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Efficacy and safety of lansoprazole 30 mg versus omeprazole for 21 days treatment of acute esophagitis
A200
AGA ABSTRACTS
Q R T - P C R C L O N I N G O F RAB3 I S O F O R M S I N D I S P E R S E D C H I E F C E L I ~ F R O M G U I N E A P I G STOMACH. R.D. Raffaniello, J. Lin, F. Wang, J-P. Raufman. Gastrointestinal Cell Biology Laboratory, Dept. of Medicine, SUNY-Health Science Center at Brooklyn, NY. Rab3 proteins are low molecular weight (LMW) (20-30 kD) GriP_ b i n d i n g proteins t h a t are expressed in neurons and other secretory cells; are localized to secretory vesicles; and, may play a role in regulating exocytosis. Presently, four highly homologous tab3 isofonns (A, B, C, D) have been identified. A role for LMW GTP-binding proteins i n the r e g u l a t i o n of pepsinogen secretion has been suggested by the observation t h a t GTPTS s t i m u l a t e s secretion independent of changes in calcium, cAMP or activation of protein klnase (JBC 268:8491, 1993). In the present study, we examined the expression of rab3 isoforms in dispersed chief cells from g u i n e a pig stomach. Chief cells were prepared from g u i n e a pig gastric mucosa by co]]agonase digestion, calcium chelation and fractionation on a Percol] density gradient. E x p r e s s i o n of rab3 proteins in chief cells was e x a m i n e d u s i n g a specific monoc]ona] antibody. A 27-kD p r o t e i n was i m m u u o p r e c i p i t a t e d from chief cell ]ysates w i t h this antibody. The rab3 antibody was also used for immunohistochemicai analysis of dispersed gastric mucosal cells. Consistent with observations in other secretory cells, rab3 was localized to the Secretory g r a n u l e s of chief cells. I m m u n o s t a i n i n g was not detected in parietal or mucous cells. To d e t e r m i n e w h i c h rab3 isoforms are expressed i n chief cells, we extracted total cellular RATA and employed reverse transcription (RT)PCR. cDNA obtained by RT was subjected to PCR u s i n g specific primers for t a b s isoforms and the resulting products (approx. 200 bp) were cloned and sequenced. The nucleotide and deduced amino acid sequences obtained were 90 and 100% homologous, respectively, to published m u r i n e rab3D sequences. We used the cloned PCR rabSD f r a g m e n t as a probe to identify rab3 transcripts. N o r t h e r n blot analysis u s i n g this probe resulted in the detection of an approx. 4-kb transcript in chief cells, but not in brain. These findings are consistent w i t h the size of m u r i n e rah3D t r a n s c r i p t s and w i t h the prior observation t h a t rabSD is poorly expressed in brain. These results indicate t h a t a rab3 isoform is associated with the secretory granules of gastric chief cells. Moreover, RT-PCR cloning revealed t h a t rab3D is the major rab3 isoform expressed in gastric chief cells.
Q RESTRICTION ENZYME SYSTEMS OF HELICOBACTER PYLORI. J.Ramakrishna~ K. Mathee, A.G. Plant, A. Wright. Departments of Pediatrics (Gastroenterology and Nutrition) and Medicine, New England Medical Center, and Department of Microbiology, Tufts University School of Medicine, Boston MA 02111. To date, the analysis of H. pylori using methods such as RFLPs, PCR and pulsed field gel electrophoresis, has revealed considerable genetic diversity from strain to strain. Due to this diversity, there are as yet no methods for classifying this organism. Also there are considerable differences in the transformability of different clinical isolates, some being easily transformable while others are not. It has therefore been postulated that the organism may have restrictionmodification systems which differ among isolates. Restriction enzymes are produced by bacteria that cleave DNA at specific recognition sequences; these bacteria therefore have a modification system to methylate their own DNA at the recognition sites to protect it from cleavage. No restriction enzymes have been described in H. pylori so far. Since restriction-modification systems are highly conserved in a given strain of a pathogenic organism, this can be a reliable method for classifying clinical isolates. We cultured eleven H. pylori isolates on Campylobacter agar Skirrow(Difco) plates supplemented with 10% defibrinated sheep blood(Remel) in microaerobic chambers. Cells were resuspended in a suitable buffer and sonieated. Protein extracts were prepared by precipitation with 60% ammonium sulfate and subsequent dialysis, and these were used for restriction digests. Preliminary digests were done using a standard DNA template, the plasmid pBR322, which has a known restriction map and DNA sequence. The patterns of these digests suggested the existence of restriction enzymes. The site specificity of the restriction enzymes was elucidated using pBR322 that was linearized and labelled at one end with 32p. Partial digests of the 32p labelled DNA with Hp extracts were fractionated by electrophoresis on polyacrylamideand agarose gels alongside appropriate radiolabelled markers. By determining the sizes of the fragments of DNA generated we were able to determine the restriction sites on pBR322. The various clinical isolates studied by us so far all have distinct restriction patterns. Two have so far been identified. The enzyme present in Hp 32 has a specificity corresponding to Sau 96I while that in Hp 64 corresponds to Ddel. Work is currently under way in our laboratory to identify the restriction enzymes in other clinical isolates. We anticipate that this will yield a pattern that will form the basis for a classification system for H. pylori strains.
