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Hepatobiliary helicobacter SPP. identified from patients with primary sclerosing cholangitis
A978 AGA ABSTRACTS
GASTROENTEROLOGY Vol. 114, No. 4 G4008
• G4006
VALIDATION OF A DISEASE SPECIFIC HEALTH-RELATED QUALITY OF LIFE (HRQOL) INSTRUMENT FOR PEDIATRIC INFLAMMATORY BOWEL DISEASE (IBD). S Forget ], R Cawdron 2, RM lssenman2, N Gold 3, EJ Irvine4. Division of Pediatric Gastroenterology 1, Montreal Children's Hospital, Mc Gill University, Montreal, and Division of Pediatric Gastroenterology 2, School of Social Work3 and Division of Gastroenterology 4, Mc Master University Medical Center, Hamilton, Canada. Background HRQOL has become an essential component of outcome assessment in health research. Disease-specific instruments need to be validated prior to their use in clinical trials. There is no validated HRQOL instrument available for children and adolescents with IBD. Aim To assess the validity and reliability of a disease-specific HRQOL instrument for pediatric patients with IBD. Methods Forty-five items describing the physical, emotional and social impact of 1BD were included in the Pediatric Inflammatory Bowel Disease Questionnaire (PEDIBDQ©), according to previously reported methods, and administered to 50 pediatric IBD patients. Total HRQOL scores (range 45-315, higher scores = better HRQOL) and subscale scores (physical, emotional, social) were correlated with other measures hypothesized to tap into HRQOL, including disease activity index (DA), the Child Depression Inventory (CDI), the Personal Adjustment and Role Skills Scale III (PARS), Harter's self-perception profile, and global assessment of disease. Reliability was assessed using Cronbach's ~t. HRQOL scores were compared between different clinical subgroups using ANOVA. Results Forty-nine patients had complete data for analysis (33 Crohn's Disease (CD) / 11 Ulcerative Colitis (UC)/5 Indeterminate colitis). Mean age was 13.8 years, 28 were male, nine had undergone surgery and two had a stoma. Correlation coefficients between total HRQOL scores, sub-scale scores and other indices, shown below, were all consistent with a-priori predictions: HRQOL
DA
CDI
Harter's
PARS
Global Assess
Total
-0.4 -0.5 -0.3 -0.18
-0.65 -0.55 -0.60 -0.46
0.35 0.29 0.34 0.24
0.30 0.28 0.21 0.37
0.81 0.80 0.80 0.33
Physical Emotional Social
The reliability coefficient (a) for the whole questionnaire was 0.89, indicating a high degree of consistency of answers across items. Age, gender, diagnosis (CD vs UC), past surgery or disease duration did not significantly impact on HRQOL scores. Conclusion The PEDIBDQ is a highly valid and reliable disease-specific HRQOL instrument for pediatric patients with IBD. Supported by the Crohn's and Colitis Foundation of Canada. G4007 CROHN'S ARTHROPATHY AND NSAIDS: ASSOCIATIONS AND THERAPEUTIC DIFFICULTIES. E H F0rrest, E Graham, R I Russell. Department of Gastroenterology, Royal Infirmary, Glasgow, Scotland, UK.
