Bovine Milk Inhibits Both Adhesion of Helicobacter pylori to Sulfatide and Helicobacter pylori-Induced Vacuolation of Vero Cells

Adhesion of Helicobacter pylori to gastricmucosal cells is an initial important step incolonization and infection. To study adhesion, weinvestigated whether milk inhibits the adhesion ofHelicobacter pylori to sulfatide, an acidicglycosphingolipid that exists in human gastric mucosaand to which Helicobacter pylori adheres. As a measureof functional significance, we also studied whether milkinhibits Helicobacter pylori-induced vacuolation of Verocells. We used sulfatide-coated polystyrene plates andstudied the effect of bovine milk on the adhesion ofHelicobacter pylori to sulfatide. We used Vero cells for Helicobacter pylori -inducedvacuolation. Bovine milk 100- to 200-fold dilutedsignificantly inhibited both adhesion of Helicobacterpylori to sulfatide and Helicobacter pylori-inducedvacuolation in Vero cells. Bovine milk significantlyinhibited adhesion of Helicobacter pylori to MKN-45cells and Lewis b antigen-coated polystyrene plates. Inaddition, these results suggest that bovine milkcontains active substances that inhibit both adhesion ofHelicobacter pylori to mucosa and vacuole formation.Bovine milk may have a protective effect on the gastricmucosa in Helicobacter pylori-associatedgastritis.     

Nonsteroidal Antiinflammatory Drugs Could Reverse Helicobacter pylori-Induced Apoptosis and Proliferation in Gastric Epithelial Cells

It remains controversial whether the harmfuleffects of Helicobacter pylori (Hp) and nonsteroidalantiinflammatory drugs (NSAIDs) are additive. We studiedthe effects of Hp (virulent and nonvirulent strains) and NSAIDs, alone or in combination, onapoptosis and proliferation of gastric epithelial cellsin nonulcer dyspepsia (NUD) patients. Forty-four (25Hppositive and 19 Hp-negative) consecutive Chinese NUD patients with rheumatoid arthritis who hadtaken continuously NSAIDs for more than three monthswere recruited for this study. Another 41 (20Hp-positive and 21 Hp-negative) NUD patients not on anyNSAIDs were included as controls. All patientsunderwent a gastroscopy examination and gastricbiopsies. Hp infection was confirmed by CLOtest, anti-HpELISA, and [¹³C]urea breath test. The CagAstatus was determined by the anti-CagA antibody assay. The degree ofgastritis, apoptosis, and proliferation indices weredetermined with H&E staining, terminal uridinedeoxynucleotidyl nick end-labeling (TUNEL), andproliferating cell nuclear antigen (PCNA) immunostainingmethods, respectively. A significantly higher apoptosiswas observed in subjects who had Hp infection or hadbeen consuming NSAIDs when compared with the controls. Unlike NSAID-treated subjects, patients with Hpinfection were shown to have significantly enhanced cellproliferation. However, the increased apoptosis andproliferation in Hp-positive subjects were reversed by also taking NSAIDs. No correlation was foundbetween apoptosis and proliferation in all the studygroups. There was no association found between CagAexpression or degree of gastritis with cellproliferation or apoptosis. It was demonstrated at thecellular level that NSAIDs could abrogate apoptosis orproliferation effects induced by Hp. Furthermore, thelatter effects appeared not to be influenced by the virulent nature of the Hp strains.     

Helicobacter pylori and Mucosal Atrophy in Patients with Gastric Cancer (A Special Study Regarding the Methods for Detecting Helicobacter pylori)

We assessed the sensitivities of several methodsfor detecting Helicobacter pylori (culture, histology,rapid urease test, and serology), and evaluated the H.pylori positivity considering the degree of atrophy in the background mucosa in 202 gastriccancer patients and 101 controls. The positivity of H.pylori determined by culture (81%) was significantlyhigher than that determined by serology (62%) in gastric cancer patients (P < 0.001). Thepositivities of H. pylori determined by biopsy and/orserology in intestinal (84%) and diffuse (95%) types ofgastric cancer were higher than that observed in controls (54%) (P < 0.001).Intestinal-type gastric cancer tended to occur in theatrophic mucosa, in which H. pylori positivity was notdifferent from that in controls after adjusting for thedegree of atrophy, whereas diffuse-type gastric cancerwas observed more often in the nonatrophic mucosa, inwhich H. pylori positivity was higher than that incontrols even after adjusting for the degree ofatrophy.     

