G. Nardone

Effect of Helicobacter pylori infection and its eradication on cell proliferation, DNA status, and oncogene expression in patients with chronic gastritis

BACKGROUND Helicobacter pylori, the main cause of chronic gastritis, is a class I gastric carcinogen. Chronic gastritis progresses to cancer through atrophy, metaplasia, and dysplasia. Precancerous phenotypic expression is generally associated with acquired genomic instability. AIM To evaluate the effect of H pylori infection and its eradication on gastric histology, cell proliferation, DNA status, and oncogene expression. METHODS/SUBJECTS Morphometric and immunohistochemical techniques were used to examine gastric mucosal biopsy specimens from eight controls, 10 patients withH pylori negative chronic gastritis, 53 withH pylori positive chronic gastritis, and 11 with gastric cancer. RESULTS All patients with chronic gastritis were in a hyperproliferative state related to mucosal inflammation, regardless of H pyloriinfection. Atrophy was present in three of 10 patients withH pylori negative chronic gastritis and in 26 of 53 with H pylori positive chronic gastritis, associated in 18 with intestinal metaplasia. DNA content was abnormal in only 11 patients with atrophy and H pylori infection; eight of these also had c-Myc expression, associated in six cases with p53 expression. Fifty three patients withH pylori positive chronic gastritis were monitored for 12 months after antibiotic treatment: three dropped out; infection was eradicated in 45, in whom cell proliferation decreased in parallel with the reduction in gastritis activity; atrophy previously detected in 21/45 disappeared in five, regressed from moderate to mild in nine, and remained unchanged in seven; complete metaplasia disappeared in 4/14, and markers of genomic instability disappeared where previously present. In the five patients in whomH pylori persisted, atrophy, metaplasia, dysplasia, and markers of genomic instability remained unchanged. CONCLUSIONS ChronicH pylori infection seems to be responsible for genomic instability in a subset of cases of H pylori positive chronic atrophic gastritis; eradication ofH pylori infection can reverse inflammation and the related atrophy, metaplasia, and genomic instability.

Reflux oesophagitis in adult coeliac disease: beneficial effect of a gluten free diet

Background: Coeliac disease patients show a number of gastrointestinal motor abnormalities, including a decrease in lower oesophageal sphincter pressure. The prevalence of endoscopic oesophagitis in these subjects however is unknown. Aim: To evaluate whether untreated adult coeliac patients had an increased prevalence of reflux oesophagitis and, if so, to assess whether a gluten free diet exerted any beneficial effect on gastro-oesophageal reflux disease (GORD) symptoms. Patients and methods: We retrospectively studied 205 coeliac patients (females/males 153/52, median age 32 years) who underwent endoscopy for duodenal biopsy and 400 non-coeliac subjects (females/males 244/156, median age 37 years) referred for endoscopy for upper gastrointestinal symptoms. Each patient was given a questionnaire for evaluation of GORD symptoms prior to and 4–12 months after endoscopy. Coeliac patients were given a gluten free diet. Oesophagitis patients of both groups, following an eight week course of omeprazole, were re-evaluated for GORD symptoms at four month intervals up to one year. Significance of differences was assessed by Fisher’s exact test. Results: Oesophagitis was present in 39/205 (19%, 95% confidence interval (CI) 13.8–25.0%) coeliac patients and in 32/400 (8%, 95% CI 5.5–11.1%) dyspeptic subjects. At the one year follow up, GORD symptoms relapsed in 10/39 (25.6%, 95% CI 13–42.1%) coeliacs with oesophagitis and in 23/32 (71.8%, 95% CI 53.2–86.2%) non-coeliac subjects with oesophagitis. Conclusion: Coeliac patients have a high prevalence of reflux oesophagitis. That a gluten free diet significantly decreased the relapse rate of GORD symptoms suggests that coeliac disease may represent a risk factor for development of reflux oesophagitis.

Tissue ghrelin level and gastric emptying rate in adult patients with celiac disease.