Crohn's arthropathy causes significant morbidity, w e assessed this problem by retrospectively reviewing 105 patients with Crohn's disease attending between 1991 and 1996. Arthropathy was identified by volunteered complaints of joint discomfort on greater than 2 occasions over a period of more than 3 months. RESULTS. In total 66.7% patients were female, and the mean age at diagnosis was 33.6 - 1.5 [SEM] years. Colonic disease was present in 83.8% patients, with 31.4% having this as the only identified site. Perianal disease was noted in 22.8%. Joint symptoms were present in 55 (52.3%) patients with 4 (3.8%) patients having diagnosed sacroiliitis or anklylosing spondylitis, and one patient having sero-p0sitive rheumatoid arthritis. The 50 (47.6%) patients with peripheral arthropathy were older at the time of diagnosis (37.6 ± 2.4 v 29.2 ± 2.0years; p<0.01) than those without any joint discomfort. Patients with peripheral arthropathy tended to be female (78% v58%; p<0.02), had a lower incidence of perianal disease (14% v 46%; p<0.001), but had a higher incidence of mucocutaneous extra - intestinal manifestations of Crohn's disease (34% v 6%; p<0.001). In total 37 patients had taken NSAIDs. Of these, 9 (24.3%) had an exacerbation of their Crohn's symptoms associated with NSAID use. Such events were more common in patients diagnosed at a younger age (28.6 ± 2.5 v 40.4 ± 3.1 years; p<0.01), and in non smokers (44.4% v 67.8%; p<0.05). Dyspeptic symptoms also occurred in 9 (24.3%) patients, only one of whom had an exacerbation of Crohn's symptoms. In total 17 (45.9%) patients had adverse gastro-intestinal side-effects from NSAID usage. CONCLUSIONS. Crohn's peripheral arthropathy may be more common than previously believed, and occurs amongst slightly older patients in association with mucocutaneous extra-intestinal manifestations. NSAIDs are associated with a high incidence of gastro-intestinal adverse effects, particularly exacerbations of Crohn's symptoms.
ANTI-ENDOTHELIAL CELL ANTIBODIES IN CROHN'S DISEASE. F.G. Foschi, G. Zoli. E. Castelli, G. Addolorato °, M. Parazza, S. Pagani, E. Loggi, G. Gasbarrini °, G.F. Stefanini* and M. Bernardi. Dipartimento di Medicina Interna Cardioangiologia Epatologia, Universit~t di Bologna. °Cattedra di Medicina Interna, Universit~ Cattolica del Sacro Cuore, Roma. *Divisione di Medicina Interna, Ospedale di Faenza (Ravenna). Background: The etiopathogenesis of Crohn's disease (CD) is still unknown, however it has been suggested that vasculitis play an important role. Antiendothelial antibodies (AECA) have been reported in sera of patients with various kind of vasculitis and other autoimmune diseases and the correlation between AECA and vasculitis damage has been recently demonstrated. Aim: to evaluate the presence of AECA in CD and their role in disease activity. Patients and Methods: Serum AECA have been assayed in 40 CD patients, 22 male and 18 female (mean age 31.9±11.8 yrs) and in 64 healthy volunteers. In patient group, disease activity was evaluated by Simple Crohn Activity Index (SCDA1). AECA were evaluated by means of ELISA, using cell cultures from human umbilical vein endothelial cells. Results were expressed as Binding Index (BI%), and the cut-off value of global test (IgA, IgG, IgM) is 35,8% (mean of healthy volunteers + 2SD). Chi-square and Spearman's tests provided statistical analysis. Results: AECA (IgA, IgG, IgM) were found in 12 patients (30%) (Bl%= 32,9±38,31) and only 3 controls (4,6%) (Bl%= 13.42±11.7) (P<0.005). Among patients AECA positive, 11 showed IgG-AECA and 7 IgM-AECA, while no patients showed IgA-AECA. A significant correlation (p<0.005) was found between AECA and SCDAI. No correlation was found between AECA presence and localization of the disease. Conclusion: Our data confirm the presence of AECA in a high percentage of CD patients and its correlation with disease activity (as far as other autoimmune diseases). It is possible that the AECA bound to endothelial cells cause a vasculitic damage promoting a series of cellular modifications considered essential for lymphocyte activation and for induction or maintenance of the inflammatory process. On the other hand, may be that inflammation induce cytokine release (i.e. Tumor Necrosis Factor ct) in CD. The cytokine could promote the neoexpression of antigens target by AECA. G4009
