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Utility of a rapid whole blood test for an initial diagnosis of Helicobacter pylori infection
GASTROENTEROLOGY Vol. 114, No. 4
A164 AGA ABSTRACTS • G0669
1-1.PYLORI
EXTRACT INHIBITS APOPTOSIS OF HUMAN NEUTROPHILS. HC Jung, JS Kim, JM Kim*, IS Song, CY Kim. Dept. of Internal Med. and Liver Research Institute, Seoul National Univ. College of Med., Dept. of Microbiology*, Hanyang Univ. College of Med., Seoul, Korea Background/Aims: H. pylori infection in human elicits marked neutrophil infiltration in gastric mucosa. The mechanisms of inflammatory trigger and perpetuation have not been clearly defined. Senescent neutrophils undergo apoptosis and phagocytosed by macrophages. Neutrophil apoptosis is likely to be important in the control of inflammatory tissue injury. To elucidate the perpetuation of gastric mucosal inflammation induced by H. pylori, we investigate regulatory function of soluble products of H. pylori on neutrophil apoptosis. Methods: Neutrophils were isolated from human peripheral blood using discontinuous density gradient centrifugation with Ficoll-Hypaque. The purity of the neutrophils was more than 98% confirmed by morphology and phenotypic analysis using FACS flow cytometry. The viability was more than 95% by trypan blue exclusion test. H. pylori water extract was concentrated from ultrafiltration and filtered through 0.2 lam to minimize the effect of water on apoptosis. The neutrophils were incubated with H. pylori water extract or E. coli lipopolysaccharide (LPS) 10 lag/mL in the culture medium for 6, 12, 24, 36 and 48 hours. Neutrophil apoptosis was determined by agarose gel electrophoresis of DNA, fluorescence microscopic observation after staining with Hoechst 33342 and TUNEL assay. CDllb/CD18 expression of neutrophils was analyzed by FACS flow cytometry. Results: Stimulation with LPS or H. pylori water extract inhibited apoptosis of neutrophils for periods up to 48 hours and enhanced the expression of CD1 lb/CD 18 on the surface of neutrophils. 12h (% apoptosis) 24h(%apoptosis) 48h(%apoptosis) 40±3.8 73±6.5 Control 12 ± 1.5 6±0.4 16±1.2 47±2.6 E.coli LPS 21±1.6 54±3.7 H.pylori extract 7±0.5 Conclusions: These results suggest that neutrophil apoptosis could be suppressed by the soluble factors released by H. pylori. Consequently, the prolongation of survival of neutrophils may coincide with persistent enhanced expression of adhesion molecules on neutrophils. These events could contribute to the perpetuation of H. pylori-induced gastric mucosal inflammation.
G0670 PREVALENCE OF HELICOBACTER PYLORI cagA GENE IN CANCER AND NON-CANCER TISSUES OF GASTRIC CANCER IN KOREA. S.A. Jung,, D.Y. Kim, H.K. Jung, H.C. Lee, S.Y. Yi, I.H. Moon. Division of Molecular Biology, Department of Internal Medicine, Ewha Womans University College of Medicine, Seoul, Korea Aim 1-1.pylori is a risk factor for the development of gastric cancer. In serologic study, cagA-positive H. pylori strain was more related to the development of gatsric cancer than cagA-negative H. pylori strain. To further understand the relationship between cagA gene and gastric cancer, the positive rates of cagA gene in cancer and non-cancer tissues were investigated separately in patients with gastric cancer. Methods The cagA gene was detected by PCR (J Clin Microbiol 1995;33;2752-6) and ureC gene was also assayed as positive controls for the presence of H. priori. Each two endoscopic biopsies were obtained from cancer and non-cancer tissues of 41 patients with gastric cancer. Results 1) The total positive rate of cagA gene in Korean patients with gastric cancer was 63.4%(26/41). 2) The positive rate of cagA gene in cancer tissues was 29.3%(12/41), which was significantly lower thanthat in non-cancer tissues(63.4%, 26141). 3) Twelve (29.3%) out of 41 were positive for cagA gene in both cancer and non-cancer tissues, 14(34.1%) in non-cancer tissues only, none(0%) in cancer tissues only and 15(36.6%) were negative in both sites. 4) The ureC were negative in cancer tissue in 12(85.7%) among 14 cases who were cagA gene negative in cancer tissues but positive in noncancer tissues. 5) There was no difference of positive rates of cagA gene according to age, stage, site and pathologic cell type. Conclusion These findings indicate that positive rate of cagA gene in cancer tissue was lower than that in non-cancer tissues and this might be related to low infection rate of H. pylori rather than the presence of cagA negative H. pylori in cancer tissues.
