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Pentoxifylline: A novel treatment for ulcerative colitis
April 1995
• THE R E L A T I O N BETWEEN FECAL BILE ACIDS AND OCCURENCE OF C O L O N D Y S P L A S I A AND CARCINOMA IN EXPERIMENTAL U L C E R A T I V E COLITIS K.Kajiura, T.Ohkusa, I.Okayasu First Department of Internal Medicine, Tokyo Medical and Dental University School of Medicine, Tokyo, Japan, and First Department of Pathology, Kitasato University School of Medicine, Kanagawa. AIM: We have reported that long term administration of dextran sulfate sodium (DSS) produces dysplasia and adenocarcinoma in the colon of hamsters and m i c e (Gut 1992;33:1521-7, Gastroenterology 1990; 98:694-70). The possible role of fecal bile acids (FBA) in colorectal cancer in human ulcerative colitis (UC) has been reported. In this study, we investigated the relation between FBA and occurence of colorectal dysplasia and carcinoma in experimental UC (EUC) induced by DSS. METHODS: A group; EUC was induced by 3%DSS (MW 54,000) to i0 CBA/J female mice, 9 wks old, for 9 wks, according to our previously reported method. B group; EUC was induced by 1% DSS to 10 mice, and azoxymethane (AZO, 7.4 mg/ kg BW) was intraperitoneal injection at the begining of the experiment. C group; Control mice were given only distilled water and AZO. Voided stool specimens were collected before and after the administration of 3%DSS or water, and were immediately freeze dried. FBA were analyzed by gas-liquid chromatography. Statistical significance was determined by paired t test and ANOVA. RESULTS: Adenocarcinoma, dysplasia and inflammatory changes were seen in all of the 10 mice of B group, and inflammatory changes were observed in all of the 10 mice of A group, while no dysplasia and inflammatory change were detected in control mice. Significant increases in cholic acid, primary FBA, and total FBA were observed in the mice of A and B group between the begining and the end of the experiment, while in the control mice, no significant changes of FBA were observed between the begining and end of the experiment. Conclusion: FBA are causally related to colorectal dysplasia and cancer with this experimantal UC as well as human UC.
Diarrhea and fecal parameters in AIDS patients. N. Kap¢l (1), Y. Benhamou (2), P. Orrico (1), O. Bouchaud(3), J,P. Coulaud (3), M. Gentilini (4), P. Opolon (2), J.G. G o b e r t (1). (1) Laboratoire de Parasitologi eCoprologie. GH Bichat-Cl. Bernar& 75018 Paris. (2) Service de Gastroent6rologie. GH Piti6-Salp~triOre. 75013 Pans. (3)Service des Maladies Infectieuses. GH Bichat-CI. Bernard. 75018 Paris. (4)Service des Maladies Intectieuses. GH Piti6-Salp6tfiere. 75013 Pans. France. Diarrhea, the main digestive symptom in HIV-infected patients, is commonly related to enteric infection. However, the influence of chronic intestinal infections on fecal parameters have been poorly studied. Objective: To analyse the influence of enteric infection on fecal parameters in HIVinfected patients. M e t h o d s : Blood and 3 consecutive days stools were collected in 49 AIDS patients i.e. patients with cryptosporidiosis [group(Gr) 1, n=20], microsporidiosis (Gr2, n=7), other enteric infections (Gr3, n=7) and no enteric infection (Gr4, n=15). Body weight loss (BWL,%) since the beginning of diarrhea was determined for the 49 AIDS patients. The following fecal parameters were studied in each group according to the absence or presence of diarrhea [mean feces weight (MFW) > 300g/24h (subgroups a and b, respectively)] : MFW (g/24h), dry weight (DW,%), lipids output (STEA, g/24h), nitrogen output (CREA,g/24h), natrium output (Na+, mmol/24h) potassium output (K+, mmol/24h) and fecal al-antitrypsin clearance (ATCl,ml/24h). Results: Mean CD4+ lymphocyte cells per gl was 24+_27.39-2-34, 17+-20 and 41+52 in Grl, 2, 3 and 4, respectively. Mean BWL (%) was 17+8, 14-+7, 18-+15 and 10+-_5in Grl, 2, 3 and 4, respectively. Fecal parameters are iven in the fnllowin table as medianl n MFW DW STEA CREA Na+ K+ ATCI no diarrhea 8 115 5 1,7 0,4 7 6 18 diarrhea 12 553 8 11,6 2,0 32 26 14 2' nodiarrhea 2 187 15 2 0,5 2 9 5 diarrhea 5 432 9 17;4 3,8 33 31 32 3 no diarrhea 4 61 29 6,4 0,8 2 7 26 . aiarrhea 3 916 4 13,5 2,1 49 12 53 4 no diarrhea 7 80 23 1,6 0,5 2 6 4 diarrhea 8 555 11 12,6 3,2 28 27 18 Conclusion: Diarrhea (as MFW was not correlated with the presence of enteric infection. Furthermore, these pathogens did not significantly impaired the fecal parameters. These results suggest that HIV-related mucosal intestinal 'alteration should play an important role in the occurence of diarrhea.