HEPATOBILIARY HELICOBACTER SPP. IDENTIFIED FROM PATIENTS WITH PRIMARY SCLEROSING CHOLANGITIS. J.G. Fox*, Z. Shen*, F. Feng*, J. Dufover +, M.M. Kaplan +, F. Dewhirst #. *Massachusetts Institute of Technology, Cambridge, MA 02139. #Forsyth Dental Center & +Tufts University, Boston, MA. Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease of unknown etiology. Pathologic lesions consist of persistent inflammation, destruction and fibrosis of intrahepatic and extrahepatic bile ducts. The high correlation of PSC and ulcerative colitis has raised the hypothesis that chronic portal bacteremia may initiate inflammation and promote subsequent hepatobiliary damage. The objective of this study was to ascertain whether Helicobacter spp. known to cause hepatobiliary disease in animals was present in PSC patients. Liver biopsies and bile were obtained from eight patients with PSC. Trypticase soy agar with 5% sheep blood, TVP and CVA medium were used for Helicobacter spp. isolation. A Rapid Prep genomic DNA kit was used to extract DNA from liver and Gene Releaser was used for bile DNA extraction. The primers chosen for PCR amplification recognized conserved regions of the 16S rRNA specific for all known Helicobacter spp. Oligonucleotide primers produced an amplified products of 1220 bp. For confirmation of the PCR-amplified fragment by Southern Blot hybridization, the PCR product was electrophoresed through a 1% agarose gel, transferred onto a nylon membrane, and subjected to hybridization with Helicobacter specific PCR-generated probe. PCRTM II vector was used for cloning of PCR products which were purified by Geneclean kit. ligated with 50 ng PCR TM II vector at 15C overnight, and transferred into INVaF' ceils. Ampicillin plates with X-gal were used to select positive clones. Plasmid DNA was isolated for E. coli by Qiagen plasmid prep kit. Helicobacter spp. were not cultured. However, Helicobacter spp. were identified by PCR amplification and southern hybridization using Helicobacter specific probe in 5 of 8 patients. In 2 of these patients a 1200 bp PCR amplified product was successfully cloned and sequenced. Analysis of the sequence indicated high homology to the 16S rRNA sequence of a cluster of Helicobacter spp. previously isolated from animals, i.e. tl. rodentium. H. rappini, and H. pullorum. Identification of Helicobacter spp. in liver and bile of patients with PSC is consistent with our recent findings of Helicobacter spp. associated hepatic disease in animals. Previous isolation of H. rappini and 1-1. pullorum in the feces of diarrheic humans is also supportive evidence. These results should stimulate studies to ascertain whether these or similar helicobacters play an important role in the pathogenesis of idiopathic hepatobiliary disease in humans.
GASTROENTEROLOGY Vol. 114, No. 4 G4008
• G4006
VALIDATION OF A DISEASE SPECIFIC HEALTH-RELATED QUALITY OF LIFE (HRQOL) INSTRUMENT FOR PEDIATRIC INFLAMMATORY BOWEL DISEASE (IBD). S Forget ], R Cawdron 2, RM lssenman2, N Gold 3, EJ Irvine4. Division of Pediatric Gastroenterology 1, Montreal Children's Hospital, Mc Gill University, Montreal, and Division of Pediatric Gastroenterology 2, School of Social Work3 and Division of Gastroenterology 4, Mc Master University Medical Center, Hamilton, Canada. Background HRQOL has become an essential component of outcome assessment in health research. Disease-specific instruments need to be validated prior to their use in clinical trials. There is no validated HRQOL instrument available for children and adolescents with IBD. Aim To assess the validity and reliability of a disease-specific HRQOL instrument for pediatric patients with IBD. Methods Forty-five items describing the physical, emotional and social impact of 1BD were included in the Pediatric Inflammatory Bowel Disease Questionnaire (PEDIBDQ©), according to previously reported methods, and administered to 50 pediatric IBD patients. Total HRQOL scores (range 45-315, higher scores = better HRQOL) and subscale scores (physical, emotional, social) were correlated with other measures hypothesized to tap into HRQOL, including disease activity index (DA), the Child Depression Inventory (CDI), the Personal Adjustment and Role Skills Scale III (PARS), Harter's self-perception profile, and global assessment of disease. Reliability was assessed using Cronbach's ~t. HRQOL scores were compared between different clinical subgroups using ANOVA. Results Forty-nine patients had complete data for analysis (33 Crohn's Disease (CD) / 11 Ulcerative Colitis (UC)/5 Indeterminate colitis). Mean age was 13.8 years, 28 were male, nine had undergone surgery and two had a stoma. Correlation coefficients between total HRQOL scores, sub-scale scores and other indices, shown below, were all consistent with a-priori predictions: HRQOL
DA
CDI
Harter's
PARS
Global Assess
Total
-0.4 -0.5 -0.3 -0.18
-0.65 -0.55 -0.60 -0.46
0.35 0.29 0.34 0.24
0.30 0.28 0.21 0.37
0.81 0.80 0.80 0.33
Physical Emotional Social
The reliability coefficient (a) for the whole questionnaire was 0.89, indicating a high degree of consistency of answers across items. Age, gender, diagnosis (CD vs UC), past surgery or disease duration did not significantly impact on HRQOL scores. Conclusion The PEDIBDQ is a highly valid and reliable disease-specific HRQOL instrument for pediatric patients with IBD. Supported by the Crohn's and Colitis Foundation of Canada. G4007 CROHN'S ARTHROPATHY AND NSAIDS: ASSOCIATIONS AND THERAPEUTIC DIFFICULTIES. E H F0rrest, E Graham, R I Russell. Department of Gastroenterology, Royal Infirmary, Glasgow, Scotland, UK.
