April 1995 Gastrointestinal Oncology A461 • QUANTITAVE MORPHOLOGICAL VALUE OF THE COLON AND RECTAL ADENOMACARCINOMA SEQUENCE. S.J. Diaz-Cano, F, Ruiz-Delgado,F. Rivera- Hueto, J.J. Rfos-Martth, D. Delgado-Bellido and J.M- Herrerfas Guti6rrez. Dept. of Pathology and GastroenterologyUniversity Hospital "Virgen Macarena" - Seville - Spain INTRODUCTION The colon and rectum carcinoma is an heterogenons illness. Clinical, biological, histopathological and experimental evidencesback up the theory of the adenoma-earcinoma sequence evidences. Among the histopathuloglcal characteristics, the variables studied are subjective, although we have recently started Usingquantifying techniques in the morphological study (9), in order to obtain more objective parameters. As far as we know, these techniques have not yet been used in this pathobiology. The present work aims to introduce the information of conventional histopathological techniques, AgNORs and morphometrieal(glandular and nuclear types) in order to put in relation these variables in the pathology of the adenoma-carcinoma sequence. MATERIAL AND METHOD We selected 100 consecutive patients, 50 with adenoma (33 with low-grade -LGA- dysplasia and 17 with high-grade -HGA- and 50 with adenocarcinoma-(ACA)- who were studied in the conventional way. From these patients we obtained representative surgical and endoscopic samples that we dyed with H-E for histopathological diagnostic and morphometricstudy purposes (Zeiss-Kontron MOPp=Videoplan image analyzer) as well as for argentic tecb-,~aes for the AgNORs quantifying. We studied glandular and nuclear variables (area, perimeter, maximumdiameter and shape factor) and the cellular proliferation (mitotic index, nuclear organisingantigen -AgNORs-) in both neoplasia types and with regard to the nuclear atypia grade (dysplasia) in the adenomas (Kruskal-Wallis tests and by-steps discrimination). RESULTS In the one variant statistical and stepwise discrimiuant survey, the variables that showed significant differences (p<0.05), were as follows: mitotic index (LGA 10,8; HGA 11,0; ACA 22,5), nuclear perimeter (media) (LGA 10,6; HGA 11,3; ACA 14,4), nuclear area (typical deviation, DT) (LGA 6,9; HGA 9,4; ACA 10,0) maximumnuclear diameter (DT) (LGA 2,6; HGA 3,0; ACA 3,8), maximum glandular diameter (media) (LGA 0,2; HGA 0,1; ACA 0,09), glandular shape factor (media) (LGA 0,7; HGA 0,7; ACA 0,6), glandular shape factor (DT) (LGA 0,1 ; HGA 0,1 ; ACA 0,2), luminal nuclens~edge distance (media) (LGA 17,1; HGA 16,1; ACA 12,0). We noticed a progressive increase of the area, maximum diameter and nuclear perimeter, mitotic index along with a gradual decrease of the glandular shape and luminal nucleus-edgedistance. CONCLUSION Therefore, the development of adenocarcinomason colon and rectum adenomas happens over increasing dysplasia grades: higher nuclear size and mitosis index, smaller and more abnormal glands together with loss of the nucleus basal polarity. STATISTICAL ANALYSIS ON COLORECTAL ADENOMAS. Role of the Lynphocitary Inflammatory Infiltrate. S.J. Dfaz-Cano, F. Ruiz- Delgado, F. Rivera-Hueto, J.J. Rios-Martfn, D. Delgado-Bellido and J.M. Herrerfas Gutitrrez. Dpt. of Pathology and Gastroenterology - University Hospital "Virgen Macarena" - Seville - Spain INTRODUCTION The malignant transformation on colorectal adenomas is mainly predicted by the grade of dysplasia. We will try to predict the presence of high-grade dysplasia ir~ rton--potyposis "related colorectai adenomas by means of a multivariate analysis. We have selected 50 consecutive patients with endoscopically resected sporadic solitary adenomas. We studied clinical parameters (age, sex, tumoral location, tumoral size and obstructive symptoms presence), hispotathologic variables (growth pattern, Leutchtenberger's bodies, type and grade of inflammation), glandular and nuclear morphometric features (area, perimeter, shape factor, maximum diameter), and proliferating-related indices (mitotic index, nucleolar organizer regions quantification) respect to grade of dysplasia. Seventeen of them contained high-grade dysplasia which was statistically related to: inflammation grade (p < 0.0001, 2/33 low-grade adenomas and 14/17 high-grade adenomas displayed severe lymphocytic infiltrate) and tumoral location (p=0.0298, 2/2 right colon adenomas were of high-grade and 6/6 of left colon adenomas were of low-grade). The remaining variables showed no significant