Browsing by Author "Ferrer, L."
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- Epithelial differentiation markers in mucucocutaneous junctions of different food-producing animalsPublication . Vala, Helena; Fondevila, D.; Ferrer, L.The distribution of keratins and the expression of cornified cell envelope proteins are well studied in epidermis and mucous membranes of human specie, laboratory species and, scarcely, in the dog. The aim of this study was to evaluate, by means of immunohistochemical techniques, the expression of these components in the nasal and labial mucocutaneous junctions from food-producing animals. Samples from two mucocutaneous junctions were collected from goat, sheep, cow and pig, fixed in formalin 10% and embedded in paraffin. For chacterization of keratins, two monoclonal antibodies (DAKO) were used. MNF116, labelling human keratins 5, 6, 8, 17 and 19 in non differentiated keratinocytes and simple epithelia and LP34, labelling human keratins 5, 6 and 18 in well-differentiated keratinocytes. The distribution of corneum envelope precursors was studied using two antibodies: monoclonal antibody anti-human involucrin (NOVOCASTRA), labelling keratinocytes of the upper stratum spinosum and stratum granulosum, and polyclonal antibody anti-human profilaggrin (ZYMED), labelling keratinocytes of the stratum granulosum. With the MNF 116 antibody, the labial and nasal mucocutaneous junctions from goat, as well as labial from sheep and nasal from cow, revealed an intense reaction located at the suprabasale stratum. The labial mucocutaneous junction from pig was negative. With the LP34 antibody, were positive all suprabasale strata from both mucocutaneous junctions in ruminant species. In the pig, both mucous membranes were negatives. With the anti-involucrin antibody, the nasal mucous membrane was positive in ruminants species, at the upper part of the stratum spinosum and in the stratum granulosum, but was negative in the pig. The labial mucous membrane was positive at the same layers in most species, except in the sheep. Both mucocutaneous junctions in all studied species were negative with the anti-profilagrin antibody. It was possible to conclude that keratins were detected in nasal and labial mucous membranes from all studied species at supra-basale layers. In both mucous membranes from goat and cow, as well as in nasal mucous membrane from sheep and labial mucous membrane from pig, was also detected involucrin, at the upper part of the stratum spinosum. Both mucocutaneous junctions from all species didn’t revealed profilaggrin expression.
- Estudio inmunohistoquimico de diferenciación de los carcinomas de células escamosas bovinosPublication . Vala, Helena; Fondevila, D.; Carvalho, T.; Pinto, C.; Peleteiro, C.; Pinho, M.; Ferrer, L.En el ganado bovino son frecuentes los carcinomas de células escamosas localizados en párpados y tercer párpado. En Portugal, este tumor se diagnostica con frecuencia en bovinos de las Azores, representa un 21% del total de neoplasias diagnosticadas en esta especie, la segunda después de los tumores de la vejiga de la orina. El objetivo de este trabajo ha sido conocer el origen y grado de diferenciación de estas neoplasias utilizando anticuerpos contra queratinas expresadas en keratinocitos basales y suprabasales, y anticuerpos considerados marcadores de diferenciación terminal como la involucrina y profilagrina.
- Expresión de una citoqueratina de amplio espectro en piel y mucosas de ovejaPublication . Vala, Helena; Fondevila, D.; Ferrer, L.
- Expresión filagrina en piel de ovejaPublication . Vala, Helena; Fondevila, D.; Ferrer, L.
