Intestinal protein nitration induced by myeloperoxidase

intake between groups was similar and provided adequatemacro- and micronutrients. There were no differences in ileal villus height between groups. Ileal crypt depth was significantly increased with SBR (control 141_+13/~m vs SBPdstandarddiet 181_+6 p,m; P
1138 Cilansetron Shows Efficacy in Non-constipated Irrirable Bowel Syndrome Patients Independent of the Definition of the Patient Population by Rome I or Rome II

Steven Caras, Solvay Pharmaceuticals, Inc, Marietta, CA; Gunter Krause, Egbert Biesheuvel, Claus Steinborn, Solvay Pharmaceuticals, 8 V, Weesp Netherlands Objective: To compare the efficacy of cilansetron administered over 12 weeks in non-constipated irritable bowel syndrome (IBS) patients defined by Rome I versus Rome II criteria. Methods: Two double-blind, placebocontrolled, randomized,multicenter, parallel-groupdoseranging studies were conducted in the U.S. (n=471; doses: 1, 2, 8, 16 mg tid) and Canada/ Europe (n = 435; doses: 1,2, 4,16 mg tid). The primary efficacy parameterwas the responder rate for adequaterelief of IBS symptoms (abdominal pain/discomfort, abnormal bowel habits). Secondaryefficacy parametersincludedassessmentof abdominalpain and measuresof bowel habits including stool consistency and frequency. Non-constipated patients were selected according to Rome I criteria. A subgroup analysis of patients meeting Rome II criteria has been performed. Results: No differences in efficacy could be seen between Rome I and Rome II patients. Also, the secondary parameters did not show any relevant differences between Rome I and Rome II patients. Conclusions: Cilansetron is efficacious in achieving adequate relief of IBS symptoms in non-constipatedIBS patients regardlessof the definition according to Rome 1 or Rome I1 criteria.

1141 Intestinal Protein Nitration Induced by Myeloperoxidase

Ka Blan, Yael Harad, Menu Zhong, Mildred Lai, Norman Welabrodt, Fetid Murad, Univ of Texas Medical Sch, Houston, TX

Comparisonof Rome I and RomeII ResponseRates U.S.StudyI Dose

Rome I

Rome II

Placebo

1 mg 2 mg

40% 62% 53%

42% 61% 55%

4 mg

--

--

8 mg

55% 62%

54% 62%

16 mg

Nitration of tyrosine residues in cellular proteins may interrupt tyrosine phosphorylation, a key process in cellular signal transduction pathways. Experiments performed on isolated ceils have delineated two major pathways for nitrotyrosine formation. One is mediated by peroxynitrite, which is formed from the rapid reaction of overproduced nitric oxide with superoxide. The other is mediated by myeloperoxidase(MPO). In previous experiments, we tested the hypothesis that activation of MPO is important in protein tyrosine nitration in the intestine by utilizing the experimentalmodel of Trichinella spiralis iTS) infected mice in which jejunal inflammation was paralleled by significant increases in MPO expression. We found that 7 to 10 days post infection, MPO expression was markedly increased.Also at that time post infection, use of a polyclonal antibody detected7 distinct bands of nitrotyrosine in jejunal homogenates. A band at 105 kDa that was present in control tissues was suppressedduring infection, whereasa band at 101 kOa increasedafter the infection. The remaining five bands (at 37, 33, 29, 28, and 26 kDa) also all increased. In the current experiments, we used a monoclonal antibody which disclosed a band at -60 kDa. Also, to determine further that the nitration was due to MPO upregulation, jejunal segments isolated from control mice were treated with 100 nM MPO. Such treatment resulted in a pattern of nitrotymsine formation similar to that seen in TS infected mice. In conclusion, changes (mostly increases) in the nitration of jejunal protein tyrosine residues were observed following TS infection and MPO incubation. In TS infected animals, the degree of change was correlated with the expression of MPO, but not with NOS2. Thus MPO may play the major role in nitrotyrosine formation, and effective suppression of MPO activity may be important in preventing nitrotymsine formation during inflammation.

