Gastric adenomas: relationship between clinicopathological findings, Helicobacter pylori infection, APC mutations and COX-2 expression.

Gastric adenomas are rare neoplastic growths characterized by localized polypoid proliferations of dysplastic epithelium that tend to progress to infiltrating adenocarcinoma. Therefore, the identification of molecular markers that could reliably recognize adenomas at risk of progression is advocated in the clinical management. In this study we investigated, in a series of gastric adenoma specimens from an area at high risk of gastric cancer, the relationship between clinicopathological characteristics of adenoma and Helicobacter pylori infection, APC mutational status, and COX-2 and the down-stream enzyme mPGES1 expression. Helicobacter pylori infection, detected in 24%, and 33% by histology and PCR analyses, respectively, did not show any relationship with growth pattern, localization, size, dysplasia grade and presence of synchronous cancer. Pathogenetic mutations of MCR region (codons 1269-1589) of the APC gene were detected only in one case corresponding to a single, small size, low grade, H. pylori-negative adenoma. The expression of COX-2 largely matched that of mPGES(1). Both were overexpressed in 79% of cases showing a relationship with high-grade dysplasia, size >10 mm and presence of a synchronous carcinoma. In conclusion, COX-2 may play a key role in the development and progression of gastric adenoma and could be an attractive target in the management of gastric adenoma at major risk of cancer development.

Rifabutin based triple therapy for eradication of H. pylori primary and secondary resistant to tinidazole and clarithromycin.

BACKGROUND: Rifabutin has been empirically used in Helicobacter pylori infections resistant to triple therapy. There are no data on primary and secondary resistance to rifabutin and its use in specific cases. AIM: To analyse the susceptibility and resistance to rifabutin in H. pylori-positive patients with or without previous H. pylori therapy and to test the efficacy of rifabutin in H. pylori resistant to clarithromycin and tinidazole. METHODS: Four hundred and twenty H. pylori-positive patients without previous exposure to triple therapy and 104 patients who had already received one course of triple therapy underwent upper endoscopy for dyspeptic symptoms and H. pylori susceptibility test. Amoxicillin, clarithromycin, tinidazole and rifabutin were evaluated for resistance and susceptibility. Forty patients with primary resistance to both clarithromycin and tinidazole and with susceptibility to amoxicillin and rifabutin, and 65 patients with secondary resistance and susceptibility to the same antibiotics were identified. All these patients received a 10-day triple therapy with pantoprazole amoxicillin and rifabutin. Treatment success was evaluated by the 13C-Urea Breath test. RESULTS: In naive patients 23% of strains were resistant to clarythromycin, 35% to tinidazole, 9% to both antibiotics, and none was resistant to rifabutin In patients already treated the percentages of resistant strains were 76, 64.4, 62.5 and 1%, respectively. With rifabutin based triple therapy eradication rates were (Per Protocol and Intention-to-Treat analysis) 100 and 87.5% in primary resistance to clarithromycin and tinidazole and 82.2 and 78.5% in secondary resistance. CONCLUSION: H. pylori primary and secondary resistances to clarithromycin and tinidazole are high in our geographic area, while resistance to rifabutin is rare. Rifabutin-based triple therapy, can be successfully used in primary and secondary resistance to clarithromycin and tinidazole according to the in vitro susceptibility test.

Role of antimicrobial susceptibility testing on efficacy of triple therapy in Helicobacter pylori eradication.

BACKGROUND: Helicobacter pylori treatment failure may be due to resistance to macrolides and 5-nitroimidazoles. AIM: To test whether a preliminary in vitro susceptibility test of H. pylori to tinidazole and clarithromycin and a consequent specific regimen could improve the eradication rate. METHODS: A total of 109 consecutive H. pylori-positive patients with dyspeptic symptoms were included. At endoscopy, biopsy from the antrum was obtained for H. pylori culture and antimicrobial susceptibility testing. Fifty-six patients were treated with omeprazole, tinidazole and clarithromycin for 10 days (group OTC) and 53 patients received therapy on the basis of the susceptibility test (group SUSC). Treatment success was evaluated by the 13C-urea breath test 1 month after the end of therapy. RESULTS: Eight patients dropped out. Overall primary resistance to clarithromycin, tinidazole and both antibiotics was 13%, 33% and 4%, respectively. In group OTC, H. pylori was eradicated in 81% and 75% of patients by per protocol and intention-to-treat analysis, respectively. Per protocol and intention-to-treat eradication rates for group SUSC were 98% and 91% (P < 0.05 vs. group OTC). CONCLUSIONS: These data show that in H. pylori infection, antibiotic therapy based on the results of culture and susceptibility testing gives, in comparison to standard therapy, a significant improvement in eradication rate.