- Expression of epithelial differentiation markers in cutaneous adnexae of food-producing animalsPublication . Vala, Helena; Fondevila, D.; Ferrer, L.One striking feature of terminal differentiation in mammalian epidermis is the deposition of a 20 nm thick, insoluble layer of protein on the cell’s inner surface, named cornified cell envelope (CE). The dermal adnexae show a different and more complex differentiation pattern than other squamous stratified epithelia. The main objective of this work was to evaluate, by means of immunohistochemical techniques, the expression and distribution of keratin filaments and cornified cell envelope components in the adnexae of different cutaneous regions from food-producing animals. Samples from eight cutaneous regions were collected from goat, sheep, cow and pig. For characterization of keratins, two monoclonal antibodies (DAKO) were used. MNF116, labeling human keratins 5, 6, 8, 17 and 19 in non differentiated keratinocytes and simple epithelia and LP34, labelling human keratins 5, 6 and 18 in well-differentiated keratinocytes. The distribution of corneum envelope precursors was studied using two antibodies: monoclonal antibody anti-human involucrin (NOVOCASTRA), labelling keratinocytes of the upper stratum spinosum and stratum granulosum, and polyclonal antibody anti-human profilaggrin (ZYMED), labelling keratinocytes of the stratum granulosum. With the MNF 116 antibody, the follicular epithelium was positive in all species, always in the outer root sheath and rarely in the inner root sheath. The cutaneous glands and ducts were also positive. With the LP34 antibody, was positive the outer root sheath from follicular infundibulum. The sebaceous ducts were also positive in some sheep and pig regions. With the anti-involucrin antibody, the follicular outer root sheath was positive in most studied regions and the inner root sheath was positive in the sheep and cow. Only small ruminants species revealed positive reaction in the glandular epithelium from some cutaneous regions. The follicular outer root sheath was positive with the anti-profilagrin antibody in the infundibula from all studied species. The epithelial components of nasal and labial glands revealed an irregular reaction. It was possible to conclude that cutaneous adnexae expresses keratins characteristics from simple epithelia with major intensity than epidermal layers and that the follicular epithelium expresses keratins characteristics from well-differentiated keratinocytes when it’s differentiation and keratinisation are similar with those from epidermis. It was also possible to conclude that the follicular outer root sheath of the studied species show involucrin and profilaggrin expression and that the inner root sheath expresses involucrin in sheep and cow. Both corneum envelope precursors were expressed in glandular components, with a variable expression between the species.
- Immunohistochemical detection of filaggrin in the skin of different speciesPublication . Bardagí, M.; Vala, Helena; Fondevila, D.; Ferrer, L.Filaggrin (FLG) is a key protein that facilitates terminal differentiation of the epidermis and formation of the skin barrier. Recent researches have demonstrated that, in humans, some variants of FLG are a major predisposing factor for atopic dermatitis. Furthermore, in immunohistochemical studies, FLG’s staining intensity was found significantly reduced in non-lesional and lesional atopic skin: FLG did not reach the horny layers and stained primarily granules in the stratum corneum. To the knowledge of the authors, the location of FLG in the skin of the domestic animals has not been investigated. Therefore, the aim of this research was to investigate, using immunohistochemistry, the expression of FLG in the skin of the dog, cat, horse, cow, pig and chicken. The long term purpose of this research line would be to investigate FLG expression in canine atopic skin. Formalin fixed, paraffin-emebedded sections were used in a ABC immunohistochemical technique. A monoclonal antibody against human FLG (Acrist Antibodies, Germany) was used as primary antibody. The results were very consistent in all species. FLG was detected clearly in the granular layer of the epidermis and in the outer root sheath of the hair follicles. In the chicken, all suprabasal layers were positive for FLG. The staining in all cases was cytoplasmic, being also the keratohyalin granules strongly stained. These results are very similar top those described in human beings and open the door to further researches regarding the FLG expression in animal skin diseases
