M1274

Diclofenac Encephalopathy,Liver and Gastrointestinal Lesions in Rat andStable Gastric Pentadecapeptide BPC 157Spomenko Ilic, Domagoj Drmic, Danijela Kolenc, Marijana Coric, Luka Brcic, RobertKlicek, Bozo Radic, Marko Sever, Viktor Duzel, Marinko Filipovic, Mihovil Ivica, AlenkaBoban Blagaic, Tomislav Ani, Ivan Zoricic, Miroslav Gjurasin, Zeljko Romic, SenkaDzidic, Sven Seiwerth, Predrag Sikiric

Combined diclofenac encephalopathy, liver and gastrointestinal lesions have not yet beenestablished in rats. The stable gastric pentadecapeptide, BPC 157 (GEPPPGKPADDAGLV,MW 1419, efficient in inflammatory bowel disease trials (PL 14736) and various woundtreatment, no toxicity reported) is an anti-ulcer peptide with hepatoprotective effects thatmay also affectmany central disturbances. Diclofenac (12.5mg/kg) was given intraperitoneallyonce daily for 3 subsequent days. BPC 157 (10μg/kg, 10ng/kg) was given either (i) intraperi-toneally immediately after diclofenac or (ii) per-orally in drinking water (0.16 μg/ml, 0.16ng/ml) up until the end of the experiment. At 3 h following the last diclofenac challenge,we evidenced severe gastric, intestinal and liver lesions, increased bilirubin, AST, ALT serumvalues, liver weight, prolonged sedation/unconsciousness (after any diclofenac challenge)and finally (hepatic) encephalopathy (Fig.1, C (control) B (BPC 157)). Brain edema wasparticularly present in the cerebral cortex and cerebellum, more in white than in gray matter,damaged (balloonized) red neurons were particularly expressed in the cerebral cortex andcerebellar nuclei, Purkinje cells and less expressed in hippocampal neurons. This wasconsistently counteracted in diclofenac-rats that received BPC 157 (μg- or ng-regimen,intraperitoneally or per-orally). In conclusion, the successful counteraction of combineddiclofenac encephalopathy, liver and gastrointestinal lesions by BPC 157 regimens meansthat besides inflammatory bowel disease, diclofenac toxicity may be a new domain forpossible BPC 157 therapy.

M1275

Ibuprofen, Hepatic Encephalopathy, Hepatomegaly, Gastric Lesion and GastricPentadecapeptide BPC 157 in RatsSpomenko Ilic, Domagoj Drmic, Danijela Kolenc, Marijana Coric, Luka Brcic, RobertKlicek, Bozo Radic, Marko Sever, Viktor Duzel, Marinko Filipovic, Mihovil Ivica, AlenkaBoban Blagaic, Tomislav Ani, Ivan Zoricic, Miroslav Gjurasin, Zeljko Romic, SenkaDzidic, Sven Seiwerth, Predrag Sikiric

Stable gastric pentadecapeptide BPC 157 (GEPPPGKPADDAGLV, MW 1419) is an orallyactive anti-ulcer peptide that may also have an effect on many central disturbances, efficientin inflammatory bowel disease trials (PL14736) and various wound treatment, no toxicityreported, which showed various hepatoprotective effects. Therefore, we challenged rats withchronic ibuprofen application (0.4g/kg intraperitoneally once daily for 4 weeks) and BPC157 therapy. We evidenced a deleterious circuit not reported in ibuprofen-rats as yet: hepaticencephalopathy, gastric lesions, hepatomegaly (Fig.1, C (control) B (BPC 157)), increasedAST and ALT serum values, with prolonged sedation/unconsciousness and weight loss. Inparticular, ibuprofen toxicity exhibited itself as brain edema and cyanosis, particularly inthe cerebellum, more so in white than in gray mater, damaged (balonized) red neurons,without any inflammatory reaction, particularly in the cerebral cortex and cerebellar nuclei(Fig. 1), less in the hippocampus, dentate nucleus and Purkinje cells. BPC 157 ( 10μg,10ng/kg) was fully effective given as an antidote after ibuprofen: (i) intraperitoneally, immedi-ately after ibuprofen or (ii) in drinking water (0.16 μg, 0.16 ng/ml) until the end of the 4week-experiment. Thus, apart from its role as an anti-ulcer peptide, efficient in inflammatorybowel disease trials (PL14736) and various wound treatment, with no toxicity reported andwhich already showed various hepatoprotective effects, either BPC 157 regimen counteractedall of ibuprofen's adverse effects, such as hepatic encephalopathy, gastric lesions, hepatomeg-aly, increased liver serum values, and BPC 157 rats showed no behavioral disturbances andmaintained normal weight gain.

