[THE CORRECTION OF THE LOCAL IMMUNE RESPONSE AT THE PATIENTS WITH HELICOBACTER PYLORI INFECTION].

INTRODUCTION: Helicobacter pylori infection is due to the high prevalence in population attracts the clinical interest of researchers in the whole World. It is well known that this microorganism not only resides in the mucosa of the gastrointestinal tract, but is also defined in the periodontal pocket of the oral cavity. THE AIM OF INVESTIGATION: to evaluate Helicobacter pylori diagnostics in the mouth and prove a method of relief of the inflammatory process by applying immunomodulator Imudon. RESULTS. On the basis of obtained results it was found that the inclusion of topical immunomodulator Imudon in the complex therapy of Helicobacter pylori-associated diseases leads to reduction of inflammatory potential through the decrease of the TNFα, IL-6 activity in saliva and to increase the protective properties of saliva as a result of increased levels of mucin, significantly reduces the frequency of relapses in the one year after therapy. CONCLUSION: It is practically important to determine the effectiveness of eradication therapy by the study of the contents of the tooth-gingival pocket for the detection of genetic material of Helicobacter pylori, as well as to include in the complex therapy of Helicobacter pylori-associated diseases of the immune modulator Imudon.

[REAL-TIME PCR IN THE COMPLEX DIAGNOSTICS OF COMBINED PATHOLOGY OF THE PERIODONTIUM AND GASTRO-DUODENAL ZONE].

Biological material of 92 patients (18-85 years old) with different severity chronic periodontitis were analyzed for bacterial pathogen colonization by using Dentofol kit (DNA-technology, Moscow). The cohort included 70 individuals with chronic gastritis, 2 patients with gastric and duodenal ulceration and 20 individuals with no gastric/duodenal pathology. The tight- est association with severity of the chronic periodontitis in the analyzed sub-cohort with the chronic gastritis was found with the prevalence of a complex T. for sythensis/T. denticola. Key contribution of this complex to progression of periodon- titis in males of the eldest group (above 55) was hypothesized. This data essentially differ from published results of other research groups where T. forsythensis and T. denticola were never reported as the principal causative agents of the chronic periodonitis in the gender/age/combined pathology normalized cohorts.

Proton pump inhibitor modifies inflammatory reaction in human gastric mucosa infected by Helicobacter pylori.

AIM: To examine whether proton pump inhibitors modify the production of oxygen-derived free radicals and related cytokines in the human gastric mucosa infected with H. pylori. METHODS: Thirty-four H. pylori-positive peptic ulcer patients (23 gastric ulcer, 11 duodenal ulcer) were enrolled. Biopsy tissue samples were obtained endoscopically from the antrum and corpus. Tissue content of neutrophil myeloperoxidase (myeloperoxidase) and IL-8 was measured by ELISA. Mucosal production of oxygen-derived free radical was measured using luminol-dependent chemiluminescence (ChL). A proton pump inhibitor (either lansoprazole 30 mg, omeprazole 20 mg, or rabeprazole 10 mg) was administered daily by mouth to all patients for 8 weeks. Endoscopic examination was then repeated, and biochemical analysis was performed. RESULTS: Antral myeloperoxidase decreased significantly after proton pump inhibitor treatment (5.23 +/- 7.00-2.76 +/- 5.11 ng/mg, P < 0.02), but corpus myeloperoxidase was unchanged. IL-8 was also modified by proton pump inhibitors and these changes were parallel to those of myeloperoxidase. Corpus ChL was significantly increased from 88.5 +/- 69.8-159 +/- 172 counts/10 s/mg after proton pump inhibitor treatment, whereas antrum ChL was not altered. H. pylori infection rate was decreased in the antrum as well as the corpus. CONCLUSIONS: Proton pump inhibitor treatment stimulated oxygen-derived free radical production in the corpus mucosa.

Oral cavity as permanent reservoir of Helicobacter pylori and potential source of reinfection.

