Falk Symposium

158

Intestinal Inflammation and Colorectal Cancer March 23–24, 2007 Hotel Meliá Sevilla

Abstracts Poster Abstracts

Abstracts of Invited Lectures Poster Abstracts

Falk Symposium 158

INTESTINAL INFLAMMATION AND COLORECTAL CANCER

Sevilla (Spain) March 23–24, 2007

Scientific Organization: C. Gasche, Vienna (Austria) M. Gassull, Badalona (Spain) J.M. Herrerίas Gutiérrez, Sevilla (Spain) E. Monterio, Lisbon (Portugal)

CONTENTS

Page Session I

Epidemiology of the villain Chair: M. Gassull, Badalona J. Schölmerich, Regensburg Intestinal inflammation and colorectal cancer: Magnitude worldwide and its implications C. Pox, W. Schmiegel, Bochum

19–20

Nutritional habits and environmental clues M. Cravo, Lisbon

21–22

IBD high risk groups F. Magro, Porto

23–25

Session II

Not all colorectal cancers are the same Chair: C.R. Boland, Dallas F. Portela, Coimbra Getting familiar with familial colon cancer J. Sabates-Bellver, E. Cattaneo, K. Heinimann, J. Jiricny, G. Marra, Zurich, Basel

29

Cytoskeletal regulation by the adenomatous polyposis coli protein in interphase and mitosis I.S. Näthke, K. Kroboth, I. Newton, Z. Li, D. Dikovskaya, P. Appleton, D. Schiffmann, L. Leung, A. Quyn, J. Zumbrunn, K. Kita, C.M. Waterman-Storer, Dundee, Allschwil, La Jolla

30

Different mechanism – Different prognosis J.M. Carethers, La Jolla

31

IBD-related cancer – Just the same as sporadic? Pro J.M. Rhodes, Liverpool

32–34 3

Con: Colitis-associated colon cancer is different from sporadic colon cancer S.H. Itzkowitz, New York

35–36

Session III

Screening and surveillance Chair: T.A. Barrett, Chicago P.A.O. Fidalgo, Lisbon Latest endoscopic guidelines: FAP, HNPCC, IBD, and general D.T. Rubin, Chicago

39–42

What new techniques will replace classical surveillance? R. Kiesslich, Mainz

43

How to deal with dysplasia and adenomatous polyps in IBD? P. Chaves, P. Borralho, Lisbon

44

Session IV

Taking dysplasia to the molecular level Chair: C. Gasche, Vienna A. Matilla, Malaga

4

Inflammation-driven carcinogenesis – The players L.J. Hofseth, Columbia

47

Cancer originating from bone marrow-derived stem cells (BMDCs) T.C. Wang, New York

48

NF-κB signaling as a link between colonic inflammation and colorectal cancer M. Karin, La Jolla

49

Translating laboratory findings into clinics: Of mice and man M.F. Neurath, Mainz

50

Session V

How can we prevent? Chair: J.M. Herrerίas Gutiérrez, Sevilla A.S. Peña, Amsterdam What to eat or not to eat M. Gassull, Badalona

53

Inner and outer environment X. Llor, Chicago

54

Aspirin, NSAIDs and COX-2 inhibitors A. Lanas, Zaragoza

55–56

Use of 5-ASA, AZA, and 6-MP and the development of colorectal cancer in IBD F. Velayos, San Francisco

57

Results of a phase I multiple-dose clinical study of ursodeoxycholic acid L.M. Hess, M.F. Krutzsch, J. Guillen, H.-H. Sherry Chow, J. Einspahr, A.K. Batta, G. Salen, M.E. Reid, D.L. Earnest, D.S. Alberts, Tucson, Newark, Buffalo

58

and Phase III trial of ursodeoxycholic acid to prevent colorectal adenoma recurrence D.S. Alberts, M.E. Martínez, L.M. Hess, J.G. Einspahr, S.B. Green, A.K. Bhattacharyya, J. Guillen, M. Krutzsch, A.K. Batta, G. Salen, L. Fales, K. Koonce, D. Parish, M. Clouser, D. Roe, P. Lance, Tucson, Newark

59

Session VI

Molecular actions of 5-ASA – The magic bullet Chair: F. Carballo, Murcia W. Kruis, Cologne Replication fidelity C. Gasche, Vienna

63

Molecular actions of 5-ASA on PPARγ P. Desreumaux, Lille

64 5

Nonsteroidal anti-inflammatory drugs and Wnt signalling in colorectal cancer M.P. Peppelenbosch, C.L. Bos, M. Comalada, D.J. Richel, Groningen, Amsterdam, Granada

65

The salicylate derivative 5-aminosalicylate (5-ASA) affects cell cycle progression in colorectal cancer cells by activating a p53- and MMR-independent, reversible replication checkpoint M.G. Luciani, C. Campregher, C. Gasche, Vienna

66

Chemopreventive role of mesalazine in IBD-associated CRC: The role of DNA methylation A. Goel, Dallas

67

Molecular actions of 5-ASA: The magic bullet on EGFR signalling D. Fina, R. Caruso, S. Bellinvia, F. Pallone, M. Fantini, Rome, Milan

68–69

Session VII

State-of-the-Art Lecture Chair: E. Monteiro, Lisbon Infection, inflammation and cancer – The future C.R. Boland, Dallas

73

Session VIII

Progression of cancer through inflammatory mechanisms Chair: P. Figueiredo, Coimbra W. Mikulits, Vienna Signaling in skin and colon cancer (No abstract) W. Birchmeier, Berlin

6

Migrating cancer stem cells – An integrated concept of malignant tumour progression T. Brabletz, Erlangen

77

The inflammatory cytokine network and cancer promotion F. Balkwill, London

78

The balance between survival and apoptosis I. Fabregat, Barcelona

79

“The tumor is surrounded by the patient” – Novel approaches to translate laboratory findings to the clinic I.P. Witz, Tel Aviv

80

List of Speakers, Moderators and Scientific Organizers

81–84

7

Poster Abstracts I

Risk Factors and Epidemiology of IBD

1.

The prevalence of celiac disease in inflammatory bowel disease S. Demir, Z. Akpinar, A. Kirci, F.O. Ari, C. Cevik, E. Alper, K. Aksoz, Z. Buyrac, G. Yoruk, N. Yazicioglu, B. Unsal (Izmir, TR)

2.

The epidemiologic and clinical features of inflammatory bowel disease patients B. Unsal, S. Demir, Z. Akpinar, A. Kirci, F.O. Ari, C. Cevik, E. Alper, K. Aksoz, Z. Buyrac, G. Yoruk, N. Yazicioglu (Izmir, TR)

3.

Abdominal tuberculosis as a mask of chronic inflammatory bowel diseases O.L. Arjamkina, L.N. Savonenkova (Ulyanovsk, R)

4.

Inflammatory bowel disease in children in Shropshire, UK – An audit against national standards N. Ayub, A. Jan, S. Abdu (Shrewsbury, GB)

5.

Frequency and risk factors of major acute hemorrhage in Crohn's disease S. Seijo-Rios, M. Barreiro-de Acosta, A. Lorenzo-González, J.E. Dominguez-Muñoz (Santiago de Compostela, E)

6.

The nutritional status in patients with nonstricturing-nonpenetrating and stricturing behaviour of Crohn disease J. Budzynski, M. Klopocka, K. Grad, M. Swiatkowski (Bydgoszcz, PL)

7.

Time of evolution for the diagnosis of the inflammatory bowel disease N.C. Schneider, M.B. Machado, L. Guedes (Porto Alegre, BR)

8.

Crohn disease staging according to the Classification of Vienna 1998: Experience in the South of Brazil M.B. Machado, N.C. Schneider, L. Guedes (Porto Alegre, BR)

9.

Hepatobiliary manifestations in 1178 patients with inflammatory bowel disease S. Disibeyaz, M. Arhan, A.S. Koksal, E. Parlak, A. Ulker (Ankara, TR)

10. Analysis of exon 3 and 10 in human DLG5 gene among Polish patients with Crohn disease A. Dobrowolska-Zachwieja, M. Kaczmarek, J. Hoppe-Golebiewska, L. Jakubowska, R. Somski (Poznan, PL) 11. Evaluation of the nutritional status in children with Lesniewski-Crohn disease in relation to the mutation of the gene NOD2/CARD15 U. Grzybowska-Chlebowczyk, H. Wos, A.L. Sieron, M. Kajor (Katowice, PL)

9

12. Mycobacterium avium subsp. paratuberculosis in ulcerative colitis and Crohn's disease and environmental factors in Poland R. Marciniak, A. Szkaradkiewicz, A. Wasilewska, I. Chudzicka-Strugala, P. Majewski, M. Drews (Poznan, PL) 13. Retrospective study on epidemiology of ulcerative colitis in children in Southern Poland K. Przybyszewska, U. Jedynak-Wasowicz, A. Wedrychowicz, K. Fyderek, M. Sladek, S. Pieczarkowski (Cracow, PL) 14. The characteristics of a selected ulcerative colitis population M. Rimbas, S. Bucurica, M. Zota, M.R. Voiosu (Bucharest, RO) 15. Nutritional habits and malnutrition: Survey concerning the consumption of nutritional antioxidants and metal elements in cases of IBD patients K. Szentmihályi, O. Dörnyei, A. Kovács, E. Székely, E. Pintér, J. Fodor, Z. May, E. Dinya, A. Blázovics (Budapest, H) 16. Non-invasive recognition of pouchitis – A comparison of different methods R. Marciniak, T. Banasiewicz, J. Walkowiak, M. Drews, K.-H. Herzig (Poznan, PL; Kuopio, SF) 17. The comparison of clinical effect of mesalazine applied orally and rectally in patients with proctitis ulcerosa K. Grad, J. Budzynski, M. Klopocka, M. Swiatkowski (Bydgoszcz, PL) 18. Comparison of infliximab impact on the course of perianal and nonstricturingnonpenetrating Crohn disease - An own experience M. Klopocka, J. Budzynski, K. Grad, M. Swiatkowski (Bydgoszcz, PL) 19. Serological determination in inflammatory bowel disease M.B. Machado, L. Guedes, N.C. Schneider (Porto Alegre, BR) 20. Evaluation of the inflammatory bowel disease activity by the color Doppler ultrasonography L. Guedes, N.C. Schneider, M.B. Machado, M. Baldisserotto, L.A. Scaffaro, L. Grillo (Porto Alegre, BR) 21. Azathioprine: Who? When? Why? N.C. Schneider, M.B. Machado, L. Guedes (Porto Alegre, BR) 22. Could be anatomic extent of colitis and disease severity predictors for pouchitis after restorative proctocolectomy? G. Ianosi, D. Mercut, D. Neagoe (Craiova, RO) 23. The use of ozonized physiologic saline in patients with ulcerative colitis N. Antonova, S. Oprea, I. Butorov, N. Bodrug (Chisinau, MOL)

10

24. Doppler ultrasound measurements of the superior mesenteric artery predicts response to infliximab in patients with Crohn's disease F. Terracciano, F.R. De Filippo, G. Biscaglia, G. Forte (Caserta, I) 25. Cytokines in children with inflammatory bowel disease O.A. Tutina, E.N. Fedulova, V.N. Kopeikin, I.V. Mayanskaya, N.I. Tolkacheva (Nizhny Novgorod, R) II

IBD-Associated Cancer and Prevention

26. Protective effect of supplemental calcium lactate vs. carbonate in experimental colon carcinogenesis S. Belbraouet, X. Pelletier, G. Debry (Moncton, CDN; Nancy, F) 27. Automated classification of colon biopsy specimen by mRNA expression profiles in combination with multivariate statistical and neural network methods B. Molnár, O. Galamb, A. Nemeth, M. Juhasz, E. Dinya, K. Toth, N. Solymosi, Z. Tulassay (Budapest, H) 28. Dysplasia in inflammatory bowel disease in Romania S. Bataga, I. Torok, D. Georgescu, T. Bataga, A. Bratu (Tirgu Mures, RO) 29. The frequency and risk factors for dysplasia in Crohn's disease R. Vadan, C. Gheorghe, G. Becheanu, L. Gheorghe, M. Dumbrava, M. Diculescu (Bucharest, RO) 30. Fistula-associated carcinoma in Crohn's disease H. Vogelsang, F. Herbst (Vienna, A) 31. Systematic administration of 5-ASA derivatives during remissions for more than 9 months/year seems to protect patients with longstanding ulcerative colitis against colorectal cancer E.F. Georgescu, M. Georgescu, M. Vasile (Craiova, RO) 32. Implication of observation of patients with ulcerative colitis considering the occurrence of colorectal carcinoma M. Konecny, J. Ehrmann, V. Prochazka, V. Kucerova, K. Vyslouzil (Olomouc, CZ) 33. Colorectal cancer prevention in inflammatory bowel disease according to evidence-based medicine D. Ledro-Cano (Pilas, E) 34. Risk for colorectal cancer in inflammatory bowel disease P. Szanto, D. Manciula, C. Cruciat, A. Seicean (Cluj-Napoca, Baia-Mare, RO) 35. Clinical features and pathogenesis in inflammatory bowel disease-associated intestinal cancer T. Molnar, I. Németh, Z. Szepes, F. Nagy, L. Tiszlavicz (Szeged, H) 11

36. In what degree is the inner and outer environment affecting the developing of the colon cancer? L. Tiurean, C.E. Rezi, R. Mihaila, M. Deac, R. Mihaila, O. Petrascu (Sibiu, RO) 37. Early premalignant and malignant lesions: Detection of colon rectosigmoideum by magnifying coloscopy and chromoendoscopy Pa. Wohl, E. Honsova, Pe. Wohl, J. Spicák (Prague, CZ) 38. Methods of improving the yield of lesions at surveillance colonoscopy C. Somesundrum, B. Papadi, K. Besherdas, N. Van Somerun, R. D´Souza (Enfield, GB) 39. Diagnosis of dysplastic lesions in cases of ulcerative colitis A. Ioffe, N. Denisov, R. Abu Shamsieh (Kiew, UKR) 40. Special endoscopic methods for tattooing and localizing early colorectal cancers before laparoscopic colon resection A. Szijártó, J. Wacha, P. Kupcsulik (Budapest, H) 41. Narrow-band imaging (NBI) differentiation of polyps in the patients with ulcerative colitis M. Szura, A. Zajac, R. Solecki, J. Kulig (Cracow, PL) 42. Nimesulid impact on the expression of colorectal cancer epithelial markers in patients with ulcerative colitis after liver transplantation for primary sclerosing cholangitis Pa. Wohl, E. Honsova, A. Lodererova, E. Sticova, Pe. Wohl, P. Halakucova, J. Spicák (Prague, CZ) 43. Oxidative DNA damage levels are increased in sporadic colorectal cancer J. Sheridan, L.-M. Wang, M. Tossetto, K. Sheahan, J. Hyland, D.P. O'Donoghue, H. Mulcahy, J. O´Sullivan (Dublin, IRL) 44. Expression, localization and systemic concentration of transforming growth factor-beta1 (TGF-β1) and its receptors in patients with ulcerative colitis A. Stadnicki, G. Machnik, A. Wolanska-Karut, B. Bojko (Sosnowiec, Katowice, PL) 45. Can the statins block in an efficient way the progression of the cellular cycle, preventing in this way the colorectal cancer? R. Mihaila, C.E. Rezi, L. Tiurean, R. Mihaila, M. Deac, O. Petrascu, L. Bera (Sibiu, RO) 46. Particular aspects of premalignant lesions in inflammatory bowel disease: Morphological study O. Fratila, A. Maghiar, C. Brisc, D. Puscasiu, M. Puscasiu (Oradea, RO)

12

III

Colorectal Cancer: Detection and Treatment

47. Effectiveness of the questionnaire investigations in the group of patients with high risk of the hereditary colorectal cancers T. Banasiewicz, P. Golusinski, M. Grochowalski, A. Niziolek (Poznan, PL) 48. Development and validation of colorectal cancer-specific quality of life questionnaire R. Harisi, G. Bodoky, P. Kupcsulik, J. Weltner (Budapest, H) 49. Alimentation particularities at patients with colorectal cancer I. Marincu, M. Cornianu, C. Oancea, L. Marincu (Timisoara, RO) 50. Screening for colorectal cancer with fecal occult blood test (FOBT) in Romania C. Pojoga, O. Pascu (Cluj-Napoca, RO) 51. Risk factors contributing to the development of colorectal cancer N.I. Potaturkina-Nesterova, S. Nesterov, A.S. Nesterov, Y.Y. Krasnoperova (Ulyanovsk, R) 52. Risk factors of colorectal cancer in Latvian Chernobyl clean-up workers J. Seleznovs, J. Pokrotnieks, G. Orlikovs, A. Skesters, T. Farbtuh (Riga, LV) 53. Colo-rectal cancer screening in Romania S. Bataga, I. Torok, A. Bratu, S. Mocanu, D. Georgescu, T. Bataga (Tirgu Mures, RO) 54. Examination of metabolizing enzymes in patients with colorectal cancer P. Varga, I. Kiss, I. Ember, R. Schnabel (Budapest, Pécs, H) 55. Liver metastases in the HBs antigen positive patient with colorectal cancer T. Banasiewicz, J. Paszkowski, P. Pyda, M. Drews (Poznan, PL) 56. How to prevent colorectal cancer - Early detection of polyposis syndromes T. Banasiewicz, J. Paszkowski, A. Plawski, R. Slomski, P. Krokowicz, M. Drews (Poznan, PL) 57. Difficulties in implementing a preventive strategy for colorectal cancer in patients with adenomatous polyps D.L. Dumitrascu, D. Dumitrascu, L. Nedelcu, L. David (Cluj-Napoca, Brasov, RO) 58. Can the long-term use of aspirin and other non-steroidal anti-inflammatory drugs protect against the colorectal cancer? C.E. Rezi, R. Mihaila, L. Tiurean, R. Mihaila, M. Deac, O. Petrascu (Sibiu, RO) 59. Endoscopic therapy of early colorectal carcinoma by EMR C. Banciu, L.M. Susan, V. Vacariu, I. Romosan (Timisoara, RO)

13

60. The importance of the fecal occult blood test in detecting colon tumors C. Banciu, L.M. Susan, O. Chirileanu, I. Romosan (Timisoara, RO) 61. Palliative endoscopic treatment of obstructive colorectal carcinoma Prospective pilot study C. Banciu, O. Chirileanu, A. Pacurari, I. Romosan (Timisoara, RO) 62. Are the polyps and cancers of the colon different under and above 50 years of age? Five years results of colonoscopy in our department T. Gyökeres, A. Szakács, J. Hamvas, K. Illés, F. Kovács, M. Varsányi, I.G. Takács, A. Pap (Budapest, H) 63. Endoscopic polypectomies of colorectal adenomatous polyps - Prospective evaluation of material resources needed for colonoscopic surveillance in a tertiary gastroenterology referral center R. Iacob, I. Bancila, B. Cotruta, D. Bucur, D. Voinea, S. Iacob, L. Gheorghe, M. Manuc, D.C. Pitigoi, T. Marinescu, M. Diculescu, C. Gheorghe (Bucharest, RO) 64. The prospective analysis of material resources needed for colonoscopic surveillance in patients with curative resection for colorectal cancer S. Iacob, R. Iacob, I. Bancila, B. Cotruta, D. Bucur, D. Voinea, L. Gheorghe, A. Croitoru, I. Ciurea, M. Manuc, D.C. Pitigoi, T. Marinescu, M. Diculescu, C. Gheorghe (Bucharest, RO) 65. The role of genetic counselling in children with familial polyposis syndromes Own material I. Kubinska, E. Czkwianianc, A. Plawski, R. Makosiej, A. Durko, E. Malecka-Panas (Lódz, Poznan, PL) 66. Postoperative surveillance of colorectal cancer: Is intensive follow-up efficient? C. Searpe, V. Sbarcea, A. Rosu, A. Gavrila, C. Sbarcea (Craiova, RO) 67. Feasibility study of colon polyp computerized detection in electronic cleansed virtual colonoscopy Z. Tarjan, A. Tombácz, J. Koloszar (Budapest, H) 68. Association between the GST-pi expression and microvessel density and their combined prognostic significance in primary colorectal carcinoma T. Vlaykova, M.V. Gulubova, D. Vlaykova, G. Cirovski, I.M. Manolova (Stara Zagora, BG) 69. Detection of endocrine cells and their prognostic significance in primary colorectal carcinoma M.V. Gulubova, T. Vlaykova, I.M. Manolova (Stara Zagora, BG) 70. The effect of mesalazine on bacterial translocation in experimental colon anastomosis A. Aslan, M. Temiz, M. Cetin, C. Tümer, H. Bozkurtoglu, E. Canbolant, R. Gonenci (Antakya-Hatay, TR) 14

71. Important bioactive small molecules of erythrocytes in colorectal cancer patients after colectomy A. Blázovics, A. Szilvas, E. Székely, G. Székely, E. Tordai, E. Sárdi (Budapest, H) 72. Serrated adenomatous colonic polyps - Morphological particularities in a series of 12 cases M. Dumbrava, G. Becheanu, M. Manuc, C. Gheorghe, A. Preda, V. Serban, M. Diculescu (Bucharest, RO) 73. Histopathology is a strong prognostic factor in colorectal cancer C. Searpe, V. Sbarcea, A. Rosu, A. Gavrila, C. Sbarcea, M. Tenovici (Craiova, RO) IV

Molecular Mechanisms and Arrays

74. Mesalazine counteracts the mutagenic effect of the intercalating agent 9-aminoacridine C. Campregher, M.G. Luciani, C. Gasche (Vienna, A) 75. Detection of p53 gene Arg72Pro polymorphism and correlation with clinical characteristics in ulcerative colitis L. Caserta, G. Riegler, M.T. Vietri, A.M. Molinari, M. Cioffi (Naples, I) 76. Involvement of proapoptotic and antiapoptotic Bcl-2 family members in colorectal cancer K. Guzinska-Ustymowicz, J. Czyzewska, J. Sieczka, M. Niksa, A. Chetnik, V. Dymicka-Piekarska, A. Kemona (Bialystok, PL) 77. Beta-catenin and c-Met expression in inflammatory bowel disease A.V. Ivanova, R. Nikolov, A. Deredjan, Z. Spasova, M. Velev, D.G. Adjarov, Z. Krastev (Sofia, BG) 78. The role of mast cells and neuropeptides on stress and barrier function of the follicle-associated epithelium in rats A.V. Keita, A.-C. Ericson, J.D. Söderholm (Linköping, S) 79. 5-Aminosalicylic acid interferes with cell cycle progression of colorectal cancer cells P.J. Koelink, M.A.C. Mieremet-Ooms, W.E. Corver, A.M. Mommaas, G. Griffioen, C.B.H.W. Lamers, D.W. Hommes, H.W. Verspaget (Leiden, NL) 80. Local IL-2 application in non-operable colon cancer patients Z. Krastev, V. Koltchakov, S. Deredjian (Sofia, BG) 81. Ghrelin - A new mediator of colonic inflammation? P.C. Konturek, T. Brzozowski, P. Gaca, G. Burnat, S.J. Konturek, E.G. Hahn (Erlangen, D; Cracow, PL) 15

82. The role of interleukin-like EMT inducer (ILEI) in liver carcinoma progression C. Lahsnig, M. Mikula, H. Huber, H. Beug, W. Mikulits (Vienna, A) 83. Peripheral blood gene expression markers of local and progressive colorectal diseases determined by whole genome mRNA array analysis B. Molnár, O. Galamb, B. Galamb, N. Solymosi, F. Sipos, S. Spisák, K. Toth, P. Miheller, T. Zagoni, L. Herszenyi, Z. Tulassay (Budapest, H) 84. Correlation between mRNA expression level and protein array results in colorectal cancer S. Spisák, O. Galamb, N. Solymosi, B. Galamb, B. Nemes, F. Sipos, A. Kovacs, A. Somoracz, Z. Tulassay, B. Molnár (Budapest, H) 85. Progression markers of early and late stage colorectal cancer determined by protein arrays S. Spisák, O. Galamb, N. Solymosi, B. Galamb, B. Nemes, F. Sipos, T. Zagoni, K. Toth, Z. Tulassay, B. Molnár (Budapest, H) 86. Highdensity microarray analysis of colitis-associated cancer - Identification of new therapeutic targets C. Neufert, C. Becker, A. Nikolaew, P.R. Galle, U. Sahin, O. Tuereci, M.F. Neurath (Mainz, D) 87. The Nod2 protein is regulated via the ubiquitin-proteasome pathway I. Schoultz, T. Jiang, M. Lerm, P. Söderkvist, J.D. Söderholm (Linköping, S) 88. A regulatory role of Th2-associated transcription factor NFATc2 in murine colitis-associated colon cancer model B. Weigmann, P.R. Galle, M.F. Neurath (Mainz, D) 89. Cancer expressions of HIF-1α associate with immunoreactivities to TGF-β1 in colorectal cancers and adjacent inflammatory infiltrates A. Wincewicz, M. Sulkowska, S. Sulkowski, L. Kanczuga-Koda, M. Koda (Bialystok, PL) 90. The effect of iNOS inhibitors treatment in experimental colitis models Z. Yesilova, C.N. Ercin, A. Korkmaz, A. Ozcan, A. Uygun, K. Dagalp (Ankara, TR) 91. Clinicopathological and gene expression examination of young colorectal tumor patients Z. Szepes, I. Németh, T. Molnár, F. Nagy, A. Vörös, L. Tiszlavicz, J. Lonovics (Szeged, H) 92. Expression of Twist1 in colo-rectal tumors correlates with malignant transformation, local invasion and metastases at diagnosis L. Laghi, P. Bianchi, A. Zecchini, E. Cattaneo, J. Sabates-Bellver, K. Villa, A. Repici, M. Roncalli, G. Marra, A. Malesci (Rozzano, I; Zurich, CH; Milano, I)

16

Session I

Epidemiology of the villain

17

Intestinal inflammation and colorectal cancer: Magnitude worldwide and its implications Christian Pox, Wolff Schmiegel Ruhr-Universität Bochum, Knappschaftskrankenhaus und Berufsgenossenschaftliche Kliniken Bergmannsheil, In der Schornau 23–25, 44892 Bochum, Germany E-mail: [email protected] Colorectal cancer is the most frequent cancer in Europe and one of the most common cancers worldwide resulting in substantial morbidity and mortality. It is estimated that more than 1 million new cases occur each year. There is wide geographic variability in incidence rates with high incidences in Europe, North America, Asia and Australia and low incidences in Africa. The reasons for this variability are multifactorial consisting of a combination of genetic and environmental factors. The 5-year survival rates also vary substantially. Interestingly higher survival rates have been reported for the US compared to Europe. Most of this difference seems to be attributable to differences in stage at diagnosis. Unfortunately more than half of the cancers are diagnosed at a late stage with either lymph node or distant metastases. This has two major consequences. One, the prognosis of these tumours is much worse compared to localized disease. Secondly, these patients require either adjuvant or palliative chemotherapy. Until recently 5-FU was the only chemotherapeutic substance available. In patients with distant metastases 5-FU was able to achieve median survival rates of 12 months. The advantage of the substance was its low cost. However the last few years have seen major advances in chemotherapeutic options for treating patients with advanced disease. Oxaliplatin, Irinotecan and more recently antibodies to the VEGF ligand and EGF-receptors were developed and licensed. Due to these new chemotherapeutic compounds the median survival time for patients with metastatic disease has increased to 24 months. Furthermore, some patients have such a good response to therapy that metastases become surgically resectable with the potential for cure. However these new chemotherapeutic substances are rather expensive and cause a major financial burden for most already strained health systems or are unaffordable to others. There is good evidence to support screening of the asymptomatic population to reduce colorectal cancer mortality. Because screening detects cancers at an earlier stage it may actually reduce the number of patients requiring palliative chemotherapy possibly even making it cost-effective. Several screening methods exist including faecal occult blood testing, sigmoidoscopy and colonoscopy as well as the more recent developments CT-colonography and genetic or immunological stool tests. The evidence level is best for the use of guaiac based FOBT. Several large randomized studies have shown a modest reduction in CRC-related mortality of 30% for this test. However the sensitivity of a one-time guaiac based FOBT is around 60%. Endoscopic screening methods are seen as an attractive screening method by some experts. They are the most sensitive method for detecting asymptomatic early stage cancers and enable cancer prevention by removal of adenomatous polyps, the precursors of cancers. Although there are no randomized trials there is a large amount of indirect evidence in favour of sigmoidoscopy or colonoscopy. Recent data suggest that the protective effect of a negative colonoscopy may last twenty years and longer. What are the factors that influence if and how colorectal cancer 19

screening should be performed? One major aspect is the disease incidence as well as the financial resources available. Another factor is the available number of health care providers performing endoscopies. In countries with a high number of endoscopic facilities offering screening colonoscopy may be feasible. Germany is the first country with a national colonoscopy program which was introduced at the end of 2002. Up to today more than two million screening colonoscopies have been performed. The cancer detection rate is 0.6% most of which are early stage tumours with a good prognosis. Moreover in 19% adenomatous polyps were detected and removed. The reported morbidity due to the exam has remained low. However, compliance has to be improved. Austria has recently introduced a similar program. Other countries in which colonoscopy screening is perfomed include the US, Italy, Luxembourg and Poland. Countries that offer FOBT as the primary screening tool include France, Finland and the United Kingdom. In Japan immunological FOBT that seem to be more sensitive than guaiac tests are routinely used. Studies show that even in countries with good financial and human resources colorectal cancer patients are often not offered optimal treatment. The introduction of centres specialized in colorectal cancer treatment is seen as one way of trying to achieve that patients are treated according to guideline recommendations. These centers have to fulfil specified criteria and undergo external accreditation and benchmarking.