Abstract  Celiac disease (CD) patients show a number of gastrointestinal motor abnormalities. Ghrelin, a gastric peptide implicated in short‐term feeding control and long‐term body weight regulation, has been recently considered a key regulator of gastric motility. The aim of this study was to evaluate the gastric emptying rate of solids and the density of ghrelin‐immunopositive cells in adult CD patients before and at least 1 year after starting a gluten‐free diet. Twenty CD patients (M 8/F 12; mean age 36 years) and 10 controls underwent endoscopy with gastric and duodenal biopsies and 13C‐octanoic acid breath test to measure gastric emptying of solids. Celiac disease patients repeated the protocol at least 1 year after starting gluten‐free diet. Ghrelin tissue levels were evaluated by immunohistochemistry on gastric mucosa specimens. Gastric emptying time was normal in all control subjects (t1/2 = 89 ± 16 min) while it was delayed in CD patients prior to gluten‐free diet (t1/2 = 252 ± 101 min; P < 0.005). The mean number of ghrelin‐positive cells/field (×400) was 14.4 ± 2.7 in controls and 25.3 ± 5.7 in CD patients respectively (P < 0.0001). Gluten withdrawal was effective in normalizing gastric emptying time in all CD patients (97 ± 14 min; P < 0.0001) and resulted in a significant reduction of the density of ghrelin‐immunopositive cells (19.8 ± 5.4; P < 0.0001). The density of ghrelin‐positive cells correlated directly with the degree of duodenal damage (P < 0.001) and inversely with the body mass index of CD patients (P < 0.0001). However, in neither CD patients nor controls, a correlation between tissue ghrelin levels and gastric emptying rate was detected. In conclusion, tissue ghrelin level does not correlate with gastric emptying rate in adult CD patients and in controls.

Up-regulation of heparin binding epidermal growth factor-like growth factor and amphiregulin expression in Helicobacter pylori-infected human gastric mucosa

BACKGROUND Host response plays a major role in pathogenesis of Helicobacter pylori-induced gastroduodenal disease including adenocarcinoma of distal stomach. Epidermal growth factor-related growth factors are important modulators of gastric homeostasis in normal and damaged gastrointestinal mucosa. AIM To evaluate expression of heparin binding epidermal growth factor and amphiregulin in antral mucosa of Helicobacter pylori-infected and non-infected dyspeptic patients and to correlate levels of heparin binding-epidermal growth factor and amphiregulin mRNA with mitogenic activity of gastric epithelial cells. METHODS A total of 10 Helicobacter pylori-infected and 15 Helicobacter pylori non-infected (10 with and 5 without gastritis) dyspeptic patients were studied. Diagnosis of Helicobacter pylori infection was based on rapid urease test and histology. Heparin binding-epidermal growth factor and amphiregulin mRNA expression in antral mucosa were assessed by reverse transcriptase-polymerase chain reaction. Protein expression and localization of both peptides were determined by immunohistochemistry. Mitogenic activity of antral gastric mucosa was assessed by determination of proliferating cell nuclear antigen labelling index by immunohistochemistry. RESULTS Heparin binding-epidermal growth factor and amphiregulin mRNA expression increased in Helicobacter pylori-infected vs Helicobacter pylori non-infected patients. Heparin binding-epidermal growth factor and amphiregulin immunostaining was more intense and deeper in gastric gland compartment in infected mucosa than in non-infected mucosa. Increase in heparin binding-epidermal growth factor and amphiregulin mRNA expression significantly correlated with increase in proliferating cell nuclear antigen labelling index. CONCLUSIONS Helicobacter pylori gastritis is associated with up-regulation of heparin binding-epidermal growth factor and amphiregulin which correlates with increased mitogenic activity of gastric mucosa. Increased heparin binding-epidermal growth factor and amphiregulin expression is postulated to contribute to reparative response of gastric mucosa to Helicobacter pylori infection.

Increased severity of dyspeptic symptoms related to mental stress is associated with sympathetic hyperactivity and enhanced endocrine response in patients with postprandial distress syndrome

Background  Mental stress (MS) may alter gastric sensory‐motor function. The aim of the study was to assess postprandial autonomic nervous system activity and stress hormones in response to acute mental stress in dyspeptic patients.

Interleukin-13 mucosal production in Helicobacter pylori-related gastric diseases

UNLABELLED A shift from Th1 (IFN-gamma) towards Th2 (IL-4)-type immune response was found in patients with gastric cancer and dysplasia. Recently, IL-13 has been described as a central mediator of Th2-dominant immune response in different inflammatory diseases. AIM AND METHODS to analyse, by Enzyme-Linked-Immuno-SPOT (ELISPOT) assay and immunohistochemistry, the IL-13 production of mononuclear cells obtained from gastric biopsies of 19 H. pylori-negative subjects and 23 H. pylori-positive patients. RESULTS By ELISPOT, we did not find any significant variation of the spot range number of IL-13, IL-4 and IFN-gamma secreting cells, irrespective of H. pylori status. After antigenic exposition, the spot range for IL-13, IL-4 and IFN-gamma significantly increased (p<.0001) only in H. pylori-positive patients. A prevalent Th1 (IFN-gamma) immunoresponse was observed in 2/23 cases with active gastritis, while a prevalent Th2 (IL-13 and IL-4) was detected in 5/23 cases all with atrophic chronic gastritis of whom two with intestinal metaplasia. By immunohistochemistry, IL-13, IL-4 and IFN-gamma were detectable in all cases directly related to the inflammatory infiltrate. In the two cases with intestinal metaplasia, IL-13 and IL-4 were localised in both inflammatory and epithelial cells. This immunopattern was confirmed in selected additional 10 cases of H. pylori-positive chronic atrophic gastritis with intestinal metaplasia and 10 cases of intestinal type gastric cancer. CONCLUSION These preliminary results suggest that IL-13 could be implicated in the different outcome of H. pylori infection.