Recent studies in developed countries showed that neither dental plaques nor dentures are important reservoir for Helicobacter pylori (Hp), whereas studies in developing countries revealed a high prevalence of Hp in dental plaques, though elsewhere the culture of bacterium or its DNA analysis by polymerase chain reaction in the material obtained from oral cavity were not successful. This study was designed to compare the incidence of Hp in oral cavity (saliva, dental plaques and gingival pockets) using Campylobacter-like organism (CLO) test and culture and in the presence of Hp in the stomach using 14C-urea breath test (UBT), CLO-test and culture (antral biopsy specimens). Hundred dyspeptic subjects with endoscopically normal gastro-duodenal mucosa and 55 symptomatic patients with active duodenal peptic ulcer (DU) were tested for the presence of Hp. Thirty of these DU patients were also examined for presence of Hp in oral cavity and the stomach just before the start and 4 weeks after the termination of one week triple therapy (Omeprazole 20 mg bd, Clarithromycin 500 mg bd and Tinidazole 500 mg bd) when the DU was found endoscopically healed. In the group of 100 dyspeptic subjects, the Hp was detected by CLO-test in saliva, dental plaques and gingival pockets in 84%, 100% and 100% of cases and by the culture in 55%, 88% and 100%, respectively. The presence of Hp, as determined by UBT in the stomach in these subjects was 60%. Using CLO-test and culture, all (100%) out of 55 DU patients, were found to be Hp positive in the oral cavity and in 95% in the stomach. Following one week triple therapy in 30 DU patients, the Hp was still detected in oral cavity by CLO-test in all patients (100%) and by culture in 27 patients (90%), whereas in the stomach, the Hp was found by UTB and culture only in one of these patients (97% Hp eradicated). We conclude that the Polish population including dyspeptic and DU patients, the mouth is permanent reservoir of Hp and that the successful Hp eradication from the stomach by systemic therapy fails the Hp status in the oral cavity that might be a potential source of gastric reinfection in these patients.

Effect of antibacterial therapy and salivary secretion on the efficacy of Helicobacter pylori eradication in duodenal ulcer patients.

OBJECTIVE: Oral health status may play a role in Helicobacter pylori eradication. Because adequate secretion of saliva promotes oral health, the aim of the study was to assess the effect of salivary secretion on the efficacy of H pylori eradication from the stomach. STUDY DESIGN: The study involved 90 H pylori-positive subjects with duodenal ulcer (68 men, 22 women, aged 20-70 years) in whom saliva was collected under basal conditions for 45 min before antibacterial treatment began. They received no drugs for at least 3 days prior to saliva collection. A 7-day course of either of 2 eradication regimens--omeprazole, amoxicillin, and tinidazole (OAT); or omeprazole, amoxicillin, and clarithromycin (OAC)--was used. The efficacy of eradication therapy was evaluated 30 days after its completion. RESULTS: The efficacy of H pylori eradication from the stomach (per protocol analysis) was 77.5% in the group of subjects treated with OAT and 81.6% with OAC. Combined analysis of both groups (OAT+OAC) showed reduced salivary secretion in subjects with eradication failure (0.395 +/- 0.266 vs 0.25 +/- 0.176 mL/min, P=.042). A similar outcome was obtained when the OAT group was analyzed separately (0.436 +/- 0.316 vs 0.211 +/- 0.216 mL/min, P=.022), but in the OAC group the difference was not significant. In the combined analysis, the efficacy of eradication therapy was lower in women than in men (52.9% vs 86.9%, P=.005). In women, it corresponded to salivary secretion (successful eradication 0.337 +/- 0.133 mL/min, unsuccessful eradication 0.180 +/- 0.144 mL/min, P=.043); whereas in men, the difference was not significant (successful eradication 0.405 +/- 0.282 mL/min, unsuccessful eradication 0.321 +/- 0.186 mL/min). CONCLUSION: Low salivary secretion may contribute to the decrease in efficacy of H pylori eradication from the stomach, at least in subjects treated with certain drug regimens.

Microbiological evidence of Helicobacter pylori from dental plaque in dyspeptic patients.

