Orientador: Julio Lopes Sequeira / Banca: Sara Maria de Carvalho e Suzano / Banca: Carlos Alberto Hussni / Resumo: O presente trabalho teve por objetivo avaliar o índice proliferativo das amostras de sarcoide equino pelo método histoquímico de AgNOR e por método imuno-histoquímico pela expressão do Ki-67 (MIB 1); a avaliar o índice apoptótico pela expressão de caspase- 3; e a expressão da p53 mutante e "wild type" no sarcoide equino por meio de método imuno-histoquímico. Além disso, procurou correlacionar esses dados com os tipos clínicos de sarcoide equino, sendo cada amostra classificada clinicamente, segundo Knottenbelt (2005), a partir das descrições recuperadas nos prontuários de registro dos animais e das fichas de diagnóstico dos casos utilizados neste projeto. As amostras utilizadas foram recuperadas do material de arquivo, sendo utilizadas também novas amostras encaminhadas durante o período de realização deste projeto. Desta forma, 41 amostras de sarcoide equino provenientes de 35 equinos. O presente estudo demonstrou que entre os animais portadores de sarcoide equino, excluindo-se os SRD, predominaram àqueles pertencentes às raças Quarto de Milha, Puro Sangue Inglês e Mangalarga, preponderando os machos em relação às fêmeas, sendo que 83% dos animais possuíam menos de dez anos de idade. A localização preferencial das neoplasias na nossa casuística foi na região da cabeça, seguida dos membros e da região peitoral, prevalecendo as lesões únicas e maiores do que quatro centímetros. Dos tipos clínicos de sarcoide equino, o mais frequente entre os animais incluídos neste estudo foi o fibroblástico, seguido dos tipos verrucoso, nodular e misto, sendo o tipo oculto pouco frequente. Os resultados não mostraram diferença significativa quanto a proliferação celular (AgNOR e Ki-67), e quanto ao tipo de marcação da p53 entre os tipos clínicos de sarcoide equino. A expressão da p53... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study has been elaborated to evaluate the proliferative index of equine sarcoid samples by AgNOR methodology and Ki67 immunohistochemical expression; to also evaluate the apoptotic index by caspase-3 immunohistochemical expression, as well as the expression of p53 mutant and "wild type" protein. Moreover, we correlate these data with clinical types of equine sarcoid, according to Knottenbelt (2005), from descriptions recovered of animals data reports and diagnosis files of case studied in this project. Tissue samples were obtained from 35 horses who had their masses submitted to the Pathology Service at São Paulo State University, Botucatu, SP, Brazil, and new clinical cases during the study. Breeds most affected were Quarter Horse, Thoroughbred and Mangalarga. Males were more affected and 83% of the animals were less than ten years old. The preferential location of the tumors in our series was in the head, followed by members and chest area, prevailing single lesions larger than four centimeters. As for the clinical types of equine sarcoid, the most frequent included fibroblast, followed by verrucous, nodular and mixed. The results showed no significant differences in cell proliferation and the type of p53 expression between the clinical types of equine sarcoid. The expression of p53 has demonstrated to be predominantly perinuclear, regardless the clinical type of sarcoid. This location may indicate the functional inactivation of this protein. The expression of caspase- 3 showed that the apoptotic index was higher in fibroblastic and nodular clinical types, and lower in verrucous clinical type. This data can be... (Complete abstract click electronic access below) / Mestre

Made available in DSpace on 2014-06-11T19:27:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-07-19Bitstream added on 2014-06-13T20:57:19Z : No. of bitstreams: 1 cremasco_acm_me_botfmvz.pdf: 1408225 bytes, checksum: 22283fa61c7ebaf789ec7232efaf4d23 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O presente trabalho teve por objetivo avaliar o índice proliferativo das amostras de sarcoide equino pelo método histoquímico de AgNOR e por método imuno-histoquímico pela expressão do Ki-67 (MIB 1); a avaliar o índice apoptótico pela expressão de caspase- 3; e a expressão da p53 mutante e “wild type” no sarcoide equino por meio de método imuno-histoquímico. Além disso, procurou correlacionar esses dados com os tipos clínicos de sarcoide equino, sendo cada amostra classificada clinicamente, segundo Knottenbelt (2005), a partir das descrições recuperadas nos prontuários de registro dos animais e das fichas de diagnóstico dos casos utilizados neste projeto. As amostras utilizadas foram recuperadas do material de arquivo, sendo utilizadas também novas amostras encaminhadas durante o período de realização deste projeto. Desta forma, 41 amostras de sarcoide equino provenientes de 35 equinos. O presente estudo demonstrou que entre os animais portadores de sarcoide equino, excluindo-se os SRD, predominaram àqueles pertencentes às raças Quarto de Milha, Puro Sangue Inglês e Mangalarga, preponderando os machos em relação às fêmeas, sendo que 83% dos animais possuíam menos de dez anos de idade. A localização preferencial das neoplasias na nossa casuística foi na região da cabeça, seguida dos membros e da região peitoral, prevalecendo as lesões únicas e maiores do que quatro centímetros. Dos tipos