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Helicobacter pylori infection and acute myocardial infarction in Northern Italy
April 1998
Esophageal, Gastric, attd Duodenal Disorders A253
treated for 2 wks than in pts treated for 1 wk [ITI': 85 out of 101 pts (84%) versus 74 out of 108 pts (68%); ~2: 6.181;p < 0.02] independently from the type of treatment (see table). No differences statically significant were observed when pts were compared by PPI or antibiotic regimens associated. pts n
L1 (%)
n
L2 (%)
n
M1 (%)
17 10
(63) (37)
1 week success failure drop-out
108 74 28 6
19 4 4
(70) (15) (15)
20 5 1
(77) (23)
2 week success failure drop-out
101 85 14 2
19 5 1
(76) (20) (4)
24 2
(92) (8)
19 3 1
(83) (13) (4)
n
M2 (%)
18 9 1
(64) (32) (4)
23 4
(85) (15)
After 6 month 63% of eradicated pts have been controlled. A higher number of HP+ pts were observed in the 1 wk vs 2 wks treated groups (9 out 42 pts vs 5 out 48 pts)) and in MET vs AMO treated groups (9 out of 43 pts vs 5 out 47 pts), but the difference does not reach the statistical significance (p=0.35 and p=0.39, respectively). Conclusions: The eradication rate of this single- blind and monocentric study does not show the high percentage reported in other studies, maybe depending on antibiotic resistence due to previous unsuccessful treatments. Two wks therapies seem to be able to obtain in these cases a more acceptable eradication rate. @ G1040 eDNA TRANSFECTION AND PROTEIN EXPRESSION IN PRIMARY CULTURES OF GASTRIC PARIETAL CELLS, J. A. Parente. Jr., S. Bernatz, X. Chen and C. S. Chew. Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, GA 30912.
Transfection of eDNA into cells is a powerful technique for the analysis of expressed proteins in established cell lines. However, transfection of terminally differentiated, non-dividing cell types such as the gastric parietal cell continues to be highly problematic. The aim of this study was to use a previously characterized, agonist-responsive primary culture model of rabbit parietal cells to establish techniques for transfection and subsequent in situ analyses of living cells. In conjunction with 13-galactosidase, GFP (green fluorescent protein) or EGFP (enhanced GFP) reporter constructs, a variety of transfection reagents were tested including calcium phosphate, DEAE dextran, and several different cationic lipids. Of these, only DMRIE-C (Gibco) and a second, novel cationic lipid, HG-1, allowed for successful transfections (10-20% efficiencies) of several different plasmids while retaining excellent cellular viability as assessed by acid secretory-related responses to either histamine or carbachol. In subsequent studies with the HG1 reagent, eDNA from a novel calcium-responsive phosphoprotein (CSPP28) was inserted in-frame into the EGFP vector then expressed in parietal cells. To define the localization of this CSPP28-EGFP fusion protein in living ceils, serial z-sections of transfected cells were acquired from the stage of a Zeiss Axiovert microscope using a Photometrics high resolution cooled CCD camera driven by Inovision Isee software. Out-of-focus fluorescence was resolved with an image deconvolution algorithm (DeltaVision). Deconvolved images were then reconstructed in 3D using Molecular Dynamics ImageSpace software running on an INDY 02 UNIX computer. Other vital dyes such as Mitotracker (Molecular Probes), which specifically stains mitochondria, were employed to localize different intracellular compartments. In cells expressing the EGFP-CSPP28 fusion protein, a reticular, well-defined pattern of fluorescence was observed in both stimulated and unstimulated ceils. This staining pattern was sub-apical and, as previously observed with fluorescent immunolocalization of CSPP28 using a monoclonal antibody, did not localize to nuclear, mitochondrial or canalicular spaces. In contrast, expression of the EGFP vector alone resulted in a diffuse cytosolic distribution. Conclusions: A reliable method for transfecting terminally differentiated parietal ceils in primary culture has been developed. In addition, we have demonstrated that specific subcellular distributions of expressed proteins in living cells can be analyzed with high resolution in real time. This powerful new technology avoids problems with antibody cross-reactivity and, at least with expressed wildtype CSPP28, does not interfere with agonist-dependent secretory responses. Supported by NIH grants R37 DK31900 and F32 DK09447. • G1041 IS THERE A CORRELATION BETWEEN VACA GENOTYPE AND PEPTIC ULCER DISEASES IN KOREAN PATIENTS? S.M. Park, J.G. Kim, J.W. Park, J.H, Cho, G.K. Yon. Dept of Internal Medicine, Chung-Ang University Hospital, Seoul, Korea Background: An important virulence determinant of H. pylori is the vacnolating cytotoxin which is encoded by the vacA gene. Mosaicism of the vacA gene consists of three different families of vacA signal sequences (sla, slb, s2) and two different families of the middle-region alleles (ml, m2). Studies showed that peptic ulcer disease was closely associated with type sl strains.