• G0671 UTILITY OF A RAPID WHOLE BLOOD TEST FOR AN INITIAL DIAGNOSIS OF HELICOBACTER PYLORI INFECTION. I. K~iiiriiiinen, E. Pulkkinen, K. Romanoff, P. Sipponen t & L. Sellin 2. Departments of Gastroenterology and :Pathology, Jorvi Hospital, Espoo, Finland and 2AstraFinland, Masala, Finland. Background. The cost and delay in the diagnosis of H. pylori infection via biopsy, serology or urea breath test have increased interest in low-cost and rapid methods which can be used for screening in a primary care environment. Aim. We have performed a validation study comparing a rapid (10 minutes) and simple whole blood test for H. pylori infection (AccuMeter ®, ChemTrak,
Sunnyvale, California, U.S.A.) to 13C-urea breath test and histology in 104 consecutive patients with abdominal symptoms and referred to endoscopy. Results. Compared to breath test and histology (modified Giemsa), which were highly concordant (99%), the AccuMeter ® was 96% sensitive and 9091% specific. In the one of 27 H. pylori-positive patients where the 13C-urea breath test was negative but H. pylori organisms were demonstrated in the biopsy specimens with histology and immunohistochemistry, the AccuMeter~ result was positive. The AccuMeter ® was slightly less likely to produce a false negative result than a false positive. The calculated predictive values of a negative result to exclude infection are, for example, 96% when the prevalence of infection is 50% and 85% when the prevalence is 20%. Correspondingly, the predictive values for a positive result to indicate infection are 91% and 73%. Conclusions. The AccuMeter* is a rapid, accurate and easyto-use whole blood test for the diagnosis of H. pylori infection and may provide a simple and potentially cost-effective screening tool in the primary care environment. This study was partially funded by Astra-Finland. • G0672 STUDY ON TItE EFFECT OF A NEW ANTI-ULCER AGENT, ECABET SODIUM, ON THE DUAL THERAPY FOR ERADICATION O F HELICOBACTER PYLORI IN RANDOMIZED PATIENTS. H~ Kagay~, M. Kato, T. Kobayashi, J. Ishizuka, K. Nishikawa, Y. Komatsu, M. Sukegawa, H. Takeda, T. Sugiyama and M. Asaka, The Third Department of the Internal Medicine, Hokkaido University School of Medicine, Sapporo, Japan Obiective: Ecabet sodium is a therapeutic agent for gastritis and gastric ulcers, which is locally distributed to the gastric mucosa and remains there for a long time to exert anti-ulcer activity. Ecabet sodium has been demonstrated to have bactericidal action in vitro not only against the standard strain of Helicobacter pylori but also against strains resistant to such drugs as metronidazole and clarithromycin. We investigated the improving effect of ecabet sodium on the eradication rate and its dose-dependency in a combination therapy with lansoprazole and amoxicillin as a method not to acquire resistance. Mcth0d~; Patients with chronic gastritis, who were confirmed to be Helicobacter pylori-positive by the rapid urease test (RUT), were randomly assigned to one of the following three groups and medicated for 2 weeks. Group I: Lansoprazole (30 mg, qd) + Amoxicillin (750 mg, bid); Group II: Lansoprazole (30 rag, qd) + Amoxicillin (750 mg, bid) + Ecabet sodium (1 g, bid); Group III: Lansoprazole (30 mg, qd) + Amoxicillin (750 mg, bid) + Ecabet sodium (2 g, bid). Patients were examined four weeks after the cessation of treatment and those who were Helicobacter pylori-negative by culture test and showed a A13CO2 < 2.5% by 13C-urea breath test were considered to be successfully eradicated. Results: The number of cases included in the study was 56 (32 males and 24 females; mean age +- SD: 56.6 -+ 10.6), out of whom 53 patients completed the study. There were no biases in the patients' backgrounds. The eradication rates per protocol patients (PP) were 42.1% (8/19, 95% C1:20.3, 66.5) in Group I, 62.5% (10/16, 95% C1:35.4, 84.8) in Group iI and 77.8% (14118, 95% C1:52.4, 93.6) in Group III (p=0.068: Group Ivs Group III), and those on the intention-to-treat basis (ITT) were 40.0% (8/20, 95% CI:19A, 63.9) in Group I, 55.5% (10/18, 95% C1:30.8, 78.5) in Group II and 77.8% (14118, 95% C1:52.4, 93.6) in Group III (p=0.049: Group I v s Group III). Side effects were observed in one patient in Group I: headache and aiternans stool (1), in three patients in Group II: diarrhea (2) and eruption (1), and in two patients in Group III: eruption (1) and hepatic insufficiency (1). Conclusion: Ecabet sodium dose-dependently improved the eradication rate in the combination therapy with lansoprazole and amoxicillin, and thus proved to be a useful agent for eradication therapy. • G0673
