Programa de Apoio aos Planos de Reestruturação e Expansão das Universidades Federais / Cancer is one of the main causes of death in dogs and cats and the mammary neoplasm is one of the most frequent in these species. Although some studies have been performed, there is little data on the epidemiology of this disease in cats. Thus, this study has, as primary goal, to determine the most prevalent tumors in cats and to associate the mammary tumors to some prognostic factors. The files from the Veterinary Pathology Laboratory of the Universidade Federal de Santa Maria were reviewed. Between the years 2000 and 2011, a total of 1427 feline cases including necropsies and biopsies were found. Based on the collected data from these cases, a relation between mammary tumors and some factors such as sex, age, breed, reproductive status, use of contraceptives, number and location of affected glands, presence of ulcerations, size of the tumor, and presence and location of metastases, was studied. Mammary tumors were the second most prevalent tumors (39,57%), after skin neoplasms (41,49%). All feline tumors were in females, mostly older and Domestic Shorthaired cats. Malignant neoplasms were the more frequently diagnosed (72,35%), followed by non-neoplastic tumors (22,58%), and benign neoplasms (5,06%). The smaller tumors were mostly carcinomas. Ulcers were observed in malignant tumors (92%) and non-neoplastic tumors as well (8%). Distant metastases were found mainly in the lungs and skin. / Câncer é uma das principais causas de morte em cães e gatos, sendo o neoplasma mamário um dos mais frequentes. Embora estudos tenham sido realizados nesta área, há, ainda, poucos dados epidemiológicos na espécie felina. Logo, esta dissertação teve como objetivos determinar os tumores mais prevalentes em gatos e relacionar os tumores mamários a alguns de seus fatores prognósticos. Os arquivos do Laboratório de Patologia Veterinária (LPV) da Universidade Federal de Santa Maria (UFSM) foram revisados e um total de 1.427 protocolos de biopsias e necropsias de felinos, entre 2000 e 2011, foi encontrado. Com base nas informações dos arquivos, foi estabelecida a relação entre os tumores mamários e alguns fatores como sexo, idade, raça, estado reprodutivo, uso de contraceptivos, número e localização das glândulas afetadas, ulcerações, tamanho do neoplasma, metástases distantes e para os linfonodos. Assim, observou-se que os tumores de mama foram o segundo diagnóstico (39,57%) mais prevalente, após os tumores de pele (41,49%). Todos os gatos com tumores mamários eram fêmeas, sendo os sem raça definida e os idosos os mais afetados. Os neoplasmas malignos foram diagnosticados com maior frequência (72,35%), seguidos pelos tumores não neoplásicos (22,58%) e pelos neoplasmas benignos (5,06%). Os tumores menores eram, na sua maioria, carcinomas (82,97%). Ulcerações estavam presentes não só em neoplasmas malignos (92%), mas também em alterações não neoplásicas (8%). Metástases distantes foram encontradas principalmente nos pulmões e na pele.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-14T14:49:54Z No. of bitstreams: 1 Kath Freire de Vasconcelos.pdf: 4349613 bytes, checksum: bc483327dfe70d238ea0a29b6b376704 (MD5) / Made available in DSpace on 2016-10-14T14:49:54Z (GMT). No. of bitstreams: 1 Kath Freire de Vasconcelos.pdf: 4349613 bytes, checksum: bc483327dfe70d238ea0a29b6b376704 (MD5) Previous issue date: 2012-02-15 / Mastitis is one of the major infection diseases in dairy herds, especially in its subclinical form, because it leads to great economic losses and public health. Your early and accurate diagnosis is essential to reduce damage and prevent clinical manifestation of disease. The objective of this research was then seeking a new assistive technology for the early diagnosis of subclinical mastitis in goats. With this purpose were collected milk samples from 38 goats, of herds of the city of Venturosa–PE, to realization of Somatic Cell Count and lactic culture plus photographs taken of the iris of the animals. The analysis was conducted in two regions of the iris: at the right iris in the level of 8:15 pm and in the left iris of 3:45 pm correspond to “Region A”; the right iris at 6:15 pm and 5:45 pm in the left iris correspond to “Region B”. The region A corresponds to the iridologic location of human mammary gland (chest) and the region B to the inguinal region. The results obtained allow us to conclude that the iridologic location of goat mammary gland does not correspond to the same iridologic location of human mammary gland. In animals with clinical and histopathologic