Canada/EuropeStudy2 Rome I Rome II

36% 47% 58% 44% -54%

38% 50% 59% 43% --

56%

1. US Study:RomeI (n=471), RomeII (n=438) 2. Canada/EuropeStudy:RomeI (n---435),RomeII (n=389) 1139 Cilansetron Shows Efficacy in Male and Female Non-consUpated Patients with Irritable Bowel Syndrome in a United States Study

Steven Caras, Solvay Pharmaceuticals, Inc, Marietta, CA; Gunter Krause, Egbert Biesheuvel, Claus Steinborn, Sotvay Pharmaceuticals,R V, Weesp Netherlands Objective:To comparethe efficacy of cilansetron 1 mg, 2 rag, 8 mg and 16 mg tid to placebo, administered over 12 weeks in non-constipated irritable bowel syndrome (IBS) patients. Methods: The study was a double-blind, placebo-controlled,randomized,multicenter, parallelgroup dose-ranging study conducted at over 50 U.S. sites. The primary efficacy paramenter was the responder rate for adequate relief of IBS symptoms (abdominal pain/discomfort, abnormal bowel habits). Secondary efficacy parameters included assessment of abdominal pain and measures of bowel habits including stool consistency and frequency. Results: Response rates for the intent-to-treat sample (n =454) were 40% for placebo,62% for 1 mg (p
1142 Pancreatic Stones Shockwave Lithotripsy Induces Oxidative Stress. A Study Of Lipoperoxidation Products In Pure Pancreatic Juice.

Cristlano Spade, Digest Endoscopy Unit, Catholic University, Rome Italy; Stefano A. Santini, Biochemistry and Clin Biochemistry, Rome Italy; FrancescaFosshia, Monica Pandolfi, Vincenzo Perri, Digest EndoscopyUnit, Catholic University, Rome Italy; Niccolo Silveri Gentiloni, Dept of Internal Medicine, Rome Italy; Bruno Giardina, Biochemistry and Clin Biochemistry, Rome Italy; Guido Costamagna,Digest EndoscopyUnit, Catholic University, Rome Italy Background:extracorporealshockwavelithotripsy (ESWL) has been shown to be an effective technique to fragment pancreatic stones and is now considered as an important part of endotherapyfor chronic pancreatitis(CP). It has beensuggestedthat increasedlipoperoxidation products in pancreaticjuice of patient with CP, comparedto normal subjects, may reflect the oxidativedamageof lipids and proteins and consumption of redoxantioxidantagentssecondary to oxigen free radicals production. Aim of this study was to investigate if ESWL may induce further increaseof lipoperoxidation products in pancreaticjuice of patients with CP. Materials and methods: five patients (3 M, 2 F), mean age 62.4 yrs) with calcified CP undergoing ESWL with nasopancreatic catheter in situ were studied. ESWL was performed with a second generation electrohydraulic system under radiological control (Lithostar plus, Siemens, Germany). On average5000 discharges with a mean power of 19 KV were applied during one session. All patients required sedative and analgesic support. Pure pancreatic juice was collected via the nasopancreaticcatheterafter secretin stimulation (Sekretolin, Hoechst, Germany) 1 cu/Kg e.v. in 3 minutesbefore ESWL,shortly alter and 24 hours after. Lipoperoxidation products -coniugates diene (CD)- were measured, in pure pancreatic juice, as absorbance 246 nm and 232 nm in the chloroform phaseusing secondderivative ultraviolet spectroscopy. Results: total CD levels in pancreatic juice before ESWL were within the range of patients with CP, i.e. significantly higher than in non CP patients (mean value 0.0459 +_ 0.0091). Total CD levels significantly increased after ESWL (mean value 0.0598 -+ 0.0075) (p< 0.05) and after 24 hours ( mean value 0.0680 _+ 0.0084) (p
1140 Regulation of Glutathione Redox Status in Adapting Ileal Mucosa After Massive Small Bowel Resection in Rats

Uang Gu, Li H. Gu, Emory University School of Medicine, Atlanta, CA; Dean P. Jones, Emory University, Atlanta, CA; Aejaz Nasir, Robert R. Pascal, Nathan R. Miller, Thomas R. Ziegler, Emory University School of Medicine, Atlanta, GA Background/Objectives: Mechanisms underlying intestinal adaptation after bowel resection are unknown. In vitro studies demonstrate that the antioxidant glutathione (GSH) increases cell proliferation and protects against apoptosis in gut cell lines. Dietary glutamine (GLN) supplementationincreasesadaptiveintestinal growth after bowel resectionin rats and upregulates tissue GSH in models of catabolic stress. Our objectiveswere to investigateGSH redox status in adapting rat ileal mucosa after massive small bowel resection (SBR), with or without di~ary supplementation with I-GLN or alanyI-GLN dipeptide. Methods: Sprague-Dawleyrats (225 g) underwent80% SBR with anastomosisof proximal jejunum to distal ileum. Following SBR, rats were fed standard rat food or pair-fed isonitrogenous, isocaloric diets supplemented with either I-GLN or alanyl-GLN (4% of diet as GLN). Transected rats receiving standard diet served as controls. After 7-days, ileal mucosal crypt depth, villus height and GSH and GSSG (oxidized GSH disulfide) concentrationswere measured.The reduction potential of the GSSG/ 2GSH pool (Eh) in ileal mucosa was calculated using the Nernst equation. Results: Dietary

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