20

Epidemiology of the environmental clues

villain

–

Nutritional

habits

and

Marilia Cravo, M.D., Ph.D. Gastroenterology Department – Instituto Português de Oncologia Francisco Gentil, Lisboa, Portugal Long-standing Inflammatory Bowel Disease (IBD) has been associated to an increased risk of malignant transformation, especially when there is total involvement of the colon and disease is present for more than 10 years. Also, the risk of colorectal cancer (CRC) has been systematically associated to higher intake of certain nutrients namely meat and saturated fat However, there are very few studies published linking nutritional habits and other environmental factors to an increased risk of CRC in patients with IBD colitis. One of the pioneer studies in this field was the study by Lashner et al., published in 1989 (1) where these authors reported that individuals with longstanding ulcerative colitis taking folate supplementation had a nonsignificant 62% lower incidence of colorectal dysplasia and cancer, compared with those not receiving folate supplementation (OR, 0.38, 95% CI, 0.12–1.20). Patients with UC are at increased risk for folate deficiency due to the frequent use of sulfasalazine, a known folate antagonist, inadequate nutritional intake and increased losses and needs from intestinal inflammation. In another study (2), these authors observed that the risk of CRC and dysplasia was found to be significantly decreased by 18% for each increase of 23 nmol/L (10 ng/mL) in red blood cell folate concentrations in patients with UC (OR, 0.82, 95% CI, 0.68–0.99). Folic acid supplementation was inversely related to the risk of colorectal neoplasia in subjects with longstanding UC in a dosedependent manner (2). Although these studies had small sample sizes and the inherent limitations associated with retrospective study designs, they strongly suggested an inverse relationship between folate status and the risk of CRC as it was later demonstrated for the sporadic model (3). To my knowledge, these are the sole studies showing direct evidence for a protective role of a specific nutrient in modulating the risk of CRC in the setting of IBD/UC. In respect to fat and risk of CRC in the sporadic model, the association with saturated fat seems quite strong and probably depends not only on the amount of total fat but on the type of fat present in the diet. The mechanisms involved are probably related to a modulation of eicosanoid production via the influence on COX activity and the suppression of cell apoptosis, which are crucial processes in the process of carcinogenesis (5–7). On the other hand, a number of experimental and clinical studies also suggest that the fatty acid composition of diet might be important for controlling disease activity in patients with IBD, namely in patients with Crohn’s disease (8). While a diet rich in omega-3 fatty might reduce disease activity and has been used to decrease relapses in high risk patients, a diet rich in omega-6 fatty acids could have a promoter effect, as it has been observed for the process of colorectal carcinogenesis (9). Thus, although a direct association between dietary fat and CRC risk in patients with IBD has not been clearly demonstrated, we might speculate whether the effect that it exerts by modulating disease activity might also influence the risk for malignant transformation in an epithelium which is already at an increased risk. 21

References: 1) Lashner BA, Heidenreich PA, Su GL, et al. Effect of folate supplementation on the incidence of dysplasia and cancer in chronic ulcerative colitis. A case-control study. Gastroenterology 1989; 97: 255–259. 2) Lashner BA. Red blood cell folate is associated with the development of dysplasia and cancer in ulcerative colitis. J Cancer Res Clin Oncol 1993; 119: 549–554. 3) Young-In Kim Role of folate in colon cancer development and progression. J Nutr 2003; 133: 3731S–3739S. 4) Sandler RS. Epidemiology and risk factors for colorectal cancer. Gastroenterol Clin North Am 1996; 25: 717–735. 5) Rao CV, Hirose Y, Indranie C, Reddy BS. Modulation of experimental colon tumorigenesis by types and amounts of dietary fatty acids. Cancer Res 2001; 61: 1927–1933. 6) Llor X, Pons E, Roca A, et al. The effects of fish oil, olive oil, oleic acid and linoleic acid on colorectal neoplastic processes. Clin Nutr 2003; 22: 71–79. 7) Bartoli R, Fernandez-Banares F, Navarro E, et al. Effect of olive oil on early and late events of colon carcinogenesis in rats: modulation of arachidonic acid metabolism and local prostaglandin E2 synthesis. Gut 2000; 46: 191–199. 8) Gassull MA, Fernandez-Banares F, Cabré E, et al. Fat composition may be a clue to explain the primary therapeutic effect of enteral nutrition in Crohn's disease: results of a double blind randomised multicentre European trial. Gut 2002; 51: 164–168. 9) Gassull MA. Nutrition and inflammatory bowel disease: its relation to pathophysiology, outcome and therapy. Dig Dis 2003; 21: 220–227.

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IBD high risk groups F. Magro Universidade do Porto, Facultade de Medicina, Servico de Gastroenterologia, Porto, Portugal The risk of colorectal cancer is increased in inflammatory bowel disease (IBD). This risk is directly linked to duration of disease, extent of inflammation, age of one-set, severity of inflammation and the presence of strictures. In general population twothirds of colorectal cancers arise in the rectum and sigmoid colon. In ulcerative colitis (UC) the cancer is more spread around the colon, even so a large proportion is within in the rectum and sigmoid colon. In a Hospital series the colorectal cancer risk in extensive colitis at 10 years from onset of colitis was 0.7%, 7.2% at 20 years, 11.6% at 25 years and 16.5% at 30 years1. Most colorectal cancers complicating UC arise in patients with extensive colitis. The risk in patients with left-side colitis was smaller than those with extensive colitis – 3.6 vs. 19.2, respectively1. Another feature related to cancer risk in UC is the childhood-onset of colitis2. Early age of onset appears to be associated with positive family history of IBD3,4. The risk of cancer in patients with extensive colitis after 35 years of diagnosis was 40% in patients in whom the colitis started at less than 15 years of age and 25% in those developing colitis between 15 and 39 years5. In long-standing extensive UC, the severity of colonic inflammation is an important determinant of the risk of colorectal neoplasia. Endoscopic and histological grading of inflammation could allow better risk stratification for surveillance programs6. UC patients with a familiarly history of colon cancer have a twice risk of cancer7. Finally, a stricture should be considered a strong risk factor for cancer, requiring intensive colonoscopic surveillance8. Primary sclerosing cholangitis (PSC) appears to be an important risk factor for development colorectal carcinoma. The risk approaches 50% for PSC patients who have had UC for 25 years9,10. The duration of disease, age of onset, and time course for progression to dysplasia is similar between the PSC/UC and UC patients, however PSC/UC patients are 5 times more likely to develop dysplasia9. The patients with PSC/UC tend to have right-sided colon cancers, where bile acid concentrations are highest in the colon, compared with UC patients without PSC, who have mainly left-sided tumors11. In addition, the risk remains even after liver transplantation with an incidence of approximately 1% per person per year12. In Crohn’s disease (CD) the cancer risk is related by the extent and duration of the disease. Is small in the first 8–10 years but rises thereafter at approximately 0.5–1% per year13–15. In patients with long-standing CD with intact colon the relative risk rose from 3.4–18.216. The location of the cancers appears to be right sided and there have been reports in endoscopically uninvolved colon in CD17. In a large series reporting colorectal cancer in both UC and CD the age at development of cancer, duration of disease, multiplicity and presence of dysplasia and overall prognosis were similar18. Dysplasia does seem to be a predictor of future malignancy. Approximately half of those with high-grade dysplasia will develop malignancy within 5 years if earlier surgery is not performed19,20 and one third are found to have otherwise unsuspected cancer in the resected colon. Low-grade dysplasisa has less prognostic significance but the risk in one series is at least 18% and 50% at five years21.

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Bibliography: 1. Gyde SN, Prior P, Allan RN, Stevens A, Jewell DP, Truelove SC, Lofberg R, Brostrom O, Hellers G. Colorectal cancer in ulcerative colitis: a cohort study of primary referrals from three centres. Gut 1988; 29: 206–217. 2. Devroede GJ, Taylor WF, Sauer WG, Jackman RJ, Stickler GB. Cancer risk and life expectancy of children with ulcerative colitis. N Engl J Med 1971; 285: 17–21. 3. Monsen U, Bernell O, Johansson C, Hellers G. Prevalence of inflammatory bowel disease among relatives of patients with Crohn's disease. Scand J Gastroenterol 1991; 26: 302–306. 4. Monsen U, Brostrom O, Nordenvall B, Sorstad J, Hellers G. Prevalence of inflammatory bowel disease among relatives of patients with ulcerative colitis. Scand J Gastroenterol 1987; 22: 214–218. 5. Ekbom A, Helmick C, Zack M, Adami HO. Ulcerative colitis and colorectal cancer. A population-based study. N Engl J Med 1990; 323: 1228–1233. 6. Rutter M, Saunders B, Wilkinson K, Rumbles S, Schofield G, Kamm M, Williams C, Price A, Talbot I, Forbes A. Severity of inflammation is a risk factor for colorectal neoplasia in ulcerative colitis. Gastroenterology 2004; 126: 451–459. 7. Nuako KW, Ahlquist DA, Mahoney DW, Schaid DJ, Siems DM, Lindor NM. Familial predisposition for colorectal cancer in chronic ulcerative colitis: a casecontrol study. Gastroenterology 1998; 115: 1079–1083. 8. Lashner BA, Turner BC, Bostwick DG, Frank PH, Hanauer SB. Dysplasia and cancer complicating strictures in ulcerative colitis. Dig Dis Sci 1990; 35: 349–352. 9. Brentnall TA, Haggitt RC, Rabinovitch PS, Kimmey MB, Bronner MP, Levine DS, Kowdley KV, Stevens AC, Crispin DA, Emond M, Rubin CE. Risk and natural history of colonic neoplasia in patients with primary sclerosing cholangitis and ulcerative colitis. Gastroenterology 1996; 110: 331–338. 10. Broome U, Lofberg R, Veress B, Eriksson LS. Primary sclerosing cholangitis and ulcerative colitis: evidence for increased neoplastic potential. Hepatology 1995; 22: 1404–1408. 11. Marchesa P, Lashner BA, Lavery IC, Milsom J, Hull TL, Strong SA, Church JM, Navarro G, Fazio VW. The risk of cancer and dysplasia among ulcerative colitis patients with primary sclerosing cholangitis. Am J Gastroenterol 1997; 92: 1285–1288.

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12. Loftus EV, Jr., Aguilar HI, Sandborn WJ, Tremaine WJ, Krom RA, Zinsmeister AR, Graziadei IW, Wiesner RH. Risk of colorectal neoplasia in patients with primary sclerosing cholangitis and ulcerative colitis following orthotopic liver transplantation. Hepatology 1998; 27: 685–690. 13. Ekbom A, Helmick C, Zack M, Adami HO. Increased risk of large-bowel cancer in Crohn's disease with colonic involvement. Lancet 1990; 336: 357–359. 14. Gillen CD, Walmsley RS, Prior P, Andrews HA, Allan RN. Ulcerative colitis and Crohn's disease: a comparison of the colorectal cancer risk in extensive colitis. Gut 1994; 35: 1590–1592. 15. Sachar DB. Cancer in Crohn's disease: dispelling the myths. Gut 1994; 35: 1507–1508. 16. Gillen CD, Andrews HA, Prior P, Allan RN. Crohn's disease and colorectal cancer. Gut 1994; 35: 651–655. 17. Greenstein AJ, Sachar DB, Smith H, Janowitz HD, Aufses AH, Jr. A comparison of cancer risk in Crohn's disease and ulcerative colitis. Cancer 1981; 48: 2742–2745. 18. Choi PM, Zelig MP. Similarity of colorectal cancer in Crohn's disease and ulcerative colitis: implications for carcinogenesis and prevention. Gut 1994; 35: 950–954. 19. Woolrich AJ, DaSilva MD, Korelitz BI. Surveillance in the routine management of ulcerative colitis: the predictive value of low-grade dysplasia. Gastroenterology 1992; 103: 431–438. 20. Bernstein CN, Shanahan F, Weinstein WM. Are we telling patients the truth about surveillance colonoscopy in ulcerative colitis? Lancet 1994; 343: 71–74. 21. Connell WR, Lennard-Jones JE, Williams CB, Talbot IC, Price AB, Wilkinson KH. Factors affecting the outcome of endoscopic surveillance for cancer in ulcerative colitis. Gastroenterology 1994; 107: 934–944.

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Session II

Not all colorectal cancers are the same

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Getting familiar with familial colon cancer Jacob Sabates-Bellver*, Elisa Cattaneo*, Karl Heinimann^, Josef Jiricny, and Giancarlo Marra*. *Institute of Molecular Cancer Research, University of Zurich, Switzerland ^Division of Medical Genetics, University of Basel, Switzerland The predisposition of humans to colorectal cancer that is attributable to inherited mutations in a single gene has been well documented in ~5% of all cancers in this organ, with Lynch syndrome representing the most frequent condition. Germline mutations occur in genes involved in DNA repair (the mismatch repair genes MSH2, MSH6, MLH1 or PMS2 in Lynch syndrome, and the base excision repair gene MYH in familial polyposis with recessive inheritance), Wnt signaling (APC gene in familial polyposis with dominant inheritance), TGF-beta/BMP signaling (SMAD4 and BMPR1A in juvenile polyposis), and mTOR signaling (PTEN in Cowden disease, and STK11 in Peutz-Jeghers syndrome). Knock-out of murine genes involved in the same pathways, such as mismatch repair genes, Apc, Smad3, Ltbp-4, PI(3)Kgamma, and Cdx2, predispose mice to intestinal tumors. Although these genes are involved in different pathways, their mutations might result in similar cellular alterations, since they are responsible for the onset of similar precancerous lesions (i. e.: adenomatous polyps in the case of MYH or APC mutations, and hamartomas in the case of STK11, PTEN or SMAD4 mutations). In our ongoing screening study, 74% (131 out of 176) of index patients referred to us during the last five years because of clinically-suspected Lynch syndrome did not show any evidence of mismatch repair deficiency. Indeed, in Western countries, a familial aggregation of colorectal cancers has been reported in an additional ~10% of all cancers in this organ, but the responsible gene(s) are unknown. Finally, more than 80% of colorectal cancers arise in the absence of family history of cancer (i. e.: “sporadic” cases). Somatic alterations in genes involved in familial conditions are detected in sporadic cancers, such as the transcriptional silencing of MLH1 in ~10% of all colorectal cancers – mostly associated with methylator phenotype –, or APC mutations in most of the neoplasms following the polypoid adenoma-carcinoma pathway of transformation. Somatic alterations in genes involved in the inflammatory response also play roles in the transformation process, because drugs that modify inflammatory pathways have beneficial effects in the prevention and progression of colorectal tumors. In an exploratory transcriptomic analysis, we investigated whether different types of colorectal lesions could be distinguished by the expression profile of genes involved in the inflammatory response biological process as defined by Gene Ontology. Based on the expression of 371 genes belonging to this category, normal mucosa, adenomatous polyps and adenocarcinoma could be perfectly segregated into different clusters, and most of the mismatch repair deficient colon cancers could be easily distinguished from their proficient counterparts. These findings suggest that inflammatory response mechanisms are differently regulated in different types of colorectal tumors. Planning of future chemo-interventional trials with colorectal lesions should take account of these differences.

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Cytoskeletal regulation by the adenomatous polyposis coli protein in interphase and mitosis Presenting author: Inke S. Näthke1 Contributing authors: Karin Kroboth1, Ian Newton1, Zhouyu Li1, Dina Dikovskaya1, Paul Appleton1, David Schiffmann1, Louie Leung1, Aaron Quyn1, Jürg Zumbrunn1,2, Katsuhiro Kita3, Clare M. Waterman-Storer3 1 Division of Cell & Developmental Biology, School of Life Sciences, University of Dundee, WTB/MSI complex, Dow Street, Dundee DD1 5EH, UK 2 current address: Polyphor Ltd, Gewerbestr. 14, CH-4123 Allschwil, Switzerland 3 Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA Mutations in the adenomatous polyposis coli protein (APC) are common to most colonic tumours. A well-characterised function of the APC protein is its ability to support the assembly of a protein complex that regulates the degradation of β-catenin in a Wnt-regulated manner. Accumulating β-catenin causes changes in the activity of TCF/Lef transcription factors. This has been implicated in the transformation produced by truncations mutations in APC that disrupt the ability of APC to support the formation of a functional β-catenin degradation complex. However, APC is also an important regulator of the cytoskeleton. Inactivating APC in cells leads to a decrease in cell migration in cultured cells and tissue. In cells, this change is accompanied by changes in post-translationally modified microtubules and cell shape. Loss of APC also disrupts mitotic spindles and compromises the spindle checkpoint. This leads to defects in cell division that produce tetra- and polyploidy in cells and tissues lacking APC. Combined with a reduction in apoptosis, another immediate consequence of APC loss, these mitotic defects produce an increase in the number of potential tumour cells. The mitotic and apoptotic defects induced by loss of APC do not require transcriptional changes induced by the accumulation of β-catenin. Thus mutations in APC, which occur extremely early during tumourigenesis, lead to the inappropriate accumulation and survival of cells with altered differentiation and increased genetic instability. I will describe our work in a number of experimental systems to illustrate the effect of APC on the cytoskeleton and discuss possible implication of these findings for tumour initiation and progression.

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Not all colorectal cancers are the same: Different mechanism – Different prognosis John M. Carethers, M.D. Professor of Medicine, Chief, Division of Gastroenterology, University of California, San Diego, Bldg. UC303, MC 0063, 9500 Gilman Drive, La Jolla, CA 92093-0068, USA Chemotherapy for colorectal cancer is offered to patients based on the surgical or radiographical staging of their cancer, and there is evidence for survival benefit with 5-fluorouracil-based (5-FU) therapy for patients with stage III disease. The pathogenesis of colorectal cancer is driven by genomic instability, with 15% to 20% of tumors demonstrating a form of genomic instability termed high-frequency microsatellite instability (MSI-H) due to lack of DNA mismatch repair function, and the remainder of colorectal tumors with retained DNA mismatch repair function and called microsatellite stable (MSS). There is several lines of evidence that indicate the form of genomic instability that is present in a patient’s colorectal cancer may predict a survival benefit from 5-FU. In particular, patients whose colorectal tumors have MSI-H do not gain a survival benefit with 5-FU as compared to patients with MSS tumors. In vitro evidence supports these findings, as MSI-H colon cancer cell lines are more resistant to 5-FU compared to MSS cell lines. More specifically by biochemical experiments, components of the DNA mismatch repair system have been shown to recognize and bind to 5-FU that becomes incorporated into DNA and which could trigger induction of cell death. The binding and subsequent cell death events would be absent in colorectal tumors with MSI-H, which have lost intact DNA mismatch repair function. These findings suggest that: (a) tumor cytotoxicity of 5-FU is mediated by DNA mechanisms in addition to well-known RNA mechanisms, and (b) patients whose tumors demonstrate MSI-H may not benefit from 5-FU therapy. An important part of determining the lack of survival benefit was to have patients with MSI-H tumors that did not receive 5-FU therapy, as initial studies compared treated patients with MSI-H tumors to those with MSS tumors, and patients with MSI-H tumors without 5-FU treatment already have an inherent survival advantage. Future studies should include a better understanding of the cellular mechanisms of the DNA recognition of 5-FU, multi-centered prospective trials investigating the survival benefit of 5-FU based on genomic instability, and the investigation of alternative chemotherapeutic regimens for patients with MSI-H tumors to improve survival.

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IBD-related cancer – Just the same as sporadic? – Pro Jonathan M. Rhodes University of Liverpool, UK 1. Although there are differences in the dysplasia-cancer sequence between IBD cancer and sporadic colorectal cancer, established IBD-cancers and sporadic cancers have remarkably similar biology and natural history The natural history of colorectal cancer in patients with IBD is similar to that for sporadic cancer with a five year survival of around 50% and IBD-cancers and sporadic colorectal cancer have a similar biology: aneuploidy and microsatellite instability occur to a similar extent in both types of cancer, aneuploidy occurring in the majority and microsatellite instability in about 20%, commonly in association with hypermethylation of the DNA mismatch repair gene hMLH1. Moreover, although IBD cancers tend to arise from flat dysplastic lesions that lack APC mutations and sporadic cancers from polypoid lesions that possess APC mutations, by the time cancer has been established there is a remarkably similar rate of APC, p53 and kRas mutation rates, implying a similar “multi-hit” molecular pathogenesis. Thus IBDcancer and sporadic cancer are similar in most clinical and biological aspects excepting age at presentation (mean approx. 50 for IBD-cancer and 65 for sporadic cancer) and the nature of the preceding dysplastic lesion. 2. Inflammation seems to be the main determinant of the increased risk for cancer in IBD and this increased risk is probably mediated via NFκB activation in the epithelium. Interactions between the epithelium and the luminal bacteria might be an initiating factor for epithelial NFκB activation and thence for dysplasia-cancer progression in both IBD-cancer and sporadic colorectal cancer The incidence of cancer in Crohn’s disease is probably similar to that in ulcerative colitis with a similar extent and duration of disease. In Crohn’s disease cancers tend to occur at sites of inflammation, even sometimes in the small intestine or in fistula tracts. This strongly suggests that inflammation is the predominant driving force and this is backed up by recent studies showing that the subsequent cancer risk in UC is much lower when there is minimal inflammation at colonoscopy. In an experimental model of IBD-cancer, selective suppression of NFκB activation in the gut epithelium has been shown to markedly reduce the risk of subsequent cancer development, implying that it is epithelial NFκB activation rather than infiltration by inflammatory leucocytes that increases the cancer risk, presumably acting via inhibition of apoptosis. There is considerable evidence that epithelial NFκB activation also occurs in sporadic colorectal cancer. Recent publications suggest that bacterial components such as DNA and flagellin may suppress NFκB activation if they interact with luminal surface Toll-like receptors but activate NFκB if they interact with receptors on the baso-lateral aspects of the epithelial cells. Disruption of the epithelial barrier could therefore be critical in development not only of inflammation but also of colorectal cancer.

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3. It is failure of apoptosis, driven by factors that include NFκB activation, increased cyclo-oxygenase activity and reduced free arachidonic acid levels, rather than DNA damage per se, that seems to be the predominant determinant of colorectal cancer development. It is accepted that about 70–80% of the risk for sporadic colorectal cancer development is environmental, chiefly dietary. It is notable that the dietary factors, such as high calorie, high red meat and low green vegetable intake, that are associated with increased risk for colorectal cancer, are not closely associated with risk for adenoma development. Thus, although adenoma development is an important precursor for the majority of sporadic colorectal cancers, in most cases these adenomas do not lead on to cancer development and it is the dysplasia-cancer progression that is quantitatively more important than the initiation of the dysplasia in determining the incidence of sporadic colorectal cancer. There is growing evidence that it is apoptosis that is the key regulator of this process and that the most important dietary factors determining colorectal cancer risk may therefore be acting on apoptosis rather than on the prior development of dysplasia. It can be argued that the factors which determine progression from dysplasia to cancer are the same in both IBD-cancer and sporadic colorectal cancer and that the factors which might reduce the risk of this progression, e. g. regular use of mesalazine, might also be the same. 4. Thus, although the processes that determine the development of dysplasia in IBD and preceding sporadic cancer may differ, the mechanisms that determine progression from dysplasia to cancer, and by inference the causative factors that determine this progression, seem likely to be remarkably similar in IBD-cancer and sporadic colorectal cancer with epithelial NFκB activation as a key event.

References: 1. Ahnen DJ. Gastrointestinal malignancies in inflammatory bowel disease. Pp 379–96 in JB Kirsner (ed.) Inflammatory Bowel Disease, Saunders, Philadelphia, 2000. 2. Choi PM, Zelig MP. Similarity of colorectal cancer in Crohn's disease and ulcerative colitis: implications for carcinogenesis and prevention. Gut 1994; 35: 950–954. 3. Collins PD, Mpofu C, Watson AJ, Rhodes JM. Strategies for detecting colon cancer and/or dysplasia in patients with inflammatory bowel disease (revised). Cochrane Database Syst Rev 2006; CD000279. 4. Collier PE, Turowski P, Diamond DL. Small intestinal adenocarcinoma complicating regional ileitis. Cancer 1985; 55: 516–521.

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5. Church JM, Weakley FL, Fazio VW, et al. The relationship between fistulas in Crohn's disease and associated carcinoma: Report of 4 cases and review of the literature. Dis Colon Rectum 1985; 28: 361–366. 6. Gilbert JM, Mann CV, Scholefield J, Domizio P. The aetiology and surgery of carcinoma of the anus, rectum and sigmoid colon in Crohn's disease. Negative correlation with human papillomavirus type 16 (HPV 16). Eur J Surg Oncol 1991; 17: 507–513. 7. Rubin CE, Haggitt RC, Burmer GC, et al. DNA aneuploidy in colonic biopsies predicts future development of dysplasia in ulcerative colitis. Gastroenterology 1992; 103: 1611–1620. 8. Fleisher AS, Esteller M, Harpaz N, et al. Microsatellite instability in inflammatory bowel disease-associated neoplastic lesions is associated with hypermethylation and diminished expression of the DNA mismatch repair gene, hMLH1. Cancer Res 2000; 60: 4864–4868. 9. Melville DM, Jass JR, Shepherd NA, et al. Dysplasia and deoxyribonucleic acid aneuploidy in the assessment of precancerous changes in chronic ulcerative colitis. Observer variation and correlations. Gastroenterology 1988; 95: 668–675. 10. Redstom MS, Papadopoulos N, Caldas C, et al. Common occurrence of APC and K-ras gene mutations in the spectrum of colitis-associated neoplasias. Gastroenterology 1995; 108: 383–392. 11. Brentnall TA, Crispin DA, Rabinovitch PS, Haggitt RC, Rubin CE, Stevens AC, Burmer GC. Mutations in the p53 gene: an early marker of neoplastic progression in ulcerative colitis. Gastroenterology 1994; 107: 369–378. 12. Cao Y, Pearman AT, Zimmerman GA, McIntyre TM, Prescott SM. Intracellular unesterified arachidonic acid signals apoptosis. Proc Natl Acad Sci USA 2000; 97: 11280–11285. 13. Agoff SN, Brentnall TA, Crispin DA, et al The role of cyclooxygenase 2 in ulcerative colitis-associated neoplasia. Am J Pathol 2000; 157: 737–745. 14. Rhodes JM, Campbell BJ. Inflammation and colorectal cancer: inflammatory bowel disease-associated cancer and sporadic cancer compared. Trends Mol Med 2002; 8: 10–16. 15. Greten FR, Eckmann L, Greten TF, Park JM, Li ZW, Egan LJ, Kagnoff MF, Karin M. IKKβ links inflammation and tumorigenesis in a mouse model of colitisassociated cancer. Cell 2004; 118: 285–296. 16. Lee J, Mo J-H, Katakura K, Alkalay I, Rucker AN, Liu Y-T, Lee H-K, Shen C, Cojocaru G, Shenouda S, Kagnoff M, Eckmann L, Ben-Neriah Y, Raz E. Maintenance of colonic homeostasis by distinctive apical TLR9 signalling in intestinal epithelial cells. Nat Cell Biol 2006; 8: 1327–1336. 34

Con: Colitis-associated colon cancer is different from sporadic colon cancer Steven H. Itzkowitz, M.D. GI Division, Box 1069, Mount Sinai School of Medicine, One Gustave Levy Place, New York, NY 10029, USA, Tel: (212) 659-9697, Fax: (212) 849-2574 E-mail: [email protected] Patients with ulcerative colitis and Crohn’s disease are at increased risk for developing colorectal cancer (CRC). To date, an underlying genetic basis has not been identified to explain colon cancer predisposition in inflammatory bowel disease (IBD). Instead, it has been assumed that some environmental factor triggers chronic inflammation which then promotes the development of colon cancer. This is supported by the fact that colon cancer risk increases with longer duration of colitis, greater anatomic extent of colitis, the concomitant presence of other inflammatory manifestations (such as primary sclerosing cholangitis), and the observation in some studies that drugs used to treat inflammation, such as 5-aminosalicylates and steroids, may prevent the development of colorectal cancer. Sporadic colon cancer (SCC) and colitis-associated colon cancer (CAC) do share several features in common. They both arise from a dysplastic precursor lesion, the risk for both is increased to a similar degree by a positive family history of CRC, the overall survival of patients with SCC is similar to that of CAC, and many of the overall molecular abnormalities are similar. However, there are enough clinical, pathological, and molecular differences between the two to warrant consideration of CACs as unique entities. Compared to SCC, CACs affect individuals at a much younger age, there can often be more than one simultaneous cancer in the colitic colon, and unlike SCC where the majority of cancers arise from a single, discrete adenomatous polyp, the dysplastic precursor lesion in colitis is often flat or even invisible, and can be multifocal. Pathologically, CACs more often display mucinous, signet ring cell and poorly differentiated histology compared to SCCs. In fact, considering these clinical and histological features, the entity of colon cancer in IBD resembles that of Lynch Syndrome even more so than sporadic CRC! Moreover, a recently described pathological variant of colon cancer, termed low-grade tubuloglandular adenocarcinoma, appears to arise directly from low-grade dysplasia, and accounts for up to 10% of CAC whereas it has not been seen in SCC. In addition, as mentioned above, CRC in IBD arises in areas of prior inflammation, whereas SCC is not believed to be due to inflammation. At the molecular level, all three major pathways of colon carcinogenesis, namely chromosomal instability, microsatellite instability, and CpG island hypermethylation, occur with rather similar frequency in CAC and SCC. However, in CAC, APC mutations or loss occur rarely and quite late in carcinogenesis, whereas they occur very early, and are considered gatekeeper events in SCC. Conversely, p53 mutations, which are late events in SCC, occur very early in CAC. MSI also occurs early in CAC and studies suggest that oxidative stress can deplete DNA mismatch repair proteins resulting in abnormal DNA repair and replication infidelity. It is hoped that in the future, new insights from molecular and genomic studies will help clarify the pathophysiology and risk of CRC in IBD.