confirmation, corresponding iridologic signs were observed, suggesting that goat mammary gland is located in the region corresponding to groin. This study affirms the potencial of iridology, but noting the need for more detailed scientific studies on the subject. / A mastite é uma das principais doenças infecto-contagiosas dos rebanhos leiteiros, especialmente na sua forma subclínica, pois leva a grandes prejuízos econômicos e à saúde pública. O seu diagnóstico preciso e precoce é fundamental para diminuir os prejuízos e evitar a manifestação clínica da doença. O objetivo desta pesquisa foi então buscar uma nova tecnologia auxiliar para o diagnóstico precoce da mastite subclínica em caprinos. Com este intuito foram colhidas amostras de leite de 38 cabras, de rebanhos do município de Venturosa–PE, para a realização da Contagem de Células Somáticas e da Lactocultura além de realizadas fotografias das íris dos animais. A análise foi realizada em duas regiões das íris: na íris direita ao nível de 8,15h e na íris esquerda 3,45 correspondem à Região A; na íris direita ao nível de 6,15h e na íris esquerda a 5,45 correspondem à Região B. A região A corresponde à localização iridológica da glândula mamária humana (tórax) e a região B à região inguinal. Os resultados obtidos nos permitem concluir que a localização iridológica da glândula mamária caprina não corresponde à mesma localização iridológica da glândula mamária humana. Nos animais dignosticados clinicamente e confirmados através dos exames histopatológicos, foram observados sinais iridológicos correspondentes, sugerindo que a glândula mamária caprina localiza-se na região iridológica correspondente a região inguinal. O presente estudo assevera o potencial da iridologia, embora observe-se a necessidade de estudos científicos mais detalhados sobre o tema.

Com o objetivo de avaliar a relação entre o exame físico da glândula mamária de ovelhas com o leite, 292 ovelhas da raça Santa Inês foram examinadas. As amostras de leite foram submetidas ao exame bacteriológico, ao CMT e a CCS (automática e microscópica). Variações especificamente identificadas por inspeção da mama apresentaram alterações de celularidade observadas pelo CMT (p<0,002), na CCS automática (p<0,006), e na CCS microscópica diferencial para células mononucleares (p<0,03). Mamas pendulosas puderam ser associadas a maior isolamento bacteriológico (p<0,0006) e maior celularidade nas CCS automática (p<0,01) e microscópica (p<0,05). Na inspeção do teto foram encontradas diferenças no exame da CCS automática (p<0,002), no CMT (p<0,004) e na CCS microscópica diferencial para células mononucleares (p<0,005). Nos tetos com soluções de continuidade, foram observadas diferenças apenas no exame bacteriológico (p<0,03). Quanto à palpação da mama, foram observadas diferenças no exame bacteriológico (p<0,001), no CMT (p<0,01) e na CCS microscópica total (p<0,02) e diferencial para células polimorfonuleares (p<0,02) e para células mononucleares (p<0,0002). Quanto à palpação do teto, foram encontradas diferenças no CMT (p<0,01) e na CCS microscópica total (p<0,002) e diferencial para células polimorfonucleares (p<0,002) e para células mononucleares (p<0,002). Associando-se a inspeção à palpação das metades mamárias, observou-se diferenças na CCS automática (p<0,0002) e na CCS microscópica total (p<0,04) e diferencial para células mononucleares (p<0,01). No exame do fundo escuro, foram observadas diferenças no CMT (p<0,0001), na CCS automática (p<0,0001), e na CCS microscópica total (p<0,0001) e diferencial para células polimorfonucleares (p<0,0001) e mononucleares (p<0,0001). Quando associadas duas categorias do exame físico da glândula mamária, a inspeção e a palpação, e o exame do fundo escuro, foram observadas diferenças no CMT (p<0,0001), na CCS automática (p<0,0001) e na CCS microscópica total (p<0,0001) e diferencial para células mononucleares (p<0,0001). Nas ovelhas acompanhadas durante a lactação foram observadas diferenças na inspeção da mama (p<0,0001) e do teto (p<0,0001), na palpação da mama (p<0,005) e do teto (p<0,003), na inspeção e palpação das metades mamárias (p<0,04), na inspeção, palpação e fundo escuro das metades mamárias (p<0,03), na CCS automática (p<0,0001) e na CCS microscópica total (p<0,02) e diferencial para células polimorfonucleares (p<0,02) e para células mononucleares (p<0,02). Foi possível concluir que há relação entre o exame físico e o perfil celular e bacteriológico, mas que a inflamação da mama foi melhor identificada pelo CMT, através da inspeção da mama e do teto, da palpação da mama e do teto, do