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Biomedical subjects

S Tzipori

Publications and source records attributed to S Tzipori.

At least 73 records · Page 4Linked to original sources

Peripheral blood granulocytes and mononuclear cell responses in monkeys with experimental shigellosis.

Changes in neutrophil response to N-formyl-methionyl-leucyl-phenylalanine (FMLP) and the phenotype of peripheral blood mononuclear cells were studied in monkeys after oral challenge with Shigellae. Monkeys were first challenged with S. dysenteriae 1 which caused shigellosis in some of the monkeys. After recovery, the monkeys were rechallenged with S. flexneri 2a. No difference in sensitivity was observed in the monkeys during shigellosis caused by either S. dysenteriae 1 or S. flexneri 2a. The optimal dose of FMLP for neutrophil polarization, a measure of early cell activation, in normal healthy monkeys was 10(-7) M when 67% of the neutrophils were polarized. Neutrophils from monkeys ill with shigellosis required higher doses of FMLP (10(-6) and 5 x 10(-7) M) for maximum polarization. As the monkeys recovered, a gradual decrease in the doses of FMLP for optimal neutrophil polarization was also observed. The percentage of CD2-positive T lymphocytes, the earliest marker for T lymphocytes in the peripheral blood, decreased when the monkeys developed shigellosis and returned to normal levels as the monkeys improved. However, there was no change in the percentage of CD20-positive peripheral blood B lymphocytes.

Animals↗

Rapid detection of Shigella dysenteriae and Shigella flexneri in faeces by an immunomagnetic assay with monoclonal antibodies.

A rapid and sensitive method for the detection of Shigella dysenteriae type 1 and Shigella flexneri serotypes in faeces based on capture of the bacteria with immunomagnetic particles is described. The particles were coated with either of two different monoclonal antibodies specific for the O-antigens of Shigella dysenteriae type 1 and Shigella flexneri serotypes. Captured bacteria were detected by an enzyme immunoassay with O-antigen specific rabbit antiserum. The whole assay required 2 to 3 hours to perform and the sensitivity limit was 10(3) cfu/ml as determined by viable cell counting. One hundred and fifty enterobacteria strains, including 100 Shigella strains from a strain collection, and 302 fresh faecal samples were used for the study. All Shigella dysenteriae type 1 and Shigella flexneri culture-positive faecal samples were positive in the immunomagnetic assay. In addition 18 of 252 culture-negative faecal samples were positive. The immunomagnetic assay was compared with latex agglutination and indirect immunofluorescence using culture as the reference method. The immunomagnetic assay had a sensitivity of 100%, latex agglutination a sensitivity of 72% with 28% false-negative results, and indirect immunofluorescence a sensitivity of 95%. The immunomagnetic assay was superior in sensitivity since it also detected Shigella in faecal samples up to two days after antibiotic therapy had been started, which both latex agglutination and indirect immunofluorescence failed to do. The high sensitivity in detecting live and dead bacteria, and the ease of performance of the immunomagnetic assay render it an attractive method for detection of Shigella.

Animals↗

The role of the eae gene of enterohemorrhagic Escherichia coli in intimate attachment in vitro and in a porcine model.

The eaeA gene of enteropathogenic Escherichia coli (EPEC) is necessary for intimate attachment to epithelial cells in vitro. Enterohemorrhagic E. coli (EHEC) strains also possess an eae gene and are capable of intimate attachment and microvillus effacement in vitro and in animal models. To assess the role of the EHEC eae gene in intimate attachment, we constructed an eae deletion/insertion mutation in wild-type EHEC O157:H7 strain 86-24 by using linear electroporation of a recombinant allele. The mutant obtained was deficient in inducing f-actin accumulation in HEp-2 cells and was incapable of attaching intimately to colonic epithelial cells in a newborn piglet model of infection. Intimate attachment in vivo was restored when the EHEC eae gene or the eaeA gene of EPEC was introduced into the mutant on a plasmid. These results indicate that the eae gene is necessary for intimate attachment of EHEC in vivo. In addition, the complementation achieved by the EPEC locus indicates that the eae gene of EHEC and the eaeA gene of EPEC are functionally homologous.

Adhesins, Bacterial↗

Evaluation of a probe hybridisation serotyping method for group A rotavirus.

Serotype-specificity and sensitivity of oligonucleotide probes to serotype human rotaviruses was assessed. Probes could detect as little as 6.3 ng of homologous RNA and none reacted with as much as 100 ng of heterologous RNA. Northern-blot analysis revealed that probes reacted with one of genomic segments 7, 8 or 9 of corresponding serotypes.

