[Detection of CagA protein in strains of Helicobacter pylori isolated from patients with gastritis and gastric ulcer disease]. / Prukaz CagA proteinu u kmenu Helicobacter pylori izolovaných od nemocných s gastritidou a vredovou chorobou zaludku.

UNLABELLED: A total of 156 Helicobacter pylori strains isolated from adult patients (81 men, 75 women) with diagnosis of gastritis (43 strains GAS) and gastric ulcer (113 strains GU) were analyzed for the production of CagA protein by quantitative and qualitative immunoblot methods. 80% of gastric ulcer group strains (84% in men, 73% in women) produced CagA protein, while in the gastritis patients group only 63% of strains were CagA protein positive (59% in men, 65% in women). Quantitative production was estimated by CagA index (mean production of CagA protein in CagA positive strains) in both groups of strains GAS and GU in men and women. Strains of GU group were shown to be higher producers (index CagA 2.42) as compared to GAS group (2.09). Index CagA was higher in strains isolated from men (index CagA 2.54) than is strains from women (index CagA 2.13). CONCLUSION: CagA production has been proved in 75% of analysed Helicobacter pylori strains. CagA positive strains were more frequent in gastric ulcer disease patients, slightly higher in the male than in the female patients. Strains isolated from men, irrespective of diagnosis (GAS and GU), are higher producers of CagA protein.

[Helicobacter pylori and diabetes mellitus]. / Helicobacter pylori a diabetes mellitus.

BACKGROUND: Helicobacter (H.) pylori participates significantly on the pathogeny of chronic gastritis, duodenal a gastric ulcer, carcinoma and lymphoma of the stomach. Attention was attracted also by some extragastric diseases, including diabetes mellitus, where the elevated antibodies levels against H. pylori was found. The aim of the work was to determine the seroprevalence in relation to the sex and type of disease in a cohort of diabetics and in a control group of blood donors. METHODS AND RESULTS: In 195 diabetic patients (type I and II) and 216 blood donors levels of IgG antibodies were determined by ELISA method. Seroprevalence in the group of diabetic patients was 27%, in males 26% and in women 28% (n.s.). No differences related to sex or diabetes type were found. Significant differences in seroprevalence were found among the group of diabetic patients and blood donors (27% vs. 51%, p < 0.001), male diabetics of both types and blood donors (26% vs. 62%, p < 0.001). No differences were found between female diabetic patients and controls (28%, 27% vs. 40%, n.s.). CONCLUSIONS: Our study has shown a lower seroprevalence of H. pylori in diabetic patients of type I and II in comparison with the healthy population. Such finding differs from the generally accepted experience of the higher sensitivity of these patients to infection. The practical significance of the observation remains unsolved.

Verocytotoxin-producing Escherichia coli in children with hemolytic uremic syndrome in the Czech Republic.

Since October 1988 till July 1995, 35 children (mean age 16 +/- 15.5 months) with classical hemolytic uremic syndrome (HUS) were examined for the presence of verocytotoxin (VT)-producing Escherichia coli (VTEC) infection. Stool samples from all patients were cultured for VTEC strains and tested for free fecal neutralizable VT. Serum samples from 18 patients taken on admission were also tested for antibodies to the lipopolysaccharide (LPS) from E. coli O157, O26, O55, O111, and O128 using the passive hemagglutination assay (PHA). Diagnosis of VTEC infection was established in 28 (80%) patients by the combined use of microbiological and serological techniques. VTEC were isolated from 16 (46%) patients, 5 of them had infection with 2 different VTEC serotypes; 11 (52%) VTEC isolates belonged to the serotypes O26: H11 (5), and O157: H7/NM (6). Free fecal VT was found in 21 (60%) patients. PHA antibodies to one or more LPS were detected in 14 (78%) of 18 patients; 9 had antibodies to O157 LPS and 6 to O26 LPS. We conclude from this study that VTEC are the important cause of pediatric HUS in the Czech Republic and the strains belonging to the serogroups O157 and O26 are the most prevalent.

Isolation and preliminary evaluation of a low-molecular-mass antigen preparation for improved detection of Helicobacter pylori immunoglobulin G antibodies.