exame do fundo escuro e da avaliação da inspeção, palpação e fundo escuro das metades mamárias. A inflamação também foi identificada pela avaliação da CCS automática e/ou da CCS microscópica total e/ou diferencial através da inspeção da mama e do teto, pendulosidade da mama, palpação da mama e do teto, inspeção e palpação das metades mamárias, exame do fundo escuro e a inspeção, palpação e fundo escuro das metades mamárias. O processo infeccioso mamário pôde ser identificado através da pendulosidade mamária, presença de soluções de continuidade no teto e pela palpação da mama. A maior celularidade ocorreu no início da lactação. No final da lactação, houve maiores proporções de alterações na inspeção da mama e do teto e na palpação do teto. Nas fases intermediárias da lactação, as alterações na inspeção e palpação associadas e na inspeção, palpação e fundo escuro associados foram menores. / The aim of this work is to evaluate the relationship between the mammary gland physical examination of ewes and their milk. 292 ewes of Santa Inês breed were evaluated, and the milk samples were submitted to bacteriological examination, to CMT (California Mastitis Test), to automatic SCC (somatic cell count) and microscopic SCC. Specifically identified variations during the mama inspection had cellular alterations, observed in CMT (p<0,002), in automatic SCC (p<0,006) and in differential microscopic count for mononuclear cells (p<0,03). Pendulous mamas could be associated to the largest bacteriological isolation (p<0,0006) and the largest cellular in automatic SCC (p<0,01) and microscopic SCC (p<0,05). In the teat inspection, differences were found in the automatic SCC (p<0,002) in the CMT (p<0,004) and in differential microscopic count for mononuclear cells (p<0,005). In the teat with lesion, differences were observed only in bacteriological examination (p<0,03). In mamma palpation, there were observed differences in bacteriological examination (p<0,001), in CMT (p<0,01), in total microscopic count (p<0,02), differences to polymorphonuclear cells (p<0,02) and to mononuclear cells (p<0,0002). In teat palpation, differences were found in CMT (p<0,01), in total microscopic count (p<0,002) and differences to polymorphonuclear cells (p<0,002) and to mononuclear cells (p<0,002). Associating the inspection and palpation of mammary gland, differences were observed in automatic SCC (p<0,0002) and in total microscopic count (p<0,04) and differences to mononuclear cells (p<0,01). In tamis exam, differences were observed in the CMT (p<0,0001), in automatic SCC (p<0,0001), and in total microscopic count (p<0,0001) and differences to polymorphonuclear cells (p<0,0001) and mononuclear cells (p<0,0001). When the two categories of mammary gland physical exam, the inspection, the palpation, and the tamis exam were associated, there were observed differences in the CMT (p<0,0001), in the automatic SCC (p<0,0001) and in the total microscopic count (p<0,0001), and differences in the microscopic SCC to mononuclear cells (p<0,0001). In the ewes followed during the lactation there were differences in mamma inspection (p<0,0001), in teat (p<0,0001), in mamma palpation (p<0,005) and in teat (p<0,003), in the inspection and palpation of mammary gland (p<0,04), in inspection, palpation and tamis exam of mammary gland (p<0,03), in automatic SCC (p<0,0001) and in total microscopic count (p<0,02), and differences for polymorphonuclear cells (p<0,02) and to mononuclear cells (p<0,02). It can be concluded that there is a relationship among the physical exam and the cellular and bacteriological profile, but the mamma inflammation was better identified by the CMT, through inspection of mamma and teat, and mamma and teat palpation, and tamis exam and the evaluation by inspection, palpation and macroscopic assessment of the milk by mammary glands. The inflammation was also identified by the evaluation of automatic SCC and/or total microscopic count through inspection of mamma and teat, pendulous mamma, palpation of mamma and teat, inspection and palpation of mammary gland, tamis exam and the inspection, palpation and tamis exam of mammary gland and milk. The infection in mammary gland can be identified by pendulous mamma, lesion in teat and by palpation of mamma. The biggest cellular was found in early lactation. And in late lactation, there were found more alterations in mamma and teat inspection and palpation of teat. In the intermediate phases of lactation, the alterations in inspection and palpation associated and the inspection, palpation and the tamis exam associated were smaller.