Diarrhea↗

DNA probe analysis of diarrhoeagenic Escherichia coli: detection of EAF-positive isolates of traditional enteropathogenic E. coli serotypes among Bangladeshi paediatric diarrhoea patients.

Escherichia coli isolates from all surveillance patients less than or equal to 20 months of age seen for diarrhoea at the Dhaka Clinical Treatment Facility of the International Centre for Diarrhoeal Disease Research, Bangladesh between March 1 and August 31, 1988, were collected and hybridized with DNA probes to assess the potential importance of diarrhoeagenic E. coli among paediatric patients in Bangladesh. Of 396 patients evaluated, 18% were infected with enteropathogenic E. coli (EPEC) adherence factor (EAF)-positive E. coli, 23% were infected with enterotoxigenic E. coli (ETEC), 9% were infected with Shiga-like toxin-positive E. coli, and 13% were infected with diffuse adhesiveness-positive E. coli. None were infected with enteroinvasive E. coli. Ten percent of patients were colonized with more than one type of potential diarrhoeagenic E. coli. The majority of EAF-positive isolates were of traditional EPEC O:H serotypes. Although this was not a case-control study, the large number of EPEC and ETEC, which are recognized enteric pathogens, suggests these organisms are important causes of diarrhoeal diseases in this pediatric population.

Bangladesh↗

A comparative study of specific gene probes and standard bioassays to identify diarrhoeagenic Escherichia coli in paediatric patients with diarrhoea in Bangladesh.

We compared the usefulness of gene probes with standard bioassays to identify diarrhoeagenic Escherichia coli amongst isolates from Bangladeshi children under 1 year of age with diarrhoea. E. coli isolates were analysed with specific gene probes for localised adhesiveness (LA), diffuse adhesiveness (DA), heat-labile toxin (LT), heat-stable toxin (ST), Shiga-like toxins (SLT I and SLT II), and enteroinvasiveness, and in bioassays for production of enterotoxins and cytotoxins, and for cell adherence. With 1136 isolates from 387 patients, there was general agreement between the two assay methods. When there was disparity, gene-probe-positive isolates gave negative results in the corresponding bioassay. In the HeLa cell adherence assay, 94% of the LA probe-positive isolates and 91.6% of the DA probe-positive isolates gave positive bioassay results for LA and DA respectively. Thirty-six of 39 LT probe-positive isolates and 73 of 86 ST probe-positive isolates gave positive results in the bioassays. Of 28 isolates that gave negative results in the suckling mouse assay but were initially positive with the probe for ST, 15 were later found to hybridize with the cloning vector for the ST probe. Addition of denatured vector DNA at a concentration of 10 micrograms/ml in the hybridisation solution eliminated these false positive results. None of the other probe-positive isolates hybridised with any of the cloning vectors used. The DNA hybridisation assay appeared to be a convenient alternative to bioassays for screening large numbers of isolates in epidemiological investigation.

Bacterial Adhesion↗

Studies with enteroaggregative Escherichia coli in the gnotobiotic piglet gastroenteritis model.

Two strains of enteroaggregative Escherichia coli of human origin fed to gnotobiotic piglets caused diarrhea or death in the majority of them. Histological examination revealed moderate hyperemia of the distal small intestine and cecum, swelling of small intestinal villi, and layers of aggregated bacteria stacked together in a mucus gel-like matrix overlying intact epithelium. These findings confirm that enteroaggregative E. coli strains produce distinctive intestinal lesions different from those caused by other major categories of diarrheagenic E. coli.

Animals↗

An ELISA for the detection of localized adherent classic enteropathogenic Escherichia coli serogroups.

An ELISA for the detection of classic enteropathogenic Escherichia coli (EPEC) serogroups was developed. It detected EPEC positive for localized adherence (LA) by HeLa cell assay and EPEC positive for EPEC adherence factor (EAF) by DNA probe assay. A specific antiserum was raised with LA+ EPEC strain E2348/69 (serotype O127:H6) by immunizing rabbits and then absorbing the antiserum with its LA- derivative, MAR20. The absorbed antiserum reacted specifically with all 90 strains of E. coli belonging to eight different EPEC serogroups that were LA+ by HeLa cell assay and EAF+ by DNA probe assay. All E. coli strains including EPEC serogroups that were LA- by HeLa cell assay and EAF- by DNA probe assay were also negative by ELISA. Thus the ELISA is 100% sensitive and specific in detecting LA+ classic EPEC serogroups.

Bacterial Adhesion↗

The association of haemagglutination and adhesion with lipopolysaccharide of Shigella dysenteriae serotype 1.