S-369 AGA Abstracts

M1276

Gastric Varices: Limitation of Treatment With Interventional Radiology (Ivr)From Study of Balloon DiameterShigeru Nakano, Yoshinori Fujituka, Teppei Matui, Katsuya Higami, Mie Shinohara,Hidenari Nagai, Koji Ishii, Yoshinori Igarashi, Yasukiyo Sumino

Background and Purpose: Balloon-occluded retrograde obliteration (BRTO) is the first-linetreatment for gastric varices. Although modern devices have been developed to enableembolization of large-diameter shunts, the outcomes of such treatment are still unclear. Weexamined long-term prognosis in patients undergoing BRTO using a new balloon catheter30 mm in diameter. Subjects and Methods: The study population consisted of 24 consecutivecases undergoing BRTO using 30-mm balloon catheters between January 2005 and March2007, when these catheters first became available. BRTO was applied for gastric varices (n=20), duodenal varices (n=1), and hepatic encephalopathy (n=3). These balloon catheterswere selected when occlusion could not be achieved using those 20 mm in diameter evenin deep vessels. Results: BRTO was indicated in 23 of 24 cases. The 30-mm balloonswere applied in 4 cases, including 3 cases of hepatic encephalopathy. The amounts of 5%ethanolamine oleate iopamidol (EOI) used and number of days required for treatment weresignificantly higher in the cases treated with the 30-mm balloons than those <20 mm indiameter (P<0.05). There we no cases of complications, such as refractory ascites. Onepatient died within two years of treatment with a balloon <20 mm in diameter. Among the4 patients treated with 30-mm balloons, two died of hepatic failure within 2 years after theoperation, and one patient with liver failure underwent living donor liver transplantation.Discussion: Patients with larger vascular diameter had shorter survival times after treatmentthan those with smaller diameter. As occlusion of the shunt was accomplished in all cases,improvement of the portal vein may not participate in prolonging survival time. Earlierocclusion may be more effective for liver reperfusion than larger vascular occlusion aftermanifestation of symptoms. Therefore, assessment of proper time for BRTO seems critical.

M1277

Is Cirrhosis a Risk Factor for Diabetes?SriLakshmi Narra, Jihad O. Arteh, Bradford Waters, Satheesh Nair

Aim: Compare the prevalence of Diabetes in patients with cirrhosis and a control group.Methods: 261 consecutive cirrhotic patients seen in the transplant clinic were categorizedas those with HCV(n=142)and those without HCV(n=119). The control group(n=204) com-prised of consecutive patients seen in the VA GI clinic. Patients with underlying liver diseaseand age>65 were excluded from the control group. Data included demographics, HgbA1cand need for insulin for glycemic control. A logistic regression was performed(age,gender,BMI,cirrhosis vs no cirrhosis as variables-stepwise forward entry)to determine if cirrhosiswas an independent risk factor for diabetes. A second logistic regression model includingonly cirrhotic patients was studied to determine if HCV was an independent risk factor fordiabetes. Results: The cirrhotic and control groups were similar in age and BMI. Controlgroup had a higher percentage of men, as expected of patients attending the VA clinic. Theoverall prevalence of Diabetes was higher in the cirrhotic group especially in non HCVcirrhosis(34.5% vs 28.3% in the control). These rates are higher compared to the estimatedprevalence of Diabetes in US population per NHANES, 10.8% in the 40-59y group and23.1% in the 60 and above group. Among diabetics, a higher percentage of cirrhotic patientsrequired insulin(46.4% in HCV cirrhotics, 55.9% in Non HCV cirrhotics vs 28.3% in thecontrol group;p value 0.03). These values are again higher compared to National Healthinterview data(in adults, Insulin alone was the modality of therapy in 14%). However, themean HbA1c was similar in all the groups. In a multiple logistic regression analysis, agealone was found to be an independent predictor for diabetes(OR 1.060 with 95% CI: 1.029-1.091, p value 0.0001). Cirrhosis was not an independent predictor. In a logistic regressionmodel, including only patients with cirrhosis, HCV was not an independent predictor forDM. Conclusion: Cirrhosis was not an independent risk factor for Diabetes in our study,with a relatively higher rate of Diabetes and Obesity. As for therapy, Insulin use was morecommon in patients with cirrhosis and may reflect difficult to control diabetes or poortolerance of oral hypoglycemic agents.