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Suggested Reading: Itzkowitz S, Harpaz N. Diagnosis and management of dysplasia in patients with inflammatory bowel diseases. Gastroenterology 126: 1634–1648, 2004. Itzkowitz S, Yio X. Inflammation and cancer: IV. Colorectal cancer in inflammatory bowel disease: the role of inflammation. Am J Physiol Gastrointest Liver Physiol, 287: G7–G17, 2004. Itzkowitz, SH. Molecular biology of colon cancer in IBD. Gastroenterol Clin N Am 35: 553–571, 2006.

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Session III

Screening and surveillance

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Latest endoscopic guidelines: FAP, HNPCC, IBD, and general David T. Rubin, M.D. University of Chicago, Section of Gastroenterology, Chicago, USA Colorectal cancer (CRC) is the second leading cause of cancer related death in Europe and North America. Variations in the etiopathogenesis of CRC in patient subpopulations are associated with differences in the presentation and timeline of neoplasia progression and thus, there exist different prevention guidelines. In all of the known CRC pathways, secondary prevention strategies take advantage of the well-described lag-time between pre-cancerous neoplasia and invasive adenocarcinoma. Existing guidelines are based on a variety of levels of evidence, and colonoscopy is utilized to identify macroscopic changes (polyps), or to obtain sequential and directed biopsies to identify dysplastic changes. The macroscopic or microscopic findings of screening and surveillance colonoscopy lead to risk stratification and plans for future prevention strategies or surgical intervention. This presentation will summarize the principles of secondary prevention and review current guidelines for familial adenomatous polyposis (FAP), hereditary nonpolyposis colorectal cancer (HNPCC), the inflammatory bowel diseases (IBD), and the general population, including those with a family history, but no identified known genetic syndrome (everyone else). (See Table summarizing guidelines.)

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TABLE: Summary of suggested endoscopic screening guidelines (from multiple references listed below)

Prophylactic surgery

Screening

Frequency

General Negative family history

Not recommended

Colonoscopy or Flexible sigmoidoscopy with FOBT

Beginning at age 50: Colonoscopy every 10 years or Flexible sigmoidoscopy every 5 years with annual FOBT

General Positive family history

Not recommended

Colonoscopy (total colon examination)

Beginning at age 40 or 10 years earlier than first family cancer (whichever is younger): Colonoscopy every 5 years

FAP Clinical diagnosis (polyposis with or without confirmed mutation)

Colectomy by age 20

While awaiting surgery: Colonoscopy

Annually from age 10–13 until surgery

EGD

Every 1–3 years

FAP First degree relative of proband with APC mutation

Not recommended without clinical disease or confirmed APC mutation

Flexible sigmoidoscopy

Annually from age 10–13 until 30 or disease expression Every 2–5 years from age 30 until 60 or disease expression

HNPCC MMR mutation or family pedigree satisfying Amsterdam or Bethesda criteria

Not recommended

Colonoscopy

Every 2 years from age 20–25 until 75 (or from 5–10 years earlier than first family cancer) Biennial starting at age 50 or 5 years younger than first gastric cancer in family

IBD Pancolitis

Colectomy with concurrent high-grade dysplasia Possible colectomy with concurrent low-grade dysplasia

Colonoscopy with random biopsies

Beginning 8–10 years after disease onset: Every 1–3 years depending on compounded risk factors and increasing age

IBD Left-sided colitis

Colectomy with concurrent high-grade dysplasia Possible colectomy with concurrent low-grade dysplasia

Colonoscopy with random biopsies

Beginning 10–15 years after disease onset: Every 1–3 years depending on compounded risk factors and increasing age

IBD Proctitis

Not recommended

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If FH gastric cancer, EGD

Colonoscopy at 8 years with biopsies to confirm disease extent

Confirmed proctitis can be followed with the average risk (non-IBD) guidelines

References: General population 1. Rex DK. American College of Gastroenterology action plan for colorectal cancer prevention. Am J Gastroenterol 2004 Apr; 99 (4): 574–577. 2. Screening for colorectal cancer: recommendation and rationale. Ann Intern Med 2002 Jul 16; 137 (2): 129–131. 3. Dunlop MG. Guidance on large bowel surveillance for people with two first degree relatives with colorectal cancer or one first degree relative diagnosed with colorectal cancer under 45 years. Gut 2002 Oct; 51 Suppl 5: V17–20. 4. Winawer S, Fletcher R, Rex D, Bond J, Burt R, Ferrucci J, et al. Colorectal cancer screening and surveillance: clinical guidelines and rationale – Update based on new evidence. Gastroenterology 2003 Feb; 124 (2): 544–560. 5. Rex DK, Johnson DA, Lieberman DA, Burt RW, Sonnenberg A. Colorectal cancer prevention 2000: screening recommendations of the American College of Gastroenterology. American College of Gastroenterology. Am J Gastroenterol 2000 Apr; 95 (4): 868–877. 6. Desch CE, Benson AB, 3rd, Somerfield MR, Flynn PJ, Krause C, Loprinzi CL, et al. Colorectal cancer surveillance: 2005 update of an American Society of Clinical Oncology practice guideline. J Clin Oncol 2005 Nov 20; 23 (33): 8512–8519. FAP 1. Jarvinen HJ. Hereditary cancer: guidelines in clinical practice. Colorectal cancer genetics. Ann Oncol 2004; 15 Suppl 4: IV127–131. 2. Galiatsatos P, Foulkes WD. Familial adenomatous polyposis. Am J Gastroenterol 2006 Feb; 101 (2): 385–398. 3. Winawer S, Fletcher R, Rex D, Bond J, Burt R, Ferrucci J, et al. Colorectal cancer screening and surveillance: clinical guidelines and rationale – Update based on new evidence. Gastroenterology 2003 Feb; 124 (2): 544–560. 4. Dunlop MG. Guidance on gastrointestinal surveillance for hereditary nonpolyposis colorectal cancer, familial adenomatous polypolis, juvenile polyposis, and Peutz-Jeghers syndrome. Gut 2002 Oct; 51 Suppl 5: V21–27. 5. Giardiello FM, Brensinger JD, Petersen GM. AGA technical review on hereditary colorectal cancer and genetic testing. Gastroenterology 2001 Jul; 121 (1): 198–213.

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HNPCC 1. Jarvinen HJ. Hereditary cancer: guidelines in clinical practice. Colorectal cancer genetics. Ann Oncol 2004; 15 Suppl 4: IV127–131. 2. Lindor NM, Petersen GM, Hadley DW, Kinney AY, Miesfeldt S, Lu KH, et al. Recommendations for the care of individuals with an inherited predisposition to Lynch syndrome: a systematic review. JAMA 2006 Sep 27; 296 (12): 1507–1517. 3. Winawer S, Fletcher R, Rex D, Bond J, Burt R, Ferrucci J, et al. Colorectal cancer screening and surveillance: clinical guidelines and rationale – Update based on new evidence. Gastroenterology 2003 Feb; 124 (2): 544–560. 4. Dunlop MG. Guidance on gastrointestinal surveillance for hereditary nonpolyposis colorectal cancer, familial adenomatous polypolis, juvenile polyposis, and Peutz-Jeghers syndrome. Gut 2002 Oct; 51 Suppl 5: V21–27. 5. Giardiello FM, Brensinger JD, Petersen GM. AGA technical review on hereditary colorectal cancer and genetic testing. Gastroenterology 2001 Jul; 121 (1): 198–213. IBD 1. Carter MJ, Lobo AJ, Travis SPL, on behalf of the IBD Section of the British Society of Gastroenterology. Guidelines for the management of inflammatory bowel disease in adults. Gut 2004; 53 (Suppl V): V1–V16. 2. Itzkowitz SH, Present DH. Consensus conference: Colorectal cancer screening and surveillance in inflammatory bowel disease. Inflamm Bowel Dis 2005 Mar; 11 (3): 314–321. 3. Kornbluth A, Sachar DB. Ulcerative colitis practice guidelines in adults (update): American College of Gastroenterology, Practice Parameters Committee. Am J Gastroenterol 2004 Jul; 99 (7): 1371–1385.

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What new techniques will replace classical surveillance? Ralf Kiesslich, MD, PhD University of Mainz, Germany Endoscopy has recently witnessed major technical improvements. Magnifying, high resolution and HDTV endoscopy systems often in conjunction with chromoendoscopy enable detailed surface analysis and help to identify areas of interest within the gut. However, chromoendoscopy is now facing electronic competition, as filter or post processing techniques such as narrow band imaging may at least partially mimic chromoendoscopy by simply pressing a button to enhance visualizing of surface vessel architecture. All of these techniques unmask a plethora of new visible details and require immediate interpretation to target biopsies to suspicious surface architecture. However, they all predict histology to some extent but histology even after targeted biopsies remains the accepted gold standard for final judgement of mucosal lesions. Endomicroscopy has recently emerged as a novel technique that allows subsurface imaging and in vivo histological assessment of mucosal changes during ongoing endoscopy. This technique allows for the first time to look below the mucosal surface and to see in vivo histology of lesions during ongoing endoscopy. Chromoendoscopy studies have impressively shown that chromoendoscopy can unmask multiple, flat growing intraepithelial neoplasias in patients with long standing ulcerative colitis. The diagnostic yield is increased 3–4.5-fold as compared to random biopsies and chromoendoscopy has thus been recently incorporated into the US guidelines. Interestingly, endomicroscopy can further enhance the diagnostic yield with significant fewer biopsies needed per patient. Endomicroscopy has a limited field of view (475 x 475 µm). Therefore, one should combine methylene blue aided pan-chromoendoscopy with subsequent targeted fluorescein aided endomicroscopy. Compared to routine surveillance colonoscopy detection of intraepithelial neoplasias could be increased 4.75-fold per patient using this approach. Whereas chromoendoscopy unmasked areas of interest (flat, circumscribed lesions), subsequent endomicroscopy confirmed or excluded the presence of neoplasias and reduced the need for biopsies. With this approach the average amount of biopsies could be decreased from about 40 per patient (random biopsies) to 20 biopsies (chromoendoscopy) and theoretically to about 4 biopsies per patient (combined approach of chromoendoscopy and endomicroscopy) with still significantly higher diagnostic yield for intraepithelial neoplasias as compared to random biopsies. With this technology, we explore the in vivo cell architecture of the whole mucosal layer at subcellular resolution which will allow functional and molecular imaging in the future.

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How to deal with dysplasia and adenomatous polyps in IBD? Paula Chaves and Paula Borralho Instituto Português de Oncologia de Lisboa, Hospital Garcia de Orta, Almada, Portugal Dysplasia is “an unequivocal neoplastic proliferation” and at present, it is the best and the most reproducible marker of malignancy risk in IBD patients. The present definition and classification were proposed in 1983 and remained unchanged until 1998 when, in Vienna, an upgraded was developed in order to resolve differences between eastern and western terminologies. The basic concepts were preserved and “dysplasia” was replaced by “non-invasive neoplasia”. As the original, the Vienna system also has management implications. Grossly, there are 2 patterns of dysplasia, “flat” (endoscopically undetectable) and “raised” (endoscopically detectable) otherwise known as “dysplasia-associated lesions or masses” (DALMs). “Raised” and “flat” lesions have distinct management according to its risk of association with invasive components. The histology of both “flat” and “raised” lesions includes the categories “negative”, “indefinite” and “positive” for low- and high-grade dysplasia/non-invasive neoplasia. In the Vienna system the high-grade non-invasive neoplasia includes “high-grade dysplasia”, “noninvasive carcinoma” (non-evident invasion) and “suspicious of invasive carcinoma” (hallmarks of invasion but no unequivocal invasive component). It also considers “intramucosal” (lamina propria or muscularis mucosae) and “submucosal invasive neoplasia”. The more important pitfalls of dysplasia are subjectivity and reproducibility. Despite its detailed description the application of the criteria for dysplasia is always subjective. The two-grade system proposed in 1983 and sustained in Vienna classification allowed a better agreement in the high-grade group but in the lowgrade vs. indefinite a disturbing interobserver variability still remains. Furthermore, it is recognised that high-grade dysplasia has a high predictive value for invasive neoplasia but data are controversial for low-grade lesions. By other way, it is now evident that IBD malignancy may evolve by others than the well-recognised “adenomatous” pathway. One of these is the called “serrated”, in which traditional dysplasia may be absent. For all these reasons the AGA and the ACG recommend that dysplasia must be confirmed by at least 2 GI pathologists. Polyps in IBD patients may be “sporadic adenomas” or DALMs and, actually there is no reliable way to make its morphological distinction. In practice, if the lesion is above the upper limit of the disease, in non-colitic mucosa, it can be regarded as a “sporadic adenoma”. If it occurs in colitic mucosa it can also be managed as a “sporadic adenoma” provided that it is completely excised endoscopically, it is unassociated with dysplasia elsewhere in the bowel and the patient is in the adenoma bearing age range.

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Session IV

Taking dysplasia to the molecular level

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Inflammation-driven carcinogenesis – The players Lorne J. Hofseth Laboratory of Inflammatory-Driven Carcinogenesis, Department of Pharmaceutical and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina, Columbia, South Carolina 29208, USA Chronic inflammation drives cancer. Evidence comes from cell culture, animal and human studies. Perhaps the most compelling evidence is that inflammation over years, in humans at specific organ sites, increases cancer risk in that organ. Examples of these ‘high cancer risk, reactive species overload diseases’ include chronic hepatitis (liver cancer), chronic esophagitis (esophageal cancer), chronic gastritis (stomach cancer) and chronic colitis (colon cancer). These diseases can be triggered by the environment or a genetic predisposition. Because many are associated with a hyperactive immune system, studies have focused on immunomodulation to control disease. Balancing effector and regulatory T cell levels and/or activity, for example, can ameliorate chronic colitis. Specific targets within innate, or adaptive immune cells, or in target mucosal cells are also beginning to emerge as potential targets for chemoprevention and therapy of reactive species overload diseases. Finding and targeting a specific molecule or molecules may equate to a biological smart bomb, and lessen the side-effects of broader spectrum drugs such as steroids. Some recognized players to target include free radicals, pro- and antiinflammatory cytokines, growth factors, transcriptional activators [e. g. nuclear factorkappa B (NF-κB) and peroxisome proliferator activated receptors (PPARs)], pattern recognition receptors [PRRs; e. g. toll-like receptors (TLRs), and nucleotide-binding oligomerisation domain (NOD) proteins], the trefoil factor family (TFF), arachidonic acid pathway enzymes and kinase signaling pathways. Additionally, tumor suppressor pathways such as the p53 and the retinoblastoma (pRb) pathways are often disrupted in reactive species overload diseases. Here, we briefly review these players and point to some specific and general mediators that have been used successfully, or have potential to be used as chemopreventive and therapeutic agents in inflammatory-mediated carcinogenesis.

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Cancer originating from bone marrow-derived stem cells (BMDCs) Timothy C. Wang M.D. Columbia University Medical Center, Department of Medicine and Irving Cancer Research Center, New York, NY, USA The link between inflammation and cancer has long been recognized but poorly understood. Epithelial cancers often arise in the setting of tissue loss and chronic inflammation, and are preceded by hyperproliferation and metaplasia. Helicobacter pylori infection, which typically induces chronic inflammation in the stomach leading to atrophy and metaplasia, has been strongly linked to gastric cancer, and as such is an excellent model for studies of carcinogenesis. Carcinogenesis is a multi-step process, and recent work suggests that the target cells for transformation are pluripotent stem cells or early progenitors. In this classic model, epithelial cancers are believed to originate from transformation of a resident tissue stem cell. However, adult stem cells exhibit plasticity and are often necessary for the repair of tissue damage, and thus a second possible source of cancer stem cells is the bone marrow derived cell (BMDC). Our group has developed the Helicobacter mouse model of gastric cancer to address the role of inflammation and the origins of epithelial cancers. Work from our group has shown the H. pylori infection can activate NF-κB pathways and cytokine production in macrophages, and that a strong T helper 1 cytokine response is necessary for progression to gastric cancer. Studies in patients that have demonstrated a link between cancer risk and polymorphisms in proinflammatory genes has been supported by transgenic models developed in our laboratory, which confirm that overexpression of pro-inflammatory cytokines is sufficient for the induction of gastric cancer. Chronic inflammation leads to mobilization of bone marrow-derived stem cells (BMDCs), which can then be recruited to and engrafted in site of chronic injury. When recruited to the stomach, for example, BMDCs take on the appearance of epithelial cells as well as activated myofibroblasts. In our lab, we used lethally irradiated mice and bone marrow reconstitution studies to determine whether regeneration of damaged epithelium was accomplished by tissue-resident or bone-marrow derived stem cells (BMDCs). Only when mice were undergoing chronic H. felis infection did BMDCs engraft in the gastric mucosa and these were shown to give rise to gastric dysplasia and cancer. Eradication of infection and/or blockade of pro-inflammatory cytokines reduced engraftment and circulating progenitors and inhibited progression to dysplasia. These findings from our laboratory suggest that epithelial cancers may originate from marrow-derived sources and have broad implications for the multistep model of cancer progression.

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NF-κB signaling as a link between colonic inflammation and colorectal cancer M. Karin, Ph.D. Professor of Pharmacology, Univ. of California, San Diego, La Jolla, CA, USA Chronic inflammatory bowel diseases such as ulcerative colitis (UC) and Crohn’s disease (CD) greatly increase the risk of colorectal cancer development. To understand the mechanisms by which inflammation affects the development of colorectal cancer we used a mouse model of colitis-associated cancer (CAC) and conditional deletion of the gene encoding the IKKβ catalytic subunit of the IκB kinase (IKK) complex, a critical mediator of NF-κB activation. We found that deletion of IKKβ in intestinal epithelial cells (IEC) decreased tumor number by 80% without affecting tumor size. Deletion of IKKβ in myeloid cells (macrophages) decreased tumor number by only 50% but also decreased tumor size. Further investigation revealed that in IEC, IKKβ-mediated NF-κB activation is mainly responsible for suppressing the apoptosis of premalignant cells through induction of Bcl-XL. In macrophages, however, the main pro-tumorigenic function of IKKβ-dependent NF-κB activation is exerted through induction of mitogens that stimulate the proliferation of transformed IEC. One of these mitogens is IL-6 and disruption of the IL-6 gene also decreases the development of CAC. Transplantation experiments suggest that the major source or pro-tumorigenic IL-6 are bone marrow-derived myeloid cells.

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Translating laboratory findings into clinics: Of mice and man Prof. Dr. Markus F. Neurath I. Medical Clinic, University of Mainz, Germany Inflammatory bowel diseases (IBD: Crohn’s disease and ulcerative colitis) are relapsing inflammations of the gastrointestinal tract not due to specific pathogens. Although the precise etiology of the diseases is still unknown, recent data from animal model strongly suggest that predisposing genetic factors, barrier defects and bacterial antigens lead to an unbalanced activation of the mucosal immune system that in turn causes chronic intestinal inflammation. There has been a growing interest in understanding the role of cytokines and cytokine signaling events in IBD models in recent years. T-bet expressing Th1 cells, GATA-3 expressing T cells producing IL-13 and THIL-17 cells are key effector cell populations with major relevance for the design of novel therapeutic approaches for IBD. Furthermore, IL-12 family cytokines such as IL-23 and IL-27 appear to play a prominent role in modulating the activity of effector T cells in experimental colitis. Finally, various proinflammatory cytokines and transcription factors in the gut have been shown to regulate the development and progression of colitis associated colon cancer in murine models. These data provide a rationale for selective targeting of cytokines and transcription factors in IBD. Such targeting has the potential advantage of targeting the activity of various cell types simultaneously rather than of a single cell type. In any case, the findings in animal models of chronic intestinal inflammation have provided new insights into the pathogenesis of IBD and are important for the development of novel immunotherapeutic approaches.

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Session V

How can we prevent?

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What to eat or not to eat Miquel A. Gassull Department of Gastroenterology and Hepatology, Hospital Universitari Germans Trias i Pujol, Health Sciences Research Institute, Germans Trias i Pujol Foundation, Badalona, Spain The etiology of colorectal cancer (CRC) involves the interaction of intracellular molecular changes and environmental factors, of particular importance are, smoking, intestinal bacteria and dietary components. In this talk we will focus in the potential role of the diet as primer or preventor of CRC. Diet is very much related to life style; however, the way as both are related to colorectal carcinogenesis remains unclear. Several risk factors are frequently mentioned in Western diets as relater to CRC, such as high concentrations of fat and animal protein, as well as low amounts of fibre, fruits and vegetables. A large number of experimental studies have found a protective effect of fiber on neoplasia induction, especially in relation to fermentable fibre. Epidemiological cohort studies have also suggested that a greater intake of vegetables, fruits, cereals and seeds is associated to a lower risk for CRC. Moreover, beneficial properties of fibre (especially from vegetable sources) were documented in more than half of case-control studies. Nevertheless, recent data from longitudinal epidemiological studies and randomized trials seem not to support this view. Future research should evaluate what sources of fibre provide effective anti-neoplastic protection, carrying out interventional studies with specific fibres for longer periods and in general rather than in high risk populations. Red meat, processed meats, and perhaps refined carbohydrates have also being blamed as increasers of CRC risk. Recommendations of decreasing red meat and fat intake are widely accepted, although the total amount and composition of specific fatty acids may have distinct roles in this setting. Current evidence favours the replacement of saturated fatty acids and n- 6 derived long-chain polyunsaturated fatty acids with long-chain n-3 or n-9 fatty acids. In addition, excess body weight and excess energy intake inducing hyperinsulinaemia have been also associated to CRC, as well as lifestyle habits such as sedentarism, high alcohol consumption, smoking and low consumption of folate and methionine. Thus, current recommendations for decreasing the risk of CRC include dietary measures such as increased plant food intake; the consumption of whole grains, vegetables and fruits; and reduced red meat and saturated fat intake. However, there is a lack of well structured long-term studies, taking into account all the confounding variables, in healthy and high risk populations, before firm recommendations can be given.

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Inner and outer environment Xavier Llor, M.D., Ph.D. Associate Professor of Medicine, University of Illinois at Chicago, Chicago, IL, USA Inflammatory bowel disease is the result of an inappropriate immune response mounted in genetically susceptible individuals as the result of an interaction among environmental factors, microbial components and the immune system. One of the most recognized complications of IBD is the development of colorectal cancer, though the risk of developing this cancer appears to be lower in the most recently reported series. While the presence of sclerosing cholangitis and a family history of colorectal cancer seem to increase the risk, factors strictly related to the integrity of the gut mucosa could be of paramount importance. In fact, disease extension and degree of inflammation likely play a significant predisposing role. Most of the identified genes involved in IBD susceptibility are involved in the maintenance of the intestinal barrier (DLG5), the transport of key molecules for the homeostasis or exclusion of toxins (SLC and MDR1), and the sensing of bacteria, both on the surface (TLR, CD14) and intra-cytoplasmatically (CARD15, CARD4, CARD8). CARD15 is a bacteria-recognizing cytoplasmic protein suggesting defective mechanisms of bacterial sensing as the link between the gut flora and the altered immune response found in IBD. Furthermore, most IBD patients have an enhanced immunological reactivity against gut bacterial antigens, such as flagellin. It has been proposed that this immune reactivity is the consequence of a loss of tolerance towards the autologous enteric flora, resulting in an inappropriate immune response in the mucosa and finally chronic inflammation. To what degree the correction of some these factors could prevent the development of colorectal cancer is yet to be determined. A variety of environmental factors have been found to influence the development of colorectal cancer in non-IBD patients. Diet, physical exercise, tobacco use would be among the most determinant environmental factors. Little is known about the effect if these factors in the development of colorectal cancer in IBD patients but some of them are also clearly related to the development of IBD itself. Therefore, some interventions could be doubly useful for the prevention of IBD and one of its most important complications.

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Aspirin, NSAIDs and COX-2 inhibitors Prof. Angel Lanas Service of Gastroenterology, University Hospital, Zaragoza, Spain It is well established that most gastrointestinal (GI) cancers develop from premalignant lesions of the digestive mucosa. Colonic polyps are one of the most frequently diagnosed pre-malignant lesions where both the pathological progression and the molecular sequence of the neoplastic transformation are well known. The process of malignant transformation of these lesions takes years, which offers an excellent opportunity for prevention strategies. Chemoprevention therapy is one of the options in these prevention strategies, but it has the inherent difficulty in developing safe and effective drugs to prevent cancer in the general population, where the risk of disease in a given year is low. For example, colorectal cancer is the most frequent GI cancer which accounts for almost 12% of all newly diagnosed cancers in the US. On average, the cumulative probability of developing colorectal cancer is 6% over full-life expectancy. Thus, if any drug therapy is to be used in the prevention of colorectal cancer within the general population, the majority of the people (94%) treated will not benefit unless the benefits extend to other areas of health outcomes. In recent years it has been established that COX-2, but not COX-1, expression is increased in several types of human neoplastic tissues. Over-expression of COX-2 has been reported in 86% of human colorectal carcinoma samples and 43% of colonic adenomas, where it is present in cancer cells, inflammatory cells, vascular endothelium and fibroblasts. Among other neoplastic and pre-malignant lesions, increased expression of COX-2 has also been detected in gastric and esophageal cancers and Barrett’s esophagus as well, where it is associated with increased cell proliferation and neoplastic progression. Different in vitro and animal experiments have shown that overexpression of COX-2 led to phenotypical malignant alterations of cells and tumor growth, which were markedly attenuated by treatment with COX-2 selective inhibitors. In humans, epidemiological data have shown that the use of aspirin and nonselective NSAIDs are associated with decreased risk of gastrointestinal cancers including colorectal, gastric and esophageal cancers. Most recent data also suggest that coxib therapy is also associated with risk reduction of colonic neoplasia. Randomized controlled trials in humans have shown that low-dose aspirin is associated with decreased recurrence of advanced colorectal neoplasia, and that the use of high-dose celecoxib and sulindac has been proved to be effective in decreasing the burden of colonic polyps in FAP patients. All these data led to investigate the effect of COX-2 selective inhibitors in the long-term prevention of recurrence of sporadic polyps. Data from the APPROVE, APC and pre-SAP trials have shown positive results in the secondary prevention of colorectal polyps with 25 mg/day of rofecoxib, 200 and 400 mg of celecoxib twice a day. However, the use of this long-term therapy was also associated with increased risk of thrombotic events, mainly non-fatal cardiovascular adverse events when compared to placebo. Based on these findings, the role of these drugs as chemopreventive agents in colorectal cancer has been questioned and no future for this type of therapy seems possible for the prevention of recurrence of sporadic colorectal adenomas. However, they may be used in patients with high risk, as those suffering from familial adenomatous polyposis, where the risk benefit balance in this younger population 55

seems clear. Therefore, at present only low-dose aspirin, which is associated with gastrointestinal side effects but has additional beneficial cardioprotective properties, may be a chemopreventive candidate in selected populations. However, more data are needed before this compound can be recommended. Research should also be focused on new agents that must share both low-GI and low-cardiovascular risk in the selected population.

General References: Giardiello FM, Offerhaus JA, Tersmette AC, Hylind LM, Krush AJ, Brensinger JD, et al. Sulindac induced regression of colorectal adenomas in familial adenomatous polyposis: evaluation of predictive factors. Gut 1996; 38: 578–581. Steinbach G, Lynch PM, Phillips RK, Wallace MH, Hawk E, Gordon GB, et al. The effect of celecoxib, a cyclooxygenase-2 inhibitor, in familial adenomatous polyposis. N Engl J Med 2000; 342: 1946–1952. Sandler RS, Halabi S, Baron JA, Budinger S, Paskett E, Keresztes R, et al. A randomized trial of aspirin to prevent colorectal adenomas in patients with previous colorectal cancer. N Engl J Med 2003; 348: 883–890. Baron JA, Cole BF, Sandler RS, Haile RW, Ahnen D, Bresalier R, et al. A randomized trial of aspirin to prevent colorectal adenomas. N Engl J Med 2003; 348: 891–899. Bresalier RS, Sandler RS, Quan H, Bolognese JA, Oxenius B, Horgan K, Lines C, Riddell R, Morton D, Lanas A, Konstam MA, Baron JA; Adenomatous Polyp Prevention on Vioxx (APPROVe) Trial Investigators. Cardiovascular events associated with rofecoxib in a colorectal adenoma chemoprevention trial. N Engl J Med 2005; 352: 1092–1102. Solomon SD, McMurray JJ, Pfeffer MA, Wittes J, Fowler R, Finn P, Anderson WF, Zauber A, Hawk E, Bertagnolli M; Adenoma Prevention with Celecoxib (APC) Study Investigators Cardiovascular risk associated with celecoxib in a clinical trial for colorectal adenoma prevention. N Engl J Med 2005; 352: 1071–1080. Baron JA, Sandler RS, Bresalier RS, Quan H, Riddell R, Lanas A, Bolognese JA, Oxenius B, Horgan K, Loftus S, Morton DG; APPROVe Trial Investigators. A randomized trial of rofecoxib for the chemoprevention of colorectal adenomas. Gastroenterology 2006; 131: 1674–1682. Arber N, Eagle CJ, Spicak J, Racz I, Dite P, Hajer J, Zavoral M, Lechuga MJ, Gerletti P, Tang J, Rosenstein RB, Macdonald K, Bhadra P, Fowler R, Wittes J, Zauber AG, Solomon SD, Levin B; PreSAP Trial Investigators.Celecoxib for the prevention of colorectal adenomatous polyps. N Engl J Med 2006; 355: 885–895.