Primary and secondary antibiotic resistance of Helicobacter pylori strains isolated in central Italy during the years 1998-2002.

BACKGROUND: Antibiotic resistance of Helicobacter pylori is the most common reason for failure in its eradication. AIM: To determine the incidence of primary and secondary resistance to tinidazole, clarithromycin and amoxycillin in Helicobacter pylori isolates from dyspeptic patients in central Italy and to evaluate the modifications of resistance over the period from 1998 to 2002. METHODS: H. pylori strains were isolated from antral biopsies taken during upper endoscopy in 406 dyspeptic patients with no previous therapy against H. pylori, and in 96 patients who had already undergone one or more triple therapies. Antibiotic susceptibility test was performed using the screening agar method and agar dilution method. RESULTS: Overall primary resistance to tinidazole, clarithromycin and amoxycillin was 36.7, 23.4 and 0.2%, respectively. Secondary resistance rates were: tinidazole 69.8%, clarithromycin 82.3% and amoxycillin 1%. Resistance to clarithromycin was often associated with tinidazole resistance and was significantly higher in female patients (p<0.05). Primary and secondary antibiotic resistance did not change during the 4 years of observation. CONCLUSIONS: The dyspeptic population with H. pylori infection in central Italy shows high levels of antibiotic resistance. Primary resistance to clarithromycin is most frequent in female patients. In patients with secondary resistance, dual resistance to clarithromycin and tinidazole is found in the majority of cases.

Analysis of extended genomic rearrangements in oncological research.

Screening for genomic rearrangements is a fundamental task in the genetic diagnosis of many inherited disorders including cancer-predisposing syndromes. Several methods were developed for analysis of structural genomic abnormalities, some are targeted to the analysis of one or few specific loci, others are designed to scan the whole genome. Locus-specific methods are used when the candidate loci responsible for the specific pathological condition are known. Whole-genome methods are used to discover loci bearing structural abnormalities when the disease-associated locus is unknown. Three main approaches have been employed for the analysis of locus-specific structural changes. The first two are based on probe hybridization and include cytogenetics and DNA blotting. The third approach is based on PCR amplification and includes microsatellite or single nucleotide polymorphism (SNP) genotyping, relative allele quantitation, real-time quantitative PCR, long PCR and multiplex PCR-based methods such as multiplex ligation-dependent probe amplification and the recently developed nonfluorescent multiplex PCR coupled to high-performance liquid chromatography analysis. Whole-genome methods include cytogenetic methods, array-comparative genomic hybridization, SNP array and other sequence-based methods. The goal of the present review is to provide an overview of the main features and advantages and limitations of methods for the screening of structural genomic abnormalities relevant to oncological research.

Gastric emptying of a solid-liquid meal measured with 13C octanoic acid breath test and real-time ultrasonography: a comparative study.

OBJECTIVES: The 13C octanoic acid breath test (OBT) has been proposed as a reliable noninvasive test to measure gastric emptying. OBT has been compared with scintigraphy; however, there are no data comparing it with gastric emptying measured with real-time ultrasonography (RUS) The aim of the study was to correlate gastric emptying of a solid-liquid meal, with OBT and RUS simultaneously evaluated in a group of normal volunteers. METHODS: A total of 14 normal subjects ingested a standard test meal (one scrambled egg with two slices of white bread, 10 g of butter, and 300 ml of tap water). The egg yolk was mixed with 0.1 ml of 13C octanoic acid. Breath samples for 13CO2 analysis were collected in breath bags and were analyzed by means of isotope-selective nondispersive infrared spectrometry (IRIS). RUS was simultaneously performed by calculating the antral area following a previous validated method. Breath samples and antral area were taken at baseline and every 15 min after the meal during the first 2 h and every 30 min for another 2 h. Lag time (Tlag) and gastric half emptying time (T(1/2)) were calculated for OBT and RUS. Data were analyzed by the Student's t test for paired data, correlation coefficient, and regression line. RESULTS: The results show a statistically significant longer Tlag and T(1/2) for OBT in comparison with RUS (p < 0.001). A significant correlation and positive regression line was computed between OBT and RUS for Tlag and for T(1/2). CONCLUSIONS: Our results show that OBT overestimates gastric emptying parameters of a solid-liquid meal in comparison with RUS. However, both techniques give data in good correlation. Because OBT is less operator-dependent than RUS, it may be useful in comparative gastric emptying studies.

