 122 strains belonging to serogroups O26, O55, O111 or O128 possessed properties that may be associated with the ability to cause human diarrhoeal disease, and similar studies are needed on strains from the other classical EPEC serogroups.
Adult↗
Effect of 5-methylcytosine as a neighboring base on methylation of DNA guanine by N-methyl-N-nitrosourea.
Effects on N-methyl-N-nitrosourea (MNU) mediated methylation of the N7 position of guanine were compared in defined sequences of DNA containing cytosine or 5-methylcytosine (5mC) using a Maxam-Gilbert sequencing technique. Cytosine methylation in 5'-CpG-3' pairs within a subcloned fragment of the 5' region of the human HPRT gene was generated with SssI methylase and S-adenosylmethionine. Cytosine methylation was demonstrated by both the inhibition of DNA restriction by methylation sensitive endonucleases and the lack of cleavage at 5-methylcytosines by hydrazine. MNU-dependent methylation of the N7 position of guanine was inhibited up to 18% when 5mC was a 5' neighboring base to guanine and was inhibited up to 36% in an alternating CpG region in which both 5' and 3' neighboring bases of guanine were enzymatically altered to 5mC. It can be concluded that 5-methylcytosine has discernible effects on MNU methylation of the N7 position of specific guanine bases in DNA.
5-Methylcytosine↗
Linoleic acid inhibition of adhesion of enteropathogenic Escherichia coli to HEp-2 cells.
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Bacterial Adhesion↗
Nearest neighbor effects on carcinogen binding to guanine runs in DNA.
A synthetic DNA fragment was constructed to determine the effect of 5' and 3' neighbors of guanine runs on the binding of chemical carcinogens. Determinations were made on the relative intensity of reactivity between aflatoxin B1 or benzo(a)pyrene and methylnitrosourea or 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea with various guanine positions in an endlabeled DNA fragment of known sequence. After reaction, the fragments were depurinated to produce strand breaks to allow Maxam and Gilbert sequencing for guanine positions. Relative reaction intensities were compared densitometrically. 3' neighbors exerted greater influence on carcinogen binding than did 5' neighbors, the influence extended only to the adjacent guanine and depended upon the chemical nature of the carcinogen. In addition, the presence of one carcinogen adduct in the guanine run influenced the formation of a subsequent adduct when the DNA was exposed to a second carcinogen, and this effect also depended on the nature of the second carcinogen. The results suggest that DNA adduct formation in the presence of multiple carcinogens is more complicated than an additive mechanism would suggest.
Aflatoxin B1↗
Identification of enteropathogenic Escherichia coli isolated in Britain as enteroaggregative or as members of a subclass of attaching-and-effacing E. coli not hybridising with the EPEC adherence-factor probe.
Strains of Escherichia coli from sporadic cases of diarrhoea and belonging to serotypes O44:H18, O55:H7, O111ab:H21, O111ab:H25 or O126:H27 were examined for virulence properties. With the exception of O111ab:H25 these are considered to be classical enteropathogenic E. coli (EPEC) serotypes. The strains had been isolated in Britain from the faeces of children less than 3 years old. Of the serotypes examined, 7 of 13 O44:H18 strains, all of 10 O111ab:H21 strains and 13 of 21 O126:H27 strains belonged to the enteroaggregative class of E. coli (EAggEC) that attached to HEp-2 cells in the characteristic aggregative pattern and hybridised with the EAggEC probe. They also caused mannose-resistant haemagglutination of rat erythrocytes, a property which may be a useful marker for their identification. Strains of O44:H18 with similar properties were also isolated from three small outbreaks in Britain, one of which involved elderly patients. EAggEC have not been considered previously as aetiological agents of diarrhoea in developed countries and have rarely been reported as belonging to EPEC serotypes. All 15 O55:H7 strains and seven of eight O111ab:H25 strains were also considered to be potentially diarrhoeagenic as they gave localised attachment (LA) to HEp-2 cells that resulted in a positive fluorescence actin-staining test. This test is considered to correlate with the attaching-and-effacing virulence mechanisms of EPEC in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Identification of Escherichia coli that produces heat-stable enterotoxin STA by a commercially available enzyme-linked immunoassay and comparison of the assay with infant mouse and DNA probe tests.
By using the infant mouse test and a commercially available competitive enzyme-linked immunosorbent assay (ELISA), 100 strains of Escherichia coli carrying STA1 or STA2 genes were shown to produce the heat-stable enterotoxin STA. An additional 100 strains were negative in both tests. The ELISA was easy to perform, and results were available within 24 h. Testing strains with an enzyme-linked DNA probe kit that incorporated both STA1- and STA2-specific oligonucleotides showed that the 100 strains positive in the mouse test and ELISA also hybridized with the mixed probe. Two strains carrying STA1 genes but negative in the mouse test and ELISA also hybridized with the mixed probe.
Animals↗
Mutational analysis of the L1 binding site of 23S rRNA in Escherichia coli.
The L11 ribosomal protein operon of Escherichia coli contains the genes for L11 and L1 and is feedback regulated by the translational repressor L1. Both the L1 binding site on 23S rRNA and the L1 repressor target site on L11 operon mRNA share similar proposed secondary structures and contain some primary sequence identity. Several site-directed mutations in the binding region of 23S rRNA were constructed and their effects on binding were examined. For in vitro analysis, a filter binding method was used. For in vivo analysis, a conditional expression system was used to overproduce a 23S rRNA fragment containing the L1 binding region, which leads to specific derepression of the synthesis of L11 and L1. Changes in the shared region of the 23S rRNA L1 binding site produced effects on L1 binding similar to those found previously in analysis of corresponding changes in the L11 operon mRNA target site. The results support the hypothesis that r-protein L1 interacts with both 23S rRNA and L11 operon mRNA by recognizing similar features on both RNAs.