Previously, immunoglobulin G (IgG) antibodies to five antigens with a relative molecular mass of between 15 and 30 kDa from Helicobacter pylori were found to be significantly more frequent in H. pylori-infected patients than in noninfected patients. In this study, these specific low-molecular-mass (LMW) antigens were separated by ultrafiltration of whole-cell sonicates. The LMW antigen preparation was evaluated by enzyme-linked immunosorbent assay with serum samples from 76 children with abdominal symptoms and 151 adults with dyspeptic symptoms. H. pylori was cultured or seen in 40 (53%) children and 83 (55%) adults. Increased antibody levels to H. pylori were found in serum from 35 (46%) children and 88 (58%) adults. Values for sensitivity, specificity, and predictive value of positive and negative results of the test were higher with LMW antigens than with the heat-stable antigen previously described. The low specificity and predictive value of a positive result were due to seropositive results for 21 persons with a negative culture for H. pylori and negative microscopy results for Helicobacter-like organisms in biopsies from gastric mucosa. Histologically, chronic gastritis was demonstrated in 43% of these persons, and 19% had peptic ulcer, indicating that they have or have had H. pylori infection. Specific antibodies to H. pylori were confirmed in all 21 patients by the Western immunoblot technique. Use of the LMW antigen improved the IgG antibody detection in patients with H. pylori infection, even though the results reflect the difficulties in establishing a true gold standard for diagnosis of H. pylori infection.

[The effect of omeprazole on healing of duodenal ulcers, Helicobacter pylori and gastritis]. / Vliv omeprazolu na hojení duodenálního vredu, Helicobacter pylori a gastritidy.

Losec (omeprazole) Astra Co. is a blocker of the proton pump of the parietal cell. It inhibits basal and stimulated HCl secretion. It is used for treatment of gastroduodenal ulcers, reflux oesophagitis and Zollinger Ellison's syndrome. In a group of 17 patients with duodenal ulcers the authors investigated the effect of omeprazole on (1) healing of duodenal ulcers and bulbitis after 2-4 weeks of therapy, (2) elimination of Helicobacter pylori in the antrum, (3) chronic antral gastritis. Ad 1. After two weeks of treatment the authors found that 5 of 17 chronic duodenal ulcers were healed in the remainder substantial regression was found. Four-week treatment led to healing of 16 from a total of 17 ulcers (P < 0.001), i. e. 94%. In subjects with ulcers and bulbitis (12 patients) the ulcer healed in 11 instances, in 7 patients residual bulbitis persisted. Ad 2. H. pylori was detected before treatment in 16 of 17 patients, after treatment only in 5 (P < 0.001). Ad 3. Chronic gastritis was recorded before treatment in all patients. Treatment reduced its activity and the presence of H. pylori.

[Determination of antibodies to Campylobacter pylori in children and adolescents with gastroduodenal disorders]. / Stanovení protilátek proti Campylobacter pylori u detí a dorostu s gastroduodenálními obtízemi.

Serological examination of antibodies against Campylobacter pylori was made in 195 boys and girls aged 9-20 years. In all on account of gastrointestinal complaints endoscopy was indicated. The results of microbiological and histological finding), in particular in the antrum and corpus of the stomach). Positive titres antrum and body of the stomach were compared with results of the complement fixation test and the ELISA reaction. Antibodies against C. pylori were detected in 64 of the examined subjects. The titres assessed by the complement fixation and ELISA test were in good agreement. Patients with positive sera had significantly more often a positive endoscopic finding (bacteria, chronic gastritis) and bacteriological finding), in particular in the antrum and corpus of the stomach). Positive titres of the complement fixation reaction persisted frequently for 15 months after the first examination. From the results ensues that serological examination of antibodies against C. pylori can be considered a sparing examination method which in patients with gastrointestinal complaints should precede endoscopic examination and moreover it can be used to follow up the course of the disease and success of therapy.

[Can epigastric pain and non-ulcerative dyspepsia in children and adolescents be Campylobacter pylori infection?]. / Mohou byt bolesti v epigastriu a neulcerózní dyspepsie u detí a dospívajících infekcním onemocnením Campylobacter pylori?

The authors examined 263 children and adolescents aged 5-20 years who suffered from so-called non-ulcerative dyspepsia or epigastric pain, because of the suspected presence of Campylobacter pylori. In 31.9% of the examined subjects direct microbiological methods revealed its presence in the gastroduodenal mucosa. The confidence limit of positive results of Campylobacter pylori in the population is between 27.58% and 38.84%, the probability being 95%. The authors proved a mutual correlation between endoscopic, histological, serological findings and detection of Campylobacter pylori by microbiological methods. The statistical significance of the correlation of campylobacter pylori and chronic gastritis B (mostly inactive) provides further support for the hypothesis of the aetiological role of Campylobacter pylori in the development of chronic gastritis B in children and adolescents. From the investigation it does not ensue, however, that colonization with Campylobacter pylori is associated with certain clinical symptoms in all instances. However, in the differential diagnosis of so-called non-ulcerative dyspepsia and epigastric pain in children and adolescents we must include infection with Campylobacter pylori among their possible causes.