Aim: To investigate the prevalence of vacA genotypes from patients with GU, DU vs simple lip gastritis. Methods: Mucosal samples were obtained from 32 patients with simple Hp gastritis and from patients who were endoscopically documented either active GU (53 patients) or DU (46 patients). Presence of vacA signal- and middleregion alleles was determined by RT-PCR amplification of specific regions of the vacA gene. Res~llts: The sl genotype was found in 20 of 32 Hp gastritis (62.5%), in 41 of 53 GU (77.4%) and 38 of 46 DU patients (82.6%). (p < 0.05)
Hp GAS (n = 32)
GU (n = 53)
DU (n = 46)
sl/ml
18 (56.3%)
37 (69.8%)
37 (80.4%)
sl/m2
2 (0.06%)
4 (0.08%)
2 (0.04%)
s2/m2
2 (0.09%)
4 (0.08%)
2 (0.04%)
Conclusion: These data supports that sl vacA genotype is strongly associated with the development of peptic ulcer disease in Koreans. G1042
HELICOBACTER PYLORI INFECTION AND ACUTE MYOCARDIAL INFARCTION IN NORTHERN ITALY. op.p. Parravicini, R. Pellicano, °R. Bigi, *N. Gandolfo, *G, Baduini, V. Arena, ***E. Aruta, G. Pugliese, D. Pellicano, **F. Bazzoli, F. Palmas, M. Rizzetto, A. Ponzetto. Dpt. of Gastroenterology, Molinette Hospital, Torino; *Cardiology, Mauriziano Hospital, Torino; ***Cardiology, S. Giovanni Bosco Hospital, Torino; °Cardiology, Morelli Hospital, Sondalo (So); **Dpt of Gastroenterology, University of Bologna; ITALIA. BACKGROUND: The classical risk factors for acute myocardial infarction (AM1) fail to explain all the epidemiological variations of the disease. New risk factors were recently reported, and among them infectious diseases appear to increase the risk of AM1, particularly acute and chronic respiratory diseases and Helicobacter pylori (H. pylori) infection. AIM: To determine the prevalence of H. pylori infection in a group patients with AMI. MATERIALS AND METHODS: We studied 137 consecutive male patients, aged 40-70 years, admitted for AMI at the Coronary Care Unit in 3 Italian hospitals. H. pylori infection was assessed by the highly specific and sensitive 13C-urea breath test (UBT) and by presence of antibodies against H. pylori in circulation. Volunteer blood donors attending our Hospital's Blood Bank served as control. RESULTS: H. pylori infection was highly prevalent among AMI patients, indeed 118/137 (86%) were shown positive, as compared to 59% (183/310) among the control population. Age distribution is reported in table: Age (years) 40-49 50-59 > 60
Patients with AMI/H. pylori+ 22/24 (92%) 42/51 (82%) 54/62 (87%)
Controls 80/142 (56%) 85/135 (63%) 18/35 (55%)
CONCLUSION: Patients admitted to the Coronary Care Unit for acute myocardial infarction had a notably higher prevalence of H. pylori infection, than the general population. G1043 PROSPECTIVE AUDIT OF INPATIENT PRESCRIBING OF PROTON PUMP INHIBITORS IN A GENERAL HOSPITAL. S.D, Parry, A.W. Harris. Department of Gastroenterology, Kent and Sussex Hospital, Tunbridge Wells, Kent TN4 8AT, UK. Introduction: Proton pump inhibitor (PPI) prescribing accounts for 5.6% of annual NHS drug expenditure and this amount is increasing each year. Aim: The aim of this audit was to