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Biomedical subjects

M J Albert

Publications and source records attributed to M J Albert.

At least 91 records · Page 5Linked to original sources

Monoclonal antibodies to the enterotoxin of Bacteroides fragilis: production, characterization, and immunodiagnostic application.

A monoclonal antibody, ICT11, specific for the toxin of enterotoxigenic Bacteroides fragilis (ETBF) neutralized the cytotoxic effect of the toxin on human colonic cell line HT-29/C1. In an evaluation using 115 diarrheal stool specimens and culture as the "gold standard," the assay showed a sensitivity of 85% and a specificity of 100%. An ICT11-based sandwich enzyme-linked immunosorbent assay showed a sensitivity of 100% and a specificity of 98% for direct detection of toxin from stool samples compared with those of culture. Thus, ICT11-based assays will be useful for screening for ETBF.

Antibodies, Bacterial↗

Lipopolysaccharide-specific antibodies in plasma and stools of children with Shigella-associated leukemoid reaction and hemolytic-uremic syndrome.

Antibody responses to the lipopolysaccharide (LPS) of shigellae were compared between children with uncomplicated and complicated Shigella dysenteriae 1 infection. One hundred fifteen children between 12 and 60 months of age with S. dysenteriae 1 infection were studied. Of these children, 42 had complications (leukemoid reaction and/or hemolytic-uremic syndrome [complicated shigellosis] and 73 had no complications (uncomplicated shigellosis). Antibodies to the LPS of S. dysenteriae 1 and Shigella flexneri Y were measured in plasma and stools, as were total immunoglobulin A (IgA) and IgG concentrations in plasma and the total IgA concentration in stool, on enrollment and 3 to 5 days later. In the plasma, the concentrations of homologous (IgG) and heterologous (IgA) LPS antibodies on enrollment were higher in children with complicated shigellosis than in those with uncomplicated shigellosis. In stool, the concentrations on enrollment were similar between the two groups of children. There was a rise in antibody concentrations in the plasma (homologous and heterologous) and stool (homologous) between the day of enrollment and 3 to 5 days later in children with uncomplicated shigellosis but not in those with complicated shigellosis. These findings suggest that systemic stimulation is more marked in children with complications, so that a subsequent rise in plasma antibody concentrations does not occur in these children. In contrast, the lack of a rise in stool antibody concentrations in children with complicated shigellosis is suggestive of a lower-level mucosal response. Because the duration of diarrhea before enrollment influenced the homologous antibody concentrations, children were further divided into three subgroups (short [3 to 5 days], medium [6 to 9 days], and long [> 9 days] diarrhea durations before enrollment). Comparisons of homologous antibody concentrations between the two groups of children following such subdivisions showed that in children with complicated shigellosis, antibody concentrations were higher in the plasma of children in the short diarrhea duration subgroup but lower in the stool children in the medium diarrhea duration subgroup. No differences in antibody concentrations were observed in children in the other diarrhea duration subgroups. Thus, complications in shigellosis are associated with an early and strong systemic stimulation without a concomitant stimulation of the mucosal antibody response.

Antibodies, Bacterial↗

Cloning and sequence of a region of Vibrio cholerae O139 Bengal and its use in PCR-based detection.

We isolated and characterized a Vibrio cholerae O139 Bengal-specific DNA region by arbitrary PCR. The fragment contains open reading frames encoding two potential glycosyltransferases possibly involved in capsular polysaccharide or lipopolysaccharide biosynthesis. In order to evaluate the possibility that this region could be used for the specific detection of V. cholerae O139 Bengal, a PCR system was established. The specificity and sensitivity of the PCR were investigated by analyzing 240 strains within the family Vibrionaceae and 178 stains of other gram-negative bacteria. All V. cholerae O139 Bengal strains tested were positive, and none of the 384 control strains were amplified. The sensitivity of the assay was 10(2) CFU/ml.

