Histochemical and immunohistochemical techniques in ulcerative colitis

Objective: To demonstrate the mucin changes in Ulcerative Colitis, to evaluate the benefit of the staining method and to demonstrate the CEA staining pattern in dysplasia complicating ulcerative colitis (U.C). Methods: Colorectal biopsies were examined for changes of U.C for which a combined PAS/Alcian blue stain was applied. CEA immunohistochemical stain was used for cases of dysplasia complicating ulcerative colitis. Results: One hundred colorectal biopsies were examined, U.C. was diagnosed in (22%) of cases, the mean age was (36.68) years, (13) were males, and (9) were females. The prevalence of dysplastic changes complicating U.C were detected in (22.7%).The rectum was the most frequent site of dysplasia complicating, U.C.. Two of the cases showed mild dysplastic changes; two showed moderate dysplasia, and only one showed severe dysplasia. The secretory activity of mucin in colorectal mucosa was tow in surface epithelium and varied in the crypts from absent or weak to moderate reduction. The site of CEA distribution seemed to be affected mainly by degree of dysplasia, it was predominantly along the apical surface of the cells and also in the cytoplasm in case of mild dysplasia, while in moderate dysplasia it was of cytoplasmic distribution, and in severe dysplasia there was intensive cytoplasmic distribution. Conclusion: There was an increase in relative frequency of U.C. Immunohistochemical study of CEA localization in dysplastic gland is helpful in detection of early malignant change in U.C. ةصلاخلا : ا ل فده : رهظملا حاضيلا ةقيرطلا ىودج مييقتو يحرقتلا نولوقلا باهتلا ضرم يف طاخملا تاريغت حيضوت ـل يغبصلا CEA يحرقتلا نولوقلا باهتللا لثحلا فعاصم يف . قئارطلا : ةغبصب يحرقتلا نولوقلا باهتلا تاريغت ددصب ميقتسملا نمو نولوقلا نم تاعزخل صحف ىرجأ Annals of the College of Medicine Vol. 34 No 1, 2008 © 2008 Mosul College of Medicine 29 (PAS/Alcian) يعانم غبص ىرجاو ،ةجودزملا ءاقرزلا ـل يواميآ يجيسن CEA قعاضملا لثحلا تلااحل يحرقتلا نولوقلا باهتللا . جئاتنلا : ىدل يحرقتلا نولوقلا باهتلا صخشو ،ميقتسم وا نولوق ةعزخ ةئام صحف يرجأ ٢٢ % لدعم تلااحلا نم ناآ رمعلا ٣٦.٦٨ ؛ ةنس ١٣ و رآذ ٩ ثانا . حلا تاريغتلا تدجو ىدل يحرقتلا نولوقلا باهتللا ةفعاضملا ةيلث ٢٢.٧ % يحرقتلا نولوقلا باهتللا فعاضملا لثحلا تلااحل رثآلاا عقوملا ميقتسملا ناآو . ةفيفخ ةيلثح تاريغت ناتلاح ترهظا ا طقف ةدحاوو ةطسوتم ةيلثح تاريغت اترهظا نايرخا ناتنثاو ظ ةديدش ةيلثح تاريغت ترج ره . ا زارفا ةيلاعف تناآ طاخمل لا جيسنلا نم ط ايافخلا يف ةطسوتم وا ةئظاو ىلا ةمودعم نيبام ،ةئطاو نولوقلاو ميقتسملل يئلا ) crypts .( راشتنا عقوم نا رهظيو CEA رثأتي ام رثآاو ،لثحلا ةجردب تلااح يف يلويهلا يفو ايلاخلل يمقلا حطسلا يف ظحول تلااحلا يف يلويهلا يف ارشتنم ناآ امنيب فيفخلا لثحلا شلا لثحلا يفيلويهلا يف ةأطول اديدش اراشتناو ةطسوتملا د ي د . جاتنتسلاا : ـلا عقوم ةساردلا نم دافتسيو يحرقتلا نولوقلا باهتلا راركت يف ةيبسن ةدايز دوجو رهظي ) CEA ( ةغبصلاب يف ةركبملا ةيناطرسلا تاريغتلا فشآ يف لثحلا فاضم اهيف يتلا ددغلل ةيواميكلا ةيجيسنلا ةيعانملا نولوقلا باهتلا ضرم

Mucin changes in ulcerative colitis: In fulminant and acute active cases of short duration, the decrease in mucosubstance is slight or moderate, while in chronic active cases it is moderate or severe and these changes tend to revert again towards normal in quiescent phase (10) Dysplasia: It is an unequivocally neoplastic but non-invasive epithelial proliferation of the colonic epithelium (11.12) .
2-Indefinite for dysplasia; is applied to epithelial changes that appear to exceed the limits of ordinary regeneration.
4-Microscopically, dysplasia is identified on the basis of combination of microscopic features, which include; U 5-1-Architectural alteration exceeding that resulting from repair in chronic colitis.
6-The distribution of CEA in normal epithelium is limited to the apical surface, while in dysplasia and carcinoma it is abundantly present in both cytoplasm and the lumen (8,13,14) .
Normally Colorectal goblet cells are containing mainly acid mucin with small amount of neutral mucin (15) .The CEA is a glycoprotein of heterogeneous composition (MW 200,000), detected in small amounts in normal adult cells and benign colorectal tumors, but is present in large amounts in carcinoma (8) .The immunocytochemical localization of CEA in colorectal tissue may have a potential value in the diagnosis of premalignant and malignant lesions (14) .staining for neutral and acidic mucin were done respectively.

Patients and methods
2. According to (Riddel et al., 1983) criteria (11) .Immunohistochemical staining for CEA was performed in cases of U.C to detect dysplastic changes by using an improved Biotin-streptavidin Amplified (BSA) detection system.The staining procedure was done according to the manufacturer instructions of staining protocol (16) .

Statistical methods:
The statistical evaluation was performed using mean, range, and chi square test.

Results
Ulcerative
2. our physicians now have high index of suspicion for U.C.

Mucin Pattern in U.C.
The study of mucin pattern in U.C.
showed that, both types of mucin were detected but in decreased amount indicating that, the change in mucin is quantitative rather than qualitative; this finding is similar to that observed by Nazar (24) .The amount of mucin decreased when the activity of disease is increased, this observation is in agreement with other workers (25,26,27) .In our study, all types of mucin present in an inactive colitis were higher in amount than that found in active colitis, this is consistent with observations of other workers (28,29) .
Special attention has been focused on had dysplastic changes and this is more or less similar to the some studies (21 %) (12,30) , and higher than that reported , by (Katran), and (Subbar), (14.7) and (6.5) composition of the goblet cell mucin varies within their level in the crypt and in different segments of colon; in the left colon sulphamucin predominates in the lower half of the crypt, whereas in the upper crypt and surface epithelium a variable proportion of sulphamucin and sialomucin are often demonstrated.

.
Distribution of CEA immunohistochemical staining in dysplasia complicating U.C: In mild dysplasia, CEA distribution was demonstrated predominantly along the apical surface of the cells, while in moderate dysplasia and in severe dysplasia there was intensive cytoplasmic staining.
pre-carcinomatous changes (dysplasia) in patients with U.C. which is considered as a histological marker for increased cancer risk, and thus, as a potential indication for colectomy in patients with U.C.In the present study, (22.7%) of cases of U.C.

Table 2 :
Sex distribution of U.C. with and without dysplasia.

Table 4 :
Predominance of CEA distribution in colonic biopsies of U.C with dysplasia