HELICOBACTER PYLORI INDUCES MORE CYTOTOXIC INFLAMMATORY RESPONSES IN THE GASTRIC MUCOSA IN GASTRIC ULCER THAN DUODENAL ULCER VIA INDUCIBLE NITRIC OXIDE SYNTHASE AND POLYMORHPONEUTROPHIL INFILTRATE. IV[. Kaise, N. Suzuki, J. Miwa, K. Iihara, M. Hijikata, Y. Ohta and K. Kanai. Toshiba General Hospital, Tokyo, Japan Background: Helicobacter pylori (11. pylori) induces inflammatory responses in the gastroduodenai mucosa and can elicit peptic ulcer in some individuals. The mechanism by which a part but not all of infected population develops peptic ulcer may be explained by the diversity of H. pylori strain and host susceptibility, but it remains controversial to date. Objective: Since a large amount of nitric oxide generated by inducible nitric oxide sythase (iNOS) acts as a potent cytotoxic molecule in tissue damage, we performed the study to elucidate the role of iNOS in the pathogenesis of peptic ulcer associated with H. pylori infection. Methods: H. pylori-positive patients with healed ulcer (DU, n=13) and gastric ulcer (GU, n=13) undergoing eradication therapy were selected at random. Biopsy samples taken from the gastric antrum and body before and I month after eradication were used for RT-PCR to test iNOS mRNA, Quantification of iNOS mRNA normalized by GAPDH mRNA was achieved by digital
A164 AGA ABSTRACTS • G0669
1-1.PYLORI
EXTRACT INHIBITS APOPTOSIS OF HUMAN NEUTROPHILS. HC Jung, JS Kim, JM Kim*, IS Song, CY Kim. Dept. of Internal Med. and Liver Research Institute, Seoul National Univ. College of Med., Dept. of Microbiology*, Hanyang Univ. College of Med., Seoul, Korea Background/Aims: H. pylori infection in human elicits marked neutrophil infiltration in gastric mucosa. The mechanisms of inflammatory trigger and perpetuation have not been clearly defined. Senescent neutrophils undergo apoptosis and phagocytosed by macrophages. Neutrophil apoptosis is likely to be important in the control of inflammatory tissue injury. To elucidate the perpetuation of gastric mucosal inflammation induced by H. pylori, we investigate regulatory function of soluble products of H. pylori on neutrophil apoptosis. Methods: Neutrophils were isolated from human peripheral blood using discontinuous density gradient centrifugation with Ficoll-Hypaque. The purity of the neutrophils was more than 98% confirmed by morphology and phenotypic analysis using FACS flow cytometry. The viability was more than 95% by trypan blue exclusion test. H. pylori water extract was concentrated from ultrafiltration and filtered through 0.2 lam to minimize the effect of water on apoptosis. The neutrophils were incubated with H. pylori water extract or E. coli lipopolysaccharide (LPS) 10 lag/mL in the culture medium for 6, 12, 24, 36 and 48 hours. Neutrophil apoptosis was determined by agarose gel electrophoresis of DNA, fluorescence microscopic observation after staining with Hoechst 33342 and TUNEL assay. CDllb/CD18 expression of neutrophils was analyzed by FACS flow cytometry. Results: Stimulation with LPS or H. pylori water extract inhibited apoptosis of neutrophils for periods up to 48 hours and enhanced the expression of CD1 lb/CD 18 on the surface of neutrophils. 12h (% apoptosis) 24h(%apoptosis) 48h(%apoptosis) 40±3.8 73±6.5 Control 12 ± 1.5 6±0.4 16±1.2 47±2.6 E.coli LPS 21±1.6 54±3.7 H.pylori extract 7±0.5 Conclusions: These results suggest that neutrophil apoptosis could be suppressed by the soluble factors released by H. pylori. Consequently, the prolongation of survival of neutrophils may coincide with persistent enhanced expression of adhesion molecules on neutrophils. These events could contribute to the perpetuation of H. pylori-induced gastric mucosal inflammation.