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Use of 5-ASA, AZA, and 6-MP and the development of colorectal cancer in IBD F. Velayos, M.D., Ph.D. Assistant Professor of Medicine, Translational Research, University of California, San Francisco, Center for Crohn’s and Colitis, San Francisco, CA 94115, USA Inflammation is an important biological risk factor for the development of colorectal cancer (CRC). Not surprisingly, the risk of CRC in inflammatory bowel disease (IBD) is six-fold higher than the background population, making IBD the third highest risk condition for CRC. Current cancer prevention strategies rely on the regular performance of surveillance colonoscopy and removal of the colon if histological dysplasia is detected on mucosal biopsies. This strategy has recognized limitations and does nothing to prevent the initiation or progression of the neoplastic process in the first place. Chemoprevention refers to the use of a medication or other substance to prevent the initiation or progression of neoplasia. Since inflammation is an important risk factor for the development of CRC in IBD and suppression of chronic inflammation is the main goal of therapy in IBD, there has been interest in determining whether use of anti-inflammatory therapy used in IBD, such as 5-aminosalicylic acid (5-ASA) and immunomodulators (6-mercaptopurine, azathioprine), can prevent CRC. 5-ASA shares several important anti-inflammatory and anti-cancer properties with aspirin and nonsteroidal anti-inflammatory medications (NSAIDs). Many, but not all, observational studies have shown that the use of 5-ASA reduces the risk of CRC in IBD. In a pooled analysis of 334 CRC-UC cases, regular use of 5-ASA reduced the risk of CRC by approximately 50% (P 450 score) were treated with 5 mg/kg of infliximab, which was applied according to the scheme at 0, 2 and 6 weeks. Patients with perianal CD were treated with mesalazine 2 x 0.5 g rectally and azathioprine orally, and the second group with corticosteroids and metronidazole intravenously, azathioprine and peptide diet orally when the first dose of infliximab was applied. Further therapy consisted of azathioprine and gradually decreasing doses of corticosteroids. Results: The decreased secretion from recto-gluteal fistula at the second week of therapy in four pts was observed, and the closure of fistula in all pts at 10 weeks after the first dose of infliximab was noted. Within one year of observation period in only one patient opening of perianal fistula at 22 weeks was found. It was closed two weeks after administration of single infliximab dose. Whereas pts with inflammatory CD responded to infliximab already within several days after the first dose of infliximab (decrease in CDAI score by 70), and the second and the third doses seemed not to affect significantly the disease course. Patients with inflammatory CD achieved remission (CDAI < 150 score) at 10 weeks after the first dose of biological therapy, and it was maintained for 30–40 weeks of follow-up period. Conclusion: 1. Infliximab improves the outcome of treatment in patients with CD. 2. Patients with inflammatory CD responded to infliximab earlier than patients with perianal CD. 3. The clinical effect of biological therapy is relatively long-lasting.

19 Serological determination in inflammatory bowel disease Machado, M.B., Guedes, L., Schneider, N.C. São Lucas Hospital, Catholic University, Porto Alegre, Brazil Base: Inflammatory bowel diseases are diseases of difficult diagnosis and we know that 10 to 15% of the patients present an uncertain form with as much discoveries of DC as of RCUI. Recently, it was found that the anti-neutrophilic cytoplasmic antibodies with pattern peri-nuclear in the immunofluorescence (p-ANCA) which are more common in RCUI and the anti-Saccharomyces cerevisiae antibodies (ASCA) were more prevalent in patients with DC. Objectives: to demonstrate the clinical importance and applicability of these new laboratorial markers in the evaluation of patients with IBD. Outline: The study was transverse and prospective. Patients: Patients belonging to the Clinic of Inflammatory Bowel Disease of the São Lucas Hospital were selected randomly. Blood samples were collected for the dosage of serological markers p-ANCA and ASCA. Results: 53 patients were studied, being 28 with DC and 25 with UR. At the moment the marker ASCA was evaluated, we found out that 46.4% of the patients with DC were positive for ASCA IgA or IgG while only 20% of the patients with UR who have had the same tests were positive. In relation to the ANCA marker, 60% of the patients with UR had this positive marker, being 26% p-ANCA. Conclusion: This study shows that the combined use of these two serological markers, ASCA and pANCA, can take us to differential and more accurate diagnosis between DC and RCUI, mainly in those patients with indeterminate colitis.

20 Evaluation of the inflammatory bowel disease activity by the color Doppler ultrasonography Guedes, L., Schneider, N.C., Machado, M.B., Baldisserotto, M., Scaffaro, L.A., Grillo, L. São Lucas Hospital, Catholic University, Porto Alegre, Brazil Objectives: To determine the inflammatory bowel disease (IBD) activity using color Doppler ultrasonography. Methods: Thirty-five patients with histological diagnosis of IBD (28 of Crohn´s disease and 7 of ulcerative colitis) classified as “active” or “in remission” disease according to the Disease Activity Index were evaluated using color Doppler ultrasonography. The Doppler criterion to quantify the activity of the disease was the number of pixels measured on the bowel wall with greater number of vessels: 0–2 pixels: none or low activity; 2–5 pixels: moderated activity; 5 or more pixels: severe activity. Results: Twenty-three patients were classified as “active” disease and twelve as “in remission” disease. In the first group, color Doppler active disease criteria were found in 22 cases (95.6%). In the second group, there were no color Doppler signs of active disease in 11 cases (91%). The sensibility and specificity were, respectively, 95% and 91% on detection of IBD activity. The kappa index was 0.873 (p < 0.01). Conclusion: The color Doppler ultrasonography is an accurate and practical method for the evaluation of inflammatory bowel disease activity.

21 Azathioprine: Who ? When? Why? Schneider, N.C., Machado, M.B., Guedes, L . São Lucas Hospital, Catholic University, Porto Alegre, Brazil Base: Azathioprine and its metabolite 6-mercaptopurine (6-MP) are immunosuppressive drugs that are used in inflammatory bowel disease (IBD). In IBD, azathioprine and 6-MP have been successfully used since 1962. Objectives: to evaluate azathioprine use among IBD outpatients consulting in HSL-PUCRS. We analysed its indications and the period between first dose and diagnosis. Outline: retrospective study conducted by a chart review. Results: About 239 patients were followed until January 2004 with IBD diagnosis. Among them, 53 were on azathioprine. The distribution between sex, mean age, indications was: 27 male (50%), 35.5 y (18–68 y), 40 patients had Crohn´s disease (75%), 32 had severe disease (60%). Forty patients were on chronic corticosteroids (75%). The mean period between diagnosis and azathioprine onset was 46 months (4–216 m). Conclusion: Immunosuppression therapy with azathioprine was observed in 22% of all patients with IBD. The mean indication was chronic corticosteroid use. The most prevalent diagnosis was Crohn´s disease and the mean period between diagnosis and azathioprine onset was 46 months (4–216 m).

22 Could be anatomic extent of colitis and disease severity predictors for pouchitis after restorative proctocolectomy? G. Ianosi, D. Mercut, Daniela Neagoe University of Medicine and Pharmacy, Craiova, Romania Introduction: Restorative proctocolectomy is the most commonly procedure in surgical treatment of ulcerative colitis. One of the most common complications following this procedure is pouchitis. The aim of this study is to evaluate if perioperative anatomic extent and severity of disease could be predictors of pouchitis. Patients and methods: We evaluated 37 patients who underwent restorative proctocolectomy in which we determine the extent of anatomical lesion before the intervention and also severity of disease. Results: 23 patients from all 37 had some form of pouchitis, but we found no association with extent or severity of the disease. There are a positive correlation between the histopathologic score and the occurrence of clinical pouchitis (p = 0.01). Median follow-up was 29 months (3–72 months). Conclusion: There are no correlation between pouchitis developed following restorative proctocolectomy and the severity of ulcerative colitis.

23 The use of ozonized physiologic saline in patients with ulcerative colitis Natalia Antonova, Stella Oprea, I. Butorov, N. Bodrug The State University of Medicine and Pharmacy N. Testemitanu, Chisinau, Moldova Introduction: Ozonized physiologic saline is immunomodulator, abates inflammation, improves microcirculation. The aim of this work is to define the worth and the efficiency of ozonized physiologic saline in complex therapy of patients with ulcerative colitis. Methods: 12 patients with ulcerative colitis have been under observation. The diagnosis was made relying on the clinical data and colonoscopy. The selected patients had moderate disorders of immune status (as cellular and humoral). The patients were divided into two groups. The first group – 6 patients received ozonized physiologic saline 200.0 ml in perfusion (concentration 2.0–2.4 mg/l) and Salofalk® 1.5 g per day and. The second group – 5 patients received Salofalk® 1.5 g per day. The course of the ozonized physiologic saline therapy lasted 14 days (one perfusion in 2 days) and Salofalk® – 5 weeks. Results: The first group – bloody stools, abdominal pain, diarrhea and rectal involvement has disappeared in 6 patients (100%). The decrease of hyperemia and edema of colon mucosa at colonoscopy and normalization of Hb, RSE and indices of the immune status – in 5 patients (83.3%). The changes at the second group are analogical but some of them were less expressed in terms of quantity. Bloody stools, abdominal pain, diarrhea, rectal involvement has disappeared in 6 patients (100%). The decrease of hyperemia and edema of colon mucosa and normalization of Hb, RSE and indices of the immune status – in 4 patients (66.6%). Conclusion: The use of ozonized physiologic saline with Salofalk® in the complex therapy of patients with ulcerative colitis leads to achievement of positive results during shorter periods of time and for a greater number of patients than the use of therapeutically schemes of treatment with Salofalk®. This is due to the immunoregulatory action of ozonized physiologic saline, its capability to improve microcirculation and abatement of inflammation.

24 Doppler ultrasound measurements of the superior mesenteric artery predicts response to infliximab in patients with Crohn’s disease F. Terracciano, F.R. De Filippo, G. Biscaglia, G. Forte Department of Gastroenterology- Hospital “San Sebastiano and Sant’Anna”, Caserta, Italy Introduction: Several literature data documented that the evaluation of superior mesenteric artery (SMA) is a method to determine Crohn’s disease (CD) activity and to predict relapse after steroid-induced remission. We assessed prospectively the possible relationship between changes in mesenteric blood flow and prognosis in active patients which need of therapy with Infliximab (IFB) Methods: Doppler ultrasound (DUS) measurements of SMA were performed in 22 patients with active CD before beginning therapy with IFB (US1) and during clinical remission (CD activity index < 150, Truelove index score I) (US2). In the SMA evaluation, anteroposterior diameter, pulsatility index (PI) and flow volume were measured. Patients were weaned from therapy with corticosteroids as soon as possible and were followed up for 12 months. Results: After 1 year, 16 patients with CD (51.6%) were in remission, whereas 23 patients had recurrent disease or had undergone surgery. Differences between the active and inactive groups but not between the inactive and control groups were statistically significant for mean SMA diameter, PI, and flow volume (P = 0.019; P < 0.001; and P < 0.001). SMA flow volume values were higher than 500 mL/min in the active group (sensitivity, 78%; specificity, 99%). A decreased SMA PI at US2 predicted clinical relapse in all patients with CD (100%; P < 0.005). Conversely, an increase of SMA PI was associated with sustained remission in the majority of CD patients (12/16 patients; 75%; P < 0.001). Discussion/Conclusion: Repeated DUS measurements of the SMA PI predict response to IFB in patients with chronic active CD.

25 Cytokines in children with inflammatory bowel diseases O.A. Tutina, E.N. Fedulova, V.N. Kopeikin, I.V. Mayanskaya, N.I. Tolkacheva Scientific Research Institute of Children’s Gastroenterology, Nizhny Novgorod, Russian Federation Introduction: Imbalance between pro- and antiinflammatory cytokines is one of determinative factors in inflammatory bowel disease (IBD) pathogenesis. Cytokine status of children hasn’t been thoroughly studied yet. Methods: 56 children with IBD from 6 to 17 years old, 41 of them with ulcerative colitis (UC), 15 with Crohn’s disease (CD). Serum cytokine value IL-1β, TNFα and IL-4 was defined by IFA. Results: It was proved that children with IBD have serum values of all studied cytokines higher than that of relatively healthy group (р < 0.05). IL-4 level of most children with CD was constantly higher than that of children with UC (286.7 ± 58.8 pg/ml and 127.2 ± 33.9 pg/ml). IL-1β of children in an active phase of disease was higher, than that of those with lower activity, in remission (р = 0.03) with UC and with CD (р = 0.04) and reflected the acuteness of the process. In the group with continuous course of UC IL-1β and IL-4 were higher than that of children with recurrent course (р = 0.04). Patients with abenteric manifestations such as hepatitis, arthritis the IL-1β and IL-4 levels were lower than that of children without them (р = 0.01). The serum cytokine value of children with IBD, who received steroid therapy, was lower (IL-1β, р = 0.04), than that of those who received 5-ASA. Children with CD, who received steroid therapy, had IL-4 definitely higher than that of those who received 5-ASA. Discussion/Conclusion: The serum cytokine value of children is connected with the character of the course of disease, its phase, activity, abenteric manifestations and the character of the therapy.

26 Protective effect of supplemental calcium lactate vs. carbonate in experimental colon carcinogenesis Slimane Belbraouet1,2, Xavier Pelletier2, Gérard Debry2 1 École de Nutrition, Université de Moncton, Canada; 2Centre de Nutrition Humaine, Université de Nancy I, France Introduction: The ability to complex bile acids into insoluble salts might be responsible for the protective effect of calcium in colon carcinogenesis. The purpose was to investigate the possible protective effect of different forms of supplemental calcium in rats fed high fat diets, by histological examination and by pH and bile acids evaluation. Methods: During 32 weeks, 90 Wistar rats fed high saturated fat diet (24%) were intra-rectally instilled with N-methyl-N-nitroso-urea. Groups 1 and 2 were supplemented with calcium (1.5% Ca++) lactate or carbonate respectively and compared to a non-calcium-supplemented (group 3). Fecal pH, at 4 and 26 weeks, was evaluated using compact pH-meter and fecal bile acids were analyzed by CPG. Results: Colon carcinoma incidence was 10%, 24%, and 25% in 1, 2, and 3 groups respectively. Calcium-supplemented groups exhibit a significantly higher fecal pH than the reference group (p < 0.001). Among the calcium-supplemented groups, the fecal pH in group 1 was lower than in group 2. Strong correlations were detected between fecal pH and deoxycholic acid concentration (r = +0.72; p < 0.001). Discussion/Conclusion: The calcium associated ion plays a key-role in the protection against colon carcinogenesis chemically-induced in Wistar rats and the calcium supplementation exhibited a protective effect depending on the chemical form used. Long-term adaptation of the colon to calcium carbonate may induce a basic fecal pH whereas calcium salts with metabolizable or fermentable components such as calcium lactate may help to maintain an acidic colonic pH and thus limit, through the 7-alpha-hydroxylase, the microbial transformations of primary bile acids to secondary ones.

27 Automated classification of colon biopsy specimen by mRNA expression profiles in combination with multivariate statistical and neural network methods B. Molnar1, O. Galamb1, A. Nemeth1, M. Juhasz1, E. Dinya2, K. Toth1, N. Solymosi1, Z. Tulassay1 1 2nd Department of Medicine, Semmelweis University, Budapest, Hungary, 2Medical Department, Egis Pharmaceutical Works, Budapest, Hungary Introduction: Whole-genome gene expression profiling using microarrays permits simultanoues analysis of multiple markers and has been used to categorize cancers into subgroups. Before automated classification of gene expression patterns for diagnostic purposes the reduction of the observed parameters are necessary. Techniques for the analysis of colon biopsy specimen by whole genome mRNA expression arrays with parameter reduction and automated evaluation are not standarized, yet. Aims and methods: To develop, combine and compare multivariate statistical and artificial intellgience techniques for the reliable classification of mRNA expression profiles of colon biopsies. Total RNA was extracted, amplified from frozen colonic biopses of 22 patients with CRC, 22 with adenoma, 22 with IBD and 1 healthy control. Genome wide expression profile was evaluated by Affymetrix HGU 133 plus 2.0 microarrays for 45866 transcript including 38000 well-known genes. After ANOVA analysis, significant genes (p < 0.001) were selected and used for cluster, discriminant and neural network analysis. 75% of the chips were used as training, 25% as test set in each group, respectively. Results: Based on the selected gene pattern one normal, one IBD, one adenoma and two CRC clusters were identified by the cluster analysis. Two IBD cases, one adenoma, 2 CRC 2 normals case were misclassified (90% accuracy). Using linear discriminant analysis the data set could be further reduced to 7 genes. Using 3 discriminant factors from these seven genes (Interferon induced prot. 1, proteosome subunit beta type 9, hyaluronon, proteoglycan link prot. 1, solute carrier family 36 member 1, and 3 hypothetical proteins) one adenoma and one carcinoma test were misclassified, only. Artificial neural networks using 7 input neurons, 23 hidden neurons in one layer and four output neurons could correctly classify the training and the test set before overlearning. Conclusion: ANOVA based significant gene selection in combination with neural network based automated interpretation of RNA expression array data from colon biopsy specimen yields safe classification. This marker set and classification method in combination with automated sample preparation, hybridisation and scanning techniques can lead to novel diagnostic alternatives.

28 Dysplasia in inflammatory bowel disease in Romania Simona Bataga, Imola Torok, Dan Georgescu, T Bataga, Ana Bratu University of Medicine and Pharmacy Tg-Mures, Romania Introduction: The aim of the study is to follow-up the incidence of the malignant changes on the patients with ulcerative colitis (UC) and Crohn’s disease (CD). Methods: 2027 patients (mean age 62.3 years, 61% males) who underwent rectosigmoidoscopies and total colonoscopies on a period of ten years entered this study. The diagnosis was put on endoscopy and histology. Results: From these patients 59 (2.91%) had IBD, 53 (2.56%) had UC and 6 (0.29%) had CD. Patients with IBD had colonoscopy every year and biopsies from 10 to 10 cm were taken. Patients with UC presented mostly mildly active disease with good response to the medical treatment: Salofalk®, only 4 cases (7.54%) of severe disease. From 59 patients in 2 patients were revealed polyps with dysplasia. Both patients had UC from long time: one patient from 20 years and the second from 12 years. Both patients were sent to surgery. From the patients with CD (4 males, mean age 34.7 years), 2 patients had small bowel CD and were diagnosed on surgery and 4 patients had Crohn’s colitis. None of the patients presented dysplasia, but all the patients have less than 10 years of evolution. Discussion/Conclusion: IBD have a low incidence in Romania (2.91%). UC represents 2.56% and the incidence of the dysplasia was 3.38% in UC after more than 10 years of evolution. CD has a low incidence in Romania (0.29%), but it seems to increase in the last years. At less than 10 years of evolution there was no dysplasia.

29 The frequency and risk factors for dysplasia in Crohn’s disease R. Vadan, C. Gheorghe, G. Becheanu, L. Gheorghe, M. Dumbrava, M. Diculescu Gastroenterology and Hepatology Center, Fundeni Clinical Institute, Bucharest, Romania Introduction: Crohn’s disease (CD) is characterized by persistent inflammation of the bowel wall and consequently is associated with an increased risk of dysplasia and carcinoma especially after long term evolution of disease. Our study aimed to describe the frequency and eventual risk factors for the occurrence of dysplasia in patients with CD. Methods: Patients admitted in our unit in the last two years and diagnosed (based on conventional endoscopic and histologic criteria) with CD were retrospectively evaluated. All patients included in the study had complete colonoscopic and radiologic small bowel follow up and biopsies were taken from inflamed colonic mucosa (diagnostic purpose) and also from all mass lesions encountered. The following data were noted: age at diagnosis, duration of disease, location, severity of current and previous disease flares, current and previous medication. Results: 212 patients were included in the study, 46.2% females and 53.8% males with a mean age at diagnosis of 39.54 ± 1.05 and mean duration of disease of 4 ± 0.43. Location of disease was colonic 63.7%, ileocolonic in 27.8% and jejunoileal in 8.5%. Dysplasia was diagnosed in 9% (19) patients: 2 (0.9%) with DALM (dysplasia associated lesions or mass), 5 (2.4%) with adenocarcinoma and 12 (5.7%) with adenomatous polyps. Duration of disease and severity of disease flares were not significantly associated with the presence of dysplasia. Discussion/Conclusion: The frequency of dysplastic lesions in Crohn’s disease is not low and the prognostic importance of dysplasia makes its recognition and diagnosis mandatory.

30 Fistula-associated carcinoma in Crohn’s disease H. Vogelsang1, F. Herbst2 1 Clinic of Internal Medicine IV, Department of Gastroenterology 2 Surgery Department, Medical University of Vienna, Austria Introduction: The risk of intestinal carcinoma is increased in Cohn’s disease although generally rather rare. Methods: We retrospectively investigated patients with Cohn’s disease (out of 2416 patients) operated with intestinal carcinoma during the last 10 years (1997–2006). Out of 591 abdominal operations in Crohn’s patients we found six patients (5 male, 1 female, median age: 50 years (range: 37–74)) with ileocolonic carcinoma. Results: One male patient (38 years) was operated with ileal adenocarcinoma and died of diffuse metastases within 1 year. Out of five patients with colonic adenocarcinoma two were not associated with fistula (53 and 74 years, adenocarcinoma of rectum and ascending colon resp.). 3 patients (37, 48, 66 years) suffered from Cohn’s disease (ileocolonic) for 13, 28, 33 years resp. and from perianal/ rectovaginal fistula for 12, 17, 28 years resp. They were under azathioprine treatment for 30, 5, 15 months resp., but only one infliximab infusion was given in one patient (3 years before carcinoma). No significant symptoms preceded rectal carcinoma, but new blood drainage from fistula in 2 patients. 2 patients underwent MRI of the pelvic region within 4 months before surgery without recognizing malignant changes. Adenocarcinoma in the region of rectal fistula was found in these 3 patients with a staging of Dukes A-C. Discussion/Conclusion: Intestinal adenocarcinomas are rare in patients with Cohn’s disease, but frequently associated with anorectal fistula (50%) in patients with longstanding Crohn’s disease (> 12 years of fistula). It occurred with minimal change of symptoms and was not visible in MRI (no differentiation of fistulous changes).

31 Systematic administration of 5-ASA derivatives during remissions for more than 9 months/year seems to protect patients with longstanding ulcerative colitis against colorectal cancer Eugen Florin Georgescu1, Marius Georgescu2, Manuela Vasile1 1 Filantropia University Hospital of Craiova, Department of Internal Medicine II 2 Emergency Departmental University Hospital of Craiova, Department of Internal Medicine II Introduction: The aim of this retrospective study was to estimate if administration of 5-aminosalicylic acid derivatives (5-ASA) on a regularly base during remissions has any benefit in prevention of colorectal carcinoma (CRC) in patients with longstanding ulcerative colitis (i.e. more than 10 years after initial diagnosis has been made). Methods: The study was performed between April 2000 and May 2006 and evaluated 406 patients with longstanding UC enrolled in our regional registry. Data collection showed that 219 of 406 patients (53.94 ± 3.15%) were taking 5-ASA derivatives every day on a regularly base for more than 9 months/year. The mean duration of therapy was 12.34 ± 3.47 years in this group and the extension of colitis has been stratified as follows: proctosigmoiditis: 43.12 ± 3.54%, left-sided colitis 34.23 ± 1.25% and pancolitis (22.25% ± 5.73%). In average, each patient received a dose of 1.645 ± 0.213 grams of 5-ASA per day. Results: At the end of study we found an overall incidence of CRC of 3.02 ± 0.23%. Lower incidences of 1.44 ± 0.13% were observed in the 5-ASA treated group, while in the non-5-ASA treated group a significantly higher incidence of 3.23 ± 0.45% has been noticed (p < 0.05). The risk for CRC was higher in patients with pancolitis in both groups but those from the 5-ASA group has a significantly lower risk with an odds ratio of 0.51 (95% CI, 0.37–0.69) compared with 0.86 (95% CI, 0.71–0.92) for patients which did not followed chemoprevention. Discussion/Conclusion: Chemoprevention with 5-ASA derivatives seems to protect patients with UC from CRC, but the results are influenced by the extension and perhaps the duration of disease.

32 Implication of observation of patients with ulcerative colitis considering the occurrence of colorectal carcinoma Konecny, M., Ehrmann, J., Prochazka, V., Kucerova, V., Vyslouzil, K. 2nd Medical Department, University Hospital, Olomouc, Czech Republic Introduction: Chronic course of idiopathic bowel disease manifests by recurring relapses with relatively high occurrence of complications, both intestinal and extraintestinal. The most serious complications include colorectal carcinoma (CRCa) in patients with long-term ulcerative colitis (UC) of extensive type. Set of patients and methodology: In 1995–2005, 71 patients with UC of extensive type lasting longer than 10 years were observed within the follow up of the 2nd Internal Clinics of the Palacky University Medical Faculty. In the whole set, colonoscopic examinations were carried out in regular two-year (or shorter) intervals with follow-up biopsy and histological examination focused on the occurrence of dysplastic changes and colorectal carcinoma. Results: In 5 patients CRCA was found as complications of long-term course of UC, in 39 patients, dysplastic changes have been detected and monitored their development and seriousness. In 27 persons, no dysplastic changes have been found during the whole period of observation. Conclusions: Regular colonoscopic examinations should be commenced within 10 years from the date when the UC of extensive type was found. If no dysplastic changes are found during biopsy, we continue in endoscopic examinations in twoyear intervals. When dysplasia of low grade is found, we perform another colonoscopy with the follow-up biopsy within one year. If the DALM type of macroscopic lesion or a stricture is found, colectomy is indicated. High-grade dysplasia is also an indication for colectomy as CRCa is found in the resectate in 36% of cases.

33 Colorectal cancer prevention in inflammatory bowel disease according to evidence-based medicine D. Ledro-Cano Hospital Universitario de Puerto Real, Pilas, Cádiz, Spain Aim: We performed a systematic review with metaanalysis of clinical studies evaluating the association between different drugs and colorectal cancer (CRC) or dysplasia among patients with inflammatory bowel disease. Patients and methods: We conducted a search of Medline. Studies were included if they reported 1) exposure to drugs in patients with ulcerative colitis, 2) CRC or dysplasia outcomes, 3) relative risks or odds ratio (OR). Results: 5-ASA: ten studies (3 cohort, 7 case-control) containing 410 cases of CRC, 140 cases of dysplasia, and a total of 20,901 subjects satisfied all inclusion criteria. Analysis showed a protective association between use of 5-aminosalicylates and CRC (OR = 0.6; 95% confidence interval [CI]: 0.4–0.86) or a combined endpoint of CRC/dysplasia (OR = 0.51; 95% CI: 0.38–0.69). 5-ASA use was not associated with a lower risk of dysplasia, although only two studies evaluated this outcome (OR = 1.18; 95% CI: 0.41–3.43). Ursodeoxycholic acid: 2 cohort studies containing 13 ulcerative colitis patients, disease, median age 43 years) in one study and 52 subjects suffering from ulcerative colitis and primary sclerosing cholangitis were followed-up for a total of 355 personyears. In the first study, no UDCA-treated patient had progression of dysplasia. In the second study, those originally assigned to receive UDCA had a relative risk of 0.26 for developing colorectal dysplasia or cancer (95% CI: 0.06–0.92; p = 0.034). Folic acid: 3 studies. The association of these two factors is significant. (effect size r = 0.124, p = 0.025). Conclusions: 5-aminosalicylates or UDCA or folate supplementation reduces CRC/dysplasia in patients with ulcerative colitis.

34 Risk for colorectal cancer in inflammatory bowel disease Szanto, Paula1, Manciula, Dorina2, Cruciat, Carmen3, Seicean, Andrada 1 rd 3 Medical Clinic, University of Medicine and Pharmacy, Cluj, Romania 2 County Hospital Baia-Mare, Romania 3 Adult Clinical Hospital, Cluj, Romania Introduction: Patients with inflammatory bowel disease (IBD), both ulcerative colitis (UC) and Crohn’s disease (CD), are at risk for the development of colorectal cancer (CRC). Extensive lesions, long duration of disease, older age at symptom onset, primary sclerosing cholangitis (PSC), and more recently obesity and the metabolic syndrome, are purposed risk factors. The aim of our study was to establish the role of this purported risk factors for CRC in both UC and CD. Methods: Twenty two IBD patients were followed-up between 1997 and 2006. There were 17 UC (10 men and 7 women), and 5 CD (2 men and 3 women) with mean age at the onset of the disease 37.98 years (limits 24–56). The mean duration of the disease was 10.22 years (limits 4–36), fourteen IBD patients having a duration of disease longer than 10 years. All patients with CD had mild to moderate forms of disease. One woman with a four years evolution of CD was identified with small ducts PSC. Between patients with UC, nine (53%) had pancolitis at the first diagnosis, and eight proctosigmoiditis, fifteen patients having moderate-to-severe disease. Ten patients with IBD (45.4%) had obesity. All the patients received maintenance therapy with aminosalicylates at least nine months\year and 18 of them with corticosteroids in acute periods. Results: None of our IBD patients developed high-grade dysplasia or CRC. Discussion/Conclusion: We concluded that risk for IBD-related colorectal cancer is very low in our area possibly due to the optimized treatment and surveillance, or to a particular risk-factor profile.

35 Clinical features and pathogenesis in inflammatory bowel disease-associated intestinal cancer T. Molnar, I. Németh, Z. Szepes, F. Nagy, L. Tiszlavicz University of Szeged, Szeged, Hungary Introduction: Patients with inflammatory bowel disease (IBD) [ulcerative colitis (UC) and Crohn’s disease (CD)] are at increased risk of developing gastrointestinal, mainly colorectal cancer (CRC). The main risk factors are long-standing duration of the disease, extensive colitis, chronic stenosis, primary sclerosing cholangitis and the family history of CRC. The estimated prevalence, the risk factor, the clinical features and the molecular pathogenetic alterations of IBD-associated intestinal carcinoma was examined retrospectively in a cohort of patients with IBD from a well-defined East Hungarian geographical area. Methods: There were 14 patients with IBD-associated intestinal cancer (10 with UC and 4 with CD; 13 CRC and 1 small bowel cancer) who were evaluated at the University of Szeged between 1987 and 2005. Medical records were reviewed retrospectively for endoscopic and histologic characteristics as well as detailed clinical phenotypes. The following parameters were investigated by tissue microarray technique in the tumor and in auto control healthy tissue: grade of differentiation, lymphovascular invasion, perineural spreading, peritumoral inflammation, necrosis, desmoplastic reaction and the gene expression profile of p53, MLH1, MSH2, β-catenin, PTEN, APC, K-ras, Cox-2, NOS, NFĸB and VEGF. Results: The estimated prevalence of IBD-associated intestinal cancer was 0.015% among all CD and UC patients during the examined period. The median age at diagnosis of IBD-related intestinal cancer was 56 years, the median duration was 23.7 years. The main risk factors were chronic activity, extension of the disease and positive familial anamnesis for CRC. 8/14 of IBD-related tumors were distal to the splenic flexure. The inflammatory markers were positive in all tumors. The molecular alterations found in IBD-associated cancers involved the same targets as in sporadic CRC. Discussion/Conclusion: Chronic uncontrolled inflammation leads to neoplastic transformation on the main molecular pathways of colon carcinogenesis in IBD patients. Thus, the central question of carcinoma prevention in IBD is the continuous control of inflammation in the whole gastrointestinal tract.