Amino Acid Sequence↗

Controlled study of cytolethal distending toxin-producing Escherichia coli infections in Bangladeshi children.

The role of cytolethal distending toxin (CDT)-producing Escherichia coli, a newly described category of E. coli, in the causation of diarrhea was studied by screening E. coli isolates from 546 children < 5 years of age with diarrhea and 215 matched controls without diarrhea by using a specific DNA probe. Although CDT-positive E. coli strains were isolated from more children with diarrhea than from healthy controls (3.1 versus 0.93%), this difference did not reach statistical significance (P = 0.082). All CDT-positive strains also possessed the virulence factors of enteropathogenic E. coli or enteroaggregative E. coli isolates.

Acute Disease↗

Demonstration of a lack of synergistic effect of rotavirus with other diarrheal pathogens on severity of diarrhea in children.

The severity of group A rotavirus (RV) diarrhea was compared with that of mixed infections of RV with diarrheagenic Escherichia coli, Vibrio cholerae O1, and Shigella species by a scoring system. The severity of mixed infections of RV and E. coli was the same as that of infections with RV alone. RV infections mixed with V. cholerae and Shigella species mimicked cholera and shigellosis, respectively.

Child↗

Phage specific for Vibrio cholerae O139 Bengal.

From the stool of a Vibrio cholerae O139 Bengal-infected patient, a phage that specifically lysed capsulated V. cholerae O139 strains only was isolated. The phage is useful for the confirmatory diagnosis of V. cholerae O139 infection and for the differentiation of variants that lack the capsule.

Bacteriophages↗

Epidemiology & molecular biology of Vibrio cholerae O139 Bengal.

The emergence of Vibrio cholerae O139 Bengal as the second aetiologic agent of epidemic cholera in October 1992 in the south Indian coastal city of Madras has shattered the long-held notion that only V. cholerae belonging to serogroup O1 are capable of causing epidemic (and pandemic) cholera. Within months of its appearance in Madras, V. cholerae O139 engulfed the entire Indian subcontinent in a series of outbreaks of cholera. It also spread to several neighbouring countries in Asia. Several western countries also reported imported cholera cases due to this organism. In the regions of the Indian subcontinent where cholera due to V. cholerae O1 is endemic, children are mostly susceptible because adults would have acquired at least some immunity due to earlier exposure. However, when V. cholerae O139 struck people in these areas, even though all age groups were affected, the disease was more prevalent in adults, which suggested that the disease is new in this population. As with O1 cholera, water and food seemed to be the vehicles of infection. Many family contracts of index cases of O139 cholera were found to be infected with V. cholerae O139, and in many of them, the infection was asymptomatic which is reminiscent of O1 EITor infection. Again as with O1 EITor infection, individuals of blood group O were more susceptible to O139 infection than those with other blood groups. In its molecular aspects, O139 vibrio resembles O1 EITor vibrio. The virulence genes encoding cholera toxin, zonula occludens toxin, accessory cholera enterotoxin and core-encoded pilin are present in a 4.5 kb 'virulence cassette' region of the chromosome as in EITor vibrios and the expression of these virulence factors, toxin coregulated pilus (TCP) and several outer membrane proteins are found to be under the control of the master regulator ToxR as in EITor vibrios. The iron-regulated genes involved in virulence are also found in the same locus as in EITor vibrios. However, the genes involved in the somatic antigen synthesis in O1 vibrios are found to be deleted in O139 vibrios and are replaced by a new region of chromosome which encodes the new surface antigen synthesis in O139 vibrios. When V. cholerae O139 emerged and caused outbreaks, the prevailing O1 EITor vibrios virtually disappeared from most of the areas. The disappearance of EITor vibrios, the rapid spread of O139 vibrios and the resemblance of O139 vibrios to EITor vibrios seemed to suggest that O139 vibrios might be the causative agent of the 'eighth' pandemic of cholera. However, after a year of its appearance, O139 vibrios are on the wane and O1 EITor vibrios have re-emerged as the predominant organism, in the Indian subcontinent. Thus, the immediate threat of a new cholera pandemic posed by V. cholerae O139 may not be as large as it first seemed. However, whether it will follow the pattern of EITor vibrio which took approximately 60 yr since its first isolation before emerging as the seventh pandemic strain of cholera, is not clear. The factor(s) contributing to the diminished isolation of O139 vibrios and the re-emergence of O1 EITor vibrios are not understood. The vibrios might have undergone changes that would have affected their ability to survive and compete in the environment.

