Histology and histopathology Vol.16, nº 1 (2001)
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- PublicationOpen AccessImmunohistochemical expression of p53 in animal tumors: a methodological study using four anti-human p53 antibodies(2001) Albaric, O.; Bret, L.; Amardeihl, M.; Delverdier, M.Mutations in the p53 tumor suppressor gene are the most common genetic alterations in human cancers. These mutations usually lead to strongly enhanced protein stabilization and allow detection by immunohistochemistry. Two monoclonal (DO-7 and PAb-240) and two polyclonal (Ab-7 and CM-l) antibodies were evaluated by standard immunoperoxidase method in domestic animal tumors, chiefly squamous cell carcinomas (Scq, and osteosarcomas as positive controls. Immunoreactivity was detected in SCC of cattle, sheep, horse and cat as well as in feline actinic keratosis, with PAb-240 and CM-l antibodies. One polyclonal antibody (Ab-7) did not give positive result at all, whereas DO-7 monoclonal antibody did not react in dogs and cats. Immunodetection of p53 protein is thus possible in all domestic species tested, especially with CM-l and PAb-240 antibodies, and p53 alterations seem to occur early in carcinogenesis of feli ne SCC as in comparable human lesions.
- PublicationOpen AccessA salvianolic acid 8-rich fraction of Sa/via miltiorrhiza induces neointimal cell apoptosis in rabbit angioplasty model(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2001) Hung, H.-H.; Chen, Y.-L.; Lin, S.-J.; Yang, S.-P.; Shih, C.-C.; Shiao, M.-S.; Chang, C.-H.Apoptosis has been suggested to participate in stabilizing cell number in restenosis. Salvia miltiorrhiza (SM) Bunge which is a Chinese herb widely used for the treatment of cardiovascular disorders contains a potent antioxidant, Salvianolic ac id B. To determ ine whether the antioxidant affects vascular apoptosis, the present study examined the frequency of apoptotic cell death in atherosclerotic plaques and in restenotic lesions of cholesterol-fed rabbits. New Zealand White rabbits were treated with a normal diet (normal), a 2% cholesterol diet (HC), a 2% cholesterol diet and endothe lial denudation (HC-ED), a 2 % cholesterol diet with 5% water-soluble extract of SM (4.8 glKg B. W./day) and endothelial denudation (HCED-SM), or with a 2% cholesterol diet containing probucol (0.6 glkg B.W./day) and endothelial denudation (HC-ED-probucol). Apoptosis and associated cell types were examined in serial paraffin sections by in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and immunohistochemistry. The expression of p53, an apoptosis-related protein, was also examined. Apoptosis was mainly detected in the neointima of the three gro ups with endothe lial denudation. The percentage of apoptotic cells in SMtreated group (68.5±5.9%) was significantly higher than that of normal (0 %), HC (1.9±1.2 %), HC-ED (46.1±5.4%), and probucol-treated (32.8±3.9%) groups. The SM treatment markedly reduced the thickness of the neointima which was mainly composed of smooth muscle cells with few macrophages. In accordance with the apoptotic cell counts, positive immunoreactivity for p53 was observed in restenotic lesions from HC-ED, SM-treated and probucol-treated groups but not in the intima of the other two groups. These results suggest that the treatment with salvianolic acid B-rich fraction of SM induces apoptosis in neointima which in turn may help prevent the neointimal thickening.
- PublicationOpen AccessStudies on the involvement Histology and Histopathology Cellular and Molecular Biology of endogenous neuropeptides in the control of thymocyte proliferation in the rat(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2001) Trejter, M.; Warchol, J. B.; de Caro, R.; Brelinska, R.; Nussdorfer, G. G.; Malendowicz, L. K.The possible involvement of endogenous vasoactive intestinal peptide (VIP), cholecystokinin (CCK) and neurotensin (NT) in the control of thymocyte proliferation ha s been investigated in vivo in the immature rat. For this task, we have studied the effects of the administration of selective antagonists of the receptors of the three neuropeptides on the mitotic index (%0 of metaphase-arrested cells after vincristin injection) of thymocytes. Both CCK- and TN-receptor antagonists were ineffective. In contrast, two VIP receptor antago nists (VIP-As) enhanced the mitotic index of thymocytes. VIP reversed the effect of VIP-As, but when administered alone it did not alter the mitotic activity of thymocytes. In light of these findings, we conclude that endogenous VIP exerts a maximal tonic inhibitory influence on the basal proliferative activity of rat thymocytes, while endogenous CCK and NT do not play a relevant modulatory role in thi process.
- PublicationOpen AccessPrognostic value of p53 protein expression and clinicopathological factors in infiltrating ductal carcinoma of the breast. A study of 192 patients(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2001) Sirvent, J. J.; Fortuño Mar, A.; Olana, M.; Ortí, A.The p53 gene is located on the short arm of chromosome 17. It encodes a 53-kd nuclear protein (p53) found in scant amounts in normal tissue. Mutations of the p53 gene have been reported in different human tumours. In breast cancer, it has been noted that the overexpression of p53 protein in the nucleus is an indicator of poor prognosis, although there is a high degree of variability, which may be due to different immunohistochemical techniques, varying assessment of results a nd the type of monoclonal antibody used. This study is an immunohistochemical analysis of p53 expression in 192 cases of infiltrating ductal carcinoma of the breast, correlating it with clinicopathological factors and the clinical course of the disease. Of all the breast-cancer tissue analysed, stains for p53 antibody were found in 87 tumours (45.3%). The results of multivariate ana lysis show that the independent predictors related to recurrence are tumour size, Iymphnode metastasis and p53, while those related to death are necrosis, lymph-node metastasis and p53. In summary, our series showed prognostic significance between the expression of p53 and shorter survival time and disease-free interval for all patients in general as well as those who prese nted lymph-node metastases at the time of diagnosis.
- PublicationOpen AccessMicroglial reaction in focal cerebral ischaemia induced by intra-carotid homologous clot injection(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2001) Ng, Y.-K.; Ling, E.-A.This study examined the microglial reaction in a simulated thrombo-embolus ischaemia in rats given an intracarotid injection of a suspension of homologous blood clot. All rats including the controls receiving vehicle injection were perfused at 5 hours, and 1, 3 and 7 days post-operation. The brains were removed and processed for immunohistochemistry using a panel of monoclonal antibodies: OX-42, OX-18 and OX-6 for labeling of microglia. In rats given saline injection OX42 immunoreactive microglial cells were observed to be distributed quite evenly throughout the whole brain. When injection of clot suspension was given, microglial cells responded vigorously, particularly in the ipsilateral hippocampus. Microglial reaction was also detected in the ipsilateral cerebral cortex, caudate as well as septal nuclei. The majority of the detected reactive microglial cells were hypertrophied showing thick or stout processes. Some rod-like and amoeboid microglia were also observed. Rarely did the reactive microglia express OX-6 immunoreactivity. All microglial cells were unreactive for OX-18. The actual mechanisms leading to the microglial activation as well as functions of reactive microglia in focal cerebral ischaemia remain speculative. In the absence of direct evidence, it could only be suggested that they may act as sensor cells for detection of subtle alterations in the microenvironment, probably in response to focal ischaemia and/or leakage of serumderived factors induced by thrombo-embolus stroke.