To evaluate the possible route of transmission of Helicobacter pylori, stomach biopsies and dental plaques were cultured from patients with dyspeptic symptoms who underwent endoscopy. A total of 31 patients were examined. Twenty patients out of thirty one (64%) were H. pylori positive in gastric biopsy. Among the microorganisms isolated in dental plaque only one sample (corresponding to a patients with duodenal ulcer H. pylori positive) showed colonies morphologically and biochemically compatible with H. pylori. Proteic patterns of whole cells and restriction endonuclease analysis with Hind III and Hae III endonucleases of DNA extracted from the strain subcultured from a stomach biopsy and from dental plaque of the same patient indicated that both sites were infected with the same strain of H. pylori.

[Detection and differentiation of Helicobacter pylori from gastric biopsy and saliva by PCR-SSCP].

In this study we detected the species-specific antigen gene of Helicobacter pylori (Hp) from gastric biopsy specimens and saliva of 48 patients by polymerase chain reaction (PCR), then adopted single strand conformation polymorphism (SSCP) to differentiate Hp strains from population and different sites of the same individual. Among patients of positive gastric biopsy results, the detection rate of saliva was 72.2% (26/36). In 12 patients of negative gastric biopsy results, 2 saliva specimens were positive. Further SSCP analysis demonstrated that in population the infected Hp strains showed 6 different band patterns. SSCP analysis of PCR products from the gastric biopsy specimen and saliva of the same individual showed no significant difference. The results showed that Hp at different sites of one individual probably belongs to the same strain.

[Methodological study of 13C-urea breath test for detection of Helicobacter pylori infection].

In this study, we investigate simple breath test for detection of Helicobacter pylori (HP) infection using 13C-urea. Thirty-nine patients (30 were HP positive, 9 were HP negative) were given three different doses (50, 100 and 150 mg) of 13C-urea at fasting, and keep sitting after mouth washing with water. Breath samples were taken before and 10, 20, 30, 45, and 60 minutes after urea administration. More than 100mg of 13C-urea was necessary for correct diagnosis of HP infection, because 2 HP positive cases were not detected by 50mg 13C-urea administration. In cases with patchy distribution of HP in the stomach, it may be necessary to change the posture to distribute urea within the whole stomach. In most of HP positive cases, peak delta 13CO2 were obtained within 30 minutes, but one HP negative case showed high delta 13CO2 at 10 minutes, which was probably caused by urease activity in the mouth. So it is appropriate to take breath sample at 20 minutes after urea administration. In this study, cut-off value for a positive test can be setted between 4 to 7 delta/1000, it is necessary to investigate much more cases to set exact cut-off value.

[Enterosorption in ulcerative lesions of the gastrointestinal tract with concomitant intestinal dysbacteriosis]. / Enterosorbtsiia pri iazvennykh porazheniiakh zheludochno-kishechnogo trakta s soputstvuiushchim disbakteriozom kishechnika.

Adjuvant use of enterosorbents in combined therapy of gastroduodenal ulcers (158 patients), nonspecific ulcerative colitis with intestinal dysbacteriosis was assessed. Enterosorption in combined therapy of gastrointestinal diseases with dysbacteriosis potentiated positive effect. The highest benefit of enterosorption occurs in intestinal affections (nonspecific ulcerative colitis, irritable bowel-syndrome, chronic enteritis, colitis). In gastroduodenal ulcer lignin sorbent (polyfepan) is preferable. Polyfepan acted positively on intestinal dysbacteriosis through eliminating Escherichia with hemolyzing properties and enterococci, by reducing lack of bifidobacteria.

Salivary IgG assay to detect Helicobacter pylori infection in an Indian adult population.