La thèse avait pour objectif de caractériser l’adaptabilité de la glande mammaire, organe support de la fonction de lactation, afin de contribuer à l’étude de la robustesse des vaches laitières et à l’identification de vaches adaptables. L’approche utilisée pour caractériser l’adaptabilité de la glande mammaire consistait à décrire les réponses inter-individuelles de production laitière lors d’une perturbation, puis à identifier des critères phénotypiques et/ou génétiques modulant les caractéristiques adaptatives. Deux modalités d’allongement de l’intervalle entre traites ont été choisies pour perturber le fonctionnement de la glande mammaire : un intervalle de traite unique de 24h (24h-MI) et 3 semaines de monotraite (ODM). Les 2 composantes de l’adaptabilité, à savoir la résistance et la résilience, ont été respectivement estimées par les pertes de lait lors de l’allongement de l’intervalle entre traites et par le taux de récupération au retour à 2 traites par jour,Quatre profils de réponse, définis par leur association des modalités de résistance et de résilience, ont été mis en évidence, dont 3 étaient communs aux 2 perturbations (24h-MI et ODM). Ces profils étaient caractérisés par la compliance de la glande mammaire (c’est-à-dire son aptitude à se distendre) et par son état inflammatoire. Les réponses de production laitière seraient des caractères héritables (h2= 0.32 pour les pertes de lait en kg/j et 0.63 pour la récupération de lait (kg/d), respectivement) ouvrant des perspectives de sélection des vaches sur ces caractères adaptatifs. Les zones du génome influant / This study aimed to enhance knowledge on dairy cows robustness and to enable the identification of adaptable cows through the characterization of the adaptability of the mammary gland, the organ responsible for lactation. The methodology used to characterize the adaptability of the mammary gland consisted in describing inter individual variability in milk yield responses profiles to a perturbation and then identifying phenotypes and/or genotypes that modulated the adaptive characteristics. The mammary gland functioning was challenged by using two different lengthening of milking interval as a perturbation on dairy cows usually milked twice-daily (TDM): a single extended milking interval of 24h (24h-MI) and a 3-wk once-daily milking (ODM). For each of them, the two components of adaptability, resistance and resilience, were estimated by milk yield loss in % when switching cows to 24h-MI or ODM and milk recovery:loss ratio when switching cows back to TDM.Four individual responses profiles, defined by the association of specific resistance and resilience modalities, were found with 3 being common between 24h-MI and ODM. These profiles were characterized by udder compliance (= ability to distend) and mammary inflammation. Milk yield responses were also found to be heritable (h2= 0.32 for milk yield losses and 0.63 for milk yield recovery (kg/d), respectively), so that it could be possible to select animals on these adaptability traits. The genomic regions related to these responses were partly different from regions controlling milk production traits during TDM, so that adapt

Le facteur nutritionnel affecte de façon significative la sécrétion et la composition des constituants du lait qui conditionnent sa qualité nutritionnelle. Dans la glande mammaire, ces processus font intervenir de nombreux gènes dont l’expression est modulée par l’alimentation, cependant les mécanismes de régulation sous-jacents ne sont pas connus. Les microARNs (miARN) sont des petits ARN non codants qui se lient sur leurs ARNm cibles pour en réguler l’expression. Ils ouvrent donc des pistes d’investigation pour la compréhension de ces mécanismes. La première partie de mon travail de thèse a consisté à obtenir une meilleure connaissance des miARN exprimés dans la glande mammaire, notamment en dressant les miRNomes de référence par séquençage haut débit chez la souris, la vache et la chèvre. Ensuite, pour la première fois, l’impact de la nutrition sur l’expression des miARN mammaires a été étudié. Deux modèles ruminants, un modèle dit « extrême » et un modèle de supplémentation lipidique proche des conditions d’élevage, ont permis d’identifier 30 et 2 miARN, respectivement, dont l’expression est nutrirégulée. L’analyse in silico des cibles des miARN nutrirégulés a révélé un rôle potentiel de ceux-ci dans le métabolisme des lipides. Certaines des cibles sont effectivement différentiellement exprimées dans ces modèles, parmi celles-ci certains gènes sont essentiels pour la lactation tels que ESR1. Enfin, une étude pilote de la fonction de trois miARN nutrigulés a été initiée in vitro dans des cellules épithéliales mammaires bovines. Ces travaux permettent donc