Grl,
Immunology, Microbiology, and Inflammatory Disorders A845
O PENTOXIFYLLINE: A NOVELTREATMENTFOR ULCERATIVE COLITIS. L. Kam, V. Casini-Raggi, F. Cominelli. USC School of Medicine, Los Angeles, CA 90033. Patients with active Ulcerative C o l i t i s (UC) have been shown to have elevated cytokine levels in the serum and stool, and elevated cytokine production levels from isolated peripheral mononuclear cells. Pentoxifylline is a xanthine derivative with anti-cytokine properties, and has been shown to increase survival in mice treated with endotoxin. We report the results of an open labelled p i l o t study investigating the efficacy and safety of Pentoxifylline as a novel treatment for mild-moderate UC. Methods: Patients with active UC involving more than lOcm from the anal verge were e l i g i b l e for enrollment in the study. The diagnosis of UC was confirmed by endoscopy and biopsy, and exclusion of infectious colitis. All patients had mild-moderately active disease {Truelove and Witts scale). After informed consent was obtained, patients, were treated with Pentoxifylline 400mg orally TID for 8 weeks. Patients undergoing treatment with prednisone, immunosuppressive drugs, or experimental medications within 30 days were excluded. Efficacy was determined by a 3 point Disease Activity Index (DAI) (Onormal, l-mild, 2-moderate, 3-severe), endoscopic severity, andhistologicevaluation. Flexible sigmoidoscopywith biopsy was done at screening and week 8. Results: Eight patients were enrolled in the study. Three patients withdrew (two after 2-4 days and one a f t e r 4 weeks of treatment) because no symptomatic improvement had occurred. One of these patients also complained of dizziness and another complained of abdominal pain. Both symptoms were present prior to study enrollment, and persisted after study termination. Four of the remaining five patients reported symptomatic improvement at the end of 8 weeks with a mean decrease in DAI by 1.4 points. Two patients were completely asymptomatic. Two patients showed endoscopic and histologic improvement at the end of 8 weeks. Except for two patients who complained of dizziness and abdominal pain, no other side effects or adverse events were noted. Results of cytokine levels in the serum and tissue will be reported later. These results are promising and support the need for a randomized, placebo controlled t r i a l of Pentoxifylline as a novel treatment for UC.
O EXPRESSION OF cagA AND cagC IN H. pylori IS INHIBITED AT LOW pH. M. Karita, M.K.R. Tummuru, J.C. Atherton, M.J. Blaser. Division of Infectious Diseases, Department of Medicine, Vanderbih University School of Medicine and Veterans Affairs Medical Center, Nashville, TN. /4. py/or/strains possessing cagA and cagC are associated with peptic ulceration, but the role of these genes in H. pylori physiology is not known. To determine regulation of expression of cagA and cagC, we created H. p.flori strains with a promoterless reporter (xyIE) inserted into these genes. First, a 2.6kb fragment with promotefless xyIE and km (with promoter) was excised from pMKQ. This fragment was ligated into a unique restriction site in cagA cloned in pBC4 to create pMK25, and in cagC cloned in pMT2 to create pMK13. The position and orientation ofxyIE contained in each plasmid was confirmed by mapping and PCR, and its expression in E. cell also was confirmed using a colorimetric assay. Natural transformations using these plasmids and H. py/or/strain CPY3401 were performed to create CPY340 i A cagA:xylE:km(3401A') and CPY3401A eagC:xylE:km(3401C-). The position and orientation of the reporter in these mutants was confirmed by PCR. Expression ofx'yIE in cagA (mean 476.2-+SD 92,2 mUnit/mg protein, n=4) was significantly higher than that in cagC (18.1±0.9) in broth culture (p
pH of
3401 A
3401C
citrate buffer 7 462.5_+120.4 22.0_+2.9 6 4 2 1 . 6 +- 134.1 * 13.6_+ 1.9* 5 320.9-+72.8* 8.6_+ 1.9"** 4 167.2_+67.4** 5 . 9 +- 1.9*** Compared to pH7, *p<0.05, **p<0.01, ***p<0.001; n = 4 determinations. W e conclude: 1, a xylE promoterless reporter works well in H. pyloN; 2, expression ofcagA is more than 20-fold greater than cagC,in broth culture; 3, expression of cagA and cagC are markedly inhibited at low pH. This is the first description of pH-related gene expression in H. pyloN.