BACKGROUND: Helicobacter pylori infection, the commonest chronic bacterial infection in humans, causes chronic gastritis, peptic ulcer, and possibly gastric carcinoma and lymphoma. Recently, investigators have focused on its role in the development of extra-gastrointestinal diseases with oral manifestations. H. pylori infection can be diagnosed by various methods. Of late, H. pylori IgG antibodies have been detected in saliva using enzyme-linked immunosorbent assay (ELISA). However, local validation of serological test is needed before implementing a test in different populations. AIMS: To detect anti H. pylori specific immunoglobulin G (IgG) antibodies in saliva of adult patients with gastrointestinal symptoms by ELISA, to diagnose H. pylori infection in such patients by histopathology, and to evaluate the diagnostic accuracy of the immunoassay as compared to histopathologic diagnosis. METHODS: The study included 40 adult patients with gastrointestinal symptoms suggestive of peptic ulcer disease. Saliva samples were analyzed for anti H. pylori IgG using EIAgen H. pylori IgG kit. Histopathologic diagnosis using gastric biopsy samples was the gold standard. RESULTS: The sensitivity and specificity of the test were 79.31% and 63.64%, respectively. The positive and negative predictive values were 85.19% and 53.85%, respectively. The accuracy of EIAgen H. pylori IgG kit for salivary detection of anti H. pylori IgG antibodies was found to be 75%. CONCLUSION: EIAgen H. pylori IgG assay is a noninvasive, moderately accurate, and sensitive method for the detection of H. pylori infection in saliva. Salivary anti H. pylori IgG test prior to endoscopy is a useful screening test for seroepidemiological studies.

Secretion of epidermal growth factor in saliva of duodenal ulcer patients; an association with Helicobacter pylori eradication and erosive esophagitis.

PURPOSE: Erosive esophagitis frequently develops after successful Helicobacter pylori eradication. Since salivary secretion of epidermal growth factor (EGF) plays a crucial role in the pathogenesis of gastroesophageal reflux disease, the current study objective was to find out whether erosive esophagitis development after Helicobacter pylori eradication is associated with the secretion of EGF in saliva. MATERIALS AND METHODS: A total of 115 H. pylori infected patients (positive results of CLO-test, histology and serology) with a duodenal ulcer were recruited for the study. Gastroscopic examinations and saliva collections were performed twice, prior to H. pylori eradication and one year after its cessation. The salivary EGF was determined using a radioimmunological method. RESULTS: Salivary EGF secretion was lower in H. pylori infected subjects with erosive esophagitis than without (0.82+/-0.66 vs 1.70+/-3.49 ng/min, p=0.021). However, a year after successful H. pylori eradication, salivary EGF did not differ between the groups (2.17+/-2.06 vs 1.79+/-2.06 ng/min); the lack of difference was due to high peptide secretion in patients who developed erosive esophagitis after eradication. CONCLUSION: Erosive esophagitis development following H. pylori eradication is not the result of decreased salivary EGF secretion.

Polymerase chain reaction based analysis of the cytotoxin associated gene pathogenicity island of Helicobacter pylori from saliva: an approach for rapid molecular genotyping in relation to disease status.

BACKGROUND AND AIMS: The genetic composition of the intricate cytotoxin associated gene pathogenicity island (cag PAI) of Helicobacter pylori is known to significantly influence the outcome of the disease. Hence, analysis of complete cag PAI of H. pylori isolated from saliva would be of immense importance in standardizing saliva as a reliable non-invasive diagnostic specimen and also to evaluate the type of H. pylori infection. The aim of the present study was to analyze the genes of cag PAI of H. pylori for their presence and correlating them with the disease status of the patients. METHODS: One hundred and twenty patients (55 duodenal ulcer [DU], 25 gastric ulcer and 40 non-ulcer dyspepsia [NUD]) were investigated for the present study. Eight pairs of oligonucleotide primers (cagA1, cagA2, cagAP1, cagAP2, cagE, cagT, LEC1 and LEC2) of five different loci; cagA, cagA promoter region, cagE which represents cagI region, cagT and LEC representing cagII were used to detect the presence of the cag PAI genes by polymerase chain reaction. RESULTS: The comprehensive analysis of the genes constituting cag PAI showed almost equivalent prevalence of all the genes between both the study groups (ulcer and NUD) included. Little significant difference was found in the percentage distribution in both the clinical groups. cagE and cagT were found in a larger proportion of the ulcer group (92.5% and 96.2%) compared with the NUD group (77.5% and 85%), respectively. CONCLUSION: Saliva could be efficiently used as a non-invasive source for H. pylori and cagT might be an important locus of the cag PAI, thus greatly influencing the disease condition of the subjects.

Salivary-specific immunoglobulin G in the diagnosis of Helicobacter pylori infection in dyspeptic patients.

