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Expression of intestinal trefoil factor in early repairing process of acute gastric mucosal damage induced, by indomethacin in rat
A208 AGA ABSTRACTS
GASTROENTEROLOGYVol. 114, No. 4
G0850
can be speculated that the oral cavity may not be an important reservoir for
EXPRESSION OF INTESTINAL TREFOIL FACTOR IN EARLY REPAIRING PROCESS OF ACUTE GASTRIC MUCOSAL DAMAGE INDUCED BY INDOMETHACIN IN RAT. W.S. Luo, H. Hiraishi, M. Tamano, Y. Kuronuma, T. Shimada, A. Terano. Second Department of Internal Medicine, Dokkyo University School of Medicine, Tochigi, Japan
H. pylori.
Backeround & Aims; Trefoil peptides such as pS2, PSP, and intestinal trefoil factor (ITF) are a relatively new group of peptides which have been shown to prevent and/or ameliorate gastric mucosal injury induced by ethanol and by NSAIDs. They may also serve to reseal mucosal wounds, influence cell proliferation, and accelerate the migration of proliferating epithelial cells in humans and mice. All are considered to be tissue-specific, and ITF is shown to be produced by goblet ceils in the normal intestinal mucosa and to be delivered with mucin glycoprotein. However, it has not been fully examined whether ITF is expressed constitutively in the stomach. Moreover, a possible role of ITF expression in repairing process of gastric mucosal injury remains undetermined. In the present study, a model of acute gastric mucosal damage in the rat was used to examine the expression of ITF which may play an active part in the healing response. Methods: Acute g~/stric mucosal damage was induced by intragastric administration of indomethacin (20 mg/kg). The expression of ITF was monitored over the next 72 hours using immunohistochernistry, and was assessed quantitatively by measuring proportion of ITF-positive area (%) to the background on the samples with immunohistochemical staining. The expression of proliferating cell nuclear antigen (PCNA) was also examined. Re$ult$: (1) In the normal gastroduodenal rnucosa, ITF immunoreactivity was present strongly at the glandular bases of the gastric body, weakly at the glandular necks, and most strongly within the duodenal glands (linked with mucin glycoprntein). No immunostaining was seen in the bulk of the gastric surface epithelium. (2) Administration of indomethacin induced hemorrhagic lesions over the body after 3 hrs, erosions and ulcerations over the antrum after 24 hrs, and reepithelialization of the ulcerations after 72 hrs. (3) Three hours after ulcer induction, enhanced immunoreactivity of ITF was not detected. Twenty four hours or later after induction, immunoreactive ITF was enhanced not only in the mucous neck cells, but also at the glandular bases. ITF-positive area at the ulcer beds was 0% after 3 and 24 hrs, and 0.8% after 72 hrs. In contrast, ITF-positive area at the ulcer edges increased in a time-related fashion (0%, 3.3% and 8.4% after 3, 24 and 72 hrs, respectively), whereas it did not change at non-ulcerated regions during the period. (4) PCNA labeling index at the ulcer edge increased, corresponding with enhanced immunoreactivity of ITF. Conclusions: The localization of ITF to the normal gastric epithelium, particularly the bases of fundic glands, raises the possibility that ITF may act as a regulator of secretory function. The pronounced expression of ITF around the ulcerations after induction of gastric injury suggests that ITF peptide may play a significant role in the repair-accelerating mechanisms, and also in the later ulcer-healing processes in the stomach. • G0851
SUGGESTION AGAINST AN ORO-ORAL ROUTE OF TRANSMISSION FOR HELICOBACTER PYLORI INFECTION: A SEROEPIDEMIOLOGICAL STUDY IN A RURAL AREA. F Luzza, M Imeneo, M Maletta, G Paluccio, S Nisticr*, M Costa, F Perticone, C Cavaliere, A Foc~t* & F Pallone. Dipartimento di Medicina Sperimentale, *Cattedra di Microbiologia, Universith di Catanzaro, Italy.
Background: The mode of transmission of H. pylori infection is still unknown. Epidemiological data support both oro-oral and feco-oral routes.
Aim: To compare the seroepidemiology of H. pylori and Epstein-Barr virus (EBV) in the same setting, using the last as a marker of oro-oral exposure.