differences. At the stepwise discriminant analysis, only the inflammation grade retains its independent predicting value. So, it was able to classify correctly the 93.9% of low- grade adenomas and the 82.4% of high-grade adenomas. In conclusion, the grade of dysplasia of colorectal adenomas may be accurately predicted by inflammatory infiltrate intensity: the presence of a severe lymphocytic infiltrate around adenomatous glands would systematically obligate to investigate the existence of coexisting high-grade dysplasia. Key words: Colorectal adenomas, high-grade dysplasia, multivariate analysis. • UPPER GASTROINTESTINAL MANIFESTATIONS OF THE ATTENUATED ADENOMATOUS POLYPOSIS COLI SYNDROME (AAPC). J.A, DiSario, S.K. Khullar, D.J. Bjorkman, M. Leppert, M. Slattery, R. White, R.W. Burt. Depts. of Medicine, Human Genetics and Family and Preventive Medicine, University of Utah, and Medical Service, Salt Lake VA, Salt Lake City, Utah. Upper gastrointestinal (UGI) polyps and periampullary carcinoma are rare in the general population. Gastric fundic gland polyps, duodenal adenomas and periampullary carcinomas, however, are common in typical adenomatous polyposis coli (APC), with a prevalence of 25%-53%, 34%- 90% and 3%, respectively. AAPC is an inherited polyposis syndrome arising from mutations of the same gene that is mutated in APC. It carries a high risk of colon cancer, but affected subjects exhibit fewer adenomas and a later onset of colon cancer compared to those with typical APC. UGI polyps have been observed in some patients with AAPC, but their frequency has not been precisely defined. AIM: To determine the frequency of UGI polyps and cancer in persons with a genetic diagnosis of AAPC from a single large kindred. METHODS: Screening EGD was performed on 21 AAPC gene carriers and ampullary biopsies were obtained in 12. The prevalence ofperiampullary cancer was determined in 122 gene carders and 727 non-gene carders by reviewing their medical histories. RESULTS: For subjects having EGD: median age was 40 yrs (21-73); 13 (62%) had UGI lesions; all had normal appearing ampullae of Vater. Gastric fundie gland polyps 11/21 (52.3%) Duodenal adenomas 3/21 (14.3%) Ampullary adenomas 5/12 (41.6%) History of periampullary ca AAPC mutation present 2/122 (1.6%) AAPC mutation absent 0/727 (0.0%) p<0.02 CONCLUSIONS: 1) UGI polyps occur frequently in AAPC, 2) persons with AAPC are at an increased risk for ampullary adenomas and periampullary cancers, and 3) ampullary adenomas are frequently present in the absence of endoscopically detectable changes. (NIH Grant CA 40641 and Huntsman Cancer Institute) ALTERATIONS OF DNA-REPEATS IN HUMAN PANCREATIC CANCER. P. Di Sebastiano, P. Vitullo, R. Palmirotta, H. Friess, P. Innocenti, R. Mariani Costantini, MW. Buechler and P. Battista. Dpt. of General Surgery and Human Pathology, University of Chieti (Italy) and Institut of Visceral and Transplantation Surgery, Universityof Bern (Switzerland). Genetic alterations in simple repeated sequences (microsatellites) constitute a newly recognizedclass of human mutationscausing disease. Microsatellites are a large number of tandem iterations of di-, tri- and telranucleotiderepeats geneticallyinstable and susceptibleto replications error (RER). Several recent reports indicate that simply sequence repeats represent a frequent target of mutations in a vadety of tumors that display genomic instability. AIMS. Based on these considerations we aimed to investigate possible microsetellite instabilityin pancreaticadenocarcinoma. MATERIALS AND METHODS. Tumor DNAs and corresponding normal DNAs were extracted from 7 patients (4 males and 3 females, median age 58 yrs.) underwent surgery for pancreatic adenocarcinoma. Further, 6 different microsatellitesmarkers were analyzed to characterizegenomic instability and mapped at the following loci: MFD 26 (D18SR4), AR (X chromosome), D2S123, 635/636 (ACTC), MFD 27 (D5S107) and MFD3 (APOA2) by using Polimerase Chain Reaction and denaturing electrophoresison sequencinggels. RESULTS. Tumor DNA samplesexamined exhibited somatic instabilityat microsatellite for dinucleotide repeats 635/636 and trinucleotide repeats AR in 2 out of 7 patients (28%). CONCLUSIONS. Our results show that microsatellite instability is a feature of a number of pancreatic adenocarcinoma. These preliminary findings suggest that genomic instability might be involved in the pathogenesisof human pancreaticcancer.