G0670 PREVALENCE OF HELICOBACTER PYLORI cagA GENE IN CANCER AND NON-CANCER TISSUES OF GASTRIC CANCER IN KOREA. S.A. Jung,, D.Y. Kim, H.K. Jung, H.C. Lee, S.Y. Yi, I.H. Moon. Division of Molecular Biology, Department of Internal Medicine, Ewha Womans University College of Medicine, Seoul, Korea Aim 1-1.pylori is a risk factor for the development of gastric cancer. In serologic study, cagA-positive H. pylori strain was more related to the development of gatsric cancer than cagA-negative H. pylori strain. To further understand the relationship between cagA gene and gastric cancer, the positive rates of cagA gene in cancer and non-cancer tissues were investigated separately in patients with gastric cancer. Methods The cagA gene was detected by PCR (J Clin Microbiol 1995;33;2752-6) and ureC gene was also assayed as positive controls for the presence of H. priori. Each two endoscopic biopsies were obtained from cancer and non-cancer tissues of 41 patients with gastric cancer. Results 1) The total positive rate of cagA gene in Korean patients with gastric cancer was 63.4%(26/41). 2) The positive rate of cagA gene in cancer tissues was 29.3%(12/41), which was significantly lower thanthat in non-cancer tissues(63.4%, 26141). 3) Twelve (29.3%) out of 41 were positive for cagA gene in both cancer and non-cancer tissues, 14(34.1%) in non-cancer tissues only, none(0%) in cancer tissues only and 15(36.6%) were negative in both sites. 4) The ureC were negative in cancer tissue in 12(85.7%) among 14 cases who were cagA gene negative in cancer tissues but positive in noncancer tissues. 5) There was no difference of positive rates of cagA gene according to age, stage, site and pathologic cell type. Conclusion These findings indicate that positive rate of cagA gene in cancer tissue was lower than that in non-cancer tissues and this might be related to low infection rate of H. pylori rather than the presence of cagA negative H. pylori in cancer tissues.
• G0671 UTILITY OF A RAPID WHOLE BLOOD TEST FOR AN INITIAL DIAGNOSIS OF HELICOBACTER PYLORI INFECTION. I. K~iiiriiiinen, E. Pulkkinen, K. Romanoff, P. Sipponen t & L. Sellin 2. Departments of Gastroenterology and :Pathology, Jorvi Hospital, Espoo, Finland and 2AstraFinland, Masala, Finland. Background. The cost and delay in the diagnosis of H. pylori infection via biopsy, serology or urea breath test have increased interest in low-cost and rapid methods which can be used for screening in a primary care environment. Aim. We have performed a validation study comparing a rapid (10 minutes) and simple whole blood test for H. pylori infection (AccuMeter ®, ChemTrak,
Sunnyvale, California, U.S.A.) to 13C-urea breath test and histology in 104 consecutive patients with abdominal symptoms and referred to endoscopy. Results. Compared to breath test and histology (modified Giemsa), which were highly concordant (99%), the AccuMeter ® was 96% sensitive and 9091% specific. In the one of 27 H. pylori-positive patients where the 13C-urea breath test was negative but H. pylori organisms were demonstrated in the biopsy specimens with histology and immunohistochemistry, the AccuMeter~ result was positive. The AccuMeter ® was slightly less likely to produce a false negative result than a false positive. The calculated predictive values of a negative result to exclude infection are, for example, 96% when the prevalence of infection is 50% and 85% when the prevalence is 20%. Correspondingly, the predictive values for a positive result to indicate infection are 91% and 73%. Conclusions. The AccuMeter* is a rapid, accurate and easyto-use whole blood test for the diagnosis of H. pylori infection and may provide a simple and potentially cost-effective screening tool in the primary care environment. This study was partially funded by Astra-Finland. • G0672 STUDY ON TItE EFFECT OF A NEW ANTI-ULCER AGENT, ECABET SODIUM, ON THE DUAL THERAPY FOR ERADICATION O F HELICOBACTER PYLORI IN