Methods: A sample of 705 (203 male, age range 1-87yrs, median 50) resident subjects who attended the outpatient medical center of the rural town of Cir~ (11.000 inhabitants, Southern Italy) for blood testing was recruited. All subjects completed a structured questionnaire dealing with current and childhood socioeconomic features. A serum sample was drawn from each subject and assayed for H. pylori IgG by a validated ELISA. IgG antibodies to EBV were measured (Alfabiotech, Pomezia, Italy) in a subgroup of 466 (163 male, age range 1-87yrs, median 49) subjects. Data were analyzed by ~2 test, multiple logistic regression analysis and ~: statistic. Odds ratios (OR) and 95% confidence intervals (CI) were given. Results: Seropositivity was found in 63%(446/705) and 92%(429/466) of the subjects for H. pylori and EBV, respectively. Cross-tabulation of data showed that 274(58.8%) subjects were seropositive and 20(4.3%) were seronegative for both infections, 17(3.6%) were seropositive for H. pylori and 155(33.3%) were seropositive for EBV (OR=2.08, CI:1.008-4.3). The agreement between H. pylori and EBV seropositivity was no better than chance (r:=0.067) also when assessed in subjects aged less than 21 years (K=0.027). Furthermore, the age-related seroprevalence curve of EBV was similar in H. pylori seropositive and seronegative subjects. Multiple logistic regression analysis showed that dyspepsia (OR=I.5, CI:1.1-2.2) and occupation (OR=0.7, CI:0.5-0.9) were independently associated with H. pylori while no variable was found to be associated with EBV. Conclusion: Findings of this study do not support the hypothesis that H. pylori and EBV share a common mode of transmission. It
G0852
THE PREVALENCE OF HELICOBACTER PYLORI INFECTION IS NOT INCREASED IN CHILDREN WITH CELIAC DISEASE, F Luzza, M Mancuso*, M Imeneo, L Mesuraca*, A Contaldo*, L Pensabene*, G Scozia*, P. Strisciuglio* S Guandalini* & F Pallone. Dipt di Medicina Sperimentale, *Cattedra di Pediatria, Universith di Catanzaro, Italy.
Background: Celiac disease is frequently associated with chronic gastritis. Helicobacter pylori is the aetiological agent in over 90% of chronic gastritis. Aim: To assess the prevalence of H. pylori in children with celiac disease.
Methods: Eighty-one children (24M, 57E, age range: 1.4-17.7 years, median 6.8) with celiac disease were studied. The diagnosis was based on the characteristic histological finding of subtotal or severe partial villous atrophy and crypt hyperplasia in duodenal and jejunal biopsy. In 30 children (9M, 21F, age range: 1.4-17.2 years, median 7) who underwent endoscopy for duodenal biopsy, 3 further gastric (antral and body) biopsy specimens were taken for histology (ematoxilin & eosin) and H. pylori detection (urease quick test and Giemsa staining). All children had a blood sample taken while undergoing biopsy. Serum samples from 81 age and sex matched non-celiac children attending the outpatient pediatric department for diagnostic investigations were obtained. Children who underwent endoscopy were classified as H. pylori positive when urease quick test and/or histology were positive. In the remaining celiac children and in the control group, H. pylori status was determined according to a serum H. pylori IgG assay (Helori-test lgG, Eurospital, Trieste, Italy) validated (sensitivity and specificity > 90%) in a local pediatric setting. Results: Overall, 15 (18.5%) children with celiac disease and 14 (17.4%) controls were positive for H. pylori. The percentage H. pylori positivity was similar (p=0.8) in children with untreated (6/32,19%) and treated (9/49, 18%) celiac disease. Gastric nodularity and/or erytema was detected at endoscopy in 217 and 2/23 H. pylori positive and negative children, respectively (p=0.2). Superficial chronic gastritis with or without follicle formation was found in 6/7 and 19/23 11. pylori positive and negative children, respectively (p=l). Conclusion: The prevalence of H. pylori infection in children with celiac disease is not higher than in controls. In these patients, the endoscopic and histological features of the gastric mucosa did not differ according to H. pylori status. G0853 VALIDATION OF THE PCR STRING TEST (ENTERO-TEST) IN THE DIAGNOSIS OF HELICOBACTER PYLORI GASTRITIS. Lynch S, Nowak TV, Nowak JA, Konieek F, Ghazanfari K, Olivera A. Dept. of Medicine, Indiana University, Indianapolis, IN; Dept. of Pathology, Illinois Masonic Medical Center, Chicago, IL. It is