determine whether PPIs are prescribed for licensed indications within this busy general hospital or whether an alternative and possibly more cost-effective anti-secretory drug (ASD) could be prescribed. Method: Two week prospective study was carded out without informing the junior or senior physicians. Medical notes, and where available, endoscopy reports, of 43 acute medical patients receiving PPIs were reviewed. Indications for use of PPIs were assessed according to the following criteria: Absolute indications-H, pylori eradication; duodenal or gastric ulcer disease; short term relief of acid related dyspepsia; endoscopic gastro-oesophageal reflux disease (GORD) and NSAID induced damage or Zollinger-Ellinson syndrome: Relative indications-history of ulcer disease on warfarin; symptomatic GORD; prophylaxis against NSAID induced damage; other i.e. in the context of this study a haematemesis and unfit for surgery: No indications-"gastritis", functional dyspepsia, irritable bowel syndrome, stress ulceration. Results: 17 of 43 (40%) patients had absolute indications, 14 (32%) relative indications and 12 (28%) had no indications for the use of PPIs. Conclusions: Only 40% of patients assessed during this audit had definite reasons to take PPIs. The majority of the patients had either no or only relative indications to take PPIs. In such cases an alternative and more cost-effective therapy could have been used. The findings of this audit highlight the problems faced by doctors in ensuring appropriate and costeffective drug therapy for patients with upper gastro-intestinal disorders.
Esophageal, Gastric, attd Duodenal Disorders A253
treated for 2 wks than in pts treated for 1 wk [ITI': 85 out of 101 pts (84%) versus 74 out of 108 pts (68%); ~2: 6.181;p < 0.02] independently from the type of treatment (see table). No differences statically significant were observed when pts were compared by PPI or antibiotic regimens associated. pts n
L1 (%)
n
L2 (%)
n
M1 (%)
17 10
(63) (37)
1 week success failure drop-out
108 74 28 6
19 4 4
(70) (15) (15)
20 5 1
(77) (23)
2 week success failure drop-out
101 85 14 2
19 5 1
(76) (20) (4)
24 2
(92) (8)
19 3 1
(83) (13) (4)
n
M2 (%)
18 9 1
(64) (32) (4)
23 4
(85) (15)
After 6 month 63% of eradicated pts have been controlled. A higher number of HP+ pts were observed in the 1 wk vs 2 wks treated groups (9 out 42 pts vs 5 out 48 pts)) and in MET vs AMO treated groups (9 out of 43 pts vs 5 out 47 pts), but the difference does not reach the statistical significance (p=0.35 and p=0.39, respectively). Conclusions: The eradication rate of this single- blind and monocentric study does not show the high percentage reported in other studies, maybe depending on antibiotic resistence due to previous unsuccessful treatments. Two wks therapies seem to be able to obtain in these cases a more acceptable eradication rate. @ G1040 eDNA TRANSFECTION AND PROTEIN EXPRESSION IN PRIMARY CULTURES OF GASTRIC PARIETAL CELLS, J. A. Parente. Jr., S. Bernatz, X. Chen and C. S. Chew. Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, GA 30912.