OBJECTIVES: Helicobacter pylori infection is arguably the most common chronic bacterial infection in humans. The high prevalence and the association with peptic ulceration and gastric cancer indicate that simple, noninvasive methods for diagnosis of the infection are needed. In this study, the accuracy of salivary diagnosis for H. pylori infection was assessed. METHODS: Saliva and serum samples of 152 dyspeptic patients were tested for H. pylori IgG and IgA by an in-house ELISA. All patients underwent gastroscopy with biopsy. RESULTS: One hundred thirty-one patients (86%) were found to be H. pylori positive on histology. Duodenal ulcer was found in 67 patients; 85 had no macroscopic lesion. Salivary and serum H. pylori IgG as well as serum H. pylori IgA titers were significantly higher in H. pylori-positive than in H. pylori-negative patients. The sensitivity and specificity of salivary H. pylori IgG were 82% and 71%, respectively; the positive and negative predictive values were 95% and 40%, respectively; and the accuracy 81%. The corresponding figures for serum H. pylori IgG were 97% and 91%; 98% and 83%; and 96%. Those for serum H. pylori IgA were 80% and 52%; 91% and 30%; and 76%. The sensitivity of salivary H. pylori IgG in detecting duodenal ulcer was 83% (56/67) that of serum H. pylori IgG was 97% (65/67) (odds ratio = 0.15; confidence interval = 0.02-0.8; p = 0.02). CONCLUSIONS: Salivary H. pylori IgG was a fairly sensitive and accurate indicator of gastric H. pylori colonization, with a high positive predictive value in our population. Data, however, suggest that salivary H. pylori IgG measurements do not compare favorably with serology.

H. pylori in the pathogenesis of duodenal ulcer: interaction between duodenal acid load, bile, and H. pylori.

OBJECTIVE: Helicobacter pylori (H. pylori) growth is inhibited by bile yet it can grow in the duodenal bulb and cause ulcer disease. The aim of this study was to test the effect of bile on H. pylori viability and growth and to determine whether acidification of bile reduces its inhibitory activity. METHODS: Fresh human bile was collected at laparotomy and tested for inhibitory activity of H. pylori using broth dilution assays. Six clinical isolates of H. pylori obtained from patients with duodenal ulcer were used for each experiment. The bile was diluted from 1:3 to 1:192; its inhibitory effect on H. pylori was tested before and after acidification, treatment with cholestyramine, or chloroform. Bile was acidified to a pH of 2-6, centrifuged at 8000 rpm for 20 min to remove precipitated bile acids, and the supernatant pH readjusted. Controls included BHI broth without bile (positive control) and bile that was acidified to pH 2 and neutralized without centrifugation. RESULTS: Human bile inhibited H. pylori growth in a dose dependent manner. Growth of all strains was supported for all strains only at a dilution of 1:192. In contrast, after acidification to pH < or =5 and centrifugation to remove precipitated bile acids, all strains grew at a bile dilution of 1:12. Neither chloroform extraction of lipids, nor acidification without centrifugation removed the inhibitory action of bile. In contrast, cholestyramine sequestration of bile acids completely removed all inhibitory activity. CONCLUSIONS: The duodenal acid load may be the critical factor to explain the ability of H. pylori to colonize the duodenal bulb by precipitating glycine-conjugated bile salts. The combination of a high duodenal acid load and H. pylori infection is likely the critical event in the pathogenesis of H. pylori-related duodenal ulcer disease.

IgA antibodies to herpes simplex virus type 1 in duodenal juice and saliva from patients with peptic ulcer and non-ulcer controls.

Fasting duodenal juice and saliva were obtained from 26 peptic ulcer patients and 28 controls. IgA antibodies to herpes simplex virus type 1 (HSV-1) were measured by the indirect enzyme-linked immunosorbent assay (ELISA). Total IgA in the samples was measured by the double-sandwich ELISA. The median and interquartile range of anti HSV-1 IgA titres in saliva were 8 (4-12) and 4 (1-8) in ulcer patients and controls, respectively (p less than 0.02). Comparative values in duodenal juice were 12 (2-16) and 4 (0-6) (p less than 0.002). The concentration of total IgA was not higher in ulcer patients than in controls, and no correlation existed between total IgA and anti HSV-1 titres. The results add further evidence that HSV type 1 is involved in the pathogenesis of peptic ulceration.