known that Helicobacter pylori (HP) inhabits the gastric mucus layer of infected individuals. Our previous investigations have shown that the density of HP organisms in the gastric mucus of untreated patients is approximately 107-108 organisms per ml (Nowak JA. Am J Clin Pathol 1997; 108:284-288). We have also shown that gastric mucus is an optimum specimen for the detection of HP in infected individuals, and that PCR-based assays of gastric mucus are significantly more sensitive than histologic testing or the CLO test for demonstration of HP infection (Nowak JA: op. cit.). The "string test" has been successfully used for many years for the recovery of upper gastrointestinal fluid to aid in the diagnosis of parasitic infection. We hypothesized that recovery of gastric juice by use of the string test and analysis of recovered fluid by PCR might be a valid test to determine infection with HP. Forty-eight adult patients with symptoms of upper gastrointestinal dysfunction were evaluated. After an overnight fast, and just prior to endoscopic examination, each patient swallowed the gelatin string test capsule (ENTERO-TEST, HDC Corporation) with several ounces of water. Each patient lay on his left side for 30-60 minutes with the proximal end of the string taped to the cheek. The string was manually removed and each patient underwent endoscopy with biopsy of the gastric mucosa. Gastric biopsies were submitted for histologic examination for HP, or for rapid urease testing (CLO test or PYLORITEK). Approximately 150-200 ul of gastric mucus were expressed from each string and 5 ul of each sample were neutralized and used for PCR. PCR was performed using primers specific for the ribosomal RNA gene or the urease gene. PCR products were analyzed by agarose gel electrophoresis and visualized under UV light after ethidium bromide staining. The presence of inhibitors in each sample was identified by the addition of a known HP laboratory strain. The undiluted specimen in many instances was found to contain substances inhibitory to PCR. A 10-fold solution was sufficient to remove the inhibition in every case. Fifteen of 48 patients (32%) had evidence of HP infection based on histologic examination or assay of gastric biopsies for urease activity. The PCR-string assay accurately detected HP in 12 of these 15 patients, for a sensitivity of 80% Thirty-three patients had no evidence of HP by histologic examination or urease activity. The PCR assay was negative in all 33 patients, a specificity of 100%. We conclude that the PCR-string test is an accurate, non-invasive, and highly specific test for the diagnosis of Helicobacter pylori infection.
GASTROENTEROLOGYVol. 114, No. 4
G0850
can be speculated that the oral cavity may not be an important reservoir for
EXPRESSION OF INTESTINAL TREFOIL FACTOR IN EARLY REPAIRING PROCESS OF ACUTE GASTRIC MUCOSAL DAMAGE INDUCED BY INDOMETHACIN IN RAT. W.S. Luo, H. Hiraishi, M. Tamano, Y. Kuronuma, T. Shimada, A. Terano. Second Department of Internal Medicine, Dokkyo University School of Medicine, Tochigi, Japan
H. pylori.
Backeround & Aims; Trefoil peptides such as pS2, PSP, and intestinal trefoil factor (ITF) are a relatively new group of peptides which have been shown to prevent and/or ameliorate gastric mucosal injury induced by ethanol and by NSAIDs. They may also serve to reseal mucosal wounds, influence cell proliferation, and accelerate the migration of proliferating epithelial cells in humans and mice. All are considered to be tissue-specific, and ITF is shown to be produced by goblet ceils in the normal intestinal mucosa and to be delivered with mucin glycoprotein. However, it has not been fully examined whether ITF is expressed constitutively in the stomach. Moreover, a possible role of ITF expression in repairing process of gastric mucosal injury remains undetermined. In the present study, a model of acute gastric mucosal damage in the rat was used to examine the expression of ITF which may play an active part in the healing response. Methods: Acute g~/stric mucosal damage was induced by intragastric administration of indomethacin (20 mg/kg). The expression of ITF was monitored over the next 72 hours using immunohistochernistry, and was assessed quantitatively by measuring proportion of ITF-positive area (%) to the background on the samples with immunohistochemical staining. The expression of proliferating cell nuclear antigen (PCNA) was also examined. Re$ult$: (1) In