d’apporter des premiers éléments pour la compréhension de la régulation de l’expression des gènes en réponse à la nutrition et de l’impact des miARN sur la lactation. / Nutrition significantly affects the secretion and the composition of milk which determine its nutritional quality. In the mammary gland, regulation of these processes involves numerous genes which expression can be affected by nutrition. However, their regulations remain unclear. MicroRNAs (miRNA) are small non coding RNA which can bind mRNAs and regulate their expression of target genes. Consequently, they offer opportunities to understand the regulation of gene expression in response to nutrition. The first step of my PhD aimed to obtain a better knowledge of miRNA expressed in the mammary gland. Mammary miRNome were established from the lactating mouse, cow and goat using high-throughput sequencing. Later, the effect of nutrition on the expression of miRNA in the mammary gland was analyzed for the first time. Two models in ruminants, a food deprivation (“extreme” model) and a lipid supplementation (model similar to breeding conditions) highlighted 30 and 2 nutriregulated miRNA, respectively. The analysis of nutriregulated miRNA’s predicted targets, in silico, revealed their potential role in lipid metabolism. Some of those target genes have been previously identified as differently expressed in the same conditions and could thus be involved in the regulation of the expression of genes essential for the mammary gland function, such as ESR1. Finally, three nutriregulated miRNA were selected and used in a preliminary study of their functions in vitro in bovine mammary epithelial cells. These works bring first evidences in understanding the nutritional regulation of gene expression in the mammary gland as well as the role of miRNA in lactation.

La recherche sur la fonction de lactation est indispensable pour la filière laitière. La glande mammaire est un organe composé de cellules épithéliales, de cellules myoépithéliales, de fibroblastes, d’adipocytes et de cellules endothéliales. L’identification et l’évolution en dynamique des populations cellulaires dans la glande mammaire bovine au cours la lactation reste encore à ce jour méconnues. Les objectifs de cette thèse ont été 1) l’obtention d’un panel des biomarqueurs pour identifier les différentes cellules dans la glande mammaire bovine et 2) le suivi de leur évolution au cours d’un cycle de lactation. Les biomarqueurs de surface (CD49f, EpCAM, CD24 et CD10) ont permis de phénotyper deux lignées mammaires bovines. Nous avons également utilisé ces biomarqueurs pour phénotyper les cellules dans la glande mammaire bovine au cours d’un cycle de lactation.Dans cet objectif, des biopsies ont été prélevées sur 5 vaches laitières primipares à quatre étapes durant la lactation. Après la digestion des biopsies, les cellules obtenues ont été marquées et phénotypées par cytométrie en flux. L’analyse des résultats montre des corrélations positives entre les populations cellulaires CD49f+ et CD49f+/CD24- avec la production laitière. Nous avons mis en évidence une population cellulaire CD49f-/EpCAM- qui augmente au cours de la lactation. Ces résultats suggèrent qu’à partir de l’utilisation de biomarqueurs il est possible d’identifier et phénotyper l’évolution de différentes cellules pendant la lactation dans la glande mammaire bovine. / Research of lactation function of the bovine mammary gland remains essential in dairy farming. The mammary gland is an organ composed of epithelial cells, myoepithelial cells, fibroblasts, adipose cells and endothelial cells. The identification and evolution of cell populations in the bovine mammary gland during lactation is currently unknown. The objectives of this thesis were 1) to obtain a panel of biomarkers capable of identifying different cells within the mammary gland 2) to follow the evolution of these cells throughout a lactation cycle. The cell surface biomarkers (CD49f, CD24, CD10 and EpCAM) allowed us to phenotype two bovine mammary cell lines. We also used these cell biomarkers to phenotype cell populations during lactation.For this objective mammary gland explants were obtained by biopsies taken on 5 primiparous lactating cows at four different times during lactation. After the biopsies were digested, the cells obtained were phenotyped by flow cytometry. Analysis of the results revealed a positive correlation between the CD49f+ and CD49f+/CD24- cell populations and milk yield. We also found evidence that one cell population (CD49f-EpCAM-) was increased during lactation. In general these results suggest that biomarker expression can be utilized to identify the phenotype and the evolution of different cell types during lactation in the bovine mammary gland.

