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Effect of nutrients on post heparin plasma diamine oxidase activity
AGAAI07
April 2000
700
702
ASCA AND PANCA IN A NEW BELGIAN IBD DATASET. Severine Vermeire, Marc Peeters, Sofie Joossens, Paul J. Rutgeerts, Univ hospitals Gasthuisberg, Leuven, Belgium; Univ Hosp Gasthuisberg, Leuven, Belgium.
CORRELATION BETWEEN mSTLOGICAL ACTIVITIES AND PHOSPHOLIPASE A 2 IIA LEVELS IN SERA OF PATIENTS WITH ULCERATIVE COLITIS. Osamu Yamaguchi, Kazuhito Sugimura, Kisei Ishizuka, Koji Suzuki, Katsuhiko Hasegawa, Kazuo Otuka, Terasu Honma, Hitoshi Asakura, Niigata Univ Sch of Medicine, Niigata, Japan. Background & Aims: In ulcerative colitis (UC), accurate disease activity is difficult to be determined by the laboratory tests. To date, it is mainly assessed by the endoscopic and histrogical examinations.Meanwhile phospholipase A2(PLA2 ) II participates in the regulation of phospholipid metabolism and eicosanoids biosynthesis, and serum level of PLA21I is suggested to referct the disease activity in patients with UC.Recently, PLA21I is divided into four subgroups (I1A, IIC, V, X). We examined the relationship between histological assessments of disease activity and serum level of PLA2IIA using a specific monoclonal antibody. Additionally, mucosal production of PLA2I1A was estimated by the immunohistochemically. Methods.Twenty-three patients with UC, who had underwent endoscopy,and controls were studied. After informed consent was approved, biopsy specimens of colonic mucosa were obtained. The histological score was determined as regared, active = I, inactive = O.Andthe total of each histological score from 10 segments of colon was assessed as disease activity. The levels of PLA2IIA in sera were measured by specific radioimmunoassay kit (S-1647, Shionogi Phamaceutical Ltd). Immunohistochemical analysis was performed with same monoclonal antibody. Data are expressed as mean +SEM. Resuls.Twenty-one of the 23 patients were active status. The serum PLA2IIA levels in patients with UC were significantly higher than those of controls (UC 9.1 :t 3.25 ng/ml, NC 2.9:t 0.13 ng/ml, P = 0.029). Serum PLA2IIA levels in UC were strongly correlated with histological activities(Pc= 0.0045, Spearmans corration). The production of PLA2I1A by the polymorphonuclear cells and epithelial cells was observed histochemically. Conclusions:Serum PLA2 IIA level is associated with thehistlogical activity of UC. PLA2 I1A can be a good candidate to evaluate the disease activity of UC by means of laboratory examination.
Introduction: Serological markers have become more valuable in inflammatory bowel disease, not only for diagnosis but also in the search for defining subgroups of patients. So far, only 2 markers have shown to be of interest for clinical utility, ASCA for Crohn's disease (CD) and UCspecific pANCA for ulcerative colitis (UC). A specific subset of CD patients also carry pANCA and are defined as DC-like CD patients. Several laboratories and commercial companies have developed assays for ASCA and pANCA. However, international agreement upon techniques is of greatest importance especially when comparing results. Therefore, we wanted to evaluate the commercial serological kit assays from Prometheus Laboratories and compare with previous results in a new IBD dataset. Methods: A group of 201 sporadic IBD patients (100 CD and 101 UC) and 100 healthy controls were tested for UC-specific pANCA and ASCA (IgA and IgG) at Prometheus Laboratories Inc (San diego, CAl. For ASCA a standardised ELISA was used, whereas for pANCA the three steps reaction as previously described was used. Samples were blinded for clinical diagnosis. Results: (table) Conclusion: We previously showed that ASCA is associated with CD in 62% of patients and pANCA with UC in 57% of patients. We tested these markers in a new group of Belgian IBD patients with the kit assays from Prometheus Laboratories, and found a similar prevalence for pANCA in UC patients (67%) and for ASCA in CD patients (59%). Of all pANCA + CD patients 7/15 (46.7%) behaved clinically as UC-like CD. This study represents the validation of the clinical utility of the Prometheus assay systems in a European cohort. Tablet CD(n=100)
UC(n=101)
Controls (n=100)
ASCA+(lgA or IgG) pANCA+
59(59%) 15(15%)
8(7,9%) 68(67,3%)
3(3%) 4(4%)
pANCA+/ASCA· pANCA+/ASCA+ pANCA·/ASCA+ pANCA·/ASCA·
7(7%) 8(8%) 51 (51%) 34(34%)
63(62,4%) 5(4,9%) 3(2,9%) 30(29,7%)
4(4%) 0(0%) 3(3%) 93(93%)