the normal gastroduodenal rnucosa, ITF immunoreactivity was present strongly at the glandular bases of the gastric body, weakly at the glandular necks, and most strongly within the duodenal glands (linked with mucin glycoprntein). No immunostaining was seen in the bulk of the gastric surface epithelium. (2) Administration of indomethacin induced hemorrhagic lesions over the body after 3 hrs, erosions and ulcerations over the antrum after 24 hrs, and reepithelialization of the ulcerations after 72 hrs. (3) Three hours after ulcer induction, enhanced immunoreactivity of ITF was not detected. Twenty four hours or later after induction, immunoreactive ITF was enhanced not only in the mucous neck cells, but also at the glandular bases. ITF-positive area at the ulcer beds was 0% after 3 and 24 hrs, and 0.8% after 72 hrs. In contrast, ITF-positive area at the ulcer edges increased in a time-related fashion (0%, 3.3% and 8.4% after 3, 24 and 72 hrs, respectively), whereas it did not change at non-ulcerated regions during the period. (4) PCNA labeling index at the ulcer edge increased, corresponding with enhanced immunoreactivity of ITF. Conclusions: The localization of ITF to the normal gastric epithelium, particularly the bases of fundic glands, raises the possibility that ITF may act as a regulator of secretory function. The pronounced expression of ITF around the ulcerations after induction of gastric injury suggests that ITF peptide may play a significant role in the repair-accelerating mechanisms, and also in the later ulcer-healing processes in the stomach. • G0851
SUGGESTION AGAINST AN ORO-ORAL ROUTE OF TRANSMISSION FOR HELICOBACTER PYLORI INFECTION: A SEROEPIDEMIOLOGICAL STUDY IN A RURAL AREA. F Luzza, M Imeneo, M Maletta, G Paluccio, S Nisticr*, M Costa, F Perticone, C Cavaliere, A Foc~t* & F Pallone. Dipartimento di Medicina Sperimentale, *Cattedra di Microbiologia, Universith di Catanzaro, Italy.
Background: The mode of transmission of H. pylori infection is still unknown. Epidemiological data support both oro-oral and feco-oral routes.
Aim: To compare the seroepidemiology of H. pylori and Epstein-Barr virus (EBV) in the same setting, using the last as a marker of oro-oral exposure.
Methods: A sample of 705 (203 male, age range 1-87yrs, median 50) resident subjects who attended the outpatient medical center of the rural town of Cir~ (11.000 inhabitants, Southern Italy) for blood testing was recruited. All subjects completed a structured questionnaire dealing with current and childhood socioeconomic features. A serum sample was drawn from each subject and assayed for H. pylori IgG by a validated ELISA. IgG antibodies to EBV were measured (Alfabiotech, Pomezia, Italy) in a subgroup of 466 (163 male, age range 1-87yrs, median 49) subjects. Data were analyzed by ~2 test, multiple logistic regression analysis and ~: statistic. Odds ratios (OR) and 95% confidence intervals (CI) were given. Results: Seropositivity was found in 63%(446/705) and 92%(429/466) of the subjects for H. pylori and EBV, respectively. Cross-tabulation of data showed that 274(58.8%) subjects were seropositive and 20(4.3%) were seronegative for both infections, 17(3.6%) were seropositive for H. pylori and 155(33.3%) were seropositive for EBV (OR=2.08, CI:1.008-4.3). The agreement between H. pylori and EBV seropositivity was no better than chance (r:=0.067) also when assessed in subjects aged less than 21 years (K=0.027). Furthermore, the age-related seroprevalence curve of EBV was similar in H. pylori seropositive and seronegative subjects. Multiple logistic regression analysis showed that dyspepsia (OR=I.5, CI:1.1-2.2) and occupation (OR=0.7, CI:0.5-0.9) were independently associated with H. pylori while no variable was found to be associated with EBV. Conclusion: Findings of this study do not support the hypothesis that H. pylori and EBV share a common mode of transmission. It
G0852
THE PREVALENCE OF HELICOBACTER PYLORI INFECTION IS NOT INCREASED IN CHILDREN WITH CELIAC DISEASE, F Luzza, M Mancuso*, M Imeneo, L Mesuraca*, A Contaldo*, L Pensabene*, G Scozia*, P. Strisciuglio* S Guandalini* & F Pallone. Dipt di Medicina Sperimentale, *Cattedra di Pediatria, Universith di Catanzaro, Italy.
Background: Celiac disease is frequently associated with chronic gastritis. Helicobacter pylori is the aetiological agent in over 90% of chronic gastritis. Aim: To assess the prevalence of H. pylori in children with celiac disease.
