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The sentinel node (SN) technique has gained a key role in breast cancer surgery, allowing for an accurate staging of the axillary status with a minimally invasive resection. In this study, we explored the implication of three proteins (E-cadherin, a- and b-catenins) that form the cadherin-catenin complex, a receptorial structure strictly involved in tumoral vascular invasion and embolization in this biologic event. We studied the immunohistochemical expression of the complex in patients with metastatic SN, matching the group with involved nonsentinel lymph nodes (NSNs) with that having free axillary NSNs. The simultaneous staining of the SN metastases for the three proteins has been considered an indicator of preserved function. Our data confirmed the lack of cadherin-catenin complex in tumors with lobular morphology even in SN metastasis, but statistical evaluation could not prove a significant relation between complex integrity and NSN involvement. Moreover, considering traditional histopathologic parameters, only vascular peritumoral embolization was related to an increased risk of metastatic spread to axillary NSNs.

In vitro uptake of bromodeoxyuridine and expression of proliferating cell nuclear antigen (PCNA) were evaluated histochemically in rectal mucosa of control subjects and subjects with colorectal neoplasia in large intestine adenomas and adenocarcinomas. Both labeling indices progressively increased along the path of tumor progression, as did the difference between them (PCNA labeling indices were always greater than those of bromodeoxyuridine). The correlation between them was fairly close in the controls and in adenomas with low-grade dysplasia, whereas no significant linear relations were noted in adenomas with high-grade dysplasia or in adenocarcinomas. The progressive increase in PCNA would thus seem to be related to both hyperproliferation and neoplastic deregulation of PCNA synthesis. In the mucosa of subjects with colorectal neoplasia, PCNA labeling revealed hyperproliferation but not the surface-wards shift of the proliferative compartment detected by bromodeoxyuridine. PCNA expression, therefore, is not a sufficiently sensitive marker of the risk of tumor transformation in the intestinal mucosa.

An in vitro study of proliferative activity as shown by immunohistochemical detection of the uptake of bromodeoxyuridine was run on rectal biopsies from 400 patients with nonfamilial large bowel neoplasia: 200 adenoma; 150 adenocarcinoma; 50 adenoma plus adenocarcinoma. The controls were 400 subjects with negative personal and family histories of colorectal neoplasia. The number and height distribution of bromodeoxyuridine positive cells were determined by dividing the crypt into five longitudinal compartments. The total labeling index and the labeling index of each compartment were higher in all three groups compared with the controls. In subjects with adenoma, total labeling index and labeling index values were correlated with tumor size and decreased in function of the duration of the polyp-free colon state. The major zone of DNA synthesis had shifted to the intermediate and surface crypt compartments in all three groups. This stage II abnormality was more marked in adenoma patients with a high degree of dysplasia and in those with adenoma plus adenocarcinoma. Hyperproliferation and the proliferative compartment shift are cytokinetic abnormalities that coexist in the flat rectal mucosa of patients with colorectal neoplasia. Nonetheless, they are independent, controlled by different factors, and are expressions of different biological aspects of large bowel carcinogenesis.

A monoclonal antibody to bromodeoxyuridine was used in tissue specimens previously incubated with bromodeoxyuridine to show S-phase cells by immunohistochemical technique. Biopsy specimens of normal mucosa (n = 10), hyperplastic polyps (n = 10), adenomas with low-grade dysplasia (n = 20), adenomas with high-grade dysplasia (n = 10), and invasive adenocarcinomas (n = 10) of the large bowel were studied. Labeling index and cell proliferative patterns were analyzed. No statistically significant difference was found in labeling index between normal mucosa and hyperplastic polyps or between adenomas with high-grade dysplasia and adenocarcinomas. The labeling index was significantly lower in normal mucosa and in hyperplastic polyps than in adenomas and adenocarcinomas (p less than 0.001). The difference in labeling index between adenomas with high-grade dysplasia and low-grade dysplasia was also statistically significant (0.01 less than p less than 0.05). In normal mucosa and in hyperplastic polyps the proliferative zone was confined to the lower two-thirds of the crypt; no kinetic activity was found in the upper portions of the crypt or in surface epithelium. In adenomas the labeled cells were either present in the upper third or scattered along the whole axis of the crypt and in the surface epithelium. Labeling patterns in invasive carcinomas were similar to those observed in adenomas with high-grade dysplasia. The difference in proliferative patterns between hyperplastic polyps and adenomas supports a different significance of the two polypoid lesions in the histogenesis of large bowel cancer; our results confirm the subsequent steps of the adenoma-carcinoma sequence. Immunohistochemical labeling patterns observed with monoclonal antibody to bromodeoxyuridine in polypoid and cancer lesions of the large bowel are similar to those described by autoradiographic studies.

The analgesic effect of etorphine was compared with that of morphine in rats with electrolytic lesions of the nucleus medianus raphe or injected intraventricularly with 6-hydroxydopamine. The effect of both compounds was markedly reduced in animals with raphe lesions, but not significantly modified in those receiving 6-hydroxydopamine. Etorphine and morphine significantly increased the forebrain levels of 5-hydroxyindolacetic acid when administered subcutaneously at doses of 5mg/kg and 2 microgram/kg, respectively. Neither drug significantly affected the forebrain levels of monoamines. It is concluded that, as for morphine the integrity of the serotoninergic system in the brain is important for the full analgesic effect of etorphine.

Thin viable slices of normal or pathological human tissues were incubated in vitro with bromodeoxyuridine (BrdU). Later, cryostatic sections and histological sections from the same samples embedded in paraffin were examined by an immunohistochemical method using a monoclonal antibody anti-bromodeoxyuridine (anti-BrdU-MAb): on both cryostatic and histological sections, the nuclei of the S-phase cells proved positive. The optimization of the technique depends on the concentration of bromodeoxyuridine in the culture medium (160 microM), the duration of incubation (not less than two h), the method of DNA denaturation (2N or 4N HCl) and the dilution of the anti-BrdU-MAb (1:50). In vitro, immunohistochemical application of the BrdU/anti-BrdU-MAb method permits a quantitative assessment of the proliferative activity of a tissue as well as the direct location of the actively replicating cells in histological sections.

The AR-3 monoclonal antibody, which defines the tumor-associated antigen CAR-3, was previously found to be able to discriminate between neoplastic cells in gastric, pancreatic, colonic, ovarian and endometrial carcinomas and their normal counterparts. In fact, it strongly reacts with carcinomatous cells at the level of both the glycocalix and the cytoplasm, while its reactivity with normal tissues is restricted to the glycocalix of few mucin-producing epithelial cells. We have now investigated the reactivity of this antibody with immunohistochemical techniques on a series of formalin-fixed paraffin-embedded specimens, from precancerous and cancerous lesions of the large bowel which were classified as adenomas with mild, moderate or severe dysplasia, adenomas with cancer and adenocarcinomas, respectively. It was found that the intensity and extent of the staining correlated with the degree of dysplasia and that the highest expression of the CAR-3 epitope was detectable in adenocarcinomas. Also the localization of the staining in the lesions displayed an increasingly complex pattern, going from linear in adenomas with mild dysplasia to a very strong intracytoplasmic and/or intraluminal expression in adenomas with severe dysplasia or adenocarcinomas.