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M1278

Depression and Sensitivity to Stress Can Be Induced by Transient GastricIrritation in the Neonatal Period: Behavioral and Molecular CorrelatesLiansheng Liu, Qian Li, Mohan M. Shenoy, Tugba Colak, Kshama R. Mehta, Pankaj J.Pasricha

Background: Patients with functional dyspepsia (FD) tend to be more depressed and anxiousthan healthy controls but what comes first is not clear. We hypothesized that psychologicalmorbidity can be secondary to abnormal gastrointestinal afferent signaling. We thereforeevaluated depression-like and anxiety-like behaviors and the sensitivity to stress in a previ-ously described rat model of FD. Methods: 10 days old male Sprague-Dawley rats received0.2 ml of 0.1% iodoacetamide (IA) in 2% sucrose daily by oral gavages for 6 days. Controlgroup received 2% sucrose. At 8-10 weeks of age, sucrose preference test (SPT) and forced-swim test (FST) were used for testing depression-like behavior and elevated plus maze (EPM)and open field test (OFT) were used to test anxiety-like behaviors. Subsequently plasmaACTH, corticosterone and oxytocin levels in response to a minor stressor (subcutaneoussaline injection) were measured. RNA was extracted from various brain regions. Results:Rats with neonatal gastric irritation (NGI) showed several abnormalities on the FST ascompared with control (Immobility: 32.4± 3.95 vs. 12.95±2.80, climbing: 7.05± 1.18 vs.18.65 ±1.9 and swimming: 20.6± 3.02 vs. 28.35± 2.1, P <0.05). Sucrose preference (definedas > 75% of total intake) was decreased in NGI rats (45% versus 80%; p <0.05). Nosignificant change between NGI and control animals was observed in EPM test. On theother hand, OFT showed that the time at center and total distance moved were significantlydecreased in NGI rats compared to control rats (16± 4. vs 30±5 and 9182±655 vs 11548±332, P <0.05). The basal level of plasma corticosterone in NGI rats was significantly higherthan that in control rats (3.05±0.74 vs. 1.41±0.25, P<0.05) although stress did not inducea further elevation. Further, while stress did not alter the plasma ACTH and oxytocinconcentration in control rats, the hormone levels were significantly increased in stressedNGI rats compared to non-stressed FD rats (49.45±11.42 vs 22.76±2.05 and 14.87±1.50vs 7.07±0.62, P<0.05). Gene array data showed significant changes in key molecules thatmodulate stress and depression including 5-HT1A, 5-HT2A, TrkB, NPYr1 and oxytocin.Conclusions: These results suggest that transient gastric irritation in the neonatal periodcan induce long lasting depressive behavior as well as sensitivity to stress, associated withspecific molecular correlates. These findings have major implications for the pathogenesisand treatment of psychological co-morbidity in patients with FD. Thus, gastric hypersensitiv-ity could be a cause rather than an effect of depression.

