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Tu2029

Does Dyssynergy on High Resolution Anorectal Manometry Correlate WithSpecific Symptoms?Daniela Jodorkovsky, Sameer Dhalla, Lisette Musaib-Ali, Ellen M. Stein, John O. Clarke

BACKGROUND: Although the pathophysiology and management of dyssynergic defecation(DD) differs from that of other types of constipation, it shares many of the same symptoms.The diagnosis of DD relies largely on anorectal manometry, defecography, or balloon expul-sion tests, which are not available in all practices. Ideally, a specific symptom from the historycould suggest DD, leading the physician to efficiently utilize these resources. METHODS: For5months, data was prospectively collected frompatients undergoing high resolution anorectalmanometry (HRARM) for an indication of constipation. Patients completed the validatedquestionnaire “patient assessment of constipation” (PAC-SYM) and a 7 item bowel habitsquestionnaire (BHQ) prior to HRARM. The BHQ was designed to further characterize theevacuation process such as need and frequency for digital extraction of stool (TABLE 1).HRARM was performed using a solid state catheter with 10 circumferential sensors spacedat 0.6cm intervals. Sensation was assessed by inflating the rectal balloon with saline at 10ccincrements. HRARMs were analyzed and mean anorectal pressures were collected. Presenceof a dyssynergy pattern (either paradoxical contraction or inadequate relaxation of theanal sphincter) was noted. Statistical analysis was performed using Spearman's correlationcoefficient and the point biserial correlation coefficient. RESULTS: Thirty patients presentingfor HRARM fully completed the questionnaires. The median age was 39, 83.4% patientswere female, 16.6% male, 70% were white, 23.3% black, and 6.7% other races. The meantotal PAC-SYM score was 21 (range 7-41) of which 0 is the minimum and 48 the maximumscore possible. The mean total BHQ score was 6.7 (range 2-12) of which 0 is the minimumand 28 the maximum score possible. On HRARM, a dyssynergy pattern was seen in 70%of the patients, of which 58.8%were characterized by paradoxical contraction of the sphincterand 41.2% by inadequate relaxation. Impaired rectal sensation was seen in 79% of patients.There was no statistically significant correlation between the total PAC-SYM score, totalBHQ score, or score from any individual question and the presence of dyssynergy. Therewas no significant correlation between the total PAC-SYM score, total BHQ score, or scorefrom any individual question and the percent anal relaxation during bear down maneuvers(TABLE 2). CONCLUSION: Even with the inclusion of a customized bowel habit question-naire that aimed totarget anorectal symptoms, a significant correlation between symptomsand dyssynergy could not be found. This highlights the importance of a focused rectal examand physiologic testing such as anorectal manometry in the diagnosis of DD.Bowel Habits Questionnaire (BHQ)

S-906AGA Abstracts

Representative sample of correlation coefficients (R) for presence of dyssynergy (point biserial)and percent anal relaxation (Spearman)

Tu2030

A Ghrelin Enhancer, Rikkunshito, Ameliorates Anorexia in Cachexia Model ofPulmonary FibrosisHironobu Tsubouchi, Shigehisa Yanagi, Koji Toshinai, Sachiko Mogami, Chihiro Yamada,Seiichi Iizuka, Masamitsu Nakazato

Background: In terminal stages of pulmonary fibrosis, dyspnea develops and transitions tocachexia associated with emaciation and muscle weakness. Weight loss is an importantprognostic factor, and treatment of cachexia is expected to improve patient outcome. Ghrelinnot only has an orexigenic effect but also has an anti-inflammatory effect and a secretoryeffect on growth hormone. Administration of ghrelin has been reported to improve musclewasting and pulmonary cachexia in patients (Chest 2005, 128:1187-1193) and in an animalmodel (Eur J Pharmacol. 2011, 672(1-3):153-158). Rikkunshito (RKT) is reported to improvethe suppressed ghrelin secretion via serotonin 2b/2c receptor antagonistic effects (Gastroen-terology 134:2004-2013) and ameliorate anorexia and muscle wasting in the cancer cachexiamodel (Biol Psychiatry 65:748-759). In this study, we examined (1) the relationship betweenghrelin and anorexia and (2) the effect of RKT on cachexia in mice with chronic respiratorydisturbance. Methods: Ten-week-old male C57BL/6J mice received endotracheal instillationof bleomycin sulfate (BLM, 3.0 mg/kg) on Day 0. Body weight and food intake were measureddaily and the weight of the gastrocnemius muscle was measured on Day 11. Plasma ghrelinlevels were measured by ELISA and compared to those of pair-fed control mice (mice givenan identical amount of food with the BLM mice) to eliminate the influence of the foodintake. On Day 11, exogenous ghrelin (300 nmo/kg, i.p.) was administered to control andBLM mice, and food intake was measured. RKT (1.0 g/kg, p.o.) was administered to BLMmice once daily (from day-3 to the day of autopsy). Results: Body weight, food intake, andgastrocnemius muscle weight was significantly decreased in the BLM mice compared to thecontrol mice on Day 11. In these mice, the plasma ghrelin concentration was significantlydecreased compared to that in the pair-fed control mice (control: 311±166 fmol/mL, BLM:126±32 fmol/mL; p<0.01). Suppression of food intake was significantly improved by exogen-ous ghrelin administration in BLM mice. RKT ameliorated the decreased body weight andfood intake and muscle weight were equivalent to levels in control mice. Plasma ghrelinlevel was significantly greater in the BLM + RKT mice than in the BLM mice. Conclusion:In this cachexia model of pulmonary fibrosis with significantly decreased plasma ghrelinlevels, RKT ameliorated the development of cachexia by increasing plasma ghrelin levels.Although the restoration by RKT was significant, the small changes in level indicate thatthe effect of RKT assumed to be also attributable to the direct serotonin 2b receptor antagon-istic effect in addition to improved ghrelin secretion.