Methods: Eighty-one children (24M, 57E, age range: 1.4-17.7 years, median 6.8) with celiac disease were studied. The diagnosis was based on the characteristic histological finding of subtotal or severe partial villous atrophy and crypt hyperplasia in duodenal and jejunal biopsy. In 30 children (9M, 21F, age range: 1.4-17.2 years, median 7) who underwent endoscopy for duodenal biopsy, 3 further gastric (antral and body) biopsy specimens were taken for histology (ematoxilin & eosin) and H. pylori detection (urease quick test and Giemsa staining). All children had a blood sample taken while undergoing biopsy. Serum samples from 81 age and sex matched non-celiac children attending the outpatient pediatric department for diagnostic investigations were obtained. Children who underwent endoscopy were classified as H. pylori positive when urease quick test and/or histology were positive. In the remaining celiac children and in the control group, H. pylori status was determined according to a serum H. pylori IgG assay (Helori-test lgG, Eurospital, Trieste, Italy) validated (sensitivity and specificity > 90%) in a local pediatric setting. Results: Overall, 15 (18.5%) children with celiac disease and 14 (17.4%) controls were positive for H. pylori. The percentage H. pylori positivity was similar (p=0.8) in children with untreated (6/32,19%) and treated (9/49, 18%) celiac disease. Gastric nodularity and/or erytema was detected at endoscopy in 217 and 2/23 H. pylori positive and negative children, respectively (p=0.2). Superficial chronic gastritis with or without follicle formation was found in 6/7 and 19/23 11. pylori positive and negative children, respectively (p=l). Conclusion: The prevalence of H. pylori infection in children with celiac disease is not higher than in controls. In these patients, the endoscopic and histological features of the gastric mucosa did not differ according to H. pylori status. G0853 VALIDATION OF THE PCR STRING TEST (ENTERO-TEST) IN THE DIAGNOSIS OF HELICOBACTER PYLORI GASTRITIS. Lynch S, Nowak TV, Nowak JA, Konieek F, Ghazanfari K, Olivera A. Dept. of Medicine, Indiana University, Indianapolis, IN; Dept. of Pathology, Illinois Masonic Medical Center, Chicago, IL. It is known that Helicobacter pylori (HP) inhabits the gastric mucus layer of infected individuals. Our previous investigations have shown that the density of HP organisms in the gastric mucus of untreated patients is approximately 107-108 organisms per ml (Nowak JA. Am J Clin Pathol 1997; 108:284-288). We have also shown that gastric mucus is an optimum specimen for the detection of HP in infected individuals, and that PCR-based assays of gastric mucus are significantly more sensitive than histologic testing or the CLO test for demonstration of HP infection (Nowak JA: op. cit.). The "string test" has been successfully used for many years for the recovery of upper gastrointestinal fluid to aid in the diagnosis of parasitic infection. We hypothesized that recovery of gastric juice by use of the string test and analysis of recovered fluid by PCR might be a valid test to determine infection with HP. Forty-eight adult patients with symptoms of upper gastrointestinal dysfunction were evaluated. After an overnight fast, and just prior to endoscopic examination, each patient swallowed the gelatin string test capsule (ENTERO-TEST, HDC Corporation) with several ounces of water. Each patient lay on his left side for 30-60 minutes with the proximal end of the string taped to the cheek. The string was manually removed and each patient underwent endoscopy with biopsy of the gastric mucosa. Gastric biopsies were submitted for histologic examination for HP, or for rapid urease testing (CLO test or PYLORITEK). Approximately 150-200 ul of gastric mucus were expressed from each string and 5 ul of each sample were neutralized and used for PCR. PCR was performed using primers specific for the ribosomal RNA gene or the urease gene. PCR products were analyzed by agarose gel electrophoresis and visualized under UV light after ethidium bromide staining. The presence of inhibitors in each sample was identified by the addition of a known HP laboratory strain. The undiluted specimen in many instances was found to contain substances inhibitory to PCR. A 10-fold solution was sufficient to remove the inhibition in every case. Fifteen of 48 patients (32%) had evidence of HP infection based on histologic examination or assay of gastric biopsies for urease activity. The PCR-string assay accurately detected HP in 12 of these 15 patients, for a sensitivity of 80% Thirty-three patients had no evidence of HP by histologic examination or urease activity. The PCR assay was negative in all 33 patients, a specificity of 100%. We conclude that the PCR-string test is an accurate, non-invasive, and highly specific test for the diagnosis of Helicobacter pylori infection.