701 EFFECT OF NUTRIENTS ON POST HEPARIN PLASMA DIA· MINE OXIDASE ACTIVITY. Armin Wollin, Michael Roy, Dept of Physiology, Coli of Med, Univ of Saskatchewan, Saskatoon, SK, Canada, Postheparin plasma diamine oxidase activity has been measured to assess mucosal damage arising from intestinal diseases. Postheparin plasma enzyme activity diminishes in patients with inflammatory bowel disease compared to healthy individuals but the response is not always consistent and considerable overlap has been observed in some studies. We have recently observed in rats that fat ingestion increases diamine oxidase release from the intestinal tissue 5-6 fold over fasting levels. Since dietary restrictions were not indicated in the clinical studies, our aim is to evaluate the effect of nutrients on the postheparin diamine oxidase activity in plasma of healthy rats. Eighteen hours before the experiment the animals received 2000 Ulkg (ip) of heparin, to release the extracellular bound diamine oxidase, and were deprived of food but not water. Following the 18 hr fast the rats were tube fed I ml of saline or olive oil or 9% sucrose or 5% albumin, Two hours after feeding, the femoral vein was cannulated for the heparin injection and blood sampling. Three to four hours after feeding a preheparin blood sample was taken, followed by an iv injection of heparin (2000 Ulkg). Blood samples were subsequently collected every 15 min for 60 min. The plasma diamine oxidase activity was measured, The enzyme activity in fasted (saline) animals and those fed olive oil, sucrose or albumin was 3.6+/-0.9,32,9+/-9.8,4.6+/-1.9 and 5.8+/-1.2 mu/ml respectively, n = 4 to 8/group. Pretreating olive oil fed animals with cycloheximide (2mglkg), a protein synthesis inhibitor, resulted in a response of 3.3+1-1.0 mulml, thus eliminating the significant (p5 times that of carbohydrates or protein. The marked increase appears to be newly synthesized diamine oxidase. Therefore, dietary restrictions prior to a postheparin diamine oxidase activity assessment for mucosal damage are necessary.
703 A POLYMORPHISM IN INTERLEUKIN·4 IS ASSOCIATED WITH CROHN'S DISEASE. Guruprasad P. Aithal, Christopher P. Day, Julian Leathart, Ann K. Daly, Mark Hudson, Univ of Newcastle, Newcastle, United Kingdom; Freeman Hosp, Newcastle, United Kingdom. Immunomodulatory cytokine genes are potential candidate genes in inflammatory bowel disease. Interleukin-4 (IL-4) is important in the T helper-2 (Th2) cell dominant immune response. Functionally significant polymorphisms have been described in the gene encoding IL-4 as well as the protein coding region of the gene encoding the IL-4 receptor and these have been associated with rhuematoid arthritis and asthma. The variant alleles are associated with increased transcriptional activity in the case of IL-4 and increased level of signal transduction in the case of the receptor. We have performed an allelic association study to determine the role of these polymorphisms in the susceptibility to ulcerative colitis (UC) and Crohn's disease (CD). Patients and Methods: 98 patients with UC and 86 patients with CD were recruited from Freeman Hospital, Newcastle. 184 healthy controls were recruited from hospital and University staff. Genotyping for the polymorphisms at position -34 in the IL-4 gene and codon 576 in the IL-4 receptor gene were performed by PCR-RFLP analysis. Results: 31% of patients with CD carried I or 2 copies of the variant T allele for IL-4 at position -34 compared with 12% of the control group (Odds Ratio: 3.49[1.83-6.64D. This polymorphism was in linkage disequalibrium with the functionally significant C-590T polymorphism. A trend towards an association of CD with the variant IL-4 receptor was detected but was not significant (OR:1.6[0.94-2.7D. Conclusions: The -34 IL-4 polymorphism is a strong determinant of susceptibility to Crohn's disease in our population. Association of Crohn's disease with an allele resulting in high IL-4 transcription suggests that Th2 cells may have a role in the pathogenesis of Crohn' s disease. No ofsubjects with atleast one variant allele in UC orCD compared with controls Groups Controls Ulcerative Colitis Pvalue for UC Crohn's Disease p value for CD
IL-4Receptor (Q576RI
IL-4(-34T]
57/184 34/94 0.42 36/86 009
21/181 12/98 0,85 27/86 0,0001
April 2000
700
702
ASCA AND PANCA IN A NEW BELGIAN IBD DATASET. Severine Vermeire, Marc Peeters, Sofie Joossens, Paul J. Rutgeerts, Univ hospitals Gasthuisberg, Leuven, Belgium; Univ Hosp Gasthuisberg, Leuven, Belgium.