• THE R E L A T I O N BETWEEN FECAL BILE ACIDS AND OCCURENCE OF C O L O N D Y S P L A S I A AND CARCINOMA IN EXPERIMENTAL U L C E R A T I V E COLITIS K.Kajiura, T.Ohkusa, I.Okayasu First Department of Internal Medicine, Tokyo Medical and Dental University School of Medicine, Tokyo, Japan, and First Department of Pathology, Kitasato University School of Medicine, Kanagawa. AIM: We have reported that long term administration of dextran sulfate sodium (DSS) produces dysplasia and adenocarcinoma in the colon of hamsters and m i c e (Gut 1992;33:1521-7, Gastroenterology 1990; 98:694-70). The possible role of fecal bile acids (FBA) in colorectal cancer in human ulcerative colitis (UC) has been reported. In this study, we investigated the relation between FBA and occurence of colorectal dysplasia and carcinoma in experimental UC (EUC) induced by DSS. METHODS: A group; EUC was induced by 3%DSS (MW 54,000) to i0 CBA/J female mice, 9 wks old, for 9 wks, according to our previously reported method. B group; EUC was induced by 1% DSS to 10 mice, and azoxymethane (AZO, 7.4 mg/ kg BW) was intraperitoneal injection at the begining of the experiment. C group; Control mice were given only distilled water and AZO. Voided stool specimens were collected before and after the administration of 3%DSS or water, and were immediately freeze dried. FBA were analyzed by gas-liquid chromatography. Statistical significance was determined by paired t test and ANOVA. RESULTS: Adenocarcinoma, dysplasia and inflammatory changes were seen in all of the 10 mice of B group, and inflammatory changes were observed in all of the 10 mice of A group, while no dysplasia and inflammatory change were detected in control mice. Significant increases in cholic acid, primary FBA, and total FBA were observed in the mice of A and B group between the begining and the end of the experiment, while in the control mice, no significant changes of FBA were observed between the begining and end of the experiment. Conclusion: FBA are causally related to colorectal dysplasia and cancer with this experimantal UC as well as human UC.
Diarrhea and fecal parameters in AIDS patients. N. Kap¢l (1), Y. Benhamou (2), P. Orrico (1), O. Bouchaud(3), J,P. Coulaud (3), M. Gentilini (4), P. Opolon (2), J.G. G o b e r t (1). (1) Laboratoire de Parasitologi eCoprologie. GH Bichat-Cl. Bernar& 75018 Paris. (2) Service de Gastroent6rologie. GH Piti6-Salp~triOre. 75013 Pans. (3)Service des Maladies Infectieuses. GH Bichat-CI. Bernard. 75018 Paris. (4)Service des Maladies Intectieuses. GH Piti6-Salp6tfiere. 75013 Pans. France. Diarrhea, the main digestive symptom in HIV-infected patients, is commonly related to enteric infection. However, the influence of chronic intestinal infections on fecal parameters have been poorly studied. Objective: To analyse the influence of enteric infection on fecal parameters in HIVinfected patients. M e t h o d s : Blood and 3 consecutive days stools were collected in 49 AIDS patients i.e. patients with cryptosporidiosis [group(Gr) 1, n=20], microsporidiosis (Gr2, n=7), other enteric infections (Gr3, n=7) and no enteric infection (Gr4, n=15). Body weight loss (BWL,%) since the beginning of diarrhea was determined for the 49 AIDS patients. The following fecal parameters were studied in each group according to the absence or presence of diarrhea [mean feces weight (MFW) > 300g/24h (subgroups a and b, respectively)] : MFW (g/24h), dry weight (DW,%), lipids output (STEA, g/24h), nitrogen output (CREA,g/24h), natrium output (Na+, mmol/24h) potassium output (K+, mmol/24h) and fecal al-antitrypsin clearance (ATCl,ml/24h). Results: Mean CD4+ lymphocyte cells per gl was 24+_27.39-2-34, 17+-20 and 41+52 in Grl, 2, 3 and 4, respectively. Mean BWL (%) was 17+8, 14-+7, 18-+15 and 10+-_5in Grl, 2, 3 and 4, respectively. Fecal parameters are iven in the fnllowin table as medianl n MFW DW STEA CREA Na+ K+ ATCI no diarrhea 8 115 5 1,7 0,4 7 6 18 diarrhea 12 553 8 11,6 2,0 32 26 14 2' nodiarrhea 2 187 15 2 0,5 2 9 5 diarrhea 5 432 9 17;4 3,8 33 31 32 3 no diarrhea 4 61 29 6,4 0,8 2 7 26 . aiarrhea 3 916 4 13,5 2,1 49 12 53 4 no diarrhea 7 80 23 1,6 0,5 2 6 4 diarrhea 8 555 11 12,6 3,2 28 27 18 Conclusion: Diarrhea (as MFW was not correlated with the presence of enteric infection. Furthermore, these pathogens did not significantly impaired the fecal parameters. These results suggest that HIV-related mucosal intestinal 'alteration should play an important role in the occurence of diarrhea.