- Immunohistochemical studies of proliferation cell markers and DNA mutation in bovine ocular squamous cell carcinomaPublication . Carvalho, T.; Vala, Helena; Fondevila, D.; Pinto, C.; Peleteiro, C.; Pinho, M.; Ferrer, L.The purpose of the present study was to immunocharacterize Bovine Ocular Squamous Cell Carcinoma neoplastic cells, in terms of proliferation index and DNA damage, using two human primary antibodies anti- Ki67 antigen of the cell cycle and anti- p53. Ki67 is a nuclear antigen expressed in dividing cells during late G1, G2, S-phase and M-phase that disapears rapidly after mitosis. p53 is a tumor supressor protein that plays a key role in the control of the DNA damage acting as a supressor of tumour cell proliferation, enhancing genetic stability. Mutation of the gene that codes for p53 provides an extended intracellular half-life of the protein and greater stability, therefore increasing its expression. Tumours for this study were obtained at the slautherhouse, in Ponta Delgada, Azores. A total of thirty samples were colected from eighteen bovine. Immunohistochemical analysis was performed in parafin embeded sections of formalin fixed material, using the Avidin-Biotin-Peroxidase method (ABC). Primary antibodies used were anti-human p53 (NCL – p53 CM-1, NOVOCASTRA, UK) and anti-human Ki67 (NCL – Ki67p, NOVOCASTRA, UK), both policlonal. Positivity for both Ki67 and p53 was evaluated quantitatively and semi-quantitatively, respectively
- Immunohistochemistry of epidermal differentiation markers in bovine ocular squamous cell carcinoma and distribution of cytokeratinsPublication . Vala, Helena; Fondevila, D.; Carvalho, T.; Pinto, C.; Peleteiro, C.; Pinho, M.; Ferrer, L.The aim of this study was to evaluate the possibility of using some antibodies as diagnostic instruments, in Bovine Ocular Squamous Cell Carcinoma (BOSCC) and establishing the relationship between immunohistochemical results and the degree of differentiation. Thirty two samples from nineteen bovine ocular tumours were collected at the Matadouro Frigorífico e Industrial of Ponta Delgada, S. Miguel Island, Azores. Material was fixed in 10% formalin, embedded in paraffin and stained for routine histopathological diagnosis for tumour classification. Diagnosis of BOSCC was confirmed in all samples. Nine carcinomas were well differentiated, five moderately differentiated and five non-differentiated. Immunohistochemical indirect technique Avidin-Biotin-Peroxidase was applied. Two antibodies against distinct keratins were used: a monoclonal antibody MNF116, anti-cytokeratins 5, 6, 8, 17 and 19 (DAKO), characteristic of non differentiated keratinocytes and simple epithelia, and a monoclonal antibody LP34, anti-cytokeratins 5, 6 and 18 (DAKO), characteristic of well-differentiated keratinocytes. Epidermal differentiation markers were also used: a monoclonal antibody anti-human involucrin, for keratinocytes of the stratum spinosum and stratum granulosum and a polyclonal antibody anti-human profilaggrin (ZYMED), for keratinocytes of the stratum granulosum. Most antibodies, without exception, reacted with the various types of BOSCC; The antibody MNF 116 was positive in all carcinomas, independently of their degree of differentiation. The antibody LP34 was positive in all carcinomas, except for one in each differentiation grade group. The anti involucrin antibody was positive in all but one non-differentiated carcinomas. The anti profilaggrin antibody was positive in only two moderately differentiated carcinomas, in all but one well-differentiated carcinomas, and in all but one non-differentiated carcinomas.
- Immunohistochemistry studies on bovine squamous cell carcinoma morphological characterization of epidermal cell proliferation and differentiation markers and characterization of cytokeratinsPublication . Vala, Helena; Fondevila, D.; Carvalho, T.; Pinto, C.; Peleteiro, C.; Pinho, M.; Ferrer, L.Bovine Ocular Squamous Cell Carcinoma (OSCC) is a general designation for a group of primary neoplasias of keratinocytes arising from ocular tissues, especially the lids and particularly the third eye lid. OSCC has been diagnosed all over the world with high prevalence, being the most common bovine tumour and the one causing the most significant economic losses (Hamir & Parry, 1980; Dennis et al., 1985, Heeney & Valli, 1985; Wilcock, 1993). In Portugal, the frequency of these tumours is particularly high in the Azores, where in S. Miguel Island a large number of cattle affected with OSCC is rejected for consumption at slaughter. OSCC is the second most frequent neoplasia after urinary bladder tumours, representing 21% of all cases of rejection due to neoplasia (Pinto et al, 1996). Several reasons have been advanced to explain this situation namely the fact that animals stay in pasture all year around, with a prolonged exposition to day light and benefiting from few shelters. The ingestion of toxic plants present the pasture could also give rise to photosensitazation problems, either primary or due to hepatic toxicity, that could generate predisposing conditions to the development of OSCC.
- Involucrin and filaggrin expression in normal dog skin and different epidermal hyperplasiasPublication . Vala, Helena; Fondevila, D.; Ferrer, L.