RANDOMIZED PATIENTS. H~ Kagay~, M. Kato, T. Kobayashi, J. Ishizuka, K. Nishikawa, Y. Komatsu, M. Sukegawa, H. Takeda, T. Sugiyama and M. Asaka, The Third Department of the Internal Medicine, Hokkaido University School of Medicine, Sapporo, Japan Obiective: Ecabet sodium is a therapeutic agent for gastritis and gastric ulcers, which is locally distributed to the gastric mucosa and remains there for a long time to exert anti-ulcer activity. Ecabet sodium has been demonstrated to have bactericidal action in vitro not only against the standard strain of Helicobacter pylori but also against strains resistant to such drugs as metronidazole and clarithromycin. We investigated the improving effect of ecabet sodium on the eradication rate and its dose-dependency in a combination therapy with lansoprazole and amoxicillin as a method not to acquire resistance. Mcth0d~; Patients with chronic gastritis, who were confirmed to be Helicobacter pylori-positive by the rapid urease test (RUT), were randomly assigned to one of the following three groups and medicated for 2 weeks. Group I: Lansoprazole (30 mg, qd) + Amoxicillin (750 mg, bid); Group II: Lansoprazole (30 rag, qd) + Amoxicillin (750 mg, bid) + Ecabet sodium (1 g, bid); Group III: Lansoprazole (30 mg, qd) + Amoxicillin (750 mg, bid) + Ecabet sodium (2 g, bid). Patients were examined four weeks after the cessation of treatment and those who were Helicobacter pylori-negative by culture test and showed a A13CO2 < 2.5% by 13C-urea breath test were considered to be successfully eradicated. Results: The number of cases included in the study was 56 (32 males and 24 females; mean age +- SD: 56.6 -+ 10.6), out of whom 53 patients completed the study. There were no biases in the patients' backgrounds. The eradication rates per protocol patients (PP) were 42.1% (8/19, 95% C1:20.3, 66.5) in Group I, 62.5% (10/16, 95% C1:35.4, 84.8) in Group iI and 77.8% (14118, 95% C1:52.4, 93.6) in Group III (p=0.068: Group Ivs Group III), and those on the intention-to-treat basis (ITT) were 40.0% (8/20, 95% CI:19A, 63.9) in Group I, 55.5% (10/18, 95% C1:30.8, 78.5) in Group II and 77.8% (14118, 95% C1:52.4, 93.6) in Group III (p=0.049: Group I v s Group III). Side effects were observed in one patient in Group I: headache and aiternans stool (1), in three patients in Group II: diarrhea (2) and eruption (1), and in two patients in Group III: eruption (1) and hepatic insufficiency (1). Conclusion: Ecabet sodium dose-dependently improved the eradication rate in the combination therapy with lansoprazole and amoxicillin, and thus proved to be a useful agent for eradication therapy. • G0673
HELICOBACTER PYLORI INDUCES MORE CYTOTOXIC INFLAMMATORY RESPONSES IN THE GASTRIC MUCOSA IN GASTRIC ULCER THAN DUODENAL ULCER VIA INDUCIBLE NITRIC OXIDE SYNTHASE AND POLYMORHPONEUTROPHIL INFILTRATE. IV[. Kaise, N. Suzuki, J. Miwa, K. Iihara, M. Hijikata, Y. Ohta and K. Kanai. Toshiba General Hospital, Tokyo, Japan Background: Helicobacter pylori (11. pylori) induces inflammatory responses in the gastroduodenai mucosa and can elicit peptic ulcer in some individuals. The mechanism by which a part but not all of infected population develops peptic ulcer may be explained by the diversity of H. pylori strain and host susceptibility, but it remains controversial to date. Objective: Since a large amount of nitric oxide generated by inducible nitric oxide sythase (iNOS) acts as a potent cytotoxic molecule in tissue damage, we performed the study to elucidate the role of iNOS in the pathogenesis of peptic ulcer associated with H. pylori infection. Methods: H. pylori-positive patients with healed ulcer (DU, n=13) and gastric ulcer (GU, n=13) undergoing eradication therapy were selected at random. Biopsy samples taken from the gastric antrum and body before and I month after eradication were used for RT-PCR to test iNOS mRNA, Quantification of iNOS mRNA normalized by GAPDH mRNA was achieved by digital