Le gène TP63 fait partie de la famille du gène suppresseur de tumeur TP53. Il code plusieurs isoformes. L’une d’entre elles, tronquée dans la région amino-terminale et appelée ΔNp63, présente des propriétés oncogéniques. Elle est impliquée dans la progression tumorale et la chimiorésistance. Sa surexpression est fréquente dans certains types de cancers. La première partie de mes travaux a consisté à identifier les facteurs de transcription impliqués dans la régulation du promoteur de ΔNp63. J’ai montré que l’expression de cette isoforme est inhibée par p53 et activée par ΔNp63 elle-même et par la β-caténine, dans des lignées de carcinome hépatocellulaire et de carcinome épidermoïde de l’œsophage. Dans des conditions physiologiques, un des types cellulaires dans lequel ΔNp63 est exprimée est la cellule basale mammaire. Il est admis que les tumeurs mammaires dites basal-like sont issues des cellules basales. Certaines de ces tumeurs présentant une surexpression de ΔNp63, nous avons émis l’hypothèse que ΔNp63 serait impliquée dans la tumorigenèse des cellules basales. Dans la deuxième partie, j’ai montré que l’expression de ΔNp63 peut être augmentée en cultivant les cellules mammaires en présence de surnageant de fibroblastes embryonnaires humains. L’identification de facteurs solubles responsables est en cours. D’autre part, j’ai caractérisé les conséquences biologiques de cette augmentation en termes de prolifération, de motilité et de survie des cellules immatures mammaires normales et tumorales. Les plus grandes modifications observées concernent (i) l’état de différenciation, les cellules surexprimant ΔNp63 présentant un phénotype plus immature ; (ii) la balance entre migration et adhésion qui penche en faveur de cette dernière. ΔNp63 semble donc être au carrefour des mécanismes de prolifération, d’adhésion, de différenciation et de motilité, processus impliqués dans la formation et l’homéostasie des tissus, mais dont l’altération peut conduire à l’initiation et à la progression tumorale ainsi qu’à la dissémination métastatique. Mes travaux apportent des informations sur le rôle de cette protéine dans ces processus et devraient, à terme, permettre de mieux comprendre la genèse de certains cancers, en particulier les carcinomes basal-like / TP63 gene belongs to the TP53 tumor suppressor gene family. It encodes several isoforms. One of these, truncated in its amino-terminal end and called ΔNp63, displays oncogenic properties. It is involved in tumor progression and chemoresistance and is overexpressed in some tumor types. The first part of my work consisted of identifying the transcription factors involved in the regulation of the ΔNp63 promoter. I have shown that ΔNp63 expression is inhibited by p53 and activated by ΔNp63 itself and by β-catenin in hepatocellular carcinoma and esophageal squamous cell carcinoma cell lines. Under physiological conditions, one of the cell types in which ΔNp63 is expressed is the mammary basal cell. The “basal-like” mammary tumor sub-type seems to stem from basal cells. As some of these tumors exhibit overexpression of ΔNp63, we hypothesized that this isoform could be involved in the genesis of basal-like tumors. In the second part, I have shown that ΔNp63 expression can be increased in mammary cells cultivated in the presence of human embryonic fibroblast supernatant. Identifying the soluble factors responsible for this increase is in progress. In parallel, I have evaluated the biological consequences of ΔNp63 overexpression in terms of proliferation, cell motility and survival of normal and malignant immature mammary cells. The main modifications relate to (i) the differentiation status, ΔNp63-overexpressing cells exhibiting a more immature phenotype; (ii) the balance between migration and adhesion that is in favor of this latter. ΔNp63 seems to be at the crossroads of proliferation, adhesion, differentiation and motility, processes implicated in tissue formation and homeostasis, but whose alteration may lead to tumor initiation and progression and to metastatic dissemination. My work provides information on the role of this isoform in these processes and should allow better understanding of the genesis of some tumor types, in particular basal-like breast carcinomas