M1279

Overlapped Heartburn/Regurgitation Amplifies Abdominal Pain andIndigestion via Common Central Serotonin Neurotransmission Disorders inPatients With Functional DyspepsiaKazunari Tominaga, Chikako Tsumoto, Suzuka Ataka, Hirohisa Machida, HirotoshiOkazaki, Hirokazu Yamagami, Tetsuya Tanigawa, Kenji Watanabe, Toshio Watanabe,Yasuhiro Fujiwara, Susumu Shiomi, Yasuyoshi Watanabe, Tetsuo Arakawa

Background & Aims:Heartburn/regurgitation often overlaps abdominal pain and indigestionand affects each other in functional dyspepsia (FD) patients. Thus, upper abdominal dyspepticsymptoms may be mediated by similar neurotransmission pathways based on the brain-gutinteraction. Central serotonergic abnormalities are associated with the pathophysiology offunctional gastrointestinal disorders or psychiatric depression and anxiety. To evaluate theroles of the cerebral serotonin (5-HT) neurotransmission systems in overlapped heartburn/regurgitation of FD patients, we examined both 5-HT transporter (5-HTT) binding potentialin the brain and correlation of 5-HTT binding potential to overlapped heartburn/regurgitationbetween FD patients and controls. Methods: Patients with FD diagnosed according to theRome III criteria (N=9, female: 6, age range 36-76 yrs) were recruited for this study. Therewere 8 healthy controls (female: 2, age range 25-61 yrs). To measure 5-HTT binding potentialin areas of the thalamus, putamen, amygdala, midbrain, and cerebellum (as a referenceregion), positron emission tomography (PET) with [11C]N,N-dimethyl-2-(2-amino-4-cyano-phenylthio) benzylamine ([11C]DASB), which binds specifically to 5-HTT, was performed.We used the Multi-linear Reference Tissue Mode method within the standard softwarepackage of PMOD Technologies for analysis of [11C]DASB with reference to the co-registeredMRI images. Abdominal symptoms reflux, abdominal pain, and indigestion were evaluatedon the Gastrointestinal Symptoms Rating Scale (GSRS). Results: All scores for heartburn/regurgitation, abdominal pain, and indigestion were higher for FD patients than for controls(p<0.01). In FD patients, the binding potential of [11C]DASB in the midbrain (p=0.041)and thalamus (p=0.031) was higher than in the corresponding areas in controls. Bindingpotential of [11C]DASB in the midbrain was correlated with reflux scores (p=0.012, r=0.568)as well as abdominal pain (p=0.050, r=0.410) scores and indigestion (p=0.029, r=0.475).In the thalamus, however, it was correlated with reflux scores (p=0.019, r=0.521) differentfrom a tendency to abdominal pain (p=0.051, r=0.401) scores and indigestion (p=0.062,r=0.384). Scores of abdominal pain and indigestion in FD patients overlappedwith heartburn/regurgitation were higher than those of FD patients without it (p<0.05). Conclusion: These

S-370AGA Abstracts

findings suggest that disorders of central 5-HT neurotransmission in the midbrain andthalamus are correlated with heartburn/regurgitation in FD patients, which may amplifyabdominal pain and indigestion symptoms.

M1280

Treating Chronic Dysphagia Post Stroke With Neurostimulation BasedInterventions: A Preliminary StudyEmilia Michou, Satish Mistry, Samantha Jefferson, Salil Singh, Shaheen Hamdy

Background:Oropharyngeal dysphagia after stroke has an incidence up to 45% and whenchronic, leads to increased risk of institutionalisation and death in the long term. PharyngealElectrical Stimulation(PES)[1],repetitive Transcranial Magnetic Stimulation(rTMS)[2]andPaired Associative Stimulation(PAS)[3]are three neurostimulation techniques developed toincrease cortical excitability of pharyngeal motor cortex, with therapeutic potential. Howeverthe effects of neurostimulation on chronic but stable dysphagia in stroke patients remainunknown. We therefore investigated the effects of these interventions in stroke patients withdysphagia persisting for more than 6 weeks.Methods:In 12 dysphagic stroke patients(69±9years old,79±25 weeks post stroke(mean±SD), 6 left, 5 right hemispheric, 1 undeter-mined)pharyngeal electromyographic responses were recorded using an intraluminal catheterafter the application of single TMS pulses over pharyngeal motor cortex, in order to measurecortico-bulbar excitability before, immediately and 30 minutes after real and sham applica-tions of either:a)PES(10 mins of 5Hz),b)rTMS(250 repeated TMS pulses at 5Hz),or c)PAS(10mins of repeated pairs of pharyngeal and TMS pulse every 20sec). In 9 subjects, swallowingperformance and safety(aspiration-penetration scores,APs) were assessed with videofluoro-scopy before and after both real and sham neurostimulation techniques.Results:Comparedto sham, the application of all neurostimulation interventions significantly increased thecortical excitability in the unaffected hemisphere by 54±17%(mean±SEM) immediately post(-repeated ANOVA,p<0.001,Time*Treatment,p=0.005) not observed in the affected hemi-sphere(p=0.12). In addition, after active neurostimulation, APs were reduced by-14±6%(mean±SEM)compared to sham(Wilcoxon's test,p=0.05). Moreover, the delaybetween the oral and pharyngeal phases of swallowing, important for swallowing safety,was shortened by -56±13%(mean±SEM)after real neurostimulation(Wilcoxon's test,p=0.01)compared to sham. There was no consistent difference in cortical or swallowing responsebetween the interventions.Conclusion:These preliminary results show that neurostimulationinterventions have beneficial neurophysiological and behavioural effects in chronic strokepatients with dysphagia, increasing cortical excitability and improving swallowing safety.These findings provide support for neurostimulation being a useful adjunct in swallowingrehabilitation, even in chronic neurologic illness, and lay the foundation for further clinicaltrials of these forms of intervention.References:1.Fraser et al,Neuron 2002,2.Jefferson etal,Gastro 2009,3.Singh et al,Gastro 2009.