CORRELATION BETWEEN mSTLOGICAL ACTIVITIES AND PHOSPHOLIPASE A 2 IIA LEVELS IN SERA OF PATIENTS WITH ULCERATIVE COLITIS. Osamu Yamaguchi, Kazuhito Sugimura, Kisei Ishizuka, Koji Suzuki, Katsuhiko Hasegawa, Kazuo Otuka, Terasu Honma, Hitoshi Asakura, Niigata Univ Sch of Medicine, Niigata, Japan. Background & Aims: In ulcerative colitis (UC), accurate disease activity is difficult to be determined by the laboratory tests. To date, it is mainly assessed by the endoscopic and histrogical examinations.Meanwhile phospholipase A2(PLA2 ) II participates in the regulation of phospholipid metabolism and eicosanoids biosynthesis, and serum level of PLA21I is suggested to referct the disease activity in patients with UC.Recently, PLA21I is divided into four subgroups (I1A, IIC, V, X). We examined the relationship between histological assessments of disease activity and serum level of PLA2IIA using a specific monoclonal antibody. Additionally, mucosal production of PLA2I1A was estimated by the immunohistochemically. Methods.Twenty-three patients with UC, who had underwent endoscopy,and controls were studied. After informed consent was approved, biopsy specimens of colonic mucosa were obtained. The histological score was determined as regared, active = I, inactive = O.Andthe total of each histological score from 10 segments of colon was assessed as disease activity. The levels of PLA2IIA in sera were measured by specific radioimmunoassay kit (S-1647, Shionogi Phamaceutical Ltd). Immunohistochemical analysis was performed with same monoclonal antibody. Data are expressed as mean +SEM. Resuls.Twenty-one of the 23 patients were active status. The serum PLA2IIA levels in patients with UC were significantly higher than those of controls (UC 9.1 :t 3.25 ng/ml, NC 2.9:t 0.13 ng/ml, P = 0.029). Serum PLA2IIA levels in UC were strongly correlated with histological activities(Pc= 0.0045, Spearmans corration). The production of PLA2I1A by the polymorphonuclear cells and epithelial cells was observed histochemically. Conclusions:Serum PLA2 IIA level is associated with thehistlogical activity of UC. PLA2 I1A can be a good candidate to evaluate the disease activity of UC by means of laboratory examination.
Introduction: Serological markers have become more valuable in inflammatory bowel disease, not only for diagnosis but also in the search for defining subgroups of patients. So far, only 2 markers have shown to be of interest for clinical utility, ASCA for Crohn's disease (CD) and UCspecific pANCA for ulcerative colitis (UC). A specific subset of CD patients also carry pANCA and are defined as DC-like CD patients. Several laboratories and commercial companies have developed assays for ASCA and pANCA. However, international agreement upon techniques is of greatest importance especially when comparing results. Therefore, we wanted to evaluate the commercial serological kit assays from Prometheus Laboratories and compare with previous results in a new IBD dataset. Methods: A group of 201 sporadic IBD patients (100 CD and 101 UC) and 100 healthy controls were tested for UC-specific pANCA and ASCA (IgA and IgG) at Prometheus Laboratories Inc (San diego, CAl. For ASCA a standardised ELISA was used, whereas for pANCA the three steps reaction as previously described was used. Samples were blinded for clinical diagnosis. Results: (table) Conclusion: We previously showed that ASCA is associated with CD in 62% of patients and pANCA with UC in 57% of patients. We tested these markers in a new group of Belgian IBD patients with the kit assays from Prometheus Laboratories, and found a similar prevalence for pANCA in UC patients (67%) and for ASCA in CD patients (59%). Of all pANCA + CD patients 7/15 (46.7%) behaved clinically as UC-like CD. This study represents the validation of the clinical utility of the Prometheus assay systems in a European cohort. Tablet CD(n=100)
UC(n=101)
Controls (n=100)
ASCA+(lgA or IgG) pANCA+
59(59%) 15(15%)
8(7,9%) 68(67,3%)
3(3%) 4(4%)
pANCA+/ASCA· pANCA+/ASCA+ pANCA·/ASCA+ pANCA·/ASCA·
7(7%) 8(8%) 51 (51%) 34(34%)
63(62,4%) 5(4,9%) 3(2,9%) 30(29,7%)