[Cytotoxic changes in cell cultures induced by newly isolated strains of Campylobacter pylori]. / Cytotoxické zmeny vyvolané cerstve izolovanymi kmeny Campylobacter pylori v bunecných kulturách.

Suspensions of primocultures of two strains of Camplyobacter pylori, isolated from bioptic materials collected from patients with nodular hyperplasia of the pyloric antrum and with gastric ulcers were inoculated to 20-hour confluent or almost confluent cell cultures line VERO, McCoy and MA 104 in test tubes and Petri dishes. Both strains of C. pylori adhered markedly to cell lines VERO and McCoy and after 72 hours led to marked morphological changes of the cells. The changes were characterized in VERO cells by the presence of polygonal to star-shaped cells with processes, a reduced cytoplasm and only in the final stages by the formation of circular cells separating from the medium. Some cells were vacuolized. Cells of the McCoy line responded to the infection with C. pylori by a similar morphological picture, changes in the cell periphery were, as compared with the VERO line, more marked in the early stages. Cells of line MA 104 did not respond to infection with C. pylori by any morphological changes, not even after six days. The intensity of morphological changes was more marked in cell cultures line VERO infected with a strain isolated from a patient with a gastric ulcer, as compared with changes caused by a strain of C. pylori isolated from a patient with nodular hyperplasia of the pyloric antrum. The possibility to induce morphological changes of some stable lines supports the idea of the pathogenic action of Campylobacter pylori on the gastric mucosa.

Purification and some properties of exotoxin from Corynebacterium ulcerans strain ATCC 9015.

Corynebacterium ulcerans strain ATCC 9015 develops during growth in environmental media an exotoxin which is lethal for guinea-pigs. The exotoxin has been purified by ion-exchange chromatography on DEAE-Sephadex A-50, affinity chromatography using immunosorbent techniques, and gel-filtration through Sephadex G-75. The exotoxin is of a protein nature and consists perhaps of two subunits. The molecular weights of the exotoxin and its subunits as estimated by SDS-PAGE were 40 000, 25 000, and 15 000, respectively. The toxic properties of exotoxin were destroyed upon heating (60 degrees C, 30 min), treatment with trypsin (10 micrograms/ml, 37 degrees C, 30 min), and by the extremes of pH (pH 4.0 or 10;0 24 hrs). Homologous antibodies readily neutralized the exotoxin while commercial diphtheria antitoxin had no effects. No immunological cross-reactivity was revealed between exotoxin of C. ulcerans and other bacterial toxins such as diphtheria toxin, enterotoxin from V. cholerae, and exotoxin A of P. aeruginosa.

Proof for the lack of diphtheria toxin in culture filtrates of Corynebacterium ulcerans strain ATCC 9015.

Extensive efforts to identify the diphtheria toxin in culture filtrates of C. ulcerans strain ATCC 9015 was unsuccessful. The culture filtrates did contain a component (called D-antigen) which formed a precipitin line with commercial diphtheria antitoxin. This component was purified by immunosorbent techniques using immobilized preparations of diphtheria antitoxin. It was shown, that D-antigen has a molecular weight of 84000, is not toxic for guinea-pigs, and has nothing in common with purified diphtheria toxin when tested in gel-diffusion tests. The lack of diphtheria toxin in form of its fragments in culture filtrates concentrated 20-fold was proved by the highly sensitive and specific viroimmunoassay.

Enterotoxigenic E. coli in humans.

E. coli strains isolated from persons having diarrhoeal disease were tested for the production of TS enterotoxin. The production of TSE was demonstrated in 2.5% in a series of 80 strains isolated from children under one year of age, having acute diarrhoea. TSE was produced by 8.4% of E. coli strains out of 59 strains isolated from patients over one year of age. Among these strains, an interesting E. coli strain was isolated from the patient T. J., which produced TSE for more than 15 months. The production of TLE was tested though not proved in all strains by experiment on an isolated intestinal loop of an adult rabbit. The test on suckling mice so far appears to be the most suitable test for the demonstration of TSE. The results were considered positive when the index (the ratio of the weight of the whole intestine to the weight of the rest of the body) was higher than 0.08 while indices up to 0.078 were considered negative. E. coli strains with indices of intermediate values and strains with temporary production of TSE, occurring particularly in very small children, deserve special attention. The height of the indices was not influenced by a 30-minute exposure at 60 degrees C, but a decrease in the values of the indices was observed after boiling for a period of 15 min. The occurrence of E. coli strains producing TSE is evidently small in humans in European countries but, without doubt, they are important in the aetiology of diarrhoeal diseases.