Short- and long-term effect of glyceryl trinitrate (GTN) ointment 0.2% and 2% on anal canal pressure in patients with chronic anal fissures.

Controversial data are available on the duration of action of glyceryl trinitrate after acute and chronic application on anal canal pressure. Our aim was to assess the effect of glyceryl trinitrate at 0.2% and 2% on anal canal pressure before and after eight weeks of treatment. Anal canal pressure was evaluated in 12 patients with chronic anal fissures with an electronic probe with three recording sites before and after the application of glyceryl trinitrate, 120 mg on the external anal verge. Six patients received glyceryl trinitrate at 0.2% and six at 2%. Glyceryl trinitrate 0.2% and 2% equally reduce basal anal canal pressure in all three recording sites (P < 0.001) with major effect on the inner site of the canal toward the rectum, for a 60-min period. Eight weeks after application, the effect of glyceryl trinitrate was unchanged. In conclusion, glyceryl trinitrate ointment at 0.2% and 2%, equally reduces anal canal pressure for 60 min and this effect is kept unchanged after eight weeks of application.

Transmission of Helicobacter pyori in an animal model.

An experimental murine model was studied to evaluate the orogastrointestinal colonization of Helicobacter pylori and the animal-to-animal transmission. Balb/C mice were infected with H. pylori and housed with uninoculated mice in cages with and without a grate on the floor. Mice were killed after 7, 14, 30, and 45 days, and samples from the esophagus, stomach, small intestine, colon, and rectum were analyzed for H. pylori by PCR and immunohistochemistry and for histological changes. Bacterial colonization was assessed also by culture from stomach samples. H. pylori was cultured by stomach samples of infected mice at 7, 14, and 30 days. Using PCR and immunohistochemistry, H. pylori was detected in inoculated and uninoculated mice in all areas examined, with an high percentage of positive samples in the esophagus and stomach. Moreover transmission was detected, without differences, regardless of whether mice were housed with or without a grate on the floor, supporting an orooral animal transmission.

Effect of hyoscine N-butylbromide on gastroesophageal reflux in normal subjects and patients with gastroesophageal reflux disease.

OBJECTIVES: Recent studies have shown that atropine reduces gastroesophageal reflux in normal subjects and patients with gastroesophageal reflux. The aim of the study has been to assess the effects of an atropine derivative, hyoscine N-butylbromide in normal subjects and patients with gastroesophageal reflux disease by recording esophageal and gastric pH-metry for a 24-h period. METHODS: Ten normal subjects and 10 patients with gastroesophageal reflux disease were evaluated. PH-metry was performed using two glass pH flexible probes with distal incorporated electrodes. The two catheters were introduced nasally under fluoroscopy. One probe was positioned in the gastric body; the other was placed 5 cm above the lower esophageal sphincter which had been evaluated manometrically before the study. Recording lasted without interruption for 48 h. Patients and normal subjects were assigned to receive hyoscine N-butylbromide (10 mg p.o. t.i.d.) for 24 h followed by a placebo for another 24 h or vice versa in a random manner. The pH was analyzed for a total number of acid refluxes and percentage of the period with pH <4 in the esophagus and the mean gastric pH in 24 h, before and after treatment with hyoscine N-butylbromide. RESULTS: The number of reflux episodes was significantly greater with hyoscine N-butylbromide in comparison with a placebo in patients with gastroesophageal reflux disease and normal subjects (p < 0.02). The percentage of time with pH <4, was also significantly greater in patients with gastroesophageal reflux disease and in controls (p < 0.05). The mean 24-h gastric pH after hyoscine N-butylbromide was not different from placebo in gastroesophageal reflux disease and controls. CONCLUSIONS: Hyoscine N-butylbromide, an anticholinergic agent, increases the total number of esophageal acid refluxes in patients with gastroesophageal reflux disease and in controls, therefore it is not recommended in the treatment of gastroesophageal reflux disease.