Tu2031

Colon Polyps and Vitamin D Supplementation; a Meta-AnalysisSailajah Janarthanan, Anna G. Taranova

Colon polyps and Vitamin D supplementation; A meta-analysis. Introduction: Adenomatouspolyps are recognized as the precursor lesions for the vast majority of colorectal cancers.Adenomas are known to lead to colorectal cancers. A number of studies have found someassociation with Vitamin D supplementation and a reduction in colorectal adenoma formationand recurrence. But the data has not always been consistent. Consolidating the study datato come to conclusion will help with adenoma prevention recommendations. Methods Weconducted a comprehensive search for studies published up to April 2010, on PubMed andMedline. Studies that looked into vitamin D supplementation and colorectal adenoma were

eligible for analysis. We excluded studies that involved the development of colon cancerand hyperplastic polyps. Reviews of each studywere conducted and references were reviewed.The most adjusted risk estimates were extracted by two independent authors and summarizedusing random effects meta-analysis with Comprehensive meta-analysis software version 2.0.Results 6 studies were eligible for analysis. They combined a randomized controlled trial,case-control and cohort studies. Most studies showed a reduction in the incidence of colorectaladenoma. The overall summary risk estimate by a random effects meta-analysis for VitaminD supplementation and adenoma formation was RR= 0.821, 95% CI 0.675 to 0.997 (P <0.001). There was evidence of publication bias among the different studies included. Theheterogeneity was relatively low as shown by an I2 value of 38.75. Conclusion Our meta-analysis results suggest that when vitamin D is taken as a supplement it acts as a protectiveagent against colorectal adenoma formation.

Tu2032

Effect of β-(1,3-1,6)-D-glucan Against Irritable Bowel Syndrome-RelatedColonic HypersensitivityTeita Asano, Tohru Mizushima

Background: IBS is a functional gastrointestinal disorder characterized by chronic and recur-rent abdominal pain and discomfort (colonic hypersensitivity) that are associated with alteredbowel habits. Although IBS highly affects the quality of life of patients, the current clinicaltreatments for IBS patients have proved unsatisfactory. This is because abdominal painassociated with IBS (visceral pain) is difficult to treat. IBS is one of the most commongastrointestinal disorders. Therefore, considering health-care cost, its treatment by methodsother than medicines (such as supplements) is beneficial. β-glucans are naturally-occurringpolysaccharides found in the cell walls of yeast, fungi, cereal plants and certain bacteria. Assuggested by the fact that various foods contain β-glucans, they are known to have littletoxic and adverse effects. We succeeded in the purification and industrial-scale productionof low-molecular-weight β-(1,3-1,6)-D-glucan from Aureobasidium pullulans (A. pullulans)GM-NH-1A1 strain (LMW β-glucan). We here tested an idea that LMW β-glucan is effectivefor IBS, using animal models for IBS. Methods: Effect of oral administration of LMW β-glucan on mental stressor- or drug-induced alteration in defecation was examined. Effectof this administration on the electrical activity of abdominal muscle (visceromotor response(VMR)) in response to repeated colorectal distension (CRD) was also examined as a standardanimal model for abdominal pain (visceral pain). We also used butyrate enema-inducedhypersensitivity to CRD as an animal model of IBS. On the other hand, we used inflamedpaw pressure test, in which yeast solution was administered to the hind paw to induceinflammation and the pressure inducing pain response (pain threshold) was determined, asnon-visceral pain test. Results: Oral administration of β−glucan suppressed restraint stress-or drug-induced increase in fecal pellet output. β-glucan also suppressed visceral painresponse against colorectal distension. Enemas of butyrate stimulates VMR to CRD and wefound that when LMW β-glucan was orally pre-administered, VMR to CRD was similar tothat in control rats. Oral administration of LMW β-glucan did not affect the paw pressurerequired for a nociception response in both presence and absence of yeast injection, suggestingthat LMW β-glucan does not affect the pain response in general but specifically affectsvisceral pain response. Conclusions: The difficulty of therapeutic treatment of IBS attributesto that both abdominal pain and bowel habit disorders should be managed. Therefore,results in this study suggest that LMW β-glucan could prove therapeutically beneficial forthe prevention and treatment of IBS, especially for diarrhea-predominant IBS.