Effect of Helicobacter pylori infection on gastric cell proliferation and genomic instablity in a paediatric population of Southern Italy

BACKGROUND The incidence of gastric cancer is high in areas with a high prevalence of Helicobacter pylori infection. Cell transformation and tumour progression occur over a long period of time and markers of genomic instability usually precede morphological changes. AIM To evaluate the effect of Helicobacter pylori infection on cell proliferation, DNA status and oncogene expression in children. PATIENTS AND METHODS Morphometric and immunohistochemical techniques were used to analyse DNA content, p53 and c-myc oncogene expression and cell proliferation on gastric biopsies of 53 children (27 Helicobacter pylori-negative and 26 Helicobacter pylori-positive). RESULTS Gastric mucosa was normal in 11% of Helicobacter pylori-positive and in 33% of Helicobacter pylori-negative subjects. Most children had chronic non-atrophic gastritis regardless of Helicobacter pylori infection, and only a minority of children affected by Helicobacter pylori had mild atrophic gastritis. Cell proliferation was significantly higher in children with Helicobacter pylori-positive gastritis than in those with Helicobacter pylori-negative gastritis. No metaplasia, dysplasia, p53 overexpression or altered DNA content was found in any child. Interestingly, 46% of children with and 29% without Helicobacter pylori infection had c-myc overexpression closely related to the cell proliferation rate. CONCLUSION Helicobacter pylori infection in children may coexist with a normal gastric mucosa, and it is not associated with genomic instability markers in cases of chronic gastritis.

Clinical nutrition practice in Italian Gastroenterology Units

BACKGROUND Nutritional status affects the course, ensuing complications and prognosis of virtually all diseases. AIMS To define the role of nutrition in Gastroenterology Units by means of two investigations that analyse: a) availability of devices for assessing nutritional status; b) nutritional treatment in clinical practice: incidence and frequency of indications for its use, together with type of treatment adopted. PATIENTS AND METHODS Two questionnaires were sent to Italian Academic and Hospital Gastroenterology Units, all with clinical wards. RESULTS Results refer to 27 Units, 22 of which took part in both parts of the analysis, enrolling 547 patients during the two-week study The first analysis shows that scales and the altimeter are not available everywhere, while more specific tools, such as skinfold calipers are available in 54% of the Units, and caloric intake can be assessed in 22-41%. The second analysis reveals that nutritional treatment was necessary in 50% of patients in the series examined, and that this was taken into account and prescribed in almost all cases (91%). Of the patients treated, 69% received dietetic supplementation and 31% artificial nutrition [12% enteral, 88% parenteral), although supportive parenteral nutrition is often contraindicated in conditions where good bowel function provides the conditions for enteral nutrition. CONCLUSION Data emerging from the investigation showed that i) artificial nutrition is commonly used in gastroenterology Units in Italy although 23% of them never consider either enteral or parenteral nutrition as medical treatment of gastrointestinal disease; ii) malnutrition is a very frequent complication (mean 27%; range 4-55%0) in Gastroenterology Unit patients albeit only 42% of malnourished patients received artificial nutrition; iii) indications for enteral and parenteral nutrition are not always respected, as there is an excessive use of parenteral nutrition and an unjustified resistance to the use of enteral nutrition; iv] nutritional treatment is often administered without adequate nutritional assessment and without a complete adherence to the standards recommended for preparation of parenteral bags, supported by suitable technology; v) only two Gastroenterology Units report admitting and following patients in a home parenteral nutrition programme; vi) this investigation probably reflects the response of those Gastroenterology Units most aware of the importance of nutritional problems. Better awareness of correct practices for nutritional support should be promoted, encouraging greater use of diagnostic and monitoring techniques and a more discerning choice of the most suitable type of artificial nutrition to be administered in gastroenterology

P.03.10 VITAMIN B12 SUPPLEMENTATION IMPROVES SUSTAINED-VIRAL-RESPONSE RATES IN HEPATITIS C VIRUS GENOTYPE 1B-INFECTED PATIENTS