Bacterial Proteins↗
Nucleotide sequencing and expression of the fadL gene involved in long-chain fatty acid transport in Escherichia coli.
The fadL gene of Escherichia coli codes for an outer membrane protein involved in long-chain fatty acid transport. Its product was purified from outer membrane proteins. We determined the nucleotide sequence of a 2.8-kb chromosomal DNA segment that contains the fadL gene. The fadL gene consists of a 1149-nucleotide coding region and contains a highly hydrophobic polypeptide of 383 amino acids with a calculated molecular weight of 42,380. We have used S1-mapping analysis to identify the transcription initiation site. A region exhibiting extensive dyad symmetry and perfect homology to the catabolite activator protein binding site was detected.
Amino Acid Sequence↗
Purification and characterization of an outer membrane-bound protein involved in long-chain fatty acid transport in Escherichia coli.
We report the purification and localization of the fadL gene product (FLP), an essential component of the long-chain fatty acid transport machinery in Escherichia coli. FLP was extracted from total membranes by differential extraction with the nonionic detergents Tween 20 and Triton X-100. This protein was further purified from a Tween 20-insoluble-Triton X-100-soluble extract by salt fractionation, gel filtration chromatography, and hydrophobic interaction chromatography. This regime results in a 95-fold purification of FLP from total membranes. The purified protein preparation was homogeneous based on silver staining and gave the characteristic behavior established for the fadL gene product in the presence of sodium dodecyl sulfate at different temperatures prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mr of 33,000 when heated at 25 degrees C and Mr of 43,000 when heated at 100 degrees C) and on two-dimensional polyacrylamide gels (pI of 4.6 and a Mr of 33,000). Purified FLP was rich in hydrophobic residues accounting for approximately 45% of the total amino acid composition. To localize FLP, antisera were raised against the purified protein and were used to probe differentially fractionated membranes by Western immunoblotting. This procedure demonstrated the presence of this protein only in the outer membrane fraction of fadL+ strains. We confirmed the outer membrane localization of FLP by measuring long-chain fatty acid transport in fadL+ and fadL strains treated with EDTA to alter outer membrane permeability and in spheroplasts generated from fadL+ and fadL strains. Both EDTA-treated cells and spheroplasts transported long-chain fatty acids at essentially the same rate regardless of whether they contained a wild-type or mutant fadL gene. These data imply that FLP is a protein in the outer membrane which is specifically involved in long-chain fatty acid transport.
Amino Acids↗
Rat heart fatty acid-binding protein is highly homologous to the murine adipocyte 422 protein and the P2 protein of peripheral nerve myelin.
The rat heart contains an abundant cytosolic protein which binds long chain fatty acids. We have determined its primary structure by Edman degradation of peptides generated from chymotryptic, tryptic, and elastase digestions. This polypeptide (Mr = 14,992) contains 134 amino acids and has a blocked (acetylated) NH2 terminus. The sequence of rat heart fatty acid-binding protein (FABP) is remarkably similar to the murine adipocyte 422 protein and the P2 protein of peripheral nerve myelin. Computer-assisted alignment of heart FABP and 422 revealed that 82 of 132 comparable residues are identical (62%). There are 77 identities out of 131 possible matches between this protein and the human myelin P2 protein (59%). Similar comparisons demonstrate that heart FABP has significant homology to several other proteins which bind hydrophobic ligands. The rank of order of similarity to heart FABP is: 422 greater than myelin P2 greater than cellular retinoic acid-binding protein greater than cellular retinol-binding protein II greater than cellular retinol-binding protein greater than intestinal FABP greater than liver FABP. These eight sequences form a family of paralogous homologues. Heart FABP has a region of internal homology involving tandemly arrayed oligopeptides spanning residues 71-100 and 101-131. This feature is not found in the 422 and P2 sequences. The endogenous ligands bound by the 422, P2, and heart FABP sequences have not been defined. Interpretation of the biological significance of their structural similarities and differences will require information about their ligand specificities and affinities.
Amino Acid Sequence↗
Fatty acid binding protein from rat heart. The fatty acid binding proteins from rat heart and liver are different proteins.
The binding of [3H]oleic acid and [3H]palmitic acid to the low molecular weight fatty acid binding proteins present in the cytosols of rat liver and heart was studied. Both fatty acids were bound by Z protein of liver, whereas only oleic acid was bound by the fraction of heart that contains the fatty acid binding protein. However, after delipidation of heart cytosolic proteins with butanol, the binding of palmitic acid to the fatty acid binding protein was detected. In contrast to a published report (Gloster, J., and Harris, P. (1977) Biochem. Biophys. Res. Commun. 74, 506-513), oleic acid was not bound by rat heart or bovine heart myoglobin. Both rat heart fatty acid binding protein and rat liver Z protein were purified to homogeneity or near homogeneity. On polyacrylamide gel electrophoresis under nondenaturing conditions, liver Z protein gave rise to three bands, none of which was identical with the single band due to heart fatty acid binding protein. Rabbit antibodies to rat liver Z protein were used to demonstrate that the purified fatty acid binding protein from rat liver (Z protein) and rat heart are immunologically unrelated and that no Z protein is present in rat heart cytosol. Taken together, these observations lead to the conclusion that the low molecular weight fatty acid binding proteins from rat liver and heart are different proteins.
Animals↗