GASTROENTEROLOGY, Vol. 108, No. 4
RELATIONSHIP OF THE MANOMETRIC "DOUBLE HUMP" TO HIATAL HERNIA & REFLUX IN GERD PATIENTS. F Raiser. N Katada, RA Hinder, CJ Filipi, R Stalzer, PJ McBride, U Arnelo, S Enguidanos. Department of Surgery, Creighton University School of Medicine, Omaha NE Patients with hiatal hernia (HH) often have a characteristic manometric "double hump" (DH) at the LES. The pressure measured at the LES is a Composite of intrinsic muscle tone, ambient abdominal pressure, and pressure exerted by the crural hiatus on the esophagus. Displacement of the muscular high pressure zone (HPZ) into the chest can create insufficiency of the LES since negative thoracic pressure lowers effective resting pressure. Thf~ pressure effect of the crural hiatus also no longer overlaps the muscular HPZ. The manometric "double hump" consists of crural pressure deflections distal to a muscular HPZ, with a return to near baseline pressure in between. We studied the relationship of these manometric findings to anatomy and antireflux function in 68 consecutive patients being evaluated for, GERD using manometry, 24hr pH analysis, contrast radiography, and EGD. A DH was seen in 17 of 46 patients with HH on contrast radiography or EGD, but in only 1 of 22 patients with no HH. Sensitivity was 37%, specificity 95%. The low sensitivity is related to the inability of manometry to resolve the crural and muscular high pressure zones of small hiatal hernias into two distinct humps. Among patients with hiatal hernias larger than 5 cm, 14 of 15 patients (93%) had a DH. Mean LES length was 6.6+0.3cm for DH patients, longer than for patients with no DH (4.3+0.2cm,p
EFFICACY AND SAFETY OF L A N S O P ~ 30 M G V~RSUS ~ FOR 21 DAYS T R E A T ~ T ~ OF ACUTE ESOPH~EITIS. P. RAMPAL 111,A" CCt~RIER( 21 ,M. L~a~REZ 131, and multicenter group. (1>Service de Gastroent~rologie, HSpltal de l'Archet, 06012 NICE C~DEX; 21Service de Gastrcent~rologie, H~pital Ban-Secours, 57000 METZ, FRANCE; C31Service de Gastroent~xolcgie, Centre Hespitalier G~I1~ral, 77305 F O N T ~ L E A U , FRANCE. Proton Immp inhibitors are very effective treaumenu of acute esophagitis. The aim of the study was ~o ccmpare cmeprazele 20m~ versus lansoprazole 30rag as short term treatment for acute esophagitis. : 97 patients with a peptic esophagitis grade IB, II, III, IV (Savary Miller) were randcmized in a double-blind controlled study comparing cmeprazole 20~g oad (Ome 20) and larlsoprazole 30rag oad (Lanso 30) . ~%doscopy was performed at DO
and D21 x 1. 75 patients bad their endoscopy registred and hereafter projected to an /ndependant grotlo of e~perts who did not know the creat[~/ic neither the d a t e of endosccpy.Esophagitis was considered as healed in case of grade 0 (I= analysis) or in case of grade 0 or IA Savary Miller (2~ analysis) . Safety was assessed by e~-aluaticn of clinical and biological adverse events Results : i s~ analysis = endoscopists assessment of esophagitis healing races (grade 0) was respectively 69% (Ome 20) and 78% (Lanso 30) = NS. Experts assessment was respectively 56.5% (Ome 20)and 86% CLanso 30) = p < 0,005. 2~ analysis .= endoscopists assessment of grade 0-IA esophagitis was respectively 79.5% (Ome 20) and 94.5% (Lanso 30) = NS. Experts assessment was 77.5% (Ome 20) and 91.5% (Lanso 30) = NS. Two benign adverse events were cc~]sidered related =o treatment b y investigators : c~le in lansoprazole 30rag group (diarrhea) and one in craeprazole 20rag grotto (plasma gastrin level above twofold upper normal value ) . Ccmclusicn : i) Lansoprazole 30n~3 and cmeprazole 20m~ are both safe treatment for acute esophagitis. 2} W1hatever analysis, lansoprazole 30m~ showed superior efficacy to omeprazole 20rag . This superiority reached statistical significance if the analySlS was restricted co ccr~plete healing and diagnosed b y independant exper~ Ccmnittee
AGA ABSTRACTS
Q R T - P C R C L O N I N G O F RAB3 I S O F O R M S I N D I S P E R S E D C H I E F C E L I ~ F R O M G U I N E A P I G STOMACH. R.D. Raffaniello, J. Lin, F. Wang, J-P. Raufman. Gastrointestinal Cell Biology Laboratory, Dept. of Medicine, SUNY-Health Science Center at Brooklyn, NY. Rab3 proteins are low molecular weight (LMW) (20-30 kD) GriP_ b i n d i n g proteins t h a t are expressed in neurons and other secretory cells; are localized to secretory vesicles; and, may play a role in regulating exocytosis. Presently, four highly homologous tab3 isofonns (A, B, C, D) have been identified. A role for LMW GTP-binding proteins i n the r e g u l a t i o n of pepsinogen secretion has been suggested by the observation t h a t GTPTS s t i m u l a t e s secretion independent of changes