36 In what degree is the inner and outer environment affecting the developing of the colon cancer? L. Tiurean1, C.E. Rezi1, R. Mihaila2, M. Deac2, R. Mihaila3, O. Petrascu1 1 County Clinical Hospital Sibiu, 2nd Medical Department, Romania 2 University of Sibiu, Faculty of Medicine, Sibiu, Romania 3 Public Health Authority Sibiu, Romania Introduction: It is known that the hypercaloric and hyperlipidic diet is involved in the appearance of the colon cancer. But, in the same time, obesity, dyslipidaemia and diabetes mellitus constitute risk factors for this neoplasm. Methods: We desired to study if the presence of the components of the metabolic syndrome (MS) can influence the appearance of the colorectal cancer. We have studied a group of 332 patients which were hospitalized in the 2nd Medical Department of the County Clinical Hospital from Sibiu during January 2002– September 2006, which had as the main diagnosis Ischemic Cardiac Disease. At this group, we have studied: age, gender, cholesterol and triglycerides level, glycaemia, the presence and the type of the MS’s components, the colorectal cancer’s diagnosis. Results: From the 332 patients which were studied, 65% were women and 35% were men. The medium age of the lot was 64.98 ± 11.54 years. We obtained the next average values: glycaemia 102.92 ± 29.98 mg/dl, cholesterolaemia 200.118 ± 57.08 mg/dl, triglyceridaemia 127.149 ± 100.46 mg/dl. 35 of the patients had the diagnosis of colorectal cancer. The relative risk (RR) for the appearance of the colorectal cancer at the patients with dyslipidaemia was 1.01, at those with diabetes mellitus – 1.12, at those with arterial hypertension – 1.39. These results suggest that these 3 conditions are risk factors for the colorectal cancer. The RR for the appearance of the colorectal cancer at the patients with obesity was only 0.43, but the patients with severe obesity (stage III) have a risk of developing the colorectal cancer twice higher than the patients which have a mild obesity (stage I), (RR = 2.25). The RR of the appearance of the colorectal cancer at the patients with MS comparing with those without MS was 2.23, which means that those with MS have a two times higher risk to develop colorectal cancer than those without this syndrome. The RR for the developing the colorectal cancer at those who have 2 or more components of the MS against those who did not have any component at all was 1.85. Discussion/Conclusion: At the patients with MS, the colorectal cancer has a higher prevalence comparing with those without MS and the risk of developing the cancer increases together with the number of components of the MS and with the stage of the obesity. The diet of these patients needs to be corrected.

37 Early premalignant and malignant lesions: Detection of colon rectosigmoideum by magnifying coloscopy and chromoendoscopy Wohl, Pavel, Honsova, E., Wohl, Peter, Spicak, J. Institute for Clinical and Experimental Medicine, Prague, Czech Republic Introduction: Non-polypoid colorectal premalignant and malignant lesions can negatively influence screening programs efficacy. Occurrence of such lesions in our population is unknown. The aim of this study is to evaluate the suitability of magnifying coloscopy and chromoendoscopy as a detection method for early carcinoma. Method: The study included patients who underwent first coloscopy-magnifying coloscopy and chromoendoscopy 0 to 30 cm from anus has been performed on them. All lesions were evaluated by Kudo’s pit pattern classification and examined histologically. Results: 57 patients participated in the study, 28 men and 29 women. The median age was 57.59 ± 7.45 years. The indications for coloscopy were screening (23), FOBT (9) and blood in the stool (6). From total of 211 detected polypoid and nonpolypoid lesions in rectosigmoideum 188 (89.1%) were visible only by indigocarmine. The median size was 2 mm (1 ± 4 mm). Pit pattern I wasn’t detected. From 188 nonpolypoid lesions pit pattern II was detected on 170 lesions (90.4) from which 126 (74.1%) were hyperplastic polyps, 37 (20.5%), nonspecific lesions, 4 inflammations and 1 xantom. Pit pattern III was detected on 17 lesions, 8 (47.05%) tubular adenomas with mild dysplasia, 8 (47.05%) hyperplastic polyps and 1 inflammation. One was of pit pattern III, a lymphatic follicle. Pit pattern IV and V weren’t detected. Cancer was detected in 2 patients with macroscopically visible lesions. Conclusion: Magnifying coloscopy and chromoendoscopy are suitable for rectosigmoid visualization. We can conclude that non-polypoid pathogenesis of colorectal carcinoma plays no significant role in our population. But we found more hyperplastic polyps compare to other studies (IGA MZ CR 7878/3)

38 Methods of improving the yield of lesions at surveillance colonoscopy Somesundrum, C., Papadi, B., Besherdas, K., Van Somerun, N., D’Souza, R. Department of Gastroenterology, Chase Farm Hospital, The Ridgeway, Enfield, UK Introduction: Multiple factors impact upon the yield of colonoscopy for the detection of neoplasia. The impact of colonoscopy withdrawal time, position change and buscopan use on the yield of lesions was assessed on a porcine model and in surveillance colonoscopies. Methods: 100 surveillance colonoscopies were reviewed. Different endoscopists were involved. The length of time for withdrawal varied from 4–12 minutes. Patients receiving buscopan or change of position during withdrawal were recorded. A porcine model with different sized buttons sewn into the lumen was used. The same endoscopist then withdrew the colonoscope at different times and the number of buttons visualised recorded. Results: Large polyps/buttons (> 1 cm) were easily detectable with shortened withdrawal times, but smaller and flat polyps/buttons (< 5 mm) were detected with longer withdrawal times (> 7 mins). The optimum withdrawal for detecting polyps was 7 minutes. Polyp detection rate was around 28%. Buscopan use seemed to detect more polyps. Changing the position increased the chance of detecting polyps compared to those operators who kept the patients in the same position. Numbers were too small to analyse statistically. Discussion/Conclusion: There is increased pressure in the health service to do more endoscopic procedures. Reducing the withdrawal time at colonoscopy would be detrimental as some of the flat lesions missed were adenocarcinomas. Change of position and buscopan on withdrawal are recommended as they increase the chance of detection of polyps. The role of dye spraying for dysplastic changes were not analysed in this study.

39 Diagnosis of dysplastic lesions in cases of ulcerative colitis A. Ioffe, N. Denisov, R. Abu Shamsieh National Medical University, Kyiv, Ukraine Introduction: The objective of study was to investigate possibilities of PDD method for reliable detection of colon mucosa dysplastic lesions using hypericin induced photosensitizer Hyperflav (Borshagiwka Pharmaceutical Co., Kyiv, Ukraine). Methods: PDD of colon mucosa was performed afterwards oral administration of encapsulated Hyperflav. A sufficient fluorescence contrast of suspicious versus normal tissue has been obtained after 6–10 hours. Hyperflav peak at 603 ± 3 nm illustrates clearly its considerable accumulation by dysplastic colon mucosa and enables discrimination of lesion margins. Results: In 2001–2006 we have investigated 86 patients (male/female 47/39; average age 47 ± 3) with ulcerative colitis using proposed technique. The biopsies taken both from areas indicated Hyperflav presence and from control areas have been histologically tested (42 + 22 biopsies of 22 patients). Histological examination results of biopsies taken as predicted routine colonoscopy and PDD are presented in Table.

Method

No. of patients

Morphology

No. of biopsies

Adenocarcinoma

Dysplasia higher stage

lower stage

Normal mucosa

Colonoscopy

86/86*

837

3

7

1

826

PDD using Hyperflav

86/22*

42 + 22**

6

25

1 + 1***

9 + 22**

* Patients tested with biopsy. ** Control biopsies taken from normal mucosa. *** Biopsies specified as false-negative results.

Discussion/Conclusion: Method demonstrated the advantages to select areas for biopsies that raised effectiveness of target biopsy (accuracy 84%, sensitivity 97%, specificity 71%). Application of PDD using Hyperflav might be effective tool in dysplasia detection in cases of ulcerative colitis.

40 Special endoscopic methods for tattooing and localizing early colorectal cancers before laparoscopic colon resection Attila Szijártó, Judit Wacha, Péter Kupcsulik Semmelweis Egyetem University, 1st Department of Surgery, Budapest, Hungary The early colorectal cancer could be developed by some new special endoscopic techniques and devices, i.e. high-resolution videoendoscopy, zoom endoscopy, chromoendoscopy. Chromoendoscopy involves the topical application of stains or pigments to improve tissue localization, characterization, or diagnosis during endoscopy. The staining results a special „pit pattern”, which refers about the degree of dysplasia in the mucosa. This method allows recognizing some special tumor, as flat adenoma, depressed adenoma and depressed type of early colorectal cancer. Some of these lesions are suitable for polypectomy or mucosectomy, but some of them require surgical intervention. In the current era laparoscopic-assisted surgical colonic resection is becoming as well accepted as primary colonoscopy. Unlike open laparotomy, the laparoscopist cannot palpate the colonic lesion. Thus, it is important to have an easily visible marker that can be seen through the telescopic lens of the laparoscope. It is our practice to inject a permanent surgical marker during the initial colonoscopy if a lesion cannot be removed or if an endoscopically removed polyp has clinical features that may require surgical resection or surveillance colonoscopy. A number of methods continue to be used for localizing lesions in the colon. However, only colonic tattooing and metal clips are consistently reliable. In this study 85 laparoscopic colon resections are presented, where in 10 cases preoperative endoscopic tumor localization was done after chromoendoscopy. We used metal clips, which were placed through the colonoscope and onto the mucosa at any location. They can assist in radiographic location of the marked segment. In our study will discuss our experience about localization of colonic lesions, while focusing on chromoendoscopy and metal clips as a permanent marker.

41 Narrow-band imaging (NBI) differentiation of polyps in the patients with ulcerative colitis Miroslaw Szura, Aleksander Zajac, Rafal Solecki, Jan Kulig 1st Department of General and GI Surgery, Jagiellonian University, Kraków, Poland Head of the Department: Prof. Jan Kulig, M.D., Ph.D. The patients with ulcerative colitis are generally more susceptible to colon cancer. In these patients colon cancer usually arises from polyps that are detectable by endoscopy. Their number frequently does not allow to perform one-step removal of all polyps. For this reason, only macroscopically suspicious polyps are removed. Narrow-band imaging is one of the latest technological achievements of the digital endoscopy. The technique is using interference filter to light surfaces in narrow bands of red, green, and blue colour that visualizes the differences in mucosa colouring and enhances contrast between the mucosa surface and submucosa vascular network. The study evaluates the possibility of using NBI for macroscopic differentiating colon polyps in the patients with ulcerative colitis. Thirty patients with endoscopically detected multiple polyps and histopathologically proven ulcerative colitis were enrolled in the study. All endoscopic examinations were performed using HDTV Olympus equipment series 180. Polyps were assessed using xenone light and NBI. Polyps suspected of developing colon cancer and unsuspected lesions were sent for histopathological verification. Both endoscopic examinations and macroscopic polyp differentiating were done by three independent experienced endoscopists based on the specially designed protocol. In thirty macroscopically suspected polyps, there was 1 colon cancer, 4 cases of high grade dysplasia, 6 low grade dysplasias, and inflammatory reactions in the remaining 19 patients. In macroscopically unsuspected lesions there was 1 case of high grade dysplasia, 5 low grade dysplasias and in the remaining patients inflammatory reactions. The endoscopists differentiated adenomas from inflammatory polyps with 76% accuracy. A new technique of NBI allows more precise preliminary assessment of polyps localized in colon and at the same time may be useful in qualifying polyps for endoscopic removal in the patients with ulcerative colitis.

42 Nimesulid impact on the expression of colorectal cancer epithelial markers in patients with ulcerative colitis after liver transplantation for primary sclerosing cholangitis Wohl, Pavel1, Honsova, E.1, Lodererova, A.1, Sticova, E.1, Wohl, Peter1, Halakucova, P.2, Spicak, J.1 1 Institute for Clinical and Experimental Medicine, Prague, Czech Republic 2 Department of Gastroenterology, Rakovnik, Czech Republic Introduction: The risk of colorectal carcinoma increases with ulcerative colitis (UC) and primary sclerosing cholangitis (PSC). The aim of this study was to evaluate the impact of nonsteroidal anti-inflammatory drugs (NSAID) on the expression of colorectal cancer epithelial markers with long term UC patients after liver transplantation (OLT) for PSC. Method: Randomized double-blind placebo controlled study with nimesulid (200 mg once a day, for 6 months). Twenty-two UC patients after OLT for PSC were included. 19 patients completed the study (1 excluded at his own request, 2 diagnosed with posttransplantation lymphoproliferative disease PTLD). Patients were randomized 2:1; Nimesulid (INT, n = 13) vs. placebo (PLB, n = 6). Biopsy specimen were processed histologically and semi-quantitatively imunohistochemically. To prove antigen presence two-grade indirect imunoperoxidase reaction using following antibodies: DCC (BD Pharmigen), APC, SMAD 4 (Santa Cruz Biotechnology), bcl-2 Oncoprotein, p53 (DakoCytomation), COX-2, COX-1 (Cayman) and immunoperoxidase polymer (Histofine Simple Stain MAX PO, Nichirei) and chromogen 3,3´ diaminobenzidin (DakoCytomation). Statistical relevance was evaluated by χ2 quadrate. Results: INT group showed statistically significant decrease in APC expression before and after nimesulid treatment (expression 2+3 to 0+1) (7.7% and 92.3% vs. 23.1% and 76.9%, p < 0.002). Decrease in COX-2 expression was not statistically significant (38.5% and 61.5% vs. 61.5% and 38.5%, NS), COX-1 expression increased statistically after Nimesulid use (69% and 30.8% vs. 69.2% and 30.8%, NS, p < 0.002). No statistical difference in expression of p53, SMAD 4, DCC and bcl-2 before and after Nimesulid treatment was found in the INT group. No statistically significant difference in expression of epithelial markers was found in PLB group. Likewise, no statistically significant difference was found in histological activity of UC in INT and PLB. Colorectal carcinoma or dysplasia was not detected. Conclusion: Nimesulid treatment led to significant decrease in APC expression, which is due to COX-2 independent mechanisms of NSAID impact. Despite the fact that COX-2 expression decrease was not statistically significant, which may be caused by the small size of the group, the trend was distinct. Nimesulid most probably influences COX dependent and independent mechanisms of colorectal carcinogenesis. (grant IGA MH CR 7878/3)

43 Oxidative DNA damage levels are increased in sporadic colorectal cancer Juliette Sheridan1, Lai-Mun Wang1, Miriam Tossetto1, Kieran Sheahan1, John Hyland1, Diarmaid O'Donoghue1, Hugh Mulcahy1, Jacintha O'Sullivan1 1 Department of Gastroenterology, St. Vincent's University Hospital, Dublin, Ireland Introduction: 8-oxodG DNA levels are a biomarker of oxidative DNA damage; 8-oxodG is a pro-mutagenic lesion as it mispairs preferentially with adenine during replication leading to transversions which are often found in abundance in mutated tumour suppressor genes. Our aim was to evaluate the extent of oxidative DNA damage in colorectal cancer patient group vs. a control group of non-cancer patients. Methods: Tissue micro-arrays were constructed from formalin-fixed paraffin embedded tissues from 143 randomly selected colorectal cases and 105 non-cancer cases. The control group consisted of patients with normal histology and normal inflammatory markers. Immunohistochemical analysis was performed on 5 µm sections using an anti 8-oxodG monoclonal antibody (Genox) and the DAKO Envision kit. The levels and localization of 8-oxodG was examined in both groups. Results: The levels of epithelial nuclear staining were significantly higher in the normal non-adjacent mucosa of the colorectal cancer group (mean 67.1) vs. the control group (mean 54.88) (p < 0.001). Age was not a factor. When looking at the cancer group alone, the levels of epithelial nuclear staining were significantly higher in the normal non-adjacent mucosa vs. the tumour mucosa (mean 65.44) vs. (mean 46.76) (p < 0.001). Cytoplasmic staining (confirmed mitochondrial oxidative DNA damage by co-localisation immunofluorescence) was significantly higher in tumour mucosa vs. normal non adjacent mucosa (mean 87.69) vs. (mean 52.18) (p < 0.001). Discussion/Conclusion: Higher levels of oxidative nuclear DNA damage found in the normal mucosa of colorectal cancer patients may cause the propagation of mutated cells leading to the malignant transformation of colonic mucosa.

44 Expression, localization and systemic concentration of transforming growth factor-beta1 (TGF-β1) and its receptors in patients with ulcerative colitis A. Stadnicki1,2, G. Machnik3, A. Wolanska-Karut4, B. Bojko5 1 Department of Basic Biomedical Sciences, Medical University of Silesia, Katowice, Poland, 2Department of Gastroenterology, St Barbara’s Hospital, Sosnowiec, Poland, 3 Department of Molecular Biology and Genetics, 4Department of Pathology, 5 Department of Physical Pharmacy, Medical University of Silesia, Katowice, Poland Introduction and aims: Transforming growth factor β1 (TGF-β1) is a multifunctional cytokine, that plays a role in homeostasis of intestinal mucosa by activating the specific three receptor’s (TGFβR I–III). In the present study we investigate the expression of genes encoding TGF-β1 and its receptors as well as level of TGF-β1 in colonic tissue and serum of patients with ulcerative colitis (UC). Methods: Biopsy samples from 36 UC patients (22 with active UC phase, and 14 with inactive UC phase) and 14 normal controls were taken during colonoscopy. The expression of genes encoding TGF-β1 and TβRI, II, III was quantitatively assessed by QRT-PCR reaction using (TaqMan). The localization of TGF-β1 protein in intestinal tissue was estimated by immunohistochemistry, the level of TGF-β1 protein in colonic samples was measured in four grading scales. In addition TGF-β1 serum concentration was determined by ELISA. Results: We found a significant (p < 0.001) increase of TGF-β1 gene expression in patients with active UC (81,065 ± 1753 of mRNA µg/total RNA) as compared with controls (17,657 ± 4679). The expression of genes encoding receptor TRβI was significantly higher (p < 0.005) in patients with active UC (22,776 ± 5665) as compared with controls (6873 ± 857). The expression of genes encoding TRβII was found to be higher in patients with active UC (88,032 ± 17,453, p < 0.05) as well as with inactive UC (116,588 ± 16,530, p < 0.002) than in normal subjects (27,616 ± 8409). No significant difference in TRβIII genes expression was observed in all examined groups. TGF-β1 protein was visualized in enterocytes and fibroblasts localized in lamina propria. The specific reaction for TGF-β1 staining was significantly higher in active UC (26 points, p < 0.005) as well as in inactive UC (15 points, p < 0.017) as compared with normal controls. Serum level of TGF-β1 was significantly higher (p < 0.01) in UC active patients (29.5 ± 5.5 ng/ml) as compared with controls (18 ± 1.6). The serum level of TGFβ-1 was significantly higher in patients with left side/total colonic extension (28.5 ± 5.0) when compared with those with rectosigmoid (23.9 ± 2.0, p < 0.03). Conclusions: The increase of TGF-β1 gene expression as well as its protein level concomitant with altered TGF-β1 receptors profile may contribute to intestinal inflammation in ulcerative colitis. Increased TGF-β1 serum concentration is related to activity and extension of UC.

45 Can the statins block in an efficient way the progression of the cellular cycle, preventing in this way the colorectal cancer? R. Mihaila1, C.E. Rezi2, L. Tiurean2, R. Mihăila3, M. Deac1, O. Petrascu1, L. Bera1 1 University of Sibiu, Faculty of Medicine, Medical Clinic II, Romania 2 County Clinical Hospital Sibiu, Medical Clinic II, Romania 3 Public Health Authority Sibiu, Romania Introduction: It was established that the statins determine the arrest of the cellular cycle and they contribute at the appearance of the apoptosis in the colon cancer cells. We proposed ourselves to study if the long time administration of the statins at the patients with ischemic cardiopathy can prevent the appearance of the colorectal cancer. Methods: We have studied 276 patients which were hospitalized in the IInd Medical Department of the County Clinical Hospital from Sibiu, during January 2002– September 2006, which had ischemic cardiopathy as the main diagnosis. They were divided in two groups: group A – which were treated on a long period of time with statins, and group B, which were not treated with statins. At them, we have analyzed the next parameters: age, gender, the presence or the absence of the colorectal cancer diagnosis, the type and the dose of the statins which were used. The results were statistically analyzed, using the relative risk. Results: From the 34 patients from group A, just 1 had colorectal cancer. Among the 242 patients from group B, we found 32 which had a colorectal cancer. The relative risk for the appearance of the colorectal cancer at the patients from group A, comparing with those from group B is 0.222, which means that, in our study, the use of these medication can be considered as a benefic factor for the health, preventing in this way the appearance of the colorectal cancer. The statins which have been used were: simvastatin (47%), atorvastatin (26.5%), lovastatin (14.7%) and fluvastatin (11.8%). Discussion/Conclusion: These results suggest that the statins have, apart from the beneficial cardiovascular effects, pleiotropic effects on the intestine, contributing at the prophylaxis of the colorectal cancer.

46 Particular aspects of premalignant lesions in inflammatory bowel disease: Morphological study O. Fratila, A. Maghiar, C. Brisc, Dana Puscasiu, M. Puscasiu 3rd Medical Clinic, University of Oradea, Oradea, Romania Introduction: Our aim was to evaluate the clinical presentation and pathologic features of polypoid dysplastic lesions and sporadic adenomas in patients with IBD. Methods: The morphologic features of 64 polyps from 48 patients undergoing colonoscopy between 1999–2006 (36 ulcerative colitis, 8 Crohn’s disease and 4 indeterminate colitis; 25 male/23 female, mean age 45 yrs, range 27–65 yrs) were evaluated and correlated with the clinical and endoscopic data. Sixty-eight adenomas from a local matched age control group without IBD were similarly analyzed. Results: Most polyps (0.2–4 cm) were sessile, whereas only 6 (9.3%) were peduncular. The majority occurred in the left colon or rectum 43 (67.1%). Most polyps developed within an area of colitis had a tubulovillous architecture 54 (84.4%), mixed normal-low grade dysplastic epithelium and increased lamina propria mononuclear inflammation 41 (64%). The polyps located outside of areas of histological proven colitis showed similar features with those in the control group. The polyps in the control group were sessile 44 (64.7%) and peduncular 24 (37.8%), 0.4–7 cm sized. They were located mainly in the left colon and rectum 45 (66.2%). Most were tubular adenomas 48 (70.6%) with mild or moderate dysplasia 58 (85.3%) or severe dysplasia (carcinoma in situ) in 10 (14.7%). Discussion/Conclusion: In both groups, almost one third of the polyps were located in transverse/right colon, recommending total colonoscopy for diagnosis and followup of colonic polyps. Several morphologic features may help to define a dysplastic lesion as a sporadic adenoma or a potential IBD-related neoplasm in a patient with IBD, providing a different management.

47 Effectiveness of the questionnaire investigations in the group of patients with high risk of the hereditary colorectal cancers Tomasz Banasiewicz, Piotr Golusiński, Marcin Grochowalski, Aleksander Niziołek, Chair and Department of General, Gastroenterological and Endocrynological Surgery K Marcinkowski University of Medical Sciences, Poznań, Poland Wojewódzki Szpital Zespolony, Poznań, Poland Colon cancer is one of the most common cancers, in 25% of cases the positive familial history is recognized. In the group of patients with hereditary colon cancers screening and presymptomatic clinical examination is very effective method in the prevention of the malignant transformation. Early detection of the high-risk patient is still not enough sufficient, one of the useful method may be questionnaire investigation. We analyzed 9000 patients hospitalized in Central Regional Hospital in Poznań in years 1996–2001. The malignant disease was recognized in 1805 patients, in 250 cases the age of diagnosis was before 50 years old. In 94 cases at least one of the family members was treated due to malignant process. The questionnaire was send to all of these 94 patients. We became 35 full-filled questionnaires (37% of the all questionnaires), after analysis of pedigree HNPCC was recognized in 8 cases (8.5% questionnaires), HNPCCsuspected in 17 cases (18% questionnaires). The patients with familial risk of the colorectal cancers were qualified to clinical control examinations. Questionnaire investigation of the patients may be very useful method of the recognition of the patient with familial risk of the colorectal cancer.

48 Development and validation of colorectal cancer-specific quality of life questionnaire Harisi Revekka, Bodoky György*, Kupcsulik Péter, Weltner János 1st Department of Surgery, Semmelweis University, *Department of Oncology, Szent Laszlo Hospital, Budapest, Hungary The aim of the current study was to construct a colorectal cancer specific quality of life questionnaire (CRC_QoL) and to test its reliability and validity. Patients and methods: 372 patients with rectal cancer were included on a voluntary basis in this prospective study. The CRC_QoL was prepared to analyze the correlation between the characteristics of the colorectal cancer and its treatment versus the physical and psychological state, somatic sensations and social connections of the patients. The questionnaire consists of 62 questions. The questions can be answered in about 20 minutes. There are 2–5 possible answers in a simple choice system. The possible scores and scales were chosen in collaboration with the patients. Bigger numbers mean worse, the smaller mean better quality of life. In an earlier pilot study we asked healthy volunteers concerning the importance of different values, and the results of these interviews resulted in the values of the different answers on the scale of each question. The scale can also be seen as a percent distribution, where – in turn – the best quality is 100. The results were evaluated by question and by patient group too. Statistical routines were administered with the use of MiniStat and statistical significance of the data was determined using the Student’s t-test. Results: Internal consistency of each factor was assessed by calculating Cronbach alpha values and was found satisfactory (Physical state indicators 0.779, General state indicators 0.844, Gastrointestinal indicators 0.445, Stool and defecation indicators 0.758, Gender specific indicators 0.940, Emotional state indicators 0.845, Social indicators 0.281). For the test-retest reliability analysis the questionnaires were re-assessed. Correlation analysis showed that the answers were consistent, Pearson values ranging from 0.991–1.00 (p < 0.01 in all cases). Discriminative validity analysis of the factors showed a significant difference in all cases which proves that these items discriminate between patients operated for colorectal cancer and the healthy population. Conclusion: This questionnaire can be used to detect changes in QoL over time to patients with colorectal cancer. Its use as an additional outcome measure in surgery, chemotherapy and radiotherapy should be encouraged.

49 Alimentation particularities at patients with colorectal cancer I. Marincu1, M. Cornianu2, C. Oancea1, L. Marincu1 1 Department of Infectious Diseases, UMF-Timisoara, Romania 2 Department of Histology, UMF-Timisoara, Romania Introduction: The human’s alimentation represents a risk factor with multiple unknown challenges in the apparition of colorectal cancer Methods: The authors have studied a group of 32 patients diagnosticated with colorectal cancer in Department of Infectious Diseases Timisoara. The positive diagnosis was based on clinical elements (abdominal pain, diarrhea with bleeding, losing weight, fatigue, alternating constipation with diarrhea, bleeding with defecation, tenesmus, etc.), biological elements (leukocytosis, ESR, CRP, CEA, hemoglobin, hematocrit, electrophoresis, fibrinogen, stool culture, etc.) and results of rectoscopy, sigmoidoscopy, ileocolonoscopy completed with anatomopathological examination of the biopsy fragment. For each patient, the alimentation particularities and associated pathology were registered. Results: 30 patients (93.75%) had a diet mostly from animal sources (pork meat, sausages, processed meats), 29 patients (90.62%) a diet high in animal fat, 30 patients (93.75%) consumed only white bread and refined carbohydrates, 26 patients (81.25%) were smokers for over 20 years, 25 patients (78.12%) were alcohol consumers, 24 patients (75%) presented physical inactivity, 16 patients (50%) had obesity and 10 patients (31.25%) were diagnosticated with diabetes mellitus; 93.75% didn’t consume fruits, cereals, vegetables, rice, beans and had slow intestinal transit. 50% presented constipation in personal history. We mention that in the geographic area very much processed pork meat, animal fats and refined carbohydrates are consumed. Discussion/Conclusion: The study of alimentational particularities associated with colorectal cancer permits the application of prophylactic measures and delimitation of population groups of risk for this disease.

50 Screening for colorectal cancer with fecal occult blood test (FOBT) in Romania Cristina Pojoga, Oliviu Pascu Third Medical Department, Cluj Napoca, Romania Introduction: FOBT detects a high proportion of less advanced colorectal cancers (CRC) in asymptomatic patients. We aimed to assess the effectiveness of FOBT screening on CRC detection in Romania. Methods: We used Hemoccult II to assess the presence of occult blood in stools. This was a part of a national program for CRC prevention and the free administration of FOBT, together with basic information about CRC, was advertised in local media. Patients with positive FOBT were referred to colonoscopy, and so were those with negative results but who asked themselves for the procedure. Results: 521 asymptomatic subjects provided stool specimens on FOBT cards. 63 had positive results (12.09%) and 60 of them underwent colonoscopy (3 refused). The findings at colonoscopy were: CRC 10 patients (16.66%) and colonic polyps 21 patients (35%). The rest had: ulcerative colitis (2 patients), haemorrhoids (25 patients), gastric cancer and erosive gastritis (1 patient each). 34 patients with negative FOBT underwent colonoscopy and 2 of them had adenocarcinoma (5.88%) and 6 (17.64) had polyps. The sensitivity, specificity, PPV and NPV were as follows: for CRC: 83.3%, 39.02%, 16.6% and 94.1% respectively, and for colorectal polyps: 77.7%, 41.7%, 35% and 82.3% respectively. Discussion/Conclusion: Our findings suggest that consideration should be given to a national program of FOBT screening in order to detect the early cases of CRC or colonic polyps. Unfortunately the population compliance to this kind of programs is still low in Romania.