Background and aim: We previously described a case of EUS-guided Nd:YAG (neodymium:yttrium-aluminium-garnet) LA of a HCC lesion located into the caudate lobe, not suitable for percutaneous approach. Aim is to confirm the feasibility of EUS-guided Nd:YAG LA of HCC in unsafe conditions for the percutaneous approach. Material and methods: Treatment was performed in 2 patients with multifocal HCC unsuitable for surgical resection or liver transplant. First one was affected by criptogenetic liver cirrhosis Child-Pugh A6 and lesion was site in the caudate lobe with huge ombelical vein and portal hypertension. The location of the lesion and the difficult ultrasonography visualization precluded percutaneous treatment. Second patient was affected by HCV and HIV cirrhosis Child-Pugh B9 with ascites and portal hypertension. Both patients underwent previous failed transarterial embolization (TACE) and RFA of lesions located into segment 1 and 3 respectively. Trans-gastric EUS-puncture was performed using a 22-gauge needle following the application of Doppler. A Nd:YAG laser with a wavelength of 1.064 nm was used. As previously described the treatment was planned taking into account the baseline volume of the lesions at EUS and the volume of necrosis that could be achieved in relation to the energy delivered. Results: Lesions were easier targeting through the lesser gastric curve. Application of Nd:YAG LA did not have any negative effect on the quality of the EUS images during the treatment and the whole treated area was occupied by an irregular and poorly defined echogenic zone at the end. The patients didn’t report any pain or abdominal discomfort after treatment and were discharged on the third postoperative day without complications. CT performed 24 hours after procedure showed the whole treated area replaced by an homogeneous, hypoattenuating, nonenhancing area. At 2 months follow-up clinical examination and blood analysis tests were normal. CT-scan showed uniform hypoattenuation without enhancement in the ablated zone and confirm the success to ablate the entire lesion. Conclusions: EUS-guided Nd:YAG LA of a HCC is feasible and safe in lesions in which the percutaneous approach is unsure. This promising results need to be confirmed in additional patients with lesions difficult to reach by conventional ablative methods or in patients whit compromised clinical conditions.

OC.07.5 HCV-ANTIVIRAL THERAPY DELAYS GASTRIC EMPTYING TIME AND MODIFIES CCK AND MOTILIN SERUM LEVELS

Background and aim: Digestive symptoms are common side effects of antiviral therapy in patients with HCV chronic hepatitis (CH). The occurrence of digestive symptoms significantly impairs quality of life and requires reduction or even suspension of the therapy in up to 15% of the cases. Recent advances in pathophisiology of functional dyspepsia indicate that delay of gastric emptying time (GET) likely depending on altered Cholecystokinin (CCK) and Motilin serum levels is implicated in the onset of symptoms. In this study we evaluated digestive symptoms, GET and CCK and motilin fasting and post-prandial serum levels in patients with HCV-related CH before, during and after standard antiviral therapy. Material and methods: Twenty-eight patients (M/F 11/17 male, age range 28-70 yrs) with histologically proven HCV-related CH and absence of digestive diseases were enrolled in the study. Baseline, during the therapy (3th month) and after a month by the end of therapy patients underwent: an oriented questionnaire evaluating digestive symptoms. 13C-octanoate breath test (13C-OBT) was performed to evaluate GET. Fasting and post-prandial CCK and Motilin serum levels were assessed ELISA. Antiviral therapy was performed according to standard protocols. Results: Baseline none of the patients complained of significant digestive symptoms. GET was normal (t/2 <120 min) in all cases but 3/28 (2%). Baseline and post-prandial CCK and Motilin levels were 0.6±0.4 and 1.57±0.5 and 5.9±5.2 and 1.7±0.9, respectively. At three-month therapy, epigastric burning, belching, epigastric pain, post-prandial fullness, early satiety, bloating, nausea and vomiting were reported by 31%, 57%, 7%, 60%, 57%, 53%, 46% and 14%, respectively. GET rate was significantly delayed in all cases (p <0.0001). CCK fasting and post-prandial serum levels significantly increased (p<0.0001) while Motilin decreased in respect to baseline values (p<0.003). Interestingly, there was a direct significant relation between GI symptom score, GET and CCK and Motilin serum level (p<0.05). After 1 month by the end of therapy, all patients were symptom-free and GET as well as CCK and Motilin serum levels returned to baseline values. Conclusions: Digestive symptoms caused by HCV-antiviral therapy depend on the delay of GET as well as deregulation of CCK/Motilin homeostasis.