In this study the ability of strains of Shigella dysenteriae serotype 1 to agglutinate mammalian erythrocytes is attributed to the polysaccharide fraction of bacterial-cell lipopolysaccharide (LPS). LPS obtained from a rough, mutant strain of S. dysenteriae serotype 1, lacking the O-antigen polysaccharide side-chain, did not agglutinate erythrocytes, clearly demonstrating a link between O-antigen polysaccharides and haemagglutinating activity (HA). Strains of S. dysenteriae serotype 1 adhered well to cultured Henle Intestinal 407 cells, whereas rough strains adhered poorly. Pre-treatment of bacteria with LPS-specific antisera inhibited both HA and binding to cultured human-intestinal cells. The contribution of the polysaccharide side-chain and its associated HA--which appear to facilitate binding to cultured cells--to bacterial attachment to colonocytes and to the pathogenesis of shigellosis in vivo needs to be confirmed in animal studies.

Bacterial Adhesion↗

Hafnia alvei, a probable cause of diarrhea in humans.

Hafnia alvei, a member of the family Enterobacteriaceae, was the only species of bacteria cultured from the stool of a 9-month-old child who was admitted with a 3-day history of watery diarrhea. The isolated strain of H. alvei failed to produce heat-labile or heat-stable enterotoxins or Shiga-like toxin I or II and did not invade HeLa cells, nor did it cause keratoconjunctivitis (determined by the Sereny test) in a guinea pig's eye. The strain, however, induced diarrhea in 8 of 12 adult rabbits with removable intestinal ties (removable intestinal tie-adult rabbit diarrhea [RITARD] assay) and in 1 of 2 orally fed animals. No diarrhea could be induced with Escherichia coli K-12 in eight RITARD assay rabbits and three orally fed rabbits, respectively. Microscopic examination of affected animals revealed moderate inflammatory cellular infiltration of the intestinal mucosa, in which bacterial attachment to the surface epithelium and loss of the microvillus border were evident in the ileum and colon. Electron microscopy demonstrated cellular modifications of the apical surface, with cupping or pedestal formation and increased terminal web density at sites of bacterial "attachment-effacement," a well-known characteristic and mechanism of diarrhea of enteropathogenic E. coli. Identical lesions were also induced by H. alvei in rabbit ileal loops, which ruled out naturally occurring rabbit enteropathogenic E. coli strains, which are known to produce similar lesions. It is concluded that at least some strains of H. alvei have the potential to cause diarrhea and that attachment-effacement is a virulence characteristic shared by bacteria other than E. coli.

Animals↗

Localized adherence and attaching-effacing properties of nonenteropathogenic serotypes of Escherichia coli.

Traditional enteropathogenic Escherichia coli serotypes demonstrate a plasmid-mediated localized adherence in cultured HeLa or HEp-2 cells and induce an attaching-effacing intestinal lesion, both of which are considered pathognomonic and causes of diarrhea. This study describes three E. coli strains from infantile diarrhea which share these properties but belong to serotypes (O2:H2, O2:H25 and O15:H2) not considered enteropathogenic.

Animals↗

Cultivation and characterization of novel human group A rotaviruses with long RNA electropherotypes, subgroup II specificities, and serotype 2 VP7 genes.

During an epidemiological study of human rotavirus infections in Bangladesh, three group A strains hybridized with a serotype 2 oligonucleotide probe, but they had long RNA electropherotypes. The three strains were collected from 8- to 20-month-old infants with acute diarrhea and moderate malnutrition. By a modified isolation procedure, two strains (T-B and T-C) were adapted in MA104 cell cultures. They were identified to be subgroup II specific by enzyme-linked immunosorbent assay with subgroup I- and II-specific monoclonal antibodies and were identified by a fluorescent focus reduction neutralization assay with hyperimmune antisera to be serotype 2 specific. Further characterization of these unusual rotavirus strains needs to be carried out.

Antibodies, Viral↗

Rotavirus-associated diarrhea in rural Bangladesh: two-year study of incidence and serotype distribution.

Stools were evaluated from 5,811 patient visits for treatment of diarrhea in Matlab, Bangladesh, between June 1987 and May 1989. The stools were analyzed to determine the distribution of serotypes of group A rotaviruses (RV). A total of 898 stool samples (15.5%) contained RV, as determined by using an enzyme-linked immunosorbent assay. RV isolates from 855 of these samples were serotyped by using serotype-specific synthetic oligonucleotide probes. A total of 558 (65.3%) could be assigned to specific serotypes: 166 (19.4%), 228 (26.7%), 39 (4.6%), and 125 (14.6%) belonged to serotypes 1 through 4, respectively; 12 (1.4%) hybridized with more than one serotype; and 285 (33.3%) failed to hybridize. RV diarrhea was evident throughout the year, with peaks in the dry winter months and in September 1988, coinciding with a major flood. RV was isolated from 46.6% of patients between 7 and 12 months old. Among children under 24 months of age with RV diarrhea, 1.2% (10 of 828) died. The corresponding percentage for children with diarrhea from all causes is 0.9% (29 of 3,301).