Transfection of eDNA into cells is a powerful technique for the analysis of expressed proteins in established cell lines. However, transfection of terminally differentiated, non-dividing cell types such as the gastric parietal cell continues to be highly problematic. The aim of this study was to use a previously characterized, agonist-responsive primary culture model of rabbit parietal cells to establish techniques for transfection and subsequent in situ analyses of living cells. In conjunction with 13-galactosidase, GFP (green fluorescent protein) or EGFP (enhanced GFP) reporter constructs, a variety of transfection reagents were tested including calcium phosphate, DEAE dextran, and several different cationic lipids. Of these, only DMRIE-C (Gibco) and a second, novel cationic lipid, HG-1, allowed for successful transfections (10-20% efficiencies) of several different plasmids while retaining excellent cellular viability as assessed by acid secretory-related responses to either histamine or carbachol. In subsequent studies with the HG1 reagent, eDNA from a novel calcium-responsive phosphoprotein (CSPP28) was inserted in-frame into the EGFP vector then expressed in parietal cells. To define the localization of this CSPP28-EGFP fusion protein in living ceils, serial z-sections of transfected cells were acquired from the stage of a Zeiss Axiovert microscope using a Photometrics high resolution cooled CCD camera driven by Inovision Isee software. Out-of-focus fluorescence was resolved with an image deconvolution algorithm (DeltaVision). Deconvolved images were then reconstructed in 3D using Molecular Dynamics ImageSpace software running on an INDY 02 UNIX computer. Other vital dyes such as Mitotracker (Molecular Probes), which specifically stains mitochondria, were employed to localize different intracellular compartments. In cells expressing the EGFP-CSPP28 fusion protein, a reticular, well-defined pattern of fluorescence was observed in both stimulated and unstimulated ceils. This staining pattern was sub-apical and, as previously observed with fluorescent immunolocalization of CSPP28 using a monoclonal antibody, did not localize to nuclear, mitochondrial or canalicular spaces. In contrast, expression of the EGFP vector alone resulted in a diffuse cytosolic distribution. Conclusions: A reliable method for transfecting terminally differentiated parietal ceils in primary culture has been developed. In addition, we have demonstrated that specific subcellular distributions of expressed proteins in living cells can be analyzed with high resolution in real time. This powerful new technology avoids problems with antibody cross-reactivity and, at least with expressed wildtype CSPP28, does not interfere with agonist-dependent secretory responses. Supported by NIH grants R37 DK31900 and F32 DK09447. • G1041 IS THERE A CORRELATION BETWEEN VACA GENOTYPE AND PEPTIC ULCER DISEASES IN KOREAN PATIENTS? S.M. Park, J.G. Kim, J.W. Park, J.H, Cho, G.K. Yon. Dept of Internal Medicine, Chung-Ang University Hospital, Seoul, Korea Background: An important virulence determinant of H. pylori is the vacnolating cytotoxin which is encoded by the vacA gene. Mosaicism of the vacA gene consists of three different families of vacA signal sequences (sla, slb, s2) and two different families of the middle-region alleles (ml, m2). Studies showed that peptic ulcer disease was closely associated with type sl strains.