in calcium, cAMP or activation of protein klnase (JBC 268:8491, 1993). In the present study, we examined the expression of rab3 isoforms in dispersed chief cells from g u i n e a pig stomach. Chief cells were prepared from g u i n e a pig gastric mucosa by co]]agonase digestion, calcium chelation and fractionation on a Percol] density gradient. E x p r e s s i o n of rab3 proteins in chief cells was e x a m i n e d u s i n g a specific monoc]ona] antibody. A 27-kD p r o t e i n was i m m u u o p r e c i p i t a t e d from chief cell ]ysates w i t h this antibody. The rab3 antibody was also used for immunohistochemicai analysis of dispersed gastric mucosal cells. Consistent with observations in other secretory cells, rab3 was localized to the Secretory g r a n u l e s of chief cells. I m m u n o s t a i n i n g was not detected in parietal or mucous cells. To d e t e r m i n e w h i c h rab3 isoforms are expressed i n chief cells, we extracted total cellular RATA and employed reverse transcription (RT)PCR. cDNA obtained by RT was subjected to PCR u s i n g specific primers for t a b s isoforms and the resulting products (approx. 200 bp) were cloned and sequenced. The nucleotide and deduced amino acid sequences obtained were 90 and 100% homologous, respectively, to published m u r i n e rab3D sequences. We used the cloned PCR rabSD f r a g m e n t as a probe to identify rab3 transcripts. N o r t h e r n blot analysis u s i n g this probe resulted in the detection of an approx. 4-kb transcript in chief cells, but not in brain. These findings are consistent w i t h the size of m u r i n e rah3D t r a n s c r i p t s and w i t h the prior observation t h a t rabSD is poorly expressed in brain. These results indicate t h a t a rab3 isoform is associated with the secretory granules of gastric chief cells. Moreover, RT-PCR cloning revealed t h a t rab3D is the major rab3 isoform expressed in gastric chief cells.
Q RESTRICTION ENZYME SYSTEMS OF HELICOBACTER PYLORI. J.Ramakrishna~ K. Mathee, A.G. Plant, A. Wright. Departments of Pediatrics (Gastroenterology and Nutrition) and Medicine, New England Medical Center, and Department of Microbiology, Tufts University School of Medicine, Boston MA 02111. To date, the analysis of H. pylori using methods such as RFLPs, PCR and pulsed field gel electrophoresis, has revealed considerable genetic diversity from strain to strain. Due to this diversity, there are as yet no methods for classifying this organism. Also there are considerable differences in the transformability of different clinical isolates, some being easily transformable while others are not. It has therefore been postulated that the organism may have restrictionmodification systems which differ among isolates. Restriction enzymes are produced by bacteria that cleave DNA at specific recognition sequences; these bacteria therefore have a modification system to methylate their own DNA at the recognition sites to protect it from cleavage. No restriction enzymes have been described in H. pylori so far. Since restriction-modification systems are highly conserved in a given strain of a pathogenic organism, this can be a reliable method for classifying clinical isolates. We cultured eleven H. pylori isolates on Campylobacter agar Skirrow(Difco) plates supplemented with 10% defibrinated sheep blood(Remel) in microaerobic chambers. Cells were resuspended in a suitable buffer and sonieated. Protein extracts were prepared by precipitation with 60% ammonium sulfate and subsequent dialysis, and these were used for restriction digests. Preliminary digests were done using a standard DNA template, the plasmid pBR322, which has a known restriction map and DNA sequence. The patterns of these digests suggested the existence of restriction enzymes. The site specificity of the restriction enzymes was elucidated using pBR322 that was linearized and labelled at one end with 32p. Partial digests of the 32p labelled DNA with Hp extracts were fractionated by electrophoresis on polyacrylamideand agarose gels alongside appropriate radiolabelled markers. By determining the sizes of the fragments of DNA generated we were able to determine the restriction sites on pBR322. The various clinical isolates studied by us so far all have distinct restriction patterns. Two have so far been identified. The enzyme present in Hp 32 has a specificity corresponding to Sau 96I while that in Hp 64 corresponds to Ddel. Work is currently under way in our laboratory to identify the restriction enzymes in other clinical isolates. We anticipate that this will yield a pattern that will form the basis for a classification system for H. pylori strains.
