Histological analysis and cytoskeletal proteins expression on ileal mucosa before and after ileal pouch for ulcerative colitis

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89 HISTOLOGICAL ANALYSIS AND CYTOSKELETAL PROTEINS EXPRESSION ON ILEAL MUCOSA BEFORE AND AFTER ILEAL POUCH FOR ULCERATIVE COLITIS. BianconeL , Palmieri G*. Lombardi A”, Colantoni A*, Tonelli F”, Del Vecchio Blanco G, Mont&one I, Vavassori P., SpagnoliLG*, PalloneF Cattedradi Gastmentcmlogia,Dipartimentodi MedicinaIntoma e “Cane&a di Anatomia Patologica,Univasith di Roma ‘Tor Vergata”, Roma; “C&extra di Chimrgw Dipartunentodi FisiopatologiaClinica,Universita di Fireme, Firenze. Univemitl Tor Vergata,Roma Background. A humoral-mediatedimmune responseis a featureof Ulcerative Colitis (UC), associatedto serum andmucosal IgG againstseveralantigens,including p-ANCA and tropmnyosins (TM). TMs are actin-bindingproteinspresent in all eukmiotic cells. A local IgGmediatedautoimmunensponse againstTMs has beenpostulatedin UC. The role of TMs expression in the developmentofpachitis is unktwvn Aims. To assess,in a pmspective loneitudinal studv. the relation betweendeveloomentof mwhitis in UC and bistoloeicalchawzes in&ding TMs &&ssion, on ileal entemcytes:Me&& Patients.35 UC undergo& ” pmctocolstomy with ileal pouch were enrolled: I5 UC were longitudinally followed up before and &I surgery, with ileal bi&ies taken during and after 6 monthsof surg& (fmm thepouch); in 15 UC ileal biopsieswere taken only during surgery, while in 5 UC ileal biopsieswere taken only at 6 months.Rectal biopsieswere taken during surgery in 13/35UC. Controls. 2 familial adenomatous polyposis (FAP) with ileal biopsiestaken during and 6 months after surgery. Histology. Formalinfixed ileal and rectal sampleswere stainedby haematoxvlinand eosin(H&E) andby immunopemxidaseusing’enti-TM5 monocldnalantibody (IgG). Results. H&E s&ing. The diagnosisof UC was confinned in all rectal samples,while the ileal surgical sampleswere normal. At 6 months, biopsies from thepouch show& marked ileal changes,including disappearanceor shortheningof thewlli and developmentof “colon-like” crypts. This event appearednot relatedto the developmentof pouchhis, observedin I/3 of UC. Ip staining.In therectum, TMs were expressedOn the epithelialcells surface, both lining the lumen andm the crypts. In the surgical ileal samples,TMs were expressedat low gmde in the basalareaandnot on the surface of the @h&al cells. Al 6 months.TMs expression on the ileal mithelial cells showed marked chances.beine expressedon epithelialcellb surface,both hning ihe lumen and in the crypts. This e-&appeared relatedto the developmentof colon-like histological features,and not of wuchitis. Similar chanpcs observed in the ilet& from FAP. Conclusions.?he idealmucosa underg~morphologicalchanger in the pouch,developingfeatures similar to the colon. This event, and not tie developmentof pouchitx, apparsto be associatedto rearrangementsof the expressionof cytoskeletal proteins.

DOPPLER ENHANCEMENT AFTER INTRAVENOUS LEVOVISTL666: INJECTION IN CROHN’S DISEASE Di SabatinoA, Fulle I, Poti L, P.icevutiL, Ciccacioppo R, Tinomti FP, Tinoui S, Campani R, Corava GR. GastroenterologyUnit and Departmentsof Radiology and Pathology,IRCCS Policlimm S. Matteo, University of Paw, Italy. Paliclinico S.Melteo Background & Aims: Although tmnsabdombwIbowel sonography (TABS) has beenproposedas a reliable tool to assessincreasedbowel wall thickness (BWT), themost common sonogmphicpattem in patientswith C&n’s disease(CD), its accuracy is limited. We thereforetried to asseggwhether theuse in colour Doppler of Levovist, B galaefose-basedintravenoussonographiccordmst went able to enhancethe arterial Doooler sienal. increasesTABS accuracv. Patients&Methods: ‘&y-one patientswith ileal CD, di&&by &doscopy and entemclys&, and20 healthy volunteers,all sex- andage-matched,were examinedwith wnventional TABS. Colour Doppler of theinnetnu~a.lentmic vessels was thenperformed beforeand afla intmvenous irjection of Lwovist,. Diseaseactivity was assessedby Cmhn’s diseaseactivity index. Results: Twenty-two CD patients hadB BWT  4 mm, and I6 ofthem presentedan active disease. Two of the remaining 9 CD patients,all with BWT ɧM); 4 mm, presentedan active disease. By meansof colour Doppler we identified six patientswith inactive disease,normal BWT, and normalbasalDoppler signal intensity, who showedan enhancedDoppler signalin intramural vessels after contrast agentbolus application.Four of thesepatients,identified only by colour Doppler after Levovistl666; injectim relapsedwithin six months. In our hands,sensitivity and specificity of TABS, integratedwith additionalstimulatedacoustic emissionmode,were 96.7% and lOO%,respectively. Conclusions: The use of Levwistl666; in colour Doppla increasesthe accuracy of ultrasonogmphy in CD diagnosisand follow-up. Levovist, hasbeen shown particularly us&d in thoseinactive CD patientswith normal BWT and with an intramural small bowel flow not consistent enoughto make it possibleto separatethe bsc~und noise from the Doppler signals by basalacoustic emissionmode. In addition, our datasuggestthat an increased intnunurel flow evaluatedby wlour Doppler a.& Levovi: injection representsa predictive factor of impendingrelapsein patientsmth quiescentCD.