Aim: To investigate the prevalence of vacA genotypes from patients with GU, DU vs simple lip gastritis. Methods: Mucosal samples were obtained from 32 patients with simple Hp gastritis and from patients who were endoscopically documented either active GU (53 patients) or DU (46 patients). Presence of vacA signal- and middleregion alleles was determined by RT-PCR amplification of specific regions of the vacA gene. Res~llts: The sl genotype was found in 20 of 32 Hp gastritis (62.5%), in 41 of 53 GU (77.4%) and 38 of 46 DU patients (82.6%). (p < 0.05)
Hp GAS (n = 32)
GU (n = 53)
DU (n = 46)
sl/ml
18 (56.3%)
37 (69.8%)
37 (80.4%)
sl/m2
2 (0.06%)
4 (0.08%)
2 (0.04%)
s2/m2
2 (0.09%)
4 (0.08%)
2 (0.04%)
Conclusion: These data supports that sl vacA genotype is strongly associated with the development of peptic ulcer disease in Koreans. G1042
HELICOBACTER PYLORI INFECTION AND ACUTE MYOCARDIAL INFARCTION IN NORTHERN ITALY. op.p. Parravicini, R. Pellicano, °R. Bigi, *N. Gandolfo, *G, Baduini, V. Arena, ***E. Aruta, G. Pugliese, D. Pellicano, **F. Bazzoli, F. Palmas, M. Rizzetto, A. Ponzetto. Dpt. of Gastroenterology, Molinette Hospital, Torino; *Cardiology, Mauriziano Hospital, Torino; ***Cardiology, S. Giovanni Bosco Hospital, Torino; °Cardiology, Morelli Hospital, Sondalo (So); **Dpt of Gastroenterology, University of Bologna; ITALIA. BACKGROUND: The classical risk factors for acute myocardial infarction (AM1) fail to explain all the epidemiological variations of the disease. New risk factors were recently reported, and among them infectious diseases appear to increase the risk of AM1, particularly acute and chronic respiratory diseases and Helicobacter pylori (H. pylori) infection. AIM: To determine the prevalence of H. pylori infection in a group patients with AMI. MATERIALS AND METHODS: We studied 137 consecutive male patients, aged 40-70 years, admitted for AMI at the Coronary Care Unit in 3 Italian hospitals. H. pylori infection was assessed by the highly specific and sensitive 13C-urea breath test (UBT) and by presence of antibodies against H. pylori in circulation. Volunteer blood donors attending our Hospital's Blood Bank served as control. RESULTS: H. pylori infection was highly prevalent among AMI patients, indeed 118/137 (86%) were shown positive, as compared to 59% (183/310) among the control population. Age distribution is reported in table: Age (years) 40-49 50-59 > 60
Patients with AMI/H. pylori+ 22/24 (92%) 42/51 (82%) 54/62 (87%)
Controls 80/142 (56%) 85/135 (63%) 18/35 (55%)
CONCLUSION: Patients admitted to the Coronary Care Unit for acute myocardial infarction had a notably higher prevalence of H. pylori infection, than the general population. G1043 PROSPECTIVE AUDIT OF INPATIENT PRESCRIBING OF PROTON PUMP INHIBITORS IN A GENERAL HOSPITAL. S.D, Parry, A.W. Harris. Department of Gastroenterology, Kent and Sussex Hospital, Tunbridge Wells, Kent TN4 8AT, UK. Introduction: Proton pump inhibitor (PPI) prescribing accounts for 5.6% of annual NHS drug expenditure and this amount is increasing each year. Aim: The aim of this audit was to determine whether PPIs are prescribed for licensed indications within this busy general hospital or whether an alternative and possibly more cost-effective anti-secretory drug (ASD) could be prescribed. Method: Two week prospective study was carded out without informing the junior or senior physicians. Medical notes, and where available, endoscopy reports, of 43 acute medical patients receiving PPIs were reviewed. Indications for use of PPIs were assessed according to the following criteria: Absolute indications-H, pylori eradication; duodenal or gastric ulcer disease; short term relief of acid related dyspepsia; endoscopic gastro-oesophageal reflux disease (GORD) and NSAID induced damage or Zollinger-Ellinson syndrome: Relative indications-history of ulcer disease on warfarin; symptomatic GORD; prophylaxis against NSAID induced damage; other i.e. in the context of this study a haematemesis and unfit for surgery: No indications-"gastritis", functional dyspepsia, irritable bowel syndrome, stress ulceration. Results: 17 of 43 (40%) patients had absolute indications, 14 (32%) relative indications and 12 (28%) had no indications for the use of PPIs. Conclusions: Only 40% of patients assessed during this audit had definite reasons to take PPIs. The majority of the patients had either no or only relative indications to take PPIs. In such cases an alternative and more cost-effective therapy could have been used. The findings of this audit highlight the problems faced by doctors in ensuring appropriate and costeffective drug therapy for patients with upper gastro-intestinal disorders.