4(4%) 0(0%) 3(3%) 93(93%)
701 EFFECT OF NUTRIENTS ON POST HEPARIN PLASMA DIA· MINE OXIDASE ACTIVITY. Armin Wollin, Michael Roy, Dept of Physiology, Coli of Med, Univ of Saskatchewan, Saskatoon, SK, Canada, Postheparin plasma diamine oxidase activity has been measured to assess mucosal damage arising from intestinal diseases. Postheparin plasma enzyme activity diminishes in patients with inflammatory bowel disease compared to healthy individuals but the response is not always consistent and considerable overlap has been observed in some studies. We have recently observed in rats that fat ingestion increases diamine oxidase release from the intestinal tissue 5-6 fold over fasting levels. Since dietary restrictions were not indicated in the clinical studies, our aim is to evaluate the effect of nutrients on the postheparin diamine oxidase activity in plasma of healthy rats. Eighteen hours before the experiment the animals received 2000 Ulkg (ip) of heparin, to release the extracellular bound diamine oxidase, and were deprived of food but not water. Following the 18 hr fast the rats were tube fed I ml of saline or olive oil or 9% sucrose or 5% albumin, Two hours after feeding, the femoral vein was cannulated for the heparin injection and blood sampling. Three to four hours after feeding a preheparin blood sample was taken, followed by an iv injection of heparin (2000 Ulkg). Blood samples were subsequently collected every 15 min for 60 min. The plasma diamine oxidase activity was measured, The enzyme activity in fasted (saline) animals and those fed olive oil, sucrose or albumin was 3.6+/-0.9,32,9+/-9.8,4.6+/-1.9 and 5.8+/-1.2 mu/ml respectively, n = 4 to 8/group. Pretreating olive oil fed animals with cycloheximide (2mglkg), a protein synthesis inhibitor, resulted in a response of 3.3+1-1.0 mulml, thus eliminating the significant (p
703 A POLYMORPHISM IN INTERLEUKIN·4 IS ASSOCIATED WITH CROHN'S DISEASE. Guruprasad P. Aithal, Christopher P. Day, Julian Leathart, Ann K. Daly, Mark Hudson, Univ of Newcastle, Newcastle, United Kingdom; Freeman Hosp, Newcastle, United Kingdom. Immunomodulatory cytokine genes are potential candidate genes in inflammatory bowel disease. Interleukin-4 (IL-4) is important in the T helper-2 (Th2) cell dominant immune response. Functionally significant polymorphisms have been described in the gene encoding IL-4 as well as the protein coding region of the gene encoding the IL-4 receptor and these have been associated with rhuematoid arthritis and asthma. The variant alleles are associated with increased transcriptional activity in the case of IL-4 and increased level of signal transduction in the case of the receptor. We have performed an allelic association study to determine the role of these polymorphisms in the susceptibility to ulcerative colitis (UC) and Crohn's disease (CD). Patients and Methods: 98 patients with UC and 86 patients with CD were recruited from Freeman Hospital, Newcastle. 184 healthy controls were recruited from hospital and University staff. Genotyping for the polymorphisms at position -34 in the IL-4 gene and codon 576 in the IL-4 receptor gene were performed by PCR-RFLP analysis. Results: 31% of patients with CD carried I or 2 copies of the variant T allele for IL-4 at position -34 compared with 12% of the control group (Odds Ratio: 3.49[1.83-6.64D. This polymorphism was in linkage disequalibrium with the functionally significant C-590T polymorphism. A trend towards an association of CD with the variant IL-4 receptor was detected but was not significant (OR:1.6[0.94-2.7D. Conclusions: The -34 IL-4 polymorphism is a strong determinant of susceptibility to Crohn's disease in our population. Association of Crohn's disease with an allele resulting in high IL-4 transcription suggests that Th2 cells may have a role in the pathogenesis of Crohn' s disease. No ofsubjects with atleast one variant allele in UC orCD compared with controls Groups Controls Ulcerative Colitis Pvalue for UC Crohn's Disease p value for CD
IL-4Receptor (Q576RI
IL-4(-34T]
57/184 34/94 0.42 36/86 009
21/181 12/98 0,85 27/86 0,0001