90 SERUM BASIC FIBROBLAST GROWTH FACTOR LEVELS CORRELATE WITH THE ULTRASOUND MEASURBMENTS OF BOWEL WALL THICKNESS (BWTI IN CROHN’S DISEASE (CD) Di SabatinoA, Ciccocioppo Q Cinque B, Fulle I, Morem R, Sivelli P, Campti R, Cifone MG, Caraza GR GestmentercdogyUnit and Departmentof Radio&y, IRCCS Policlinica S. Matteo, Univewty of Pavih Depts of Internal and ExperimentalMedicine,University of L’Aquila, Itiy. Policlinicn S.Matteo Sackgmund & Aims: Sines basic fibmbbwt gmwth factor (bFGF), ahepain-binding protein known to pmmote tissue repair in inflammation,can stimulatefibmblast proliferation, which may potentially play a mls in sbictwr. formation,we measuredserum bFGF levels in CD patients and correlatedthem to the BWT, assessedby ultwonog.mphy (US), to detrrmine whether szn,n, levels reflect fibmtic ectivity in CD. Patients& Methods Serumsample were obraincxlfrom 20 patients with ilcal CD, 20 !xtieots with ulcerative colitis fU0 and20 b&hv volunteera.all sex- andacematched.D&no& of CD andUC was establishedb> conventioml~clinicaland’bistopathological criteria Semm bFGF levels wex measuredby ELISA assay utilizing the Oumtikine HS Human bFGF immunoassay(R&D Systems,Minne&lis, Mbmes&), and&p&&d as paticnts were examinatedwith ~nventional US (Al&a SSD.1700;convex probe 5 MHZ). BWT up to nun wes consideredas normal. Diseaseactivih, was asses& bv C&n’s diseasea&vim index (CDAI). Results: In CD patientsserum bFGF Ieve& were sign&&y higher (mean52.2 p&l) competedto UC patients(18.6 p&nl, p
pglml. AllCD

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IMPAIRED TRANSGLUTAMINASE ULCERATIVE COLITIS

ACTIVITY

IN PATIENTS

WITH

G D’Argenio. N Della Valle, ‘G Di Malteo. V Cosenza, ‘P Giorgio, IF Jori. ‘G Salvatore, *G Peluso. G Mazzacca. Cattedra di Gastroenterolcgia e ‘Istolcga, Universitd Federico II, Napoli. ’ Sefvizio Endoscopia Digestwa. IRCCS S. De Sellis. Castellana Grow ‘IRCCS G. Pascale Napoli. Background 6 Aims. Transglutaminases are a family of enzymes that catalyze the extracellular matrix protein cross-linking and are involved in wound healing process. We previously demonswated a decreased activity of transglutaminase in colon (tTG) with local recruitment of plasma TG in rats with induced c&is inversely related to the histological grading. In the present study we evaluated plasma and colonic TO activity and fTG expression and dishibution in colon of healthy control and patients with ulcerative colitis (UC) Patiants 6 Methods. In 13 patients with UC (5 active and 6 inactive) and in 10 normal controls undergoing colonoscopy. endoscopic and histologic indices were studied. In plasma and colon biopsies. fTG adivity was evaluated by a radioenzymatic method. fTG protein content and distribution in colon were raspactively evaluated by Western blot analysis and immunoisbxhemishy using a specific anti-flG antibody. Mucosal fTG mRNA was analysecl by semiquantitative RT-PCR standardized with the internal control gene HPRT. Results. ffG activity was SignificantI; lower in both plasma and colon of patients with actw UC than those inactive and controls (‘pcO.05): I COntrOlS 1 InaCtiVe UC 1 ActiveUC 1 9.622.2 Plasma TG (mWml) I 12.1+1.7 1 4.2+2.4’ ColonTG (mu/g) 1 987+345 1 734+320 j 2Sg+107* Western blot analysis showed reduced fTG content in patients with active UC compared to those wi?h inactive disaase and controls; fTG lmmunostaining was mainly localized in damaged araas. In contrast. RTPCR showed upregulation of fTG mRNA expression in active UC (320 56 compared to controls). Conclusions. In active UC, the enhanced ITG mRNA expression, the flG staining of damaged areas and the reducad activity in plasma, are in keeping with the attempt of mucosal repair. In contrast. the reduction of protein content in colon suggest a post-transcriptional defect leading to impaired fTG activity and delayed healing. We postulate that therapies effective on the transcriptional regulation of this enzyme may contribute to improve the reparative response of the colonic mucosa in UC

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