Tu2033

Electroacupunture Improves Glucose Tolerance in Mice With InsulinResistanceJieyun Yin, Nicola Abate, Jiande Chen

Background: Our previous studies showed electroacupuncture (EA) improved glycemiccontrol in a rodent model of diabetes. Ecto-nucleotide pyrophosphate phosphodiesterase(ENPP1) over-expression in adipocytes is an important mechanism of adipose tissue dysfunc-tion leading to adipocyte insulin resistance. It's unknown whether EA is able to improveglucose tolerance in a mouse model of insulin resistance. Aims: to investigate the effectsand mechanism of EA at CV4 and CV12 on intra-peritoneal glucose tolerance (IPGTT) andinsulin sensitivity in mice with ENPP1 over-expression. Methods: Sixteen wild type (WT)and eight adipose ENPP1 transgenic (TG) mice were divided into four groups: A: WT micefed with regular chow (n = 6); B: WT mice fed with high fat diet freely (n = 6); C: WTmice fed with high fat diet paired with TG mice (n = 4); D: TG mice fed with high fat dietpaired with group C (n = 8). The study was started after 12-wk feeding and at age of 20weeks old. All experiments were performed under anesthesia (1% isoflurane inhalation).IPGTT and insulin resistant test (ITT) were performed without EA and with EA (stimulatingfor 2 hours). IPGTT was conducted by intra-peritoneal injecting of 20% glucose then theblood glucose level was measured at 0, 15, 30, 60, 90, 120 min respectively. ITT followedthe same protocol except that insulin (0.5U/kg) was injected prior to the glucose injection.EA was performed using continuous pulses: 15Hz, 0.5ms, 6mA. Results: 1. In Group A,EA had no effects on glucose tolerance although there was a trend to decrease blood glucoseat 60min and 90 min (P = 0.08). 2. WT mice fed with high fat diet freely showed insulinresistance. In the control session ITT, the glucose level increased to 77.3 ± 7.3% at 30min,62.8±12.8% at 90min and 73.1±24.0% at 120min. EA significantly reduced glucose levelduring IPGTT from 90 min to 120min; the percentage of glucose increase was reduced from87.2±9.4% to 45.3±15.1% at 90min and from 79.8±37.4% to 37.4±19.2% (P < 0.04respectively). In addition, EA increased insulin sensitivity from 15min to 120min, representedby a substantial decrease of glucose level during ITT; the reduction was from 50% to over100%, P < 0.02 respectively. 3. Similar findings were observed in the group of WT micepaired fed with high fat diet. 4. EA reduced blood glucose in TG mice from 90min to120min; however the reduction was smaller than what was found in the WT mice withhigh fat diet. The reduction was 34% at 60min and 38% at 90 min (P <0.01). However,EA had no effects on insulin sensitivity in TG mice (P > 0.2). Conclusions: EA at CV4 andCV12 improves glucose tolerance in mice with insulin resistance induced by high fat diet.

S-907 AGA Abstracts

The inhibitory effect of EA on blood glucose is probably attributed to the improvement ininsulin sensitivity. Further studies are needed to explore the mechanisms.

Tu2034

Berberine Inhibits the Growth of Human Pancreatic Cancer Cells In Vitro andIn VivoKrisztina Kisfalvi, James Sinnett-Smith, Steven H. Young, Guido Eibl, Enrique Rozengurt