clínicos de sarcoide equino, o mais frequente entre os animais incluídos neste estudo foi o fibroblástico, seguido dos tipos verrucoso, nodular e misto, sendo o tipo oculto pouco frequente. Os resultados não mostraram diferença significativa quanto a proliferação celular (AgNOR e Ki-67), e quanto ao tipo de marcação da p53 entre os tipos clínicos de sarcoide equino. A expressão da p53... / This study has been elaborated to evaluate the proliferative index of equine sarcoid samples by AgNOR methodology and Ki67 immunohistochemical expression; to also evaluate the apoptotic index by caspase-3 immunohistochemical expression, as well as the expression of p53 mutant and wild type protein. Moreover, we correlate these data with clinical types of equine sarcoid, according to Knottenbelt (2005), from descriptions recovered of animals data reports and diagnosis files of case studied in this project. Tissue samples were obtained from 35 horses who had their masses submitted to the Pathology Service at São Paulo State University, Botucatu, SP, Brazil, and new clinical cases during the study. Breeds most affected were Quarter Horse, Thoroughbred and Mangalarga. Males were more affected and 83% of the animals were less than ten years old. The preferential location of the tumors in our series was in the head, followed by members and chest area, prevailing single lesions larger than four centimeters. As for the clinical types of equine sarcoid, the most frequent included fibroblast, followed by verrucous, nodular and mixed. The results showed no significant differences in cell proliferation and the type of p53 expression between the clinical types of equine sarcoid. The expression of p53 has demonstrated to be predominantly perinuclear, regardless the clinical type of sarcoid. This location may indicate the functional inactivation of this protein. The expression of caspase- 3 showed that the apoptotic index was higher in fibroblastic and nodular clinical types, and lower in verrucous clinical type. This data can be... (Complete abstract click electronic access below)

Sarcoid-like tumours have been reported in Cape mountain zebra (Equus zebra zebra) in two South African game parks recently. These tumours caused severe distress to the animals and also made them unsightly for tourists visiting the parks. The aim of this investigation was to identify and characterize the infectious agent considered to be involved in the aetiology of sarcoid in the Cape mountain zebra. Bovine papillomaviruses (BPV) -1 and -2 deoxyribonucleic acid (DNA) were detected by the polymerase chain reaction (PCR) in sarcoid tumour tissue, but not from blood specimens or unaffected skin. Differentiation between BPV-1 and -2 was made by using the restriction endonuclease BstXI on PCR products of the E5 open reading frame (ORF). A hybridization probe real-time assay was developed for the specific and sensitive detection and differentiation of BPV-1 and -2 DNA in blood, skin and sarcoid tumour samples. For the specific detection of BPV-1, an increase in fluorescence was detected at 640 nm and of BPV-2 at 705 nm. The test is extremely sensitive and able to detect 1.5 genome copies/reaction. The presence of BPV-1 and -2 DNA could be demonstrated in the blood of sarcoid-affected and -unaffected zebras even in the blood of zebras from parks where sarcoids have never been observed. The phylogenetic relationships of the papillomaviruses detected in tumours in the Cape mountain zebra in comparison with a broad selection of papillomavirus sequences available in GenBank were compiled. The papillomavirus sequences retrieved from the zebras were identified as variants of either BPV-1 or BPV-2. The age of the most recent common ancestor for BPV-1 variants is estimated to be 1.40 million years (Mya) and for BPV-2 variants, 0.55 Mya. The age of the most recent common ancestor of BPV-1 and BPV-2 is estimated to be 5.34 Mya. Certain major histocompatibility (MHC) haplotypes are associated with increased risk of sarcoid tumours in horses. The zebras in these parks may have become inbred for the MHC region with increased prevalence for a haplotype, conferring increased risk for sarcoid tumours. Therefore typing system was developed to determine whether or not a high prevalence of sarcoids among zebras is associated with a MHC haplotype. Single strand conformational polymorphism was used to assess the genetic variation in MHC class II genes. The use of DQB and DRB genes demonstrated that genetic variation and sarcoids in the zebras could not be attributed to a specific haplotype. The developed real-time PCR technique was also applied in the detection of cutaneous papillomavirus in two giraffes (Giraffa camelopardalis) which were manifesting cutaneous papillomatosis, in the Kruger National Park and in a fibropapilloma in a sable antelope (Hippotragus niger), on a game farm in the Kimberley district, South Africa. In conclusion, this was the first study to confirm the presence of BPV-1 and -2 DNA in the sarcoid tumours, healthy skin and blood of sarcoid-affected and healthy free-roaming zebras from sarcoidaffected parks. The presence of BPV-1 and -2 DNA in the blood of zebras from parks where sarcoids have not been previously observed was a significant finding. / Thesis (PhD)--University of Pretoria, 2010. / Veterinary Tropical Diseases / unrestricted