51 Risk factors contributing to the development of colorectal cancer N. Potaturkina, S. Nesterov, A. Nesterov, Y. Krasnoperova Cancer care center of Ulyanovsk region, Russia Ulyanovsk State University, Ulyanovsk, Russia Implementing effective methods of colorectal cancer treatment without studying risk factors contributing to its development. The total number of cancer care recipients is 877. It has been found that one of the most significant risk factors for developing colorectal cancer is the patient’s age. Colorectal cancer was mostly characteristic of patients aged over 50 (73.1%), with incidence of the disease gradually decreasing in the population group aged over 70 (19.6%). Despite its prevalence among the elderly, the disease was found in 7.2% of younger patients (25–50 years old). Colorectal cancer was almost equally typical of both male and female patients. Out of the total number of male patients, 48.35% were diagnosed with blind and middle intestine cancer and 51.7% with rectal cancer, while these figures for female patients were 56% and 44% respectively. It has been revealed that those suffering from chronic colon diseases, especially ulcerative colitis, run a higher risk of developing colorectal cancer. This particular condition was characteristic of 60% of the patients who had had ulcerative colitis for 25–30 years. Another substantial risk factor is Crohn’s disease, which has been found in 21.4% of the patients. Out of the total number of patients 194 stated that cancer ran in the family. Familial cancer syndrome, a result of autosomal dominant inheritance, is especially characteristic of multiple adenocarcinoma of the middle intestine. Two types of familial cancer syndrome that have been found are Lynch I and II syndromes. Tests administered as part of the treatment have revealed various disimmunities in most of the patients, such as malignant tumors affecting other organs, chronic dermatosis, respiratory allergosis and others.

52 Risk factors of colorectal cancer in Latvian Chernobyl clean-up workers J. Seleznovs, J. Pokrotnieks, G. Orlikovs, A. Skesters, T. Farbtuh, Paula Stradin Clinical University Hospital, Riga, Latvia Introduction: It is known that colon cancer (CRC) risk increases with longer duration of ulcerative colitis and Crohn's disease. The incidence and relative risk of CC in Latvian clean-up workers (CCW) was elevated in 2.5 times in recent five years. The goal of trial is determining to risk factors (radiation dosage, activity of Ca2+ATPase, antioxidant defence [AD] and selenium) of CRC by CCW inhabitants of Latvia. Methods: The study group comprised 101 CCW (men, aged 35–70, dose of radiation 0.18 ± 0.14 Gy received in Chernobyl 1986–1990) and control group (20 healthy men, aged 25–50). We used chemiluminescence for the detection of plasma oxidazability (Sox), hydroperoxydes concentration (Hmax). We have investigated, antioxidant enzymes (glutathione peroxidase (GSH-Px) in plasma, Cu,Zn-superoxide dismutase (SOD) and catalase in blood cells) and concentration of selenium in blood. Results: We detected that Sox and Hmax was elevated in four times, activity of catalase was increased (22%), GSH-Px was considerably reduced (37%), activity of Ca2+ATPase was increased in 2.4 times, although activity of SOD was unchanged in CCW accordingly comparing to healthy Latvians. A low selenium level (16–43 mkg/L) is registred in CCW as well as in Latvians too. Conclusions: The dosage of radiation received by CCW has no influence on AD. CCW have diminished AD associated with pronounced selenium deficiency and high activity of Ca2+ATPase. Chronic oxidative stress and AD disorder one of possible risk factors of developing CRC.

53 Colo-rectal cancer screening in Romania Simona Bataga, Imola Torok, Ana Bratu, Simona Mocan, Dan Georgescu, Tiberiu Bataga Department of Gastroenterology, University of Medicine and Pharmacy Tg-Mures, Romania Introduction: Screening for colo-rectal cancer (CCR) in Romania has started recently, since 2004. In the last years the incidence of CCR has increased, so from 2004 it was promoted a national Programme from CCR prevention 2.2. Methods: The study is following-up the results of the screening for the colo-rectal cancer in our region from Transylvania, Romania. Between 2004 and 2006 1015 patients entered the screening for CCR, 623 men (61.37%), mean age 57.4 years. From these patients: 273 patients were with medium and high risk of CCR (relatives with CCR, FAP, IBD, operated with CCR etc.), 457 patients were with CCR symptoms and 285 were patients without symptoms, above 50 years. Results: Advanced neoplasia was found in 19 patients (6.95%) from the risk CCR, 15 (3.28%) within the symptomatic group and in 4 (1.40%) from the patients without symptoms. All the patients were sent to surgery. The incidence of the polyps was in 89 in 71 patients from the whole groups, that means 6.99 %. In all patients polypectomy was done. In 2 patients were diagnosed neoplasia in the polyps and surgery was done. Discussion/Conclusion: We detected advanced neoplasia at a significantly higher rate in patients within the risk categories and with symptoms: 34 cancers (4.65%). In total 38 colon cancers (3.74%) and 89 polyps (6.99%) were diagnosed, so we can conclude that the CCR screening is efficient.

54 Examination of metabolizing colorectal cancer

enzymes

in

patients

with

Varga, P.1, Kiss, I.2, Ember, I.2, Schnabel, R.1 1 Gyula Hospital and Policlinic, II. Department of Medicine, Budapest, Hungary 2 Medical School of Pécs, Department of Public Health, Pécs, Hungary Introduction: Exposures such as cigarette smoking and meat containing a variety of procarcinogens, which are thought to play a role in elevation of risk for colorectal cancer. These procarcinogens (including heterocyclic amines and polycyclic aromatic hydrocarbons) are metabolized by a variety of polymorphic enzymes including N-acetyltransferases, cytochrome P450 enzymes, or glutathione S-transferases. Cytochrome P-450 CYP1A1 is involved in the metabolism of polycyclic aromatic hydrocarbons (PAHs) that are derived from meat intake and tobacco smoking. Expression of the CYP1A1 gene is induced by compounds present in cruciferous vegetables. The glutathione S-transferases play a central role in the detoxification of carcinogens, including PAHs. Aim: We hypothesized that genetic variation in the encoding genes NAT2, CYP1A1, GSTM1, GSTT1is associated with risk of colorectal cancer and interacts with cigarette use or meat intake to modify risk of colorectal cancer. We examined the role of these genes in a clinic-based study of 31 Hungarian patients with colorectal cancer. Results: CYP1A1 Ile/Val polymorphism 16/31 (51%), NAT2 rapid acetylator 21/31 (67%), GSTM1 0 phenotype 12/31 (38%), GSTT1 0 phenotype 7/31 (22%). There were no patients without polymorphism. Simple polymorphism: 8/31 (25%), double: 12/31 (38%), triple 9/31 (29%), fourfold 2/31 (6%). Risk factors: tumours in the family 20/31 (64%), cigarette smoking 20/31 (64%), alcohol 12/31 (38%), smoked, red meat 6/31 19%. Conclusions: We found that 74% of the cases were multiple polymorphisms. 64% of the patients are smoking regularly. These results indicate that cigarette smoking and metabolizing enzymes are associated with risk of colorectal cancer. Our plan is to extend the ongoing study.

55 Liver metastases in the HBs antigen positive patient with colorectal cancer T. Banasiewicz, J. Paszkowski, P. Pyda, M. Drews Chair and Department of General, Gastroenterological and Endocrynological Surgery K Marcinkowski University of Medical Sciences, Poznań, Poland Purpose: About 15–40% of patients operated due to neoplasm of the gastrointestinal tract were found to have liver metastases. Little is known about the mechanism giving rise to such metastases and secondary tumour growth in the liver. Material and method: We correlated the frequency of liver metastases in two groups of patients operated due to colon and rectum cancer in our department between 1983 and 1998. The first group was comprised of HBs antigen positive patients (n = 36) while the second of HBs antigen negative patients (n = 867). Results: In the first group we observed that four patients (11.1%) had metastatic changes in the liver and in the second, metastases in 204 patients operated (23.5%). Metastatic tumour was rare in the HBsAg positive group (p = 0.08). Conclusions: This study is a retrospective analysis of material gathered over a period of 15 years. On the basis of the analysed material we can state that in patients who are HBs antigen positive (within the limits of statistical significance) the formation of liver metastases resulting from large bowel neoplasma is rare. This phenomenon is still very much an unresolved clinical and scientific problem even though we possess knowledge that several known factors could be responsible.

56 How to prevent colorectal cancer – Early detection of polyposis syndromes Banasiewicz, Tomasz; Paszkowski, Jacek; *Pławski, Andrzej; *Słomski, Ryszard; Krokowicz, Piotr; Drews, Michał Chair and Department of General, Gastroenterological and Endocrynological Surgery K Marcinkowski University of Medical Sciences *Department of Human Genetics, Polish Academy of Sciences, Poznań, Poland Clinical and genetical screening plays an important role in presymptomatic detection of Familial Adenomatous Polyposis coli (FAP). FAP is caused by germ line mutation in the APC gene. In all members of registered in our Department 102 families, genetic analysis and clinical investigation were done. The APC gene was screened for mutations in Polish population by the use of heteroduplex and SSCP methods. The methods used for visualization of the mutation (HD and SSCP) differed in relative effectiveness, depending on the localization of the mutation within the PCR-product. In our study HD was more sensitive as compared to SSCP analysis, however the use of both methods increased the rate of detection of molecular changes. Clinical investigation (sigmoidoscopy, gastroscopy, USG) was performed in all family members. In our study 39 mutations were found in APC gene (6 previously not described: 5 deletions and 1 substitution in region 5’). Until now, we have diagnosed FAP in 58 asymptomatic patients (in 31 cases genetic examination, in 27 cases during rectosigmoidoscopy). All were treated surgically, none had malignant transformation. Prophylactic and systematic clinical control is effective and necessary in the group of patients with familiar adenomatous polyposis. Genetic examination plays an important and increasing role in presymptomatic diagnosis of FAP.

57 Difficulties in implementing a preventive strategy for colorectal cancer in patients with adenomatous polyps Dan L. Dumitrascu1, Diana Dumitrascu1, Laurentiu Nedelcu2, Liliana David1 1 University Iuliu Hatieganu, 3rd Medical Department, Cluj, Romania 2 University Transilvania, Department of Internal Medicine, Brasov, Romania Introduction: Adenomatous polyps represent a main condition with potential evolution toward colorectal cancer. We analysed the difficulties of a preventive programme implemented in patents with resected adenomatous polyps. Methods: Adult patients with nonfamilial colonic adenoma were recruited after endoscopic polypectomy for a long-term preventive therapy with aspirin 75 mg/day and calcium carbonate 1250 mg/day. Exclusion criteria: upper digestive lesions or known intolerance to NSAID; previous colorectal cancer, use of anticoagulants, contraindications to study drugs. The acceptance of this prevention strategy and the adherence in these patients was analysed. Results: 50 consecutive patients with 88 polyps were asked to enter in this prospective open trial. Only 28 agreed to give the informed consent and to participate in this project (17 F, 11 M, aged 55 ± 12 years). But after reading the side effects of aspirin only 20 of them accepted to continue (40%). During a mean follow-up of 12 months only 13 of them still remained compliant (65%). Main reason for withdrawal was epigastric pain. Two patients required colonoscopy in the follow-up period because of the anxiety and of concomitant functional symptoms. Discussion/Conclusion: Implementing a preventive programme for colorectal adenomatous polyps is difficult due to large rate of inacceptance, side effects and nonadherence.

58 Can the long-term use of aspirin and other non-steroidal antiinflammatory drugs protect against the colorectal cancer? C.E. Rezi1, R. Mihaila2, L. Tiurean1, R. Mihăila3, M. Deac2, O. Petrascu2 1 County Clinical Hospital Sibiu, 2nd Medical Department, Romania 2 University of Sibiu, Faculty of Medicine, Sibiu, Romania 3 Public Health Authority Sibiu, Romania Introduction: There are some studies which sustain that aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) may reduce the risk of colorectal cancer by inducing cell arrest and apoptosis. Methods: We proposed ourselves to study if the use of aspirin and NSAIDs by the patients with ischemic cardiac disease can protect against the appearance of the colorectal cancer. We studied 332 patients which were hospitalized in the 2nd Medical Department of the County Clinical Hospital from Sibiu, during January 2002–September 2006, which had, as the main diagnosis Ischemic Cardiac Disease. They were divided in two groups: group A – which were treated on a long period of time with aspirin and other NSAIDs, and group B – which did not use these medication. We have studied the next parameters: age, sex, the colorectal cancer’s diagnosis, the dose of aspirin. The results were statistically analyzed using the relative risk. Results: Among the 63 patients from group A, 3 had colorectal cancer. From the 269 patients from group B, 32 patients had colorectal cancer. The relative risk of the appearance of this disease at the patients from group A, comparing with those from group B was 0.39, which suggests that, in our study, the usage of these drugs can be considered as a favourable factor for the health, preventing the appearance of the colorectal cancer. The benefit appeared to be related to the type of medication they have used: compared with the patients who reported no use, the relative risks for cancer were 0.26 for those who used aspirin and 0.68 for those who used other NSAIDs. The dose of aspirin they had been used, at least 5 days a week, was: 62.5% used 75 mg/day; 15.6% used 125 mg/day; 18.9% used 250 mg/day and 3.1% used 500 mg/day. Discussion/Conclusion: These results suggest that aspirin and other NSAIDs have, apart from the cardio-vascular effects, other pleiotropic effects on the intestine, contributing in this way at the colorectal cancer’s prophylaxis. Aspirin appears to be more protective than other NSAIDs.

59 Endoscopic therapy of early colorectal carcinoma by EMR Chr. Banciu, Lelia Susan, Violetta Vacariu, I. Romosan IV. Dept. of Internal Medicine, University of Medicine and Pharmacy Timisoara, Romania Introduction: The incidence of colorectal carcinoma (CRC) is very high throughout the world, thus early detection and treatment seems to be justified. Aim of study: to establish prospectively the use and utility of endoscopic mucosal resection (EMR) as curative therapy of early CRC (limited to the colonic mucosa). Material and methods: 39 pts who were diagnosed with early CRC by colonoscopy underwent EMR. Regarding the aspect of the tumor: flat carcinoma limited to the colonic mucosa (14 pts), adenomatous polyps with severe dysplasia (19 pts), and recurrence after surgery (4 pts). Localization of CRC: rectum and r-s junction (18 pts), sigmoid colon (13 pts), descending colon (8 pts). Technique of EMR: injection of saline and methylene blue for elevation, polypectomy in piece-meal fashion for flat large lesions and big polyps and as a one time procedure for stalked dysplastic polyps. At the end of the procedure India ink was injected at the resection site for follow-up. Results: EMR was successful in all pts. No severe complications were observed: little-moderate bleeding which stopped spontaneously or after injection of sclerosant agents/clips (6 pts), no perforation. The overall success rate of EMR at 6 months follow-up was over 92%, a recurrence of CRC was observed in 3 pts. One pt was referred to surgery; other two pts were retreated by EMR. Conclusion: EMR should represent the standard procedure for early CRC limited to the mucosa. In experienced hands the risk of complications is low. For future localization of the resection site it is important to spot it with India ink.

60 The importance of the fecal occult blood test in detecting colon tumors Chr. Banciu, Lelia Susan, O. Chirileanu, I. Romosan IV. Department of Internal Medicine, University of Medicine and Pharmacy Timisoara, Romania. Aim: The fecal occult blood test (FOBT-“Hemocult”) has been used as a screening method in detecting colonic tumors in early stages in an unselected population. Material and methods: The FOBT was carried out in 476 patients (pts) without intestinal symptoms. In some pts. FOBT was performed only once (three samples), in other pts. was repeated, according to the precautions of the producer. For pts with positive tests complementary investigations were performed: flexible sigmoidoscopy, colonoscopy, DCBE, and gastroscopy. Results: The FOBT was positive in 28 pts (5.88%). The investigations performed in these pts revealed the cause of bleeding: rectal and colonic polyps in 9 pts, hemorrhagic proctitis in 4 pts, rectal or colon cancer in 7 pts; the rest of 8 pts FOBT was due to an upper GI cause (hiatal hernia, erosive gastritis, peptic ulcer). At colonoscopy the tumors were localized in early stages at rectal level (2 pts), descending colon (4 pts) and in the ascending colon (1 pt). Although only few pts presented some upper GI symptoms, it might be stated that the diseases diagnosed by the FOBT were more or less ignored, this fact being a rather frequent situation in the pathology of colonic tumors. Conclusion: the present study confirms the efficacy of the FOBT as a screening method in tracking down pts liable to develop colonic tumors, some of which can be detected in early stages; these pts have to undergo more complex GI investigations and endoscopic procedures, subsequently, in order to obtain all the necessary information for adequate treatment of the colonic tumors.

61 Palliative endoscopic treatment of obstructive colorectal carcinoma – Prospective pilot study Chr. Banciu, O. Chirileanu, Alina Pacurari, I. Romosan IV. Dept. Of Internal Medicine, Univ. Of Medicine and Pharmacy Timisoara, Romania Introduction: Obstructive colorectal carcinomas (CRC), usually with distant metastases, are seen more frequently in daily practice. Our objective is to insert colonic stents in patients (pts) who refuse surgery or where surgery is not recommended. Material and methods: 7 pts with obstructive CRC underwent stenting of the stricture. Gender distribution: 2 females, 5 males. Localizations of stricture: r-s junction (3 pts), sigmoid colon (2 pts), stenosis after surgery (recurrence of CRC – 2 pts). Sigmoidoscopy was performed in all pts, followed by balloon dilation and stent insertion (Wallstent Enteral Endoprosthesis). Results: Relief of colon obstruction and resume of intestinal transit was obtained in all pts. Complications during stent placement: in one pt a small perforation after balloon dilation was observed, which was resolved after stent placement and with intravenous administration of antibiotics; in 6 pts bleeding from the dilated tumor which stopped spontaneously after stent insertion. The pts could better accept chemotherapy; two pts were prepared for elective colon resection without colostomy, and in the two pts with recurrence of obstructive CRC after surgery bowel transit and decompression was reestablished. Conclusion: Stent placement in obstructive CRC has a palliative effect and results in decompression of large bowel. It is a useful technique for relief of obstruction and for further therapies (chemotherapy, elective surgical colonic resection), as well as for restenosis of the colon due to recurrence of CRC after surgery. The Wallstents also improved the quality of life of all pts.

62 Are the polyps and cancers of the colon different under and above 50 years of age? Five years results of colonoscopy in our department Gyökeres, T., Szakács, A., Hamvas, J., Illés, K., Kovács, F., Varsányi, M., Takács, I.G., Pap, A. MAV Hospital, Budapest, Hungary Introduction: Colon cancer is the second leading cause of death in our country. There has been lot of efforts done to start the screening in the population at 50 years of age. Aim: To compare the properties of polyps and cancers under and above the predeterminated age of screening. Methods: Conventional colonoscopy and polypectomy was performed in patients (pts) with suspected colon tumors or other large bowel abnormalities. Patients: During a five year period we have performed 4914 colonoscopies. We had 1027 pts with polyps and 454 pts with colon cancer. Results: In 104 pts under 50 years (group I) we have found 159 polyps (the number of polyps/patient was 1.52 in average). In 923 pts above 50 years (group II) we had 1555 (1.68 polyps/patient) polyps. The histology was tubular adenoma (61.6% vs. 56.9%), villous adenoma (17.6 vs. 25.2%) and carcinoma in polypo (1.25% vs. 1.47%) in the two groups. In group I 21/159 (13.2%) polyps were located in the right, 138/159 (86.8%) in the left side of the colon. In the group II 387/1555 (24.9%) was in the right and 1168/1555 (75.1%) in the left colon. We had 19 pts with colon cancer under 50 years and 435 pts above 50 years. The cancers were located in the left side in 14/19 (73.7%) under, vs. 276/435 (63.4%) above 50 years, while proximal localisation was 5/19 (26.3%) vs. 159/435 (36.6%), respectively. In summary: In the patients above 50 years of age the rate of more dangerous villous polyps, the proximal localisation of both polyps and tumours are more frequent. Therefore the importance to get into the coecum is increasing with age.

63 Endoscopic polypectomies of colorectal adenomatous polyps – Prospective evaluation of material resources needed for colonoscopic surveillance in a tertiary gastroenterology referral center Iacob, R., Bancila, I., Cotruta, B., Bucur, D., Voinea, D., Iacob, S., Gheorghe, L., Manuc, M., Pitigoi, D., Marinescu, T., Diculescu, M., Gheorghe, C. Gastroenterology and Hepatology Center, Fundeni Clinical Institute, Bucharest, Romania Introduction: Adenoma is the most valuable risk marker for colorectal cancer (CCR), the leading cause of illness and death in the Western countries. Methods: During 01.06.2005–01.03.2006, we evaluated the patients included for surveillance after endoscopic polypectomies and the costs of performing the scheduled colonoscopies. The material resources necessary have been evaluated by multiplying half of the programmations with the cost of one surveilance colonoscopy dividing the result to the median time in which the programmations are covered according to the Kaplan Meier curve. Results: 407 patients have been included in our registry of which 111 (27.2%) for surveillance after endoscopic polypectomies (48.6% females; 51.4% males; mean age of 60.4 ± 12.0 years). The number of colorectal adenomatous polyps endoscopically resected for each patient was 1 to 6, mean 1.8 ± 1.2 polyps/patient and the size of the largest polyp variated between 5–35 mm, mean 13.6 ± 8.6 mm. The analysis of Kaplan Meier curves of programmation of colonoscopic controls indicates a median time interval for the scheduled procedures of 9.4 months. 75% of programmations are covered in 28.6 months and 90% in 31 months from the baseline date. The cost analysis indicates a necessary of 2290 Euro in 9 months for the activity of colonoscopic surveillance after endoscopic polypectomies for colorectal polyps. Conclusions:The median time interval for control colonoscopies is 9 months, thus suggesting that the procedures are performed earlier than 3 years in many patients for a number of reasons (flat lesions, large polyps, multiple polyps, bad preparation of the colon).

64 The prospective analysis of material resources needed for colonoscopic surveillance in patients with curative resection for colorectal cancer Iacob, S., Iacob, R., Bancila, I., Cotruta, B., Bucur, D., Voinea, D., Gheorghe, L., Croitoru, A., Ciurea, I., Manuc, M., Pitigoi, D., Marinescu, T., Diculescu, M., Gheorghe, C. Gastroenterology and Hepatology Center, Fundeni Clinical Institute, Bucharest, Romania Introduction: Screening and surveillance for prevention of colorectal cancer (CCR) is cost-effective and reduces mortality. Methods: During 01.06.2005–01.03.2006, we evaluated the patients included for surveillance after curative colonic resection for CCR and the costs of performing the scheduled colonoscopies. The material resources necessary have been evaluated by multiplying half of the programmations with the cost of one surveillance colonoscopy dividing the result to the median time in which the programmations are covered according to the Kaplan Meier curve. Results: 407 patients were included in our registry of which 243 (58.3%) for surveillance after curative colonic resection for CCR The distribution of age categories on sexes differs statistically significant (p = 0.0004), in favor of > 60 years category in males and of 40–60 years category in females. 118 patients (48.6%) have been included for newly diagnosed tumors and 125 patients (51.4%) have been evaluated at follow-up controls. The analysis of percentiles of the Kaplan Meier curve for the colonoscopic control shows a median duration for the scheduled procedures of 7.3 months. 75% of surveillance colonoscopies are performed in 13.7 months and 90% will be covered in 32.4 months from the baseline date of scheduled list. The cost analysis indicates a necessary of 4208 Euro in 7 months for the activity of colonoscopic surveillance after surgery. Conclusions: The median duration of scheduled colonoscopies is of 7.3 months and the cost of this activity in a referral center with 180 beds capacity is of 4208 Euro during this time interval.

65 The role of genetic counselling in children with familial polyposis syndromes – Own material Izabela Kubinska1, Elzbieta Czkwianianc1, Andrzej Plawski2, Ryszard Makosiej1, Alina Durko1, Ewa Malecka- Panas1,3 1 Department of Gastroenterology, Polish Mother's Memorial Hospital, Lodz, Poland 2 Institute of Human Genetics, Polish Academy of Science, Poznan, Poland 3 Department of Digestive Tract Diseases, Medical University, Lodz, Poland Introduction: Familial adenomatous polyposis (FAP) and juvenile generalized polyposis (GJP) are genetically determined disorders, autosomally dominant inherited. FAP is caused by mutation of APC gene, whilst GJP is connected with mutations of BMPR1 or SMAD4 genes. Both predispose to colon cancer. Methods: The cases of seven children aged 2–14 from families with familial polyposis history were analysed. Children have undergone endoscopy and were screened for mutation in the APC gene. DNA was screened with heteroduplex analysis, single strand conformational methods (SSCP) and DNA sequencing. Results: Three children were diagnosed with FAP. In one child the results of clinical and endoscopic examination were inconclusive, but mutation in the APC gene was confirmed. In one child GJP was endoscopically diagnosed, but mutations of BMPR1 or SMAD4 genes were not found. In two cases clinical FAP observation was negative and molecular screening either. Discussion/Conclusion: High coincidence between clinical observation and molecular screening was observed. Genetic counselling might play pivotal role in diagnostic process and in planning treatment of patients with familial polyposis syndromes.

66 Postoperative surveillance of colorectal cancer: Is intensive follow-up efficient? C. Searpe, V. Sbarcea, A. Rosu, A. Gavrila, C. Sbarcea Railway University Hospital, Department of Internal Medicine, Craiova, Romania Introduction: After the surgical treatment for colorectal cancer, local or distant recurrence of the disease is very important for patient long-term prognosis. The aim of our study was to evaluate the efficiency of intensive vs. conventional postoperative follow-up. Methods: In our clinic 64 patient were diagnosed with colorectal cancer stage I to III TNM between 1999 and 2001. All of them underwent surgical treatment and were enrolled into a 5 years postoperative surveillance program: 33 (group A) had clinical examination, biologic samples, FOBT and abdominal ultrasound every 6 months, yearly barium enema and chest x-rays for 5 years; 31 (group B) had colonoscopy at 6 months after surgery and then yearly plus yearly chest and abdominal CT-scan for 5 years in addition to the investigations underwent by the patients from group A. Results: The 5 years postoperative survival was: 24 (72.7%) patients from group A and 25 (80.6%) patients from group B. The recurrence rate was almost identical in both groups: 12 (36.4%) patients from group A and 11 (35.5%) patients from group B. Also, we found polyps in 5 (15.2%) patients in group A and in 7 (22.6%) patients in group B. Discussion/Conclusion: Although the recurrence rate was almost identical in both groups, the significant survival improvement (7.9%) was due to the early diagnosis of the recurrent disease in the intensive follow-up group.

67 Feasibility study of colon polyp computerized detection in electronic cleansed virtual colonoscopy Zs. Tarján, A. Tombácz, J. Koloszár Semmelweis University, Department of Diagnostic Radiology and Oncotherapy, Budapest, Hungary Purpose: Fecal tagging in CTC with/without colonic cleansing facilitates the recognition of remaining stool for human observers on 2D based reading, but complicates 3D based reading and the use of a CAD. Method and Materials: Sixty-four patients (35 female, 31 male, age: 45–82 years, mean 54 years) with suspected colonic polyps underwent CTC and optic colonoscopy within the same week. Forty-three patients had cathartic preparation and fecal tagging (group A), and 21 patients had tagging only (group B). Group B patients had no food restrictions or colonic cleansing in the days before the CT examination was performed, and only 10 times 20 ml of ionated radiologic contrast material (not containing meglumin) was administered orally, 48 hours prior to the examination. Native supine low-dose 1–2.5 mm collimated scans were obtained on 4–16 channel scanner. The data were evaluated using an academic software (ColVis) equipped with electronic cleansing and CAD. The results generated by the CAD and that of a radiologist using 2D and 3D based reading were compared. Results: In group A 41 in B 2 patients had diarrhea caused by the preparation. The CAD technique could detect 19/19 polyps > 10 mm, and 21/27 polyps between 5 and 10 mm with 18.4 + 12.7 (mean, SD) false positives/case. The radiologist on 2D based reading could detect 18/19 polyps > 10 mm and 23/27 polyps between 5 and 10 mm with 2.3 (mean) false positives/case. On 3D based reading with electronic cleansing 19/19 polyps > 10 mm and 23/27 polyps between 5 and 10 mm with 7.4 + 5.3 (mean, SD) false positives/case. Conclusion: CAD combined with electronic cleansing can detect large and small polyps similarly to a radiologist. The CAD detects much more false positive polyp candidates than the radiologist that uses 2D slices, much more than the same CAD in a clean colon, and some more than a radiologist on an electronically cleansed colon. 3D reading of cleansed CTC data improved polyp detection, but increased false positive rate too of the radiologist. 3D based reading can not be used for tagged CTC data unless electronic cleansing is done. Electronic cleansing makes artifacts that increase false positives for both 3D based reading and the CAD.