Crohn's arthropathy causes significant morbidity, w e assessed this problem by retrospectively reviewing 105 patients with Crohn's disease attending between 1991 and 1996. Arthropathy was identified by volunteered complaints of joint discomfort on greater than 2 occasions over a period of more than 3 months. RESULTS. In total 66.7% patients were female, and the mean age at diagnosis was 33.6 - 1.5 [SEM] years. Colonic disease was present in 83.8% patients, with 31.4% having this as the only identified site. Perianal disease was noted in 22.8%. Joint symptoms were present in 55 (52.3%) patients with 4 (3.8%) patients having diagnosed sacroiliitis or anklylosing spondylitis, and one patient having sero-p0sitive rheumatoid arthritis. The 50 (47.6%) patients with peripheral arthropathy were older at the time of diagnosis (37.6 ± 2.4 v 29.2 ± 2.0years; p<0.01) than those without any joint discomfort. Patients with peripheral arthropathy tended to be female (78% v58%; p<0.02), had a lower incidence of perianal disease (14% v 46%; p<0.001), but had a higher incidence of mucocutaneous extra - intestinal manifestations of Crohn's disease (34% v 6%; p<0.001). In total 37 patients had taken NSAIDs. Of these, 9 (24.3%) had an exacerbation of their Crohn's symptoms associated with NSAID use. Such events were more common in patients diagnosed at a younger age (28.6 ± 2.5 v 40.4 ± 3.1 years; p<0.01), and in non smokers (44.4% v 67.8%; p<0.05). Dyspeptic symptoms also occurred in 9 (24.3%) patients, only one of whom had an exacerbation of Crohn's symptoms. In total 17 (45.9%) patients had adverse gastro-intestinal side-effects from NSAID usage. CONCLUSIONS. Crohn's peripheral arthropathy may be more common than previously believed, and occurs amongst slightly older patients in association with mucocutaneous extra-intestinal manifestations. NSAIDs are associated with a high incidence of gastro-intestinal adverse effects, particularly exacerbations of Crohn's symptoms.