ABO Blood-Group System↗

Production, characterization and immunodiagnostic application of a monoclonal antibody to Shiga toxin.

A mouse monoclonal antibody (MAb ICT7) that is specific for Shiga toxin was produced. The MAb neutralises the cytotoxic effects of both purified Shiga toxin and culture extracts of Shigella dysenteriae type 1 in HeLa cells. Using MAb ICT7 and polyclonal rabbit antiserum, a sandwich ELISA was developed. This test detects Shiga toxin in both S. dysenteriae type 1 bacterial extracts and in stools of patients with S. dysenteriae type 1 infection. The ELISA also detects toxin in enterohaemorrhagic Escherichia coli (EHEC) strains positive for Shiga-like toxin I. The test could detect a minimum of 100 pg of purified Shiga toxin. Furthermore, the ELISA did not detect toxin in non-S. dysenteriae type 1 Shigella species or Shiga-like toxin II produced by EHEC strains.

Animals↗

Differentiation of Vibrio cholerae O1 isolates with biochemical fingerprinting and comparison with ribotyping.

The Phene Plate (PhP) system is a commercially available typing system based on the measurements of kinetics of selected biochemical reactions of bacteria grown in liquid medium in 96-well microplates. The system uses numerical analysis to identify biochemical phenotypes among the tested strains. In the present study, a set of 16 discriminatory tests were used to differentiate 117 strains of Vibrio cholerae O1 from MExico and Bangladesh. The stability of PhP types of 16 isolates under different storage temperatures and after repeated subcultures were also evaluated. The PhP system had a reproducibility of 95%. Storage either at +4 degrees C or -70 degrees C, did not affect the reactions of the isolates, whereas 4 strains (25%) stored at room temperature and 5 strains (31%) subjected to 30 consecutive subcultures, exhibited minor changes in their biochemical reactions. Endemic isolates of V. cholerae O1 from Bangladesh were more diverse (diversity index = 0.84 to 0.93) than epidemic isolates from Mexico (diversity index = 0.73). Using a collection of 33 heterogeneous isolates of classical biotype of vibrios, PhP typing and ribotyping were compared. PhP typing discriminated more types (n = 23) than ribotyping (n = 5), whereas a combination of both yielded 27 types. The PhP system appears to be a simple, reliable and highly discriminating method for typing of V. cholerae, and may prove especially useful as a first screening method in epidemiological studies of V. cholerae.

Bacterial Typing Techniques↗

Genetic rearrangements in the rfb regions of Vibrio cholerae O1 and O139.

The recent emergence of a pathogenic new non-O1 serotype (O139) of Vibrio cholerae has led to numerous studies in an attempt to identify the origins of this new strain. Our studies indicate that O139 strains have clear differences in the surface polysaccharides when compared with O1 strains: the lipopolysaccharide can be described as semi-rough. Southern hybridization with the O1 rfb region demonstrates that O139 strains no longer contain any of the rfb genes required for the synthesis of the O1 O-antigen or its modification and also lack at least 6 kb of additional contiguous DNA. However, O139 strains have retained rfaD and have a single open reading frame closely related to three small open reading frames of the O1 rfb region. This region is closely related to the H-repeat of Escherichia coli and to the transposases of a number of insertion sequence elements and has all the features of an insertion sequence element that has been designated VcIS1. Transposon insertion mutants defective in O139 O-antigen (and capsule) biosynthesis map to the same fragment as VcIS1. Preliminary sequence data of complementing clones indicate that this DNA encodes a galactosyl-transferase and other enzymes for the utilization of galactose in polysaccharide biosynthesis. We propose a mechanism by which both the Ogawa serotype of O1 strains and the O139 serotype strains may have evolved.