One-Week Triple Therapy with Lansoprazole, Clarithromycin, and Metronidazole to Cure Helicobacter pylori Infection in Peptic Ulcer Disease in Korea

The efficacy and acceptability of classicalbismuth triple therapy may be limited by poor patientcompliance and adverse effects. It is widely agreed thatimproved, simpler, and reliable therapies are needed to cure Helicobacter pylori infection andfoster patient compliance. We evaluated the efficacy andside effects of a Bazzoli triple therapy substitutinglansoprazole for omeprazole for H. pylori infection in active peptic ulcer in Korea (30 mg oflansoprazole, 250 mg of clarithromycin, and 400 mg ofmetronidazole, all twice daily). H. pylori status wasevaluated by rapid urease test, histology, and culture at entry and four or more weeks after endingantimicrobial therapy. Fifty-eight patients (mean age:43 years) with gastric (N = 30) or duodenal ulcer (N =28) and H. pylori infection were studied. H. pylori was cured in 47 (81%, 95% CI 69-90%). Mild sideeffects, including vomiting, diarrhea, and itching, wereobserved in four patients (7%). Compliance averaged 95%.Fifty-five ulcers (95%) were healed. Pretreatment pylorobulbar deformity was observed in 49patients (85%), and in 43 (88%) the deformitydisappeared after treatment. Pretreatment metronidazoleand clarithromycin resistance was observed in 87% and 2% of patients, respectively. The cure rate of H.pylori infection was significantly higher in patients>50 years of age than those <50. Treatment withlow-dose one-week lansoprazole, clarithromycin, and metronidazole resulted in a relatively lowcure rate, but was well tolerated. Studies to define theoptimal duration, dose, and dosing interval of thiscombination therapy in Korea are needed.     

Halitosis and Helicobacter pylori: A Possible Relationship

With the aim of investigating a possiblerelationship between objective halitosis(established by sulfide levels in the breath) andHelicobacter pylori, we performed a study in 58dyspeptic patients reported to suffer from "badbreath." Furthermore, we evaluated the effects onhalitosis of eradication therapy (only for H.pylori-positive patients) and chlorhexidine antisepticmouth rinses (in all patients). Sulfide compoundassay indicated objective halitosis in 52/58 patients,30 of whom were positive and 22 negative for H. pylori.In 19/30 eradication by double therapy provoked a decrease to below the cutoff value of sulfidelevels in 15. In the other 11 of the 30 subjects, inwhom H. pylori positivity persisted, halitosisparameters did not change. Chlorexidine reduced sulfides to below the cutoff value in 16/22 H.pylori-negative patients, but did not provoke any changein the 11 unsuccessfully treated H. pylori-positivesubjects. In these, objective halitosis disappeared only after a successful eradication by tripletherapy (9/11). Our results show a possible associationbetween halitosis and H. pylori since bacterialeradication may resolve the symptom. Antisepticmouthwashes may be effective only in absence of H. pylori,when halitosis may be due to oral putrefactive microbialactivity. In a small number of subjects the cause andtreatment of halitosis need to be clarified.     

Effects of Inflammatory Cytokines Induced by Helicobacter pylori Infection on Aminopyrine Accumulation in Parietal Cells Isolated from Guinea Pigs

The effects of inflammatory cytokines induced byHelicobacter pylori infection on acid secretion have notbeen well defined. The purpose of this study was toinvestigate the direct effects of these cytokines on parietal cells isolated from guinea pigs. Weexamined the effects of human recombinant IL-1(0.05-100 ng/ml), IL-8 (2-256 ng/ml), and TNF-(0.625- 80 ng/ml) on acid secretion stimulated by three secretagogues (10^-4 M histamine,10^-4 M carbachol, and 10^-5 Mtetragastrin) and on basal acid secretion from isolatedparietal cells, which was measured by the aminopyrineaccumulation method. None of three cytokines showed any significant effects onstimulated or basal acid secretion from isolated guineapig parietal cells. We concluded that inflammatorycytokines induced by Helicobacter pylori infection may affect acid secretion through mechanismsother than direct actions on parietal cells.     

Ammonia Inhibits Proliferation and Cell Cycle Progression at S-Phase in Human Gastric Cells

Helicobacter pylori (Hp) has strong ureaseactivity and produces a large amount of ammonia in thestomach. In animal studies, ammonia was shown toaccelerate cell kinetics of gastric mucosa, andlong-term exposure of the stomach to ammonia leads tomucosal atrophy. To understand this process, we examinedthe effects of ammonia on the growth and cell cycleprogression of human gastric cancer cell lines (HGC-27, MKN1, MKN45) using flowcytometric analysis. Ineach cell line, ammonia inhibited the cell growth in adose-dependent manner and caused significantaccumulation of S-phase cells at a cytostatic dose. DNA synthesis of HGC-27 cells treated with ammoniawas also suppressed to about 50% of that of theuntreated cells. Similar effects were observed onaddition of ammonium chloride at the same concentration, while adjusting the pH of the media with NaOHalone to that with the cytostatic dose of ammonia didnot affect the cell cycle progression. Theseobservations indicate that ammonia induces S-phasearrest in gastric cells independently of pH.     