Bangladesh↗

Outbreak of keratoconjunctivitis due to Salmonella weltevreden in a guinea pig colony.

The purpose of this report is to demonstrate that the ability to produce keratoconjunctivitis (KC) is a property found in Salmonella weltevreden. This observation is contrary to previous reports that Salmonella spp. do not produce KC. An outbreak of KC due to S. weltevreden occurred in a guinea pig colony, and the animals carried the organism in the intestinal tract. The same Salmonella serotype that caused an epidemic of diarrhea in humans and a routine laboratory isolate also possessed the ability to induce KC. Unlike Shigella spp. (the prototype organisms positive for KC), S. weltevreden induced KC and bound Congo red dye even when grown at 30 degrees C. It invaded HeLa cells in culture but did not hybridize with a DNA probe for invasiveness of Shigella spp. and enteroinvasive Escherichia coli even though it harbored plasmids. It was susceptible to all the antibiotics tested, was hydrophobic, and showed mannose-sensitive hemagglutination. It did not have enterotoxic or cytotoxic activities.

Animals↗

Enteroaggregative Escherichia coli infections in Bangladeshi children: clinical and microbiological features.

Escherichia coli isolated from 389 children, less than 1-year of age with diarrhoea, were analyzed for the presence of Enteroaggregative E. coli (EAggEC) by their pattern of adherence to HeLa cells. EAggEC were isolated from 58 (14.8%) children either as a sole pathogen or in combination with other enteric pathogens. In 60% of these children EAggEC infection occurred in the second half of infancy (7-12 m). Thirty-eight of 47 children having EAggEC as a sole pathogen had watery diarrhoea along with vomiting (87%) and dehydration (74%). In contrast, 9 of the 47 cases had mucoid diarrhoea with infrequent vomiting and dehydration and frequent abdominal pain. Children infected with EAggEC were successfully rehydrated with oral rehydration solution (ORS) alone. Seventy-one percent of the EAggEC strains were resistant to more than three antibiotics. It was evident by phage pattern that various EAggEC strains were present in the population. The results indicated that infections with EAggEC may have a role in the development of diarrhoea among children less than 1-year of age in Bangladesh.

Bacterial Adhesion↗

Zymodemes of Entamoeba histolytica in Dhaka, Bangladesh.

Stool samples containing Entamoeba histolytica were obtained from two sources: an urban hospital and an urban slum. Using the enzyme patterns of three enzymes (E.C.2.7.1.1 hexokinase, E.C.5.3.1.9 glucose phosphate isomerase and E.C.2.7.5.1 phosphoglucomutase) stained after cellulose acetate electrophoresis, four zymodemes (I, II, XIV and XVI) were identified in 71 isolates. Zymodemes considered to be pathogenic (II and XIV) were identified from 31 of 34 isolates from the urban hospital, and these zymodemes were strongly associated with dysentery and trophozoites containing ingested red blood corpuscles. Blood visible to the naked eye was more commonly seen in stools from which zymodeme XIV was isolated than in those containing zymodeme II (P less than 0.001). Zymodemes considered to be non-pathogenic (I and XVI) were identified in 34 of 37 isolates from slum dwellers and were not associated with blood in the stools.

Animals↗

Treatment with bovine hyperimmune colostrum of cryptosporidial diarrhea in AIDS patients.

Cryptosporidium parvum may cause severe, debilitating diarrhea in patients with AIDS. Recent anecdotal reports have suggested that hyperimmune bovine colostrum may be effective. We conducted a double-blind, controlled pilot study of hyperimmune bovine colostrum for diarrhea due to cryptosporidiosis in five AIDS patients. The patients were randomized to receive either hyperimmune or control colostrum by continuous nasogastric infusion for 10 days. All stools were collected, graded, and weighed, and the concentration of oocysts excreted was determined daily. One of the three patients treated with hyperimmune colostrum had a reduction in diarrhea and in the concentration of oocysts excreted. A second treated patient had a modest decrease in the concentration of oocysts excreted. Two patients who received control colostrum also had decreases in the volume of diarrhea but no change in the concentration of oocysts excreted. We conclude that hyperimmune colostrum with high titers of specific anti-Cryptosporidium antibody could be effective in treating patients with cryptosporidiosis. However, more studies of cow colostral immunoglobulin need to be performed so that the efficacy of this treatment can be assessed more thoroughly.

Acquired Immunodeficiency Syndrome↗