GASTROENTEROLOGY, Vol. 108, No. 4
RELATIONSHIP OF THE MANOMETRIC "DOUBLE HUMP" TO HIATAL HERNIA & REFLUX IN GERD PATIENTS. F Raiser. N Katada, RA Hinder, CJ Filipi, R Stalzer, PJ McBride, U Arnelo, S Enguidanos. Department of Surgery, Creighton University School of Medicine, Omaha NE Patients with hiatal hernia (HH) often have a characteristic manometric "double hump" (DH) at the LES. The pressure measured at the LES is a Composite of intrinsic muscle tone, ambient abdominal pressure, and pressure exerted by the crural hiatus on the esophagus. Displacement of the muscular high pressure zone (HPZ) into the chest can create insufficiency of the LES since negative thoracic pressure lowers effective resting pressure. Thf~ pressure effect of the crural hiatus also no longer overlaps the muscular HPZ. The manometric "double hump" consists of crural pressure deflections distal to a muscular HPZ, with a return to near baseline pressure in between. We studied the relationship of these manometric findings to anatomy and antireflux function in 68 consecutive patients being evaluated for, GERD using manometry, 24hr pH analysis, contrast radiography, and EGD. A DH was seen in 17 of 46 patients with HH on contrast radiography or EGD, but in only 1 of 22 patients with no HH. Sensitivity was 37%, specificity 95%. The low sensitivity is related to the inability of manometry to resolve the crural and muscular high pressure zones of small hiatal hernias into two distinct humps. Among patients with hiatal hernias larger than 5 cm, 14 of 15 patients (93%) had a DH. Mean LES length was 6.6+0.3cm for DH patients, longer than for patients with no DH (4.3+0.2cm,p
EFFICACY AND SAFETY OF L A N S O P ~ 30 M G V~RSUS ~ FOR 21 DAYS T R E A T ~ T ~ OF ACUTE ESOPH~EITIS. P. RAMPAL 111,A" CCt~RIER( 21 ,M. L~a~REZ 131, and multicenter group. (1>Service de Gastroent~rologie, HSpltal de l'Archet, 06012 NICE C~DEX; 21Service de Gastrcent~rologie, H~pital Ban-Secours, 57000 METZ, FRANCE; C31Service de Gastroent~xolcgie, Centre Hespitalier G~I1~ral, 77305 F O N T ~ L E A U , FRANCE. Proton Immp inhibitors are very effective treaumenu of acute esophagitis. The aim of the study was ~o ccmpare cmeprazele 20m~ versus lansoprazole 30rag as short term treatment for acute esophagitis. : 97 patients with a peptic esophagitis grade IB, II, III, IV (Savary Miller) were randcmized in a double-blind controlled study comparing cmeprazole 20~g oad (Ome 20) and larlsoprazole 30rag oad (Lanso 30) . ~%doscopy was performed at DO
and D21 x 1. 75 patients bad their endoscopy registred and hereafter projected to an /ndependant grotlo of e~perts who did not know the creat[~/ic neither the d a t e of endosccpy.Esophagitis was considered as healed in case of grade 0 (I= analysis) or in case of grade 0 or IA Savary Miller (2~ analysis) . Safety was assessed by e~-aluaticn of clinical and biological adverse events Results : i s~ analysis = endoscopists assessment of esophagitis healing races (grade 0) was respectively 69% (Ome 20) and 78% (Lanso 30) = NS. Experts assessment was respectively 56.5% (Ome 20)and 86% CLanso 30) = p < 0,005. 2~ analysis .= endoscopists assessment of grade 0-IA esophagitis was respectively 79.5% (Ome 20) and 94.5% (Lanso 30) = NS. Experts assessment was 77.5% (Ome 20) and 91.5% (Lanso 30) = NS. Two benign adverse events were cc~]sidered related =o treatment b y investigators : c~le in lansoprazole 30rag group (diarrhea) and one in craeprazole 20rag grotto (plasma gastrin level above twofold upper normal value ) . Ccmclusicn : i) Lansoprazole 30n~3 and cmeprazole 20m~ are both safe treatment for acute esophagitis. 2} W1hatever analysis, lansoprazole 30m~ showed superior efficacy to omeprazole 20rag . This superiority reached statistical significance if the analySlS was restricted co ccr~plete healing and diagnosed b y independant exper~ Ccmnittee