Background: Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease, with overall5-year survival rate of only 3-5%. As the current therapies offer very limited survival benefits,novel therapeutic strategies are urgently required to treat this aggressive disease. Recentstudies have elicited intense interest in metformin, the most widely used drug in the treatmentof type-2 diabetes, in the prevention and treatment of PDAC and other GI cancers. Metforminindirectly activates AMP kinase (AMPK), which negatively regulates mTORC1, but otherAMPK-independent targets have been proposed. If AMPK activation mediates the inhibitoryeffects of metformin on mitogenic signaling in PDAC cells, structurally unrelated compoundsthat induce AMPK activation should be expected to inhibit mTORC1 activation and prolifera-tion in these cancer cells. Berberine is a natural alkaloid found in many plants of the Berberisfamily and has been safely administered to humans for lowering blood sugar levels in Type2 diabetic patients in Australia and Asia. Berberine has been shown to activate AMPK inmany cell types. Here, we examined the hypothesis whether berberine has any inhibitoryeffect on human pancreatic cancer. Results: Treatment of PDAC cells (Panc-1, MiaPaCa-2)with increasing doses of berberine (0.05-5mg/ml) for 24 h decreased mitochondrial mem-brane potential, intracellular ATP levels and induced AMPK activation, as shown by immuno-blotting using antibodies that detect the phosphorylated state of acetyl-CoA carboxylase(ACC) at Ser79, a marker of AMPK activity within intact cells. Berberine dose-dependentlyinhibited p70S6K (a marker of mTORC1) activity. This natural compound also reduced theDNA synthesis in PDAC cells stimulated with insulin, neurotensin or 1-5% fetal bovineserum (FBS) and the anchorage-dependent proliferation of PDAC cells stimulated by insulinand neurotensin or 1-2% FBS for 5 days. In order to test the effect of berberine In Vivo,MiaPaCa-2 cells were implanted into the flanks of nu/nu mice. After the tumors reached a2mm diameter animals were treated with daily intraperioneal injections of 5mg/kg berberineor vehicle (n=10 for each group). The sizes of the subcutaneous tumors were measuredevery 4 days. Berberine treatment strinkingly reduced tumor growth (p=0.001) After 28days, the final volumes of the tumors were 824±158 and 242±38 mm3 in the control andberberine-treated groups, respectively. There were no evident macroscopic toxic effectsproduced by berberine treatment. Conclusion: Berberine significantly inhibits human pan-creatic cell growth In Vitro and In Vivo, in mice. The mechanism of action of berberineappears to involve the activation of AMPK and inhibition of mTORC1. As berberine hasbeen safely used in humans it could be a potential new candidate for the treatment strategiesagainst pancreatic cancer.

Tu2035

High-Fat Feeding Increases Recruitment of ZO-1+Ocldn+CD11c+ DendriticCells to the Lamina Propria and Accelerates Disease Onset in a Murine Modelof Crohn's Disease-Like IleitisLisa Gruber, Dirk Haller

Background & Aim. Recent scientific findings link reduced gut barrier function to obesity-associated inflammatory co-morbidities, but there is little evidence linking high-fat diet tointestinal inflammation per se. The aim of this study was to elucidate the connection betweendiet-induced obesity, intestinal barrier functions and inflammatory processes in the intestine.We investigated the effect of high-fat feeding on the pathogenesis of intestinal inflammationin a mouse model of Crohn's Disease-like inflammatory bowel disease, the TNFΔARE/Wtmouse. Methods. TNFΔARE/Wt and wildtype BL/6 littermates were fed either a high-fatdiet (48kcal% from fat) or a control diet (12kcal% from fat) from weaning until the age of8 and 12 weeks. Histopathological analysis of distal ileum was performed and expressionof markers related to barrier integrity and immune cell recruitment was investigated bywestern blot and qPCR analysis of primary ileal epithelial cells. Immunofluorescence tech-niques were used to study tight junction protein expression of epithelial cells and of leukocytesin the lamina propria of the small intestine. Results. Expression of the tight junction proteinOccludin (Ocldn) was dramatically reduced in the ileal epithelium of non-inflamed BL/6wildtype mice fed the high-fat diet. Consistent with increased expression levels of the DCchemokine CCL20 and the adhesion molecule ICAM-1 in ileal epithelial cells, recruitmentof ZO-1+Ocldn+CD11c+ dendritic cells into the lamina propria and epithelium was increasedunder high-fat feeding conditions. Furthermore, high-fat compared to control diet acceleratedileal inflammation in TNFΔARE/Wt mice at 8 and 12 weeks of age independent of significantoverweight or changes in fat tissue morphology. Tissue pathology correlated with the recruit-ment of ZO-1+Ocldn+CD11c+ dendritic cells into the inflamed lamina propria and epithelium.Conclusion. High-fat feeding reduced the expression of the barrier protein Ocldn in epithelialcells and induced the recruitment of ZO-1+Ocldn+CD11c+ dendritic cells to the laminapropria. Furthermore, the high-fat feeding facilitated disease initiation in the TNFΔARE/Wtmodel of Crohn's Disease-like ileitis. These findings point out that diet rich in fat mayaggravate intestinal inflammation and that this correlates to alterations of the antigen pre-senting cell populations present in the lamina propria and their possible interaction withintestinal epithelial cells.

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