Grl,
Immunology, Microbiology, and Inflammatory Disorders A845
O PENTOXIFYLLINE: A NOVELTREATMENTFOR ULCERATIVE COLITIS. L. Kam, V. Casini-Raggi, F. Cominelli. USC School of Medicine, Los Angeles, CA 90033. Patients with active Ulcerative C o l i t i s (UC) have been shown to have elevated cytokine levels in the serum and stool, and elevated cytokine production levels from isolated peripheral mononuclear cells. Pentoxifylline is a xanthine derivative with anti-cytokine properties, and has been shown to increase survival in mice treated with endotoxin. We report the results of an open labelled p i l o t study investigating the efficacy and safety of Pentoxifylline as a novel treatment for mild-moderate UC. Methods: Patients with active UC involving more than lOcm from the anal verge were e l i g i b l e for enrollment in the study. The diagnosis of UC was confirmed by endoscopy and biopsy, and exclusion of infectious colitis. All patients had mild-moderately active disease {Truelove and Witts scale). After informed consent was obtained, patients, were treated with Pentoxifylline 400mg orally TID for 8 weeks. Patients undergoing treatment with prednisone, immunosuppressive drugs, or experimental medications within 30 days were excluded. Efficacy was determined by a 3 point Disease Activity Index (DAI) (Onormal, l-mild, 2-moderate, 3-severe), endoscopic severity, andhistologicevaluation. Flexible sigmoidoscopywith biopsy was done at screening and week 8. Results: Eight patients were enrolled in the study. Three patients withdrew (two after 2-4 days and one a f t e r 4 weeks of treatment) because no symptomatic improvement had occurred. One of these patients also complained of dizziness and another complained of abdominal pain. Both symptoms were present prior to study enrollment, and persisted after study termination. Four of the remaining five patients reported symptomatic improvement at the end of 8 weeks with a mean decrease in DAI by 1.4 points. Two patients were completely asymptomatic. Two patients showed endoscopic and histologic improvement at the end of 8 weeks. Except for two patients who complained of dizziness and abdominal pain, no other side effects or adverse events were noted. Results of cytokine levels in the serum and tissue will be reported later. These results are promising and support the need for a randomized, placebo controlled t r i a l of Pentoxifylline as a novel treatment for UC.
O EXPRESSION OF cagA AND cagC IN H. pylori IS INHIBITED AT LOW pH. M. Karita, M.K.R. Tummuru, J.C. Atherton, M.J. Blaser. Division of Infectious Diseases, Department of Medicine, Vanderbih University School of Medicine and Veterans Affairs Medical Center, Nashville, TN. /4. py/or/strains possessing cagA and cagC are associated with peptic ulceration, but the role of these genes in H. pylori physiology is not known. To determine regulation of expression of cagA and cagC, we created H. p.flori strains with a promoterless reporter (xyIE) inserted into these genes. First, a 2.6kb fragment with promotefless xyIE and km (with promoter) was excised from pMKQ. This fragment was ligated into a unique restriction site in cagA cloned in pBC4 to create pMK25, and in cagC cloned in pMT2 to create pMK13. The position and orientation ofxyIE contained in each plasmid was confirmed by mapping and PCR, and its expression in E. cell also was confirmed using a colorimetric assay. Natural transformations using these plasmids and H. py/or/strain CPY3401 were performed to create CPY340 i A cagA:xylE:km(3401A') and CPY3401A eagC:xylE:km(3401C-). The position and orientation of the reporter in these mutants was confirmed by PCR. Expression ofx'yIE in cagA (mean 476.2-+SD 92,2 mUnit/mg protein, n=4) was significantly higher than that in cagC (18.1±0.9) in broth culture (p
pH of
3401 A
3401C
citrate buffer 7 462.5_+120.4 22.0_+2.9 6 4 2 1 . 6 +- 134.1 * 13.6_+ 1.9* 5 320.9-+72.8* 8.6_+ 1.9"** 4 167.2_+67.4** 5 . 9 +- 1.9*** Compared to pH7, *p<0.05, **p<0.01, ***p<0.001; n = 4 determinations. W e conclude: 1, a xylE promoterless reporter works well in H. pyloN; 2, expression ofcagA is more than 20-fold greater than cagC,in broth culture; 3, expression of cagA and cagC are markedly inhibited at low pH. This is the first description of pH-related gene expression in H. pyloN.