68 Association between the GST-pi expression and microvessel density and their combined prognostic significance in primary colorectal carcinoma 1

Vlaykova, T., 2Gulubova, M., 3Vlaykova, D., 4Cirovski, G., 5Manolova, I. Department of Chemistry and Biochemistry, 2Department of General and Clinical Pathology, 3Under-graduate student, 4Department of General Surgery, Medical Faculty, 5Laboratory of Clinical Immunology, University Hospital, Trakia University, 11 Armeiska Str, BG-6000 Stara Zagora, Bulgaria, E-mail: [email protected] 1

Introduction: Angiogenesis plays an important role in growth, progression, and metastasis of tumours. It is shown to be induced by various angiogenic factors produced by the neoplastic and/or non-malignant stromal cells. The glutathione-Stransferases (GSTs) comprise a supergene family of phase II xonobiotic metabolizing enzymes that catalyze a variety of reduced glutathione-dependent reactions with compounds with electrophilic centres. They also play important role in several cellular processes. The aim of the present study was to analyze the association of GST-pi expression tumour angiogenesis in biopsies of patients with colorectal carcinoma. Methods: The GST-pi and tumour angiogenesis were assessed by the means of immunohistochemistry in tumour specimens of 101 patients with primary colorectal carcinoma (53 with colonic and 48 with rectal cancer). Results: The mean “hot spots microvessel density (MVD) located in the invasion front of the tumors was significantly higher in the cases with strong (++) immunohistochemically detectable GST-pi expression (n = 29, 83.85 ± 42.4 vessels/mm2) than that negative or with weak expression (n = 72, 64.12 ± 36.5 vessels/mm2, p = 0.021). Patients with simultaneous high MVD and strong GST-pi expression had the worse prognosis after surgery than those with high MVD/weak GST-pi or low MVD/strong GST-pi expression and especially than patients with both low level of MVD and GST-pi expression (p = 0.007, Log-rank test). Discussion/Conclusion: In conclusion, we suggest a role of GST-pi in stimulation of tumour-associated angiogenesis and consider the combined high angiogenesis and strong GST-pi expression as an unfavourable prognostic marker for patients with colorectal carcinoma. Key words: colorectal carcinoma, angiogenesis, GST-pi, immunohistochemistry, prognosis This work is done with the financial support of research project VU-L-05, funded by the Ministry of Education and Science, Bulgaria.

69 Detection of endocrine cells and their prognostic significance in primary colorectal carcinoma 1

Gulubova, M., 2Vlaykova, T., 3Manolova, I. 1 Department of General and Clinical Pathology, 2Department of Chemistry and Biochemistry, Medical Faculty, 3Laboratory of Clinical Immunology, University Hospital, Trakia University, 11 Armeiska Str, BG-6000 Stara Zagora, Bulgaria, E-mail: [email protected] Introduction: Neuroendocrine differentiation in colorectal carcinoma has been observed but its prognostic significance is uncertain. The aim of the current study was to determine the presence of chromogranin A- (CHA), serotonin- (SER) and synaptophysin- (SYN) positive endocrine cells (ECs) in 100 primary colorectal cancers and to evaluate their prognostic significance for 4 years follow-up period Methods: For assessment of ECs we applied light microscopical and ultrastructural immunohistochemistry and immunogold staining. Results: Thirty-six percent of the samples contained CHA+ ECs, whereas only in 24% and 7% of the samples SER+ and SYN+ ECs, respectively, were observed. Patients with ECs in tumour specimens had shorter survival after the surgical treatment, as this association was statistically significant for SER+ ECs (p = 0.006, Log-rank test). By the ultrastructural investigation there were found changes in ECs in the tumour: these cells were larger and with irregular shape compared to normal ECs in mucosa; their nuclei were with irregular form and some were indented; their granules were distributed diffusely in the cytoplasm and some of them had bright areas in the centre. CHA+, SER+ and SYN+ DAB reaction product was found over the granules in tumours ECs. 10 nm gold particles, which detect the presence of CHA and SER were dispersed over granules and cytoplasmic vesicles in ECs Discussion/Conclusion: These results indicate that the presence of neuroendocrine cells may influence prognosis in primary colorectal carcinoma. Key words: endocrine cells, colorectal cancer, chromogranin A, serotonin, synaptophysin, immunogold, immunohistochemistry

70 The effect of mesalazine on experimental colon anastomosis

bacterial

translocation

in

Ahmet Aslan*, Muhyittin Temiz*, Meryem Cetin**, Cemil Tümer***, Hakan Bozkurtoglu*, Elif Canbolant*, Ramazan Gonenci**** Mustafa Kemal University, Faculty of Medicine, Department of General Surgery*, Department of Microbiology**, Department of Physiology***, Veterinary Faculty**** Antakya-Hatay, Turkey Objective: Mesalazine is an antibacterial and antiinflammatory agent used especially in the treatment of IBD. We aimed to investigate the effect of mesalazine on bacterial translocation in experimental colon anastomosis. Methods: After a colon resection (1 cm) anastomosis was performed from 2 cm proximally to the peritoneal reflection with 5/0 prolen. Wistar albino rats were divided into 5 groups containing 8 Wistar rats in each. Group 1 was the control group, group 2 was the anastomosis group with no treatment, group 3 was given oral mesalazine, group 4 was given rectal mesalazine and group 5 was given oral + rectal mesalazine for 1 week. After this period rats were sacrificed and samples obtained from blood, mesentery LN, liver, spleen and stool underwent microbiologic analysis. Results: There was bacterial translocation in one rat in group 1, all rats in group 2, 5 rats in group 3 and 4, 4 rats in group 5. A total of 71 microorganisms were found (more than one in 8 cultures). The determined microorganisms were Escherichia coli in 40 (56.3%), Enterobacter spp. in 15 (21.2%), Staphylococcus spp. in 11 (15.5%), Proteus in 2 (2.8%), Clostridium in 2 (2.8%), Pseudomonas in 1 (1.4%). The most translocation was seen in the MLN cultures (23/40). The number of the colonies determined in the stool cultures were significantly lower in groups 3, 4, 5 in contrast to group 2. Conclusions: We concluded that bacterial translocation developed in experimental colon anastomosis and that mesalazine reduced this.

71 Important bioactive small molecules of colorectal cancer patients after colectomy

erythrocytes

in

A. Blázovics, A. Szilvás, E. Székely, G. Székely, E. Tordai, E. Sárdi Semmelweis University, 2nd Department of Medicine, Budapest, Hungary Bioactive small molecules like formaldehyde and protoporphyrine are in connection with redox reactions. These molecules play an important role with free radicals in the biological system. HCHO can be formed in transmethylating reactions. HCHO can be observed mainly in a hydroxymethyl form, and reacts with nucleofil agents e. g. glutathione. 1O2 is produced while the reaction of HCHO and H2O2. Data show the important role of HCHO in proliferative and apoptotic processes. Free protoporphyrine can be detected near the Zn-protoporphyrine during the disease, especially cancerous processes, and it has pro- and antioxidant forms depending of its concentrations. Aims: to determine the HCHO and protoporphyrine concentrations of erythrocytes in colorectal cancer after colectomy and to establish the therapeutic effectiveness in the tumour. Methods: 32 adult patients after 5–10 years of their colectomy and 9 healthy volunters were drawn into this study. Tumour markers (CEA, CA 19-9, AFP), redox parameters (total scavenger capacity, H-donating ability, reducing power, free SH-group, HbA1c) HCHO and protoporphyrine concentrations were measured. Results: HCHO concentration was significantly lower in colectomized patients. Protoporphyrine concentration was very low in patients without metastasis, but in metastasis its concentration was significant. HbA1c level correlated significantly with induced free radical level and decreased antioxidant status of erythrocytes. Conclusion: Significant changes in erythrocyte function can be observed in cancerous processes. HCHO and protoporphyrine concentrations of erythrocytes are very important indexes in cancer process. ETT 012/2006 & NKFP 1B/047 Projects.

72 Serrated adenomatous colonic polyps particularities in a series of 12 cases

–

Morphological

Dumbrava, M., Becheanu, G., Manuc, M., Gheorghe, C., Preda, A., Serban, V., Diculescu, M. Gastroenterology and Hepatology Clinic – Fundeni Clinical Institute, Bucharest, Romania Introduction: Classically, serrated polyps are classified as hyperplastic (without epithelial dysplasia) and as serrated adenoma – serrated architecture and epithelial dysplasia. The incidence of serrated adenomas is low, under 1% in recent studies (Batts et al., 2002). Sessile serrated adenoma (Torlakovik et al., 2003) is an adenomatous polyp with particular morphologic features – preferential localization in the right colon, big size and sessile grossly feature. Methods: The aim of the present study is to analyze a serial of 12 cases of serrated adenomas with size between 5–20 mm, localized in caecum and ascending colon (5 cases), transverse colon (1 case), descending colon (1 case) and sigmoid/rectum (5 cases). Results: Grossly they were classified as one flat polyp, 7 sessile and 4 pediculated polyps. Microscopically, there were classified in: serrated adenoma (4 cases), sessile serrated adenoma (6 cases) and 2 mixed adenomatous/hyperplastic polyps. In both cases with mixed polyps we identified malign areas with histological aspect of well differentiated tubular adenocarcinoma. Discussion/Conclusion: Serrated glandular architecture impose an exhaustive histological exam for identification of epithelial dysplasia and classification of the polyps in the group of serrated adenomas, lesions with malign potential with a particular biological pathway (decreased apoptosis, DNA hypermethylation, hMLH-1 inactivation, microsatellites accumulation – microsatellite instability).

73 Histopathology is a strong prognostic factor in colorectal cancer C. Searpe, V. Sbarcea, A. Rosu, A. Gavrila, C. Sbarcea, M. Tenovici Railway University Hospital, Department of Internal Medicine, Craiova, Romania Introduction: In this study we tried to identify the most important histological patterns associated with 5 years postoperative prognosis in colorectal cancer patients. Methods: Between 1999 and 2001, 95 patients were diagnosed with colorectal cancer stage I to IV TNM in our clinic. Of them, only 86 patients underwent surgical treatment, they consisting our study group. After surgery the resected tumor was sent to pathology department for histological interpretation: histological type, TNM staging, grading, mitotic index, vascular invasion, inflammatory reaction, and vascular density. Results: In our study group 5 patients (5.8%) were in stage I TNM, 25 patients (29.1%) in stage II, 34 patients (39.5%) in stage III and 22 patients (25.6%) in stage IV. The overall 5 years survival rate was 61.6% with the following TNM staging distribution: 100% survival in stage I, 80% in stage II, 70.6% in stage III and 18.2% in stage IV. The histopathologic pattern of the surviving patients was: protrusive type (58.5%), TNM stage I to III (92.4%), adenocarcinoma (94.3%), low and medium grade tumors (71.7%), low mitotic index (66%), no vascular invasion (77.4%) and low-medium vascular density (73.6%). Discussion/Conclusion: The histopathologic pattern has a significant prognostic value in colorectal cancer, selecting patients for an early aggressive treatment in order to achieve a better survival.

74 Mesalazine counteracts the mutagenic intercalating agent 9-aminoacridine

effect

of

the

Campregher, Christoph; Luciani, Maria Gloria; Gasche, Christoph Medicine 4, Medical University of Vienna, Vienna, Austria Introduction: Mesalazine (5-ASA) improves replication fidelity at poly(CA) tracts (Gasche, Cancer Res 2005) and activates a replication checkpoint (Luciani, Gastroenterology 2007). The exact mechanism, however, is unknown. It has been proposed that 5-ASA’s anti-mutagenic activity is explained by its excellent oxygen scavenging properties. 9-Aminoacridine (9-AA) is an intercalating mutagen. In this study we tested the ability of 9-AA to increase mutations in colon epithelial cells and to prevent such mutations with 5-ASA. Methods: Clone A3.7 of HCT116+ch3, an hMLH1-corrected colorectal cell line, that had been previously transfected with pIREShyg2-EGFP/CA13, a EGFP-based plasmid [in which the (CA)13 shifts the EGFP reading frame into a +2 position (Gasche, PNAS 2003)] and harbors 2 integrated copies of the plasmid was seeded in 10 cm plates. 9-AA (0–10 µM) was added on day 2. After 24 hours, 9-AA was removed and 5-ASA (2.5 µM) was added for another 7 days. The total cell number (c) and the EGFP-positive fraction [mutant fraction (MF)] were analyzed by flow cytometry. The mutation rate [m/(CA)13/generation (gen)] was estimated by m = MF/(gen+1) and gen = log2(c/1000 x cloning efficiency). Results: 9-AA caused a dose dependent decrease of c and increase of MF and m in hMLH1-/+ cells. The best increase in MF and m was observed at 5 µM (about 10-fold). 5-ASA had little effect on c but reduced MF and m (by about 45%; table). A similar effect (for the total cell number, MF, and m) was observed when 5-ASA was added to 9-AA treated cells (table). Simultaneous treatment with 5-ASA and 9-AA (for 24 h) did not show such an effect. Discussion/Conclusion: In our model, 5-ASA reduces both the spontaneous and 9-AA-induced mutation rate. These data support the concept that 5-ASA improves replication fidelity at a poly(CA) tract independent of its anti-inflammatory properties. 9-AA (µM) 0 0 5 5

5-ASA (µM) 0 2.5 0 2.5

c (E+05) 86.0 (1.4) 57.9 (6.7) 8.1 (0.0) 6.8 (0.2)

MF (E-03) 0.18 (0.03) 0.10 (0.42) 1.65 (0.17) 1.04 (0.19)

m (E-05) 1.8 (0.3) 1.0 (0.3) 24.4 (2.4) 16.0 (2.8)

75 Detection of p53 gene Arg72Pro polymorphism and correlation with clinical characteristics in ulcerative colitis Caserta, Luigi1, Riegler, Gabriele1, Vietri, Maria Teresa2, Molinari, Anna Maria2, Cioffi, Michele2 1 Gastroenterology Unit – Second University of Naples, Italy 2 Department of General Pathology – Second University of Naples, Italy Introduction: p53 gene mutations are the most common alteration in human cancer. Several p53 polymorphisms has been described. Arg72Pro polymorphism in exon 4 induces changes in the p53 protein structure resulting biological differences about transcriptional machinery and activity and apoptosis ability. This polymorphism seems to be associated to susceptibility with various cancers. p53 gene alterations occur earlier in patients with ulcerative colitis (UC)-associated colorectal cancer (CRC) than in that with sporadic CRC. The aim of the study was to evaluate p53 codon 72 polymorphism in UC patients. Methods: We studied 81 consecutive UC outpatients (46 males with median age of 43 yrs, range 10–74 yrs and 35 females 43.2% with median age of 36.5 yrs, range 19–67 yrs). As control, we used peripheral blood from 71 donors (M = 32, F = 39, median age 35.7 yrs, range 4–79 yrs). Retrospective data on clinical history of the patients were obtained. Clinical characteristics of UC as well as familial cases of IBD and colorectal cancer history were collected. Genomic DNA was extracted from peripheral leukocytes by using a Wizard Genomic DNA Purification Kit and genotyping was conduced by PCR-confronting two-pair primers (CTPP). Results: In controls, the genotype frequency Arg/Arg, Arg/Pro and Pro/Pro was 52.1%, 39.4% and 8.5%, respectively. In UC patients, the distribution of 3 genotypes Arg/Arg, Arg/Pro and Pro/Pro was 50.6%, 39.5% and 9.9%, respectively. The frequency of arginine and proline alleles was 70.4% and 29.6%, while in control group was of 71.8% for arginine and of 28.2% for proline. There were no statistically significant differences. 62.5% of UC patients with Pro/Pro monozygosis showed a continuous clinical course compared to 9.8% and 6.3% of others genotypes (p < 0.001). Discussion/Conclusion: Our results suggest that codon 72 Pro/Pro alleles are statically associated with continuous clinical course and a persistent inflammation of mucosa. Detection of p53 Arg72Pro polymorphism could facilitate physicians to identify the patients at higher risk of CRC who have to undergo more intense endoscopic surveillance.

76 Involvement of proapoptotic and antiapoptotic Bcl-2 family members in colorectal cancer Guzinska-Ustymowicz Katarzyna, Czyzewska Jolanta, Sieczka Justyna, Niksa Michał, Chętnik Adam, Dymicka-Piekarska Violetta, Andrzej Kemona Department of General Pathomorphology, Medical University of Białystok, Poland Introduction: Bcl-2 gene family, consists of the anti-apoptotic genes Bcl-3, Bcl-xl, and proapoptotic genes Bax and Bak. The aim of this study was to clarify the relationship between the expression of these proteins in main mass of tumor and lymph node involvement. Methods: The immunohistochemical expression of these proteins in 55 patients with colorectal adenocarcinomas in correlation with parameters of tumor metastasis. Results and conclusion: The positive expression for Bcl-2, Bcl-xl (> 30% neoplastic cells) was observed in 33 and 43 cases respectively, where the expression for Bax and Bak (> 20% of neoplastic cells) was found in 22 and 23 patients with colorectal adenocarcinomas. We have observed statistically significant correlation between expression of Bcl-2, Bcl-xl in primary tumor and lymph node metastasis. But no such correlation was found for Bax and Bak. However, we note strong association between Bcl-2 expression in primary tumor and expression of Bax in the same localization. Expression of anti-apoptotic protein Bcl-xl was also correlated with expression of Bak and Bax in primary tumor. Corespondence to: Katarzyna Guzińska-Ustymowicz Department of General Pathomorphology Medical University of Bialystok Ul. Waszyngtona 13 PL-15-026 Białystok Poland E-mail: [email protected]

77 Beta-catenin and c-Met expression in inflammatory bowel disease A. Ivanova1, R. Nikolov1, S. Deredjan1, Z. Spasova1, M. Velev2, D. Adjarov1, Z. Krastev1 1 “St. Ivan Rilski” University Hospital, Sofia, Bulgaria 2 5th City Hospital, Sofia, Bulgaria Introduction: Dysregulation of the cell-cell adhesion molecule beta-catenin and protooncogene c-Met correlate with tumorgenesis. We investigated the frequency of the expression of beta-catenin and c-Met as markers for dysplastic changes in patients with inflammatory bowel disease. Methods: Specimens taken during colonoscopy from inflamed and noninflamed mucosa of 48 patients with ulcerative colitis and 10 patients with Crohn's disease, as well as from 11 control individuals were processed immunohistochemically for expressions of beta-catenin and c-Met using polyclonal antibody. Results: Beta-catenin was expressed in non-inflamed mucosa in 24/48 patients with ulcerative colitis, while in inflamed one it was established in only 7/48 (Pχ2 < 0.001). No statistical difference was found (Pχ2 > 0.05) between beta-catenin expression in their non-affected mucosa (28/48) and the control samples (7/11). Beta-catenin expression in patients with Crohn's disease was similar in inflamed (8/10) and noninflamed mucosa (9/10). A c-Met expression was absent in control persons. It was established in only 4 (8.3%) patients with ulcerative colitis being visible in inflamed and non-inflamed tissues. We were not able to observe co-expression of beta-catenin and c-Met. Conclusions: 1. The difference in frequency of expression of both molecular markers suggests different pathogenetic mechanisms and, probably, different phenotype of ulcerative colitis. 2. Future follow-up of the studied patients is needed for better evaluation of prognostic value of above markers.

78 The role of mast cells and neuropeptides on stress and barrier function of the follicle-associated epithelium in rats Åsa V. Keita1, Ann-Charlott Ericson2 and Johan D. Söderholm1 Department of Biomedicine and Surgery, Faculty of Health Sciences, University Hospital, Linköping, Sweden, 1Division of Surgery, 2Division of Cell Biology Introduction: Stress affects the course of inflammatory bowel disease, but the mechanisms are unknown. The earliest sign of Crohn’s disease is microscopic erosions over the follicle-associated epithelium (FAE). The FAE provides a route of entry for microorganisms, and animal studies have shown increased uptake in FAE after stress. Our aim was to evaluate the role of mast cells, corticotropin-releasing hormone (CRH), substance P (SP) and acetylcholine (Ach) on stress and barrier function of the FAE in rats. Methods: Forty rats were injected intraperitoneally with mast cell blocker doxantrazole or receptor antagonists for CRH, SP or Ach. Twenty of the rats were submitted to acute water avoidance stress. Ileal FAE segments were dissected and permeability was studied in Ussing chambers by measuring fluxes of the antigen horseradish peroxidase (HRP), 51Cr-EDTA and Escherichia (E.) coli K-12. Results: Stress increased the uptake of HRP and E. coli K-12, however, the increases were significantly abolished by blocking the mast cells and by CRH and Ach receptor antagonists, whereas the SP receptor antagonist had no effect. In contrast, 51Cr-EDTA permeability was significantly decreased by blocking the SP receptor, while unchanged by mast cell blocker and CRH and Ach receptor antagonist. Discussion/Conclusion: Stress affects the FAE barrier to antigen and bacteria with mechanisms involving mast cells, CRH, SP and Ach. Our results indicate that CRH and Ach are involved in uptake of antigen and bacteria while SP affects only paracellular transport. These findings may have implications for the pathogenesis of Crohn’s disease.

79 5-Aminosalicylic acid interferes with cell cycle progression of colorectal cancer cells P.J. Koelink1, M.A.C. Mieremet-Ooms1, W.E. Corver2, A.M. Mommaas3, G. Griffioen1, C.B.H.W. Lamers1, D.W. Hommes1, H.W. Verspaget1 Departments of Gastroenterology-Hepatology1, Pathology2, Molecular Cell Biology3, Leiden University Medical Center, Leiden, The Netherlands Introduction: The cancer-chemopreventive potential of 5-ASA was previously confirmed by us, as illustrated by the inhibition of proliferation and induction of apoptosis of various colorectal cancer cell lines and in colorectal cancer patients. Further studies were performed to elucidate the mechanisms involved. Methods and results: Flow cytometry analysis, including cyclin A/B1 and phosphohistone H3, showed that 5-ASA induced cell cycle arrest in the S and G2/M phase of HT29 cells, whereas Caco-2 cells showed an arrest in the G0/G1 phase. Microscopic analysis confirmed the mitotic arrest. Aberrant mitotic spindles, leading to features of “mitotic catastrophe” or “mitotic death”, i.e., giant cells containing multiple micronuclei, were also found by fluorescence- and electronmicroscopy. A dosedependent increase in apoptosis by 5-ASA was found, as assessed by Annexin V flow cytometry, active caspase-3 western-blotting, and immunohistochemical detection of the caspase-cleavage product of cytokeratin 18 (M30). Western-blot analysis also showed that the survivin and p21 pathways do not contribute to the 5-ASA effector mechanisms. The anti-inflammatory action of 5-ASA has been reported to be dependent on the peroxisome proliferator-activated receptor-gamma (PPAR-gamma). GW9662, a specific PPAR-gamma antagonist, was not able to antagonize 5-ASA’s effects on cell proliferation, indicating that its growth inhibitory effects are PPAR-gamma independent. Discussion/Conclusion: Thus, our experiments demonstrate that 5-ASA interferes in the cell cycle progression of colorectal cancer cells in vitro and induces apoptosis. These responses are time and concentration/dosage dependent, but PPAR-gamma independent. These observations give insight into the mechanisms involved in the chemopreventive properties of 5-ASA.

80 Local IL-2 application in non-operable colon cancer patients Krastev, Z., V. Koltchakov, S. Deredjian University Hospital “St. Ivan Rilski” Sofia, Bulgaria Background: IL-2 is autocrine and paracrine acting immunoregulatory cytokine synthesized by the activated T-helpers CD4+ (Th) subpopulation Th1. The main effects of this cytokine are induction of activated Th-cells and NK-cells, promoting the Tc-cells function and stimulation of B-cells proliferation. Aim: Our objective was to use safe, with no side effects method in increasing the quality of life (physical, functional, social aspects) and self-confidence in patients with non resectable colon cancer. Methods and patients: In three non-operable patients suffering colon cancer IL-2 was applied locally (into the tumor mass) with endoscopic technique once a week in four subsequent weeks. Afterwards, IL-2 was applied again locally once a month in six subsequent months. Results: Any side effects of the local IL-2 implication were not observed. Remarkable improvement in laboratory findings (increase in albumin, hemoglobin, maintaining normal complete blood cell count), increase in appetite, gaining weight, and increase in Karnofski score, work efficiency and better quality of life were achieved. Conclusions: Local IL-2 therapy in non operable cases with colon cancer is a safe method with no side effects that improves the quality of life, self-confidence and work efficiency in these patients.

81 Ghrelin – A new mediator of colonic inflammation? *P.C. Konturek, **T. Brzozowski, *P. Gaca, *G. Burnat, *S.J. Konturek, **E.G. Hahn *First Department of Medicine, University Erlangen-Nuremberg, Germany and **Institute of Physiology, University of Cracow, Poland Background and aims: Ghrelin is a novel growth hormone (GH)-releasing and orexigenic peptide with anti-inflammatory activities. However, the role of ghrelin in the colonic inflammation is still controversial. The aim of the present study was: 1) to examine the expression of ghrelin mRNA in the inflamed colonic mucosa of patients with ulcerative colitis (UC), 2) to analyze the effect of exogenous ghrelin on the healing of TNBS-induced colitis in rats, and 3) to assess the effect of ghrelin treatment on expression of mRNA for PPARγ, COX-2 and iNOS. Material and methods: 20 patients with UC and 20 controls were enrolled in this study. Expression of TNFα and ghrelin was assessed by quantitative RT-PCR in the colonic biopsies from UC patients and controls. In addition, the effect of exogenous ghrelin on TNBS colitis was tested in Wistar rats without or with capsaicin (125 mg/gk s.c.) to induce the functional ablation of sensory nerves. The mucosal PGE2 generation and NO release into colonic lumen was determined by RIA and ELISA, respectively. Results: Patients with UC showed a significant upregulation of ghrelin and TNFα in colonic mucosa as compared to that observed in controls. The expression of ghrelin correlated with the grade of inflammation and expression of TNFα. The exogenous ghrelin at dose 20 µg/kg i.p. daily significantly accelerated the healing of TNBS colitis and this effect was accompanied by increase in COX-2 and iNOS mRNA expression and a significant rise in the mucosal generation of PGE2. The PPARγ mRNA which was down-regulated in rat colonic mucosa after exposure to TNBS as compared to intact colonic mucosa was not significantly influenced by ghrelin treatment. Conclusion: 1) Patients with UC show an increased mucosal expression of mRNA for ghrelin in the colon; 2) Ghrelin accelerates healing of colonic lesions in animal model of colitis via increased release of NO and PGE2 due to an increase in iNOS and COX-2 expression and activity and sensory neuropeptides such as CGRP released from sensory afferent endings.

82 The role of interleukin-like EMT inducer (ILEI) in liver carcinoma progression Christian Lahsnig, Mario Mikula1, Heidemarie Huber, Hartmut Beug1 and Wolfgang Mikulits Department of Medicine I, Division: Institute of Cancer Research, Medical University of Vienna, Borschke-Gasse 8a, A-1090 Vienna, and 1Research Institute of Molecular Pathology, Dr. Bohr-Gasse 7, A-1030 Vienna, Austria Introduction: The synergy of transforming growth factor (TGF)-beta and Ras causes an epithelial to mesenchymal transition (EMT) which is known to play a major role during cancer progression and metastasis formation. Most recently, a novel key regulator of epithelial plasticity, referred to as interleukin-like EMT inducer (ILEI), has been identified by expression profiling of translationally regulated genes during the EMT of mammary epithelial cells. ILEI has been demonstrated to be sufficient for a gain in malignancy and execution of tumor dissemination independently of TGF-beta. Methods: In this study, we employed an established in vitro and in vivo model of hepatocellular carcinoma (HCC) progression in order to investigate the role of ILEI in hepatocarcinogenesis Results: We show that hepatocytes expressing both hyperactive Ras and exogenous ILEI are capable to augment tumor formation as compared to those expressing oncogenic Ras on its own. On the molecular level, murine neoplasia derived from the cooperation of Ras and ILEI closely correlated with the upregulation of PDGF receptor and ligand which has recently been described to activate Stat3 and nuclear beta-catenin accumulation. Further analyses focused on human HCC samples and aimed at examining the cellular distribution of ILEI in neoplastic tissues of different stagings. Most intriguingly, ILEI has been localized in granular compartments of normal liver tissues and differentiated tumors, whereas a cytoplasmic distribution has been detected in less differentiated and metastatic HCCs. Discussion/Conclusion: These data suggest that ILEI might crucially contribute to the progression in late stage liver tumorigenesis, and furthermore, might serve as a valuable marker for the diagnostic evaluation of human HCCs.

83 Peripheral blood gene expression markers of local and progressive colorectal diseases determined by whole genome mRNA array analysis Bela Molnar, Orsolya Galamb, Barnabás Galamb, Norbert Solymosi, Ferenc Sipos, Sándor Spisák, Kinga Toth, Pal Miheller, Tamas Zagoni, Laszlo Herszenyi, Zsolt Tulassay, M.D., D.Sc. 2nd Department of Medicine, Semmelweis University, Budapest, Hungary Introduction: Gene expression analysis of peripheral blood using high-density oligonucleotide microarray can contribute to the determination of distant blood markers of local pathophysiological alterations in colorectal diseases. With the help of whole genomic microarrays circulating tumor cell related mRNA markers can be detected and verified, as well. Methods: 8 ml peripheral blood was collected, immediately lysed, stabilised in Paxgene tubes then total RNA was extracted, amplified after β-globin reduction and biotinylated from 8 patients with colorectal cancer, 8 with villous adenoma (> 1 cm), 8 with active IBD and 8 healthy normal controls. Genome-wide gene expression profile was evaluated by HGU133 Plus2.0 microarrays. RMA background correction, quantile normalization and median polish summarization were used. Kendall’s rank correlation was performed for quantification of association between the expression level and the disease stages. Results: Cytokeratin 20 was elevated in IBD samples as compared to normal ones (2.63 ± 1.01/2.42 ± 1.15). Classification of benign and malignant tumor markers in comparison with IBD and normals could be performed using downregulated genes like albumin, coeruloplasmin, doublesex and mab-3 related transcription factor 2 (DMRT2), SNF2 histone linker PHD RING helicase, SWI/SNF related regulator of chromatin, and upregulated like C7 open reading frame 38, CYP2C18, Chemokine receptor 8, CD36. Carcinoma cases could be classified from adenomas by upregulated specrin beta 4, translocase pf inner mitochondrial membrane, synapsin 1, microtubule associated monooxygenase, max-associated gene and downregulated CDC20 cell division cycle 20 homolog b, solute carrier family 34, CYP11A1, tropomyosion 1. CRC could be classified from all of the other observed groups by upregulated tropomyosin 3, Adaptor related protein complex1, E2F7 transcription factor, Amyloid precursor, MADS box transcription factor 2, and downregulated EP400N terminal like protein, Nudix type motif 4 pseudogen1, Spouty related EVH1 domain. Albumin, CCDC7 colied-coil domain containing 7, DMRT2, SHB showed sequential changes in normal, adenoma and CRC. Fold changes were at all of these genes in significant difference to the reference normal group by (p < 0.001). Discussion/Conclusion: CK20 circulating tumor cell marker proved to be elevated in IBD patients. New multivariate expression markers were identified using mRNA while genomic expression arrays and peripheral blood. These markers and technology can lead to alternative screening procedures.