M1281

Chronic Repeated Stress Restores Impaired Gastric Motility in Wild TypeMice, but Not Oxytocin Knockout MiceReji Babygirija, Jun Zheng, Mehmet Bulbul, Kirk A. Ludwig, Toku Takahashi

Background: Stress is highly associated with common gastrointestinal disorders. The non-apeptide oxytocin is known to facilitate anxiolysis and to attenuate stress responses viainhibiting corticotrophin-releasing factor (CRF) expression of the hypothalamus. We haverecently shown that impaired gastric motility observed in acute restraint stress was restoredfollowing chronic repeated stress in mice. Chronic repeated stress upregulates oxytocinmRNA expression at the hypothalamus, resulting in downregulation of CRFmRNA expression(Am J Physiol, in press). Oxytocin knockout mice (OXT-KO) have been widely used tostudy the central oxytocin signaling pathways in response to various stressors. Previousstudies have shown that OXT-KO had higher corticosterone release and greater anxietybehaviours, compared to the wild type mice (Prog Brain Res. 170, 53-64, 2008). Westudied the effects of acute stress and chronic repeated stress on solid gastric emptying andhypothalamic CRFmRNA expression in wild type andOXT-KOmice.Methods: Heterozygousparents (B6; 129S-Oxt tm1Wsy/J mice) were obtained from Jackson lab (Bar Harbor, Maine)and were bred in our animal facility. The off springs were genotyped using DNA preparedfrom tail extracts using polymerase chain reaction (PCR). Male OXT-KO, wild type andheterozygous littermates were used for the study. Solid gastric emptying was measuredfollowing acute restraint stress (for 90 min) or chronic repeated restraint stress for 5 consecut-ive days. The expression of CRF mRNA in the paraventricular nucleus (PVN) and supraopticnucleus (SON) was measured using real time RT-PCR. Results: There were no significantdifferences of gastric emptying in wild type (68.4±4.1%, n= 6), heterozygous (71.8±3.1%, n=6) and OXT-KO (70.6±3.1%, n=6) mice in non-stressed conditions. Acute stress significantlydelayed gastric emptying in these mice. The degree of delayed emptying was significantlylower in OXT-KO mice (29.8±1.2%, n=6, P<0.01) than that of wild type (40.8±0.8%, n=6) and heterozygous mice (35.8±1.2%, n=6). Following chronic repeated stress loading,gastric emptying was significantly restored in wild type (67.3±2.4%, n=6) and heterozygousmice (66.6±4.2%, n=6). In contrast, delayed gastric emptying was still observed in OXT-KO mice (34.7±1.3%, n=6) following chronic repeated stress. The increase in CRF mRNAexpression at the PVN and SON was much pronounced in OXT-KO mice, compared to thewild type mice following chronic repeated stress. Conclusion: These findings suggest thatcentral oxytocin attenuates CRF expression and plays a pivotal role in mediating the adapta-tion mechanism following chronic stress in mice.