Amino Acid Sequence↗

Structure of the capsular polysaccharide of Vibrio cholerae O139 synonym Bengal containing D-galactose 4,6-cyclophosphate.

The capsular polysaccharide (CPS) of Vibrio cholerae O139 synonym Bengal, which is thought to carry determinants of O-specificity, was isolated by phenol/water extraction followed by delipidation of the contaminating lipopolysaccharide at pH 4.2 and gel-permeation chromatography. The CPS contained D-galactose, 3,6-dideoxy-L-xylo-hexose (colitose, Col), 2-acetamido-2-deoxy-D-glucose, 2-acetamido-2,6-dideoxy-D-glucose (N-acetyl-D-quinovosamine, D-QuiNAc), D-galacturonic acid (D-GalA), and phosphate. The CPS was studied by NMR spectroscopy, methylation analysis, and selective degradations, including partial acid hydrolysis at pH 3.1 and dephosphorylation with aqueous 48% hydrofluoric acid, which both resulted in complete cleavage of Col. It was concluded that the CPS is built up of hexasaccharide repeating units containing inter alia D-galactose 4,6-cyclophosphate and having the following structure [structure: see text] These data basically confirm the structure of the V. cholerae CPS proposed on the basis of an NMR study [L. M. Preston et al. (1995) J. Bacteriol. 177, 835-838] and specify exactly the absolute configurations of the constituent monosaccharides and the position of the cyclic phosphate.

Carbohydrate Sequence↗

Structural elucidation of the O-antigen lipopolysaccharide from two strains of Plesiomonas shigelloides that share a type-specific antigen with Shigella flexneri 6, and the common group 1 antigen with Shigella flexneri spp and Shigella dysenteriae 1.

Sugar and methylation analyses of native polysaccharides together with one-dimensional 1H- and 13C-NMR spectroscopy revealed that the two polysaccharides from strains 22074 and 12254 of Plesiomonas shigelloides are identical. The structure of the polysaccharide from strain 22074 was deduced from a uronic acid degradation and by NMR spectroscopy where heteronuclear multiple bond connectivity and two-dimensional nuclear Overhauser effect spectroscopy experiments established the pentasaccharide repeating unit as-->4)-alpha-D-GalpA-(1-->3)-alpha-D-GlcpNAc-(1-->3)-alpha-L- Rhap-(1-->2)-alpha-L-Rhap-(1-->2)-alpha-L-Rhap-(1-->.

Antigens, Bacterial↗

Moraxella septicemia in children with diarrheal disease.

We report 10 cases of Moraxella septicemia associated with diarrheal disease. Their clinical presentations and outcomes are discussed. Recognition of the pathogenicity of these microorganisms in appropriate clinical setting should result in prompt and specific therapy.

Anti-Bacterial Agents↗

Septicemia due to Vibrio cholerae 0139 Bengal.

We describe septicemia due to Vibrio cholerae 0139 Bengal, the newly described causative agent of cholera, in a child who also had simultaneous intestinal infection with Shigella boydii. Because V. cholerae 0139 is capsulated, its propensity to cause extraintestinal infection is stressed.

Bacteremia↗

Neonatal diarrhea caused by Vibrio cholerae 0139 Bengal.

Cholera rarely occurs in children under 2 years of age. We describe diarrhea due to Vibrio cholerae 0139 Bengal, the newly described etiologic agent of cholera in a 4-day-old breast-fed baby. However, the diarrhea was mild and was successfully treated with rehydration therapy and erythromycin.

Anti-Bacterial Agents↗