84 Correlation between mRNA expression level and protein array results in colorectal cancer Sandor Spisak1, Orsolya Galamb1, Norbert Solymosi1, Barnabas Galamb1, Balazs Nemes2, Ferenc Sipos1, Annamaria Kovacs1, Aron Somoracz1, Zsolt Tulassay1 and Bela Molnar1 1 2nd Department of Medicine, Semmelweis University, Budapest, Hungary 2 Department of Transplantation and Surgical Clinic, Semmelweis University, Budapest, Hungary Introduction: Time of the genome screening and whole genome mRNA arrays significantly raised available. However, we have very few information about the correlation between the mRNA and protein expression levels determined by array technologies. Our aims were to identify the correlation between expressed proteins and transcripts in parallel array experiments in normal and colorectal cancer (CRC). Methods: From 6 Dukes stage D colorectal cancer patient, malignant and normal specimen were collected and fresh frozen immediately after surgery. In parallel from the same sample total RNA and protein were isolated. After the labelling the samples were hybridized to the Clontech AB500 antibody microarrays and Affymetrix HGU133 plus 2.0 microarrays. From the Bioconductor programme packages Limma and affy were chosen for data analysis in R statistical environment. RMA and normexp background reduction method and Quantile normalization methods were applied. Significantly differently expressed genes were identified by T test. For the statistic analysis Empiric Bayes Model was used, which takes the variance in the groups into consideration, by this means creates possibility for better evaluation. Results: Comparing normal and DUKE’S D samples in case of 447 genes we do not expire expression level changing mRNA level neither protein level. It means positive correlation if R2 > 0.9. In protein level we found expression change in case of 53 genes. From these 35 down regulated and 18 up regulated. Compare to the results of the two platform, we received the following result: In the cases of 13 genes the decrease of mRNA level was followed by the reduction of protein level. (B-cell CLL/lymphoma 2, caspase 9). In the cases of 9 genes the protein level increased with the increase of mRNA level. (p53 binding protein, tyrosine kinase 2, nestin, interleukin 1 beta). In 31 cases the two levels were changing in the opposite way. (v-raf-1 murine leukemia viral oncogene homolog 1, annexin A6, DNA topoisomerase I). The normal and CRC samples could be distinguished according to both of mRNA and protein expression markers. Discussion/Conclusion: The difference between the protein and mRNA expression results could be justified by the possible mRNA splicing alternatives, half life, other type of modification and protein or mRNA digestion in the sample preparation process. However this high correlation between protein and mRNA array results supports the further application of mRNA expression arrays in progression marker discovery.

85 Progression markers of early and late stage colorectal cancer determined by protein arrays Sandor Spisak1, Orsolya Galamb1, Norbert Balazs Nemes2, Ferenc Sipos1, Tamas Zagoni1, Bela Molnar1 1 2nd Department of Medicine, Semmelweis 2 Department of Transplantation and Surgical Budapest, Hungary

Solymosi1, Barnabas Galamb1, Kinga Toth1, Zsolt Tulassay1 and University, Budapest, Hungary, Clinic, Semmelweis University,

Introduction: Colorectal cancer progression markers could be important diagnostic and therapeutic targets. The protein chip technology could be utilised for the parallel evaluation of large number of proteins at the same time. Methods: Patient and methods: From 10 Dukes B and 6 Dukes stage D colorectal cancer patient, malignant and normal specimen were collected and fresh frozen immediately after surgery. Proteins were purified by Alumina (Sigma) from homogenized collected tissue and were labelled by monoreactive Cy3 and Cy5 fluorescent die (Amersham). The labelled samples were hybridized by die swap labelled technique on AB500 antibody array (BD) and scanned by a Genepix scanner (Molecular Devices). R statistical software and Bioconductor limma and marray programme packages were used for data analysis. The background reduction was performed by RMA and normexp normalization methods. Quantile normalisation method was applied. Significantly differently expressed genes were identified by T test. The MA value or fold change of differentially expressed proteins was determined. Results: The normal and CRC samples could be distinguished according to protein expression markers. Examining these two groups we found 53 differentially expressed genes, from which 23 (tumor protein p73, caldesmon 1, heat shock 90kDa protein 1 alpha) was down regulated and 30 (DNA topoisomerase I, cyclin A1) was up regulated in tumour samples. To separate normal and Dukes ’B group 74 genes were applicable, from which 27 (interleukin 13, syntaxin 8) was down regulated and 47 (heat shock 60kDa protein, nuclear mitotic apparatus protein 1) was up regulated at protein level. Examining normal and Dukes’D groups we found 68 differentially expressed protein, where 27 (TNF receptor-associated factor 2, B-cell CLL/lymphoma 2) was down regulated and 41 (tyrosine kinase 2, cyclin D3) was up regulated. We managed to separate the two groups of tumour with the evaluation of 12 differentially expressed genes, where 5 was down regulated (syntaxin 8, sequestosome 1) and 7 was up regulated (cyclin A1 CDC-like kinase 1). In some cases we found signs of HPV 16 infection in tumour samples. The expression in cases of C-myc oncogene, Caveolin 1, HPV 16, COX-2, TGF-B was confirmed on tissue microarrays composed from the paraffin embedded surgical samples by immunhistochemistry. Discussion/Conclusion: Protein arrays can identify progression and diagnostic markers colorectal surgical specimen. The found new markers can be introduced into immunhistochemical protocols and evaluated by further therapeutic studies as potential targets.

86 Highdensity microarray analysis of colitis-associated cancer – Identification of new therapeutic targets C. Neufert, C. Becker, A. Nikolaew, P.R. Galle, U. Sahin, O.Tuereci, and M.F. Neurath I. & III. Department of Medicine, University of Mainz, Germany Introduction: Molecular mechanisms giving rise to colitis associated cancer – a lifethreatening complication of ulcerative colitis and Crohn’s disease – are still poorly understood. We have previously shown that colitis associated cancers can be studied by an experimental mouse model based on the inflammatory agent dextran sodium sulfat (DSS) in combination with azoxymethane (AOM) which promotes colonotropic mutagenicity. In order to investigate key factors that drive the growth of colitis associated cancers, we have profiled the gene expression patterns of such tumors. Methods: For the murine AOM-DSS model, a single dose of AOM was administered to C57BL/6 mice intraperitoneally and followed by 3 cycles of DSS in drinking water. Tumors and tumor-free colon epithelium were harvested after 80 days. Whole genome analysis was performed by highdensity microarrays (Affymetrix). Validations of gene expressions were carried out by RT-PCR and immunohistochemistry. Results: We have identified more than 800 genes that were differentially expressed with a significance level of p < 0.05 and showed an induction or repression of > 2-fold. Interestingly, markers of cell proliferation, and genes reported to influence tissue remodeling ranked among the most upregulated transcripts. In contrast, many genes involved in cell differentiation were markably reduced. The analysis of signaling cascades showed a frequent dysregulation of genes related to the Wnt/Beta-Catenin-pathway and TGFbeta signaling, respectively. Discussion/Conclusion: Our whole genome expression analysis has revealed numerous genes differentially expressed in colitis associated cancers, and we have identified dysregulated signaling cascades that might play key roles for the development of such tumors. Thus, our data shed light upon the pathobiology giving rise to colitis associated cancers, and can help to develop new therapeutic options for this life-threatening complication of inflammatory bowel diseases.

87 The Nod2 protein is regulated via the ubiquitin-proteasome pathway Ida Schoultz1*, Tieshan Jiang3*, Maria Lerm2, Peter Söderkvist3 and Johan D. Söderholm1 1 Department of Biomedicine and Surgery, Division of Surgery and 3Division of Cell Biology, 2Department of Molecular and Clinical Medicine, Division of Medical Microbiology, Faculty of Health Sciences, Linköping University, Linköping, Sweden *Both authors contributed equally to this work Introduction: Crohn’s disease (CD) has long been an enigma. Predisposing mutations have been mapped to the CARD15/Nod2 locus, which encodes a cytoplasmic receptor recognising bacterial muramyl dipeptide (MDP). This recognition event triggers an activation of nuclear factor-κB (NFκB). Despite active research, little is still known about how Nod2 is regulated. Methods: Immunoprecipitation experiments were performed in colorectal cancer cell line SW480 with a polyclonal Nod2 antibody. IκBα, known to be degraded following uibquitination, was used as a positive control. Co-immunoprecipitation with ubiquitin was confirmed by immunoblotting with a monoclonal ubiquitin antibody. The turnover of Nod2 was measured through immunoprecipitation following inhibition of the protein synthesis with cyclohexamide. The regulation of Nod2 was also studied by using the proteasome inhibitor MG-132. The interaction between ubiquitin and Nod2 was further confirmed in HEK297 cells co-transfected with vectors carrying hemagglutinin (HA) tagged wild type Nod2 and histidine tagged ubiquitin. Results: Experiments showed that wild type Nod2 has a rapid turn over, with 50% of the protein being degraded after 30 minutes, and co-immunoprecipitates with ubiquitin. Immunoprecipitation assays also reviled that wild type Nod2 accumulates in cells treated with MG-132. Co-transfection experiments confirmed that wild type Nod2 does bind ubiquitin. Conclusions: Our results show that Nod2 is indeed regulated via the ubiquitinproteasome pathway and give important additional information on our current understanding of Crohn’s disease and how the disease is emerging.

88 A regulatory role of Th2-associated transcription factor NFATc2 in murine colitis-associated colon cancer model B. Weigmann1, L. Glimcher2, P.R. Galle1, M.F. Neurath1 1 I. Med. Klinik, Johannes Gutenberg Universität Mainz, Germany 2 Department of Immunology, Harvard School of Public Health, Boston, USA. Introduction: Colorectal cancer is one of the most malignancies. However, the molecular pathogenesis of colorectal cancer is poorly understood. In order to investigate the functional role of the transcription factor NFATc2 (nuclear factor of activated T-cells), which is described for the induction of IL-4 expression, in colon carcinogenesis, we used a previously established murine colon carcinoma model based on the mutagenic agent azoxymethan (AOM). Methods: Accordingly, mice were treated with AOM followed by three consecutive cycles of orally administrated dextran sulfate sodium (DSS) over a period of 7 days. To monitor tumorigenesis in mice in vivo, we used a mini-endoscopic system. By using this system together with methylene blue, we were able to detect aberrant crypt foci in DSS plus AOM-treated wild-type mice at early time point before macroscopically visible lesions were seen. Small visible lesions first appeared around day 20, which were followed by the development of large tumors until day 80. Results: In our model control mice developed colitis-similar symptoms with tumors whereas NFATc2 deficient mice are protected. The possibility, that T cells play a regulatory role in the development of colon tumors led us to perform a screening of the expression of T-cell-derived cytokines in colon tissues and tumors tissues of AOM/DSS-treated wild-type and NFATc2 deficient mice. Discussion/Conclusion: Our data encourage further work of NFAT signaling in colon cancer to establish a novel mechanism for regulation of tumor cell growth.

89 Cancer expressions of HIF-1α associate with immunoreactivities to TGF-β1 in colorectal cancers and adjacent inflammatory infiltrates Wincewicz, A., Sulkowska, M., Sulkowski, S., Kanczuga-Koda, L., Koda, M. Departments of Pathomorphology, Collegium Pathologicum, Medical University of Bialystok, Waszyngtona 13, PL-15-269 Bialystok, Poland E-mail: [email protected] Introduction: HIF-1α is hypoxia upregulated subunit of transcriptional factor HIF-1, that helps the cancer cell survive during oxygen deficiency. During colorectal carcinogenesis TGF-β undergoes an amazing functional change from suppressor of cancer cell proliferation to inhibitor of T cell mediated anti-cancer immunity. HIF-1 contributed to activation of TGF-β3 gene in human trophoblast. Moreover, stromal expression of TGF-β-R1 tended to correlate negatively with cancer immunoreactivity to HIF-1α in breast cancers. HIF-1 and TGF-β co-stimulated fibrosis inducer – Cftg gene. The co-operation of HIF-1 and TGF-β seemed to be so strong that inhibition of TGF-β resulted in blocking of HIF-1 transcriptional activity. Methods: Therefore we evaluated expressions of TGF-β1 and HIF-1α by immunohistochemistry and first compared them in 108 colorectal cancers. Results: HIF-1 was richly expressed in paranuclear pattern predominantly in cytoplasm of malignant cells. As expected not only tumour cancer cells accumulated TGF-β1 abundantly and diffusely in cytoplasm but also there was cytoplasmic, granular immunoreactivity to TGF-β1 in single scattered cancer cells and stromal inflammatory cells. Cancer immunoreactivities to TGF-β1 and HIF-1α correlated with each other in all colorectal cancers (p < 0.0001, r = 0.5160) and subgroups of different clinico-pathological traits. TGF-β1 expressions of adjacent inflammatory infiltrates also correlated with immunoreactivities to HIF-1α of tumour cells (p = 0.008, r = 0.2544); particularly in node positive and deeply growing cancers. Discussion/Conclusion: Thus, we suggest that HIF-1 could co-operate with TGF-β1 and TGF-β1 might mediate cross-talk between inflammatory environment and tumour in promotion of spreading of colorectal cancer.

90 The effect of iNOS inhibitors treatment in experimental colitis models Zeki Yesilova1, Cemal Nuri Ercin1, Ahmet Korkmaz2, Ayhan Ozcan3, Ahmet Uygun1, Kemal Dagalp1 1 Department of Gastroenterology, 2Department of Physiology, 3Department of Pathology, Gulhane Military Medical Academy, Ankara, Turkey Our aim was to investigate the effectiveness of aminoguanidine (AMG), an iNOS inhibitor treatment on experimental colitis model, resembling ulcerative colitis. We induced colitis by applying 4% acetic acitis (AA) in transrectal route to 18 Sprague-Dawley rats, weighing 200–300 gr. Rats were divided into 2 groups. In control group (n = 9), we applied 2 ml. serum physiologic in intraperitoneal route daily for seven days. In AMG group (n = 9), 2 hours after induction, 100 mg/kg AMG was applied intraperitonally in two hours twice a day, for seven days. At the end of 7 days, all the rats were sacrified and the distal 10 cm part of colon were examined macroscopically and scored. In microscopic examination, edema, inflammation, ulceration, crypt hyperplasia, crypt distortion and loss of goblet cells were scored. NO levels in urine were measured by Gries method. The histologic findings, showing the severity of colitis were reduced by AMG, application. The improvement in histologic findings were significant in AMG group comparing the control group (p = 0.001). Weight loss was significantly lower in AMG (p < 0.001). In colitis induced by AA, we showed that nitric oxide activities were depressed by AMG (p < 0.001). In rats, colitis induced by AA, treatment reduced weight loss and colonic inflammation and depressed NO levels.

91 Clinicopathological and gene expression examination of young colorectal tumor patients Z. Szepes1, I. Németh2, T. Molnár1, F. Nagy1, A. Vörös2, L. Tiszlavicz2, J. Lonovics1 University of Szeged, First Department of Medicine1, Institute of Pathology2, Szeged, Hungary Introduction: In the colorectal tumorigenesis various genetic pathways are considered important in different age groups. In the view of prognosis, VEGF, GRP78 protein positivity among others seem to have overriding importance. We compared the gene expression profile of colorectal tumor patients under the age of 55 with data from literature. We also try to find the connection between the gene expression profile and the tumor invasion. Methods: In the period of 1995–2005 71 colorectal tumor cases were suitable for tissue microarray technique. The following parameters were investigated in the tumor and the auto control healthy tissue: grade of differentiation, tumor staging, lymphovascular invasion, perineural spreading, peritumoral inflammation, necrosis, desmoplastic reaction and the gene expression profile of p53, MMS, β-catenin, estrogen, cyclin D1, Cox-2 and VEGF. Results: We found the occurrence of β-catenin nuclear immunopositivity higher in the young colorectal tumor patients than in the literature (30% vs. 50%). The incidence of the other gene expression markers was the same in our young and in the old population. β-catenin positivity showed a significant correlation with the tumor progression and lymphatic spreading. We revealed microsatellite instability in 3 cases. We could not detect estrogen receptor positivity. Marked positivity of p53 was found in tumor specimens, which refers high gene instability. Discussion/Conclusion: Our study also shows that in young patients the dominating colorectal carcinogenesis pathway is APC mediated. On the other hand, the incidence of microsatellite instability is under our expectations. These findings support the prognostic value of β-catenin nuclear immunopositivity in young patients.

92 Expression of Twist1 in colo-rectal tumors correlates with malignant transformation, local invasion and metastases at diagnosis Luigi Laghi1,2, Paolo Bianchi2, Andrea Zecchini2, Elisa Cattaneo3, Jacob SabatesBellver3, Katia Villa2, Alessandro Repici1, Massimo Roncalli4,5, Giancarlo Marra3, Alberto Malesci2,6 1 Department of Gastroenterology, 2Research Laboratory and 4Department of Pathology, IRCCS Istituto Clinico Humanitas, Rozzano-MI, Italy; 3Institute for Molecular Cancer Research, University of Zurich; Department of 5Pathology and of 6 Internal Medicine, University of Milan, Italy Introduction: Colorectal cancers (CRC) with microsatellite instability (MSI) are less metastatic than microsatellite-stable (MSS) CRC. It is not known whether CRC express Twist1, a mediator of epithelial-to-mesenchymal transition (EMT) and metastatis. We assessed whether MSI and MSS CRC of different stages express Twist1. Methods: Twist1 mRNA was investigated in 32 colorectal adenomas/normal mucosa, 13 MSS CRC, and 18 CRC cell-lines. In 119 primary CRC Twist1 expression was studied by immunohistochemistry (expression scores: -, +, ++). Results: Normalized Twist1 mRNA levels were low in normal mucosa (mean 0.95 ± 0.38), adenomas (0.81 ± 0.35), and cell-lines (mean 0.98 ± 2.08), but increased by 5-fold in CRC (3.6 ± 2.2; p < 0.001 vs. other subsets). Twist1 was expressed in 55 (58.5%) MSS and in 3 (12.5%) MSI CRC (p < 0.0001). Among MSS CRC, Crohn’s disease ++Twist1 expression increased with the depth of tumor invasion (2/13 T1, 7/15 T2, 31/50 T3, and 10/16 T4; p = 0.015), OR 3.5 (p = 0.008) for CRC invading beyond the muscularis propria. The ++Twist1 pattern was more frequent advancing from stage I to IV, OR 2.3 (p = 0.03) for metastatic cases. Discussion/Conclusion: In colon tumors, Twist1 mRNA increases with malignant transformation, likely due to microenvironment interactions rather than to clonal alterations, as suggests the expression higher in CRC than in cell-lines. The type of genetic instability correlates with Twist1 expression, and possibly with EMT, a difference that might contribute to reduce the metastatic potential of MSI CRC. In MSS tumors, the expression mirrors tumor invasion and metastasis, indicating that Twist1 participates in the progression of most CRC.

Author Index to Poster Abstracts (Name - Poster Number) Abdu, S. Abu Shamsieh, R. Adjarov, D.G. Akpinar, Z. Aksoz, M.K. Alper, E. Antonova, N. Arhan, M. Ari, F.O. Arjamkina, O.L. Aslan, A. Ayub, N. Baldisserotto, M. Banasiewicz, T.

4 39 77 1, 2 1, 2 1, 2 23 9 1, 2 3 70 4

Bancila, I. Banciu, C. Barreiro, M. Bataga, S. Bataga, T. Becheanu, G. Becker, C. Belbraouet, S. Bera, L. Besherdas, K. Beug, H. Bianchi, P. Biscaglia, G. Blázovics, A. Bodoky, G. Bodrug, N. Bojko, B. Bozkurtoglu, H. Bratu, A. Brisc, C. Brzozowski, T. Bucur, D. Bucurica, S. Budzynski, J. Burnat, G. Butorov, I. Buyrac, Z.

20 16, 47, 55, 56 63, 64 59, 60, 61 5 28, 53 28, 53 29, 72 86 26 45 38 82 92 24 15, 71 48 23 44 70 28, 53 46 81 63, 64 14 6, 17, 18 81 23 1, 2

Campregher, C. Canbolant, E. Caserta, L.

74 70 75

Cattaneo, E. Cetin, M. Cevik, C. Chetnik, A. Chirileanu, O. Chudzicka-Strugala, I. Cioffi, M. Cirovski, G. Ciurea, I. Cornianu, M. Corver, W.E. Cotruta, B. Croitoru, A. Cruciat, C. Czkwianianc, E. Czyzewska, J. Dagalp, K. David, L. Deac, M. Debry, G. Demir, S. Denisov, N. Deredjan, A. Deredjian, S. Diculescu, M. Dinya, E. Disibeyaz, S. Dobrowolska-Zachwieja, A. Dominguez-Muñoz, J.E. Dörnyei, O. Drews, M. D´Souza, R. Dumbrava, M. Dumitrascu, D. Dumitrascu, D.L. Durko, A. Dymicka-Piekarska, V. Ehrmann, J. Ember, I. Ercin, C.N. Ericson, A.-C.

92 70 1, 2 76 60, 61 12 75 68 64 49 79 63, 64 64 34 65 76 90 57 36, 45, 58 26 1, 2 39 77 80 29, 63, 64, 72 15, 27 9 10 5 15 12, 16, 55, 56 38 29, 72 57 57 65 76 33 54 90 78

Farbtuh, T. Fedulova, E.N. Filippo, F.R. De Fodor, J. Forte, G. Fratila, O. Fyderek, K.

52 25 24 15 24 46 13

Gaca, P. Galamb, B. Galamb, O.

81 83, 84, 85 27, 83, 84, 85 Galle, P.R. 86, 88 Gasche, C. 74 Gavrila, A. 66, 73 Georgescu, D. 28, 53 Georgescu, E.F. 31 Georgescu, M. 31 Gheorghe, C. 29, 63, 64, 72 Gheorghe, L. 29, 63, 64 Golusinski, P. 47 Gonenci, R. 70 Grad, K. 6, 17, 18 Griffioen, G. 79 Grillo, L. 20 Grochowalski, M. 47 Grzybowska-Chlebowczyk, U. 11 Guedes, L. 7, 8, 19, 20, 21 Gulubova, M.V. 68, 69 Guzinska-Ustymowicz, K. 76 Gyökeres, T. 62 Hahn, E.G. Halakucova, P. Hamvas, J. Harisi, R. Herbst, F. Herszenyi, L. Herzig, K.-H. Hommes, D.W. Honsova, E. Hoppe-Golebiewska, J. Huber, H. Hyland, J.

81 42 62 48 30 83 16 79 37, 42 10 82 43

Iacob, R. Iacob, S. Ianosi, G. Illés, K.

63, 64 63, 64 22 62

Ioffe, A. Ivanova, A.V.

39 77

Jakubowska, L. Jan, A. Jedynak-Wasowicz, U. Jiang, T. Juhasz, M.

10 4 13 87 27

Kaczmarek, M. Kajor, M. Kanczuga-Koda, L. Keita, A.V. Kemona, A. Kirci, A. Kiss, I. Klopocka, M. Koda, M. Koelink, P.J. Koksal, A.S. Koloszar, J. Koltchakov, V. Konecny, M. Konturek, P.C. Konturek, S.J. Kopeikin, V.N. Korkmaz, A. Kovács, A. Kovács, F. Krasnoperova, Y.Y. Krastev, Z. Krokowicz, P. Kubinska, I. Kucerova, V. Kulig, J. Kupcsulik, P. Laghi, L. Lahsnig, C. Lamers, C.B.H.W. Ledro-Cano, D. Lerm, M. Lodererova, A. Lonovics, J. Lorenzo-González, A. Luciani, M.G. Machado, M.B. Machnik, G. Maghiar, A.

10 11 89 78 76 1, 2 54 6, 17, 18 89 79 9 67 80 33 81 81 25 90 15, 84 62 51 77, 80 56 65 33 41 40, 48 92 82 79 33 87 42 91 5 74 7, 8, 19, 20, 21 44 46

Majewski, P. Makosiej, R. Malecka-Panas, E. Malesci, A. Manciula, D. Manolova, I.M. Manuc, M. Marciniak, R. Marincu, I. Marincu, L. Marinescu, T. Marra, G. May, Z. Mayanskaya, I.V. Mercut, D. Mieremet-Ooms, M.A.C. Mihaila, R. Miheller, P. Mikula, M. Mikulits, W. Mocanu, S. Molinari, A.M. Molnár, B. Molnár, T. Mommaas, A.M. Mulcahy, H. Nagy, F. Neagoe, D. Nedelcu, L. Nemes, B. Nemeth, A. Németh, I. Nesterov, A.S. Nesterov, S. Neufert, C. Neurath, M.F. Nikolaew, A. Nikolov, R. Niksa, M. Niziolek, A. Oancea, C. O'Donoghue, D.P. Oprea, S. Orlikovs, G. O´Sullivan, J.

12 65 65 92 34 68, 69 63, 64, 72 12, 16 49 49 63, 64 92 15 25 22 79 36, 36, 45, 45, 58, 58 83 82 82 53 75 27, 83, 84, 85 35, 91 79 43 35, 91 22 57 84, 85 27 35, 91 51 51 86 86, 88 86 77 76 47 49 43 23 52 43

Pacurari, A. Pap, A. Papadi, B. Parlak, E. Pascu, O. Paszkowski, J. Pelletier, X. Petrascu, O. Pieczarkowski, S. Pintér, E. Pitigoi, D.C. Plawski, A. Pojoga, C. Pokrotnieks, J. Potaturkina-Nesterova, N.I. Preda, A. Prochazka, V. Przybyszewska, K. Puscasiu, D. Puscasiu, M. Pyda, P.

61 62 38 9 50 55, 56 26 36, 45, 58 13 15 63, 64 56, 65 50 52 51 72 33 13 46 46 55

Repici, A. Rezi, C.E. Riegler, G. Rimbas, M. Romosan, I. Roncalli, M. Rosu, A.

92 36, 45, 58 75 14 59, 60, 61 92 66, 73

Sabates-Bellver, J. Sahin, U. Sárdi, E. Savonenkova, L.N. Sbarcea, C. Sbarcea, V. Scaffaro, L.A. Schnabel, R. Schneider, N.C. Schoultz, I. Searpe, C. Seicean, A. Seijo-Rios, S. Seleznovs, J. Serban, V. Sheahan, K. Sheridan, J. Sieczka, J. Sieron, A.L. Sipos, F.

92 86 71 3 66, 73 66, 73 20 54 7, 8, 19, 20, 21 87 66, 73 34 5 52 72 43 43 76 11 83, 84, 85

Skesters, A. Sladek, M. Slomski, R. Söderholm, J.D. Söderkvist, P. Solecki, R. Solymosi, N. Somesundrum, C. Somoracz, A. Somski, R. Spasova, Z. Spicák, J. Spisák, S. Stadnicki, A. Sticova, E. Sulkowska, M. Sulkowski, S. Susan, L.M. Swiatkowski, M. Szakács, A. Szanto, P. Székely, E. Székely, G. Szentmihályi, K. Szepes, Z. Szijártó, A. Szilvas, A. Szkaradkiewicz, A. Szura, M. Takács, I.G. Tarjan, Z. Temiz, M. Tenovici, M. Terracciano, F. Tiszlavicz, L. Tiurean, L. Tolkacheva, N.I. Tombácz, A. Tordai, E. Torok, I. Tossetto, M. Toth, K. Tuereci, O. Tulassay, Z. Tümer, C. Tutina, O.A.

52 13 56 78, 87 87 41 27, 83, 84, 85 38 84 10 77 37, 42 83, 84, 85 44 42 89 89 59, 60 6, 17, 18 62 34 15, 71 71 15 35, 91 40 71 12 41 62 67 70 73 24 35, 91 36, 45, 58 25 67 71 28, 53 43 27, 83, 85 86 27, 83, 84, 85 70 25

Ulker, A. Unsal, B. Uygun, A.

9 1, 2 90

Vacariu, V. Vadan, R. Van Somerun, N. Varga, P. Varsányi, M. Vasile, M. Velev, M. Verspaget, H.W. Vietri, M.T. Villa, K. Vlaykova, D. Vlaykova, T. Vogelsang, H. Voinea, D. Voiosu, M.R. Vörös, A. Vyslouzil, K.

59 29 38 54 62 31 77 79 75 92 68 68, 69 30 63, 64 14 91 33

Wacha, J. Walkowiak, J. Wang, L.-M. Wasilewska, A. Wedrychowicz, A. Weigmann, B. Weltner, J. Wincewicz, A. Wohl, Pa Wohl, Pe Wolanska-Karut, A. Wos, H.

40 16 43 12 13 88 48 89 37, 42 37, 42 44 11

Yazicioglu, N. Yesilova, Z. Yoruk, G. Zagoni, T. Zajac, A. Zecchini, A. Zota, M.

1, 2 90 1, 2 83, 85 41 92 14