Comparison of serum, salivary, and rapid whole blood diagnostic tests for Helicobacter pylori and their validation against endoscopy based tests.

BACKGROUND: A rapid, reliable, and accurate test for the diagnosis of infection with Helicobacter pylori is needed for screening dyspeptic patients before referral for endoscopy. AIM: To compare a new rapid whole blood test (Helisal rapid blood, Cortecs), two serum enzyme linked immunosorbent assays (ELISAs; Helico-G, Shield and Helisal serum, Cortecs), and a salivary assay (Helisal saliva, Cortecs), with slide biopsy urease, 13C-urea breath test, and histology. METHODS: Three hundred and three consecutive dyspeptic patients attending for gastroscopy underwent two antral biopsies for histology, and one for rapid slide biopsy urease test for assessment of H pylori status. Blood and saliva were also collected. One hundred of the patients also underwent a 13C-urea breath test. Gold standard positives were defined as those with at least two positive tests among slide urease, breath test, or histology, and gold standard negatives as those with all these (or two when the breath test was not done) negative. RESULTS: Of 300 patients (median age 63, range 28-89) eligible for analysis, 137 (46%) were gold standard positives, of which Helisal rapid blood identified 116, Helico-G 129, Helisal serum 130, and Helisal saliva 120; 137 (46%) were gold standard negatives of which the number falsely identified as positive was 30 by Helisal rapid blood, 45 by Helico-G, 41 by Helisal serum, and 41 by Helisal saliva. Sensitivities and specificities were: for the whole blood test 85% and 78% respectively; for Helico-G 94% and 67%, for Helisal serum 95% and 70%, and for Helisal saliva 84% and 70%. CONCLUSIONS: If endoscopy had been undertaken only on patients with positive tests two of 16 duodenal ulcers would have been missed if the Helisal rapid blood test was used, and one if any of the ELISA tests were used. None of the blood tests would have missed any of six gastric ulcers, but the salivary test would have missed one.

Mucosal expression and luminal release of epidermal and transforming growth factors in patients with duodenal ulcer before and after eradication of Helicobacter pylori.

BACKGROUND: Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF alpha) are potent gastric acid inhibitors and stimuli of mucosal growth and protection but their involvement in Helicobacter pylori associated duodenal ulcer has been little examined. AIM: To assess gastric acid secretion, plasma gastrin concentrations, mucosal content of EGF and TGF alpha, and mucosal expression of these peptides and their receptor (EGFr) as well as salivary and gastric luminal release of EGF under basal conditions and after pentagastrin stimulation in 10 healthy subjects and in 25 H pylori positive patients with duodenal ulcer before and after two weeks of triple anti-H pylori therapy and four weeks after the termination of this therapy. RESULTS: Pentagastrin stimulation caused a significant increase in salivary and gastric release of EGF both in healthy controls and patients with duodenal ulcers but in the patients, the eradication of H pylori resulted in several fold higher gastric luminal (but not salivary) EGF release than before the anti-H pylori therapy. Mucosal contents of immunoreactive EGF and TGF alpha and mucosal expression of EGF, TGF alpha, and EGFr in H pylori positive patients with duodenal ulcer were significantly higher than those in healthy H pylori negative controls and this increase persisted after eradication of H pylori. Basal plasma gastrin was significantly reduced after two weeks of triple therapy and four weeks after the H pylori eradication all ulcers were completely healed. CONCLUSIONS: (1) H pylori infection in patients with duodenal ulcer was accompanied by enhanced plasma gastrin and increased mucosal content and expression of TGF alpha, EGF, and EGFr; (2) H pylori eradication resulted in ulcer healing, reduction in plasma gastrin, and enhancement of gastric (but not salivary) luminal release of EGF, particularly after pentagastrin stimulation; and (3) enhanced mucosal content and expression of TGF alpha, EGF, and EGFr and increased luminal release of EGF may contribute to ulcer healing after eradication of H pylori.