Changes in Gastrin and Serum Pepsinogens in Monitoring of Helicobacter pylori Response to Therapy

The aims of this study in 50 patients with H.pylori infection and duodenal ulcer were to examine theeffect of eradication therapy on the serum levels ofgastrin, pepsinogen I, and pepsinogen II and to investigate whether monitoring of the serumchanges in these peptides after treatment could predictpatient outcome. H. pylori status was assessed at entryand one and six months after therapy by culturing and microscopic analysis of the gastric mucosaand by [¹⁴C]urea breath test. Significantdecreases were observed in the serum levels of gastrin(–11.4 ± 3%), pepsinogen I (–28.9± 4%), and pepsinogen II (–40.4 ±3%) in the 45 patients whose infection was eradicated,but not in the patients without eradication. Serumvalues of these peptides were unchanged in an additionalgroup of 10 patients that only received omeprazol, none of whom had H. pylorieradicated. The best cutoff point of the percentage ofeach peptide to predict patient outcome was 10% forgastrin and pepsinogen I, and 15% for pepsinogen II. Apepsinogen II decrease >15% resulted in the best markerof H. pylori clearance, accurately identifying patientoutcome 86.6% of the time, whereas the diagnosticaccuracy of gastrin and pepsinogen I was 61.7% and76.6%, respectively. Significant correlations werefound between the bacterial load assessed by histologywith the serum concentrations of pepsinogen I and II andwith the urease activity as measured by the amount of ¹⁴CO₂ excreted. Inconclusion, eradication of H. pylori infection isfollowed by a significant drop in serum levels ofgastrin, pepsinogen I, and pepsinogen II. Changes in thelatter are the most uniform and may be used as an indirect tool to predicttreatment outcome.     

Clinical Application of Gastric Histology to Monitor Treatment of Dual Therapy in H. pylori Eradication

This preliminary study attempted to test whetherpretreatment gastric histology of H. pylori infectionmay affect the success of dual therapy and to identifywhich parameter of gastric histology could be improved after dual therapy. One hundredforty-five dyspeptic patients with H. pylori infectionreceived a two-week course of dual therapy (Amoxicillin500 mg every 6 hr plus omeprazole 20 mg twice a day). In each patient, three pairs of gastricbiopsies, sampled from the antrum, lower body, and upperbody near the cardia, were collected before treatmentand four weeks after completion of dual therapy. The density of H. pylori (score 1-5) and parametersof the modified Sydney system were applied to test theseverity of H. pylori-related gastric histology in eachspecimen. The total bacterial load (score 1-15) was a sum of the density of H. pylorisampled from three biopsies. The overall rate of H.pylori eradication rate by dual therapy is 73.1%(106/145). Univariate analysis of parameters inpretreatment histology disclosed that the presence ofmucosal atrophy (P < 0.01), lymphoid follicles (P< 0.005), and higher-density H. pylori (P < 0.001)predisposed to dual therapy failure. Multivariateanalysis by stepwise logistic regression further confirmed that boththe density of bacteria and the presence of lymphoidfollicles are the two major factors related to theoutcome of dual therapy (P < 0.001). Four weeks after dual therapy was completed, only patients withsuccessful eradication significantly improved in thesegastric histology parameters: acute activity, chronicinflammation, eosinophil infiltration, and mucosal atrophy. However, the lymphoid follicle andintestinal metaplasia were not significantly improvedduring the study period. The eradication rates amongthree subgroups with different total bacterial loads (group I: 1-5; II: 6-10; III: 11-15) discloseda downward trend (I: 89.1%; II: 73%; III: 52.7%). It isconcluded that dual therapy could improve gastrichistology especially among patients with successful eradication of H. pylori. Evaluatingpretreatment histologic parameters, including thedensity of H. pylori and the presence of lymphoidfollicles, is valuable in predicting the success of dualtherapy.     

Helicobacter pylori Infection and Gastric Cancer (A Nested Case-Control Study in a Rural Area of Japan)

We conducted a seroepidemiological nested case-control study to determine the association of gastriccancer with Helicobacter pylori infection and atrophicgastritis. A cohort of 2858 participants in an annual multiphasic health check-up werefollowed for eight years. Data for 45 gastric cancercases and 225 sex-, age-, and address-matched controlsubjects were analyzed. Helicobacter pylori infectionwas determined by IgG antibodies, and atrophicgastritis was diagnosed by both serum pepsinogen I level(70 ng/ml) and the pepsinogen I/II ratio (3.0).Univariate analysis showed that Helicobacter pylori and atrophic gastritis were significantlyassociated with gastric cancer. In a multivariateanalysis, atrophic gastritis was associated withsignificantly increased risk of cancer (odds ratio,3.38; 95% confidence interval, 1.54-7.42); however,Helicobacter pylori was not associated with cancer (oddsratio, 1.84; 95% confidence interval, 0.59-5.72). Theseresults suggest that Helicobacter pylori infection alone is not directly associated with gastriccarcinogenesis but has an indirect relation to gastriccancer through the development of atrophicgastritis.