ANTI-ENDOTHELIAL CELL ANTIBODIES IN CROHN'S DISEASE. F.G. Foschi, G. Zoli. E. Castelli, G. Addolorato °, M. Parazza, S. Pagani, E. Loggi, G. Gasbarrini °, G.F. Stefanini* and M. Bernardi. Dipartimento di Medicina Interna Cardioangiologia Epatologia, Universit~t di Bologna. °Cattedra di Medicina Interna, Universit~ Cattolica del Sacro Cuore, Roma. *Divisione di Medicina Interna, Ospedale di Faenza (Ravenna). Background: The etiopathogenesis of Crohn's disease (CD) is still unknown, however it has been suggested that vasculitis play an important role. Antiendothelial antibodies (AECA) have been reported in sera of patients with various kind of vasculitis and other autoimmune diseases and the correlation between AECA and vasculitis damage has been recently demonstrated. Aim: to evaluate the presence of AECA in CD and their role in disease activity. Patients and Methods: Serum AECA have been assayed in 40 CD patients, 22 male and 18 female (mean age 31.9±11.8 yrs) and in 64 healthy volunteers. In patient group, disease activity was evaluated by Simple Crohn Activity Index (SCDA1). AECA were evaluated by means of ELISA, using cell cultures from human umbilical vein endothelial cells. Results were expressed as Binding Index (BI%), and the cut-off value of global test (IgA, IgG, IgM) is 35,8% (mean of healthy volunteers + 2SD). Chi-square and Spearman's tests provided statistical analysis. Results: AECA (IgA, IgG, IgM) were found in 12 patients (30%) (Bl%= 32,9±38,31) and only 3 controls (4,6%) (Bl%= 13.42±11.7) (P<0.005). Among patients AECA positive, 11 showed IgG-AECA and 7 IgM-AECA, while no patients showed IgA-AECA. A significant correlation (p<0.005) was found between AECA and SCDAI. No correlation was found between AECA presence and localization of the disease. Conclusion: Our data confirm the presence of AECA in a high percentage of CD patients and its correlation with disease activity (as far as other autoimmune diseases). It is possible that the AECA bound to endothelial cells cause a vasculitic damage promoting a series of cellular modifications considered essential for lymphocyte activation and for induction or maintenance of the inflammatory process. On the other hand, may be that inflammation induce cytokine release (i.e. Tumor Necrosis Factor ct) in CD. The cytokine could promote the neoexpression of antigens target by AECA. G4009
HEPATOBILIARY HELICOBACTER SPP. IDENTIFIED FROM PATIENTS WITH PRIMARY SCLEROSING CHOLANGITIS. J.G. Fox*, Z. Shen*, F. Feng*, J. Dufover +, M.M. Kaplan +, F. Dewhirst #. *Massachusetts Institute of Technology, Cambridge, MA 02139. #Forsyth Dental Center & +Tufts University, Boston, MA. Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease of unknown etiology. Pathologic lesions consist of persistent inflammation, destruction and fibrosis of intrahepatic and extrahepatic bile ducts. The high correlation of PSC and ulcerative colitis has raised the hypothesis that chronic portal bacteremia may initiate inflammation and promote subsequent hepatobiliary damage. The objective of this study was to ascertain whether Helicobacter spp. known to cause hepatobiliary disease in animals was present in PSC patients. Liver biopsies and bile were obtained from eight patients with PSC. Trypticase soy agar with 5% sheep blood, TVP and CVA medium were used for Helicobacter spp. isolation. A Rapid Prep genomic DNA kit was used to extract DNA from liver and Gene Releaser was used for bile DNA extraction. The primers chosen for PCR amplification recognized conserved regions of the 16S rRNA specific for all known Helicobacter spp. Oligonucleotide primers produced an amplified products of 1220 bp. For confirmation of the PCR-amplified fragment by Southern Blot hybridization, the PCR product was electrophoresed through a 1% agarose gel, transferred onto a nylon membrane, and subjected to hybridization with Helicobacter specific PCR-generated probe. PCRTM II vector was used for cloning of PCR products which were purified by Geneclean kit. ligated with 50 ng PCR TM II vector at 15C overnight, and transferred into INVaF' ceils. Ampicillin plates with X-gal were used to select positive clones. Plasmid DNA was isolated for E. coli by Qiagen plasmid prep kit. Helicobacter spp. were not cultured. However, Helicobacter spp. were identified by PCR amplification and southern hybridization using Helicobacter specific probe in 5 of 8 patients. In 2 of these patients a 1200 bp PCR amplified product was successfully cloned and sequenced. Analysis of the sequence indicated high homology to the 16S rRNA sequence of a cluster of Helicobacter spp. previously isolated from animals, i.e. tl. rodentium. H. rappini, and H. pullorum. Identification of Helicobacter spp. in liver and bile of patients with PSC is consistent with our recent findings of Helicobacter spp. associated hepatic disease in animals. Previous isolation of H. rappini and 1-1. pullorum in the feces of diarrheic humans is also supportive evidence. These results should stimulate studies to ascertain whether these or similar helicobacters play an important role in the pathogenesis of idiopathic hepatobiliary disease in humans.