Objetivou-se identificar parâmetros bioquímicos, hematológicos e imunológicos, sistêmicos e locais, associados a modelo de risco de mastite bovina, da secagem ao final do período de transição da lactação subsequente, de vacas submetidas a antimicrobiano e selante na secagem. Foram utilizadas 34 vacas da raça Holandesa, divididas em três grupos: grupo controle (GC; n = 13); grupo antimicrobiano (GA; n = 10) composto animais que receberam antimicrobiano intramamário; grupo selante (GS; n = 11), composto por animais que receberam selante intramamário. Foram coletadas amostras de leite e sangue onde avaliou-se hemograma, metabolismo oxidativo de neutrófilos (NBT), imunoglobulinas, bioquímica, e CMT, contagem de células somáticas, isolamento microbológico do colostro e leite e imunoglobulinas do leite nos 60 dias que antecederam o parto (M1), no dia do parto (M2), três, sete, 15, 21, 30 dias pós-parto (M3, M4, M5, M6 e M7). Os dados foram analisados pelo software Stata utilizando um modelo de regressão logística. As variáveis foram analisadas primeiro individualmente para verificar o significado e em seguida, em combinação, para avaliar o efeito das variáveis individuais. Na primeira fase da análise, empregou-se modelo logístico para cada variável relacionada com os achados clínicos, com valores de P inferiores a 0,20 foram considerados como uma variável selecionada e passada para a próxima fase da análise. Dentro o modelo final, foram usadas variáveis selecionadas na primeira fase para desenvolver um modelo logístico multivariado em que as variáveis com P&le;0.05 foram retidas no modelo final. Foi possível identificar que alterações nos parâmetros, proteína, albumina e fibrinogênio, servem de biomarcadores associados à mastite bovina. Alterações metabólicas específicas e características do período de transição, nos parâmetros sanguíneos, sobrepõem o efeito dos protocolos de secagem empregados e, sugere-se que o perfil sistêmico da vaca o periparto está associado à mastite / Objective identify biochemical, hematological and immunological systemic and local, associated with Bovine Mastitis risk model, drying at the end of the transitional period of the subsequent lactation cows treated with antimicrobial and sealant to dry. 34 cows were used the Dutch race, divided into three groups: control group (GC; n = 13); antimicrobial Group (GA; n = 10) animals receiving antimicrobial intramamário; sealant Group (GS; n = 11), composed of animals that received intramamário sealant. Milk samples were collected and evaluated blood where blood count, neutrophil oxidative metabolism (NBT), immunoglobulins, biochemistry, and CMT, somatic cell count, microbológico isolation of colostrum and milk and milk immunoglobulins in the 60 days preceding childbirth (M1) on the day of delivery (M2), three, seven, 15, 21, 30 days postpartum (M3, M4, M5, M6 and M7). Data were analyzed by the software Stata using a logistic regression model. The variables were analyzed individually to check the meaning and then, in combination, to evaluate the effect of individual variables. In the first phase of the analysis, logistic model was used for each variable related to the clinical findings, with P values less than 0.20 were considered as a variable selected and passed to the next stage of analysis. In the final model, selected variables were used in the first phase to develop a multivariate logistic model in which the variables with P &le; 0.05 were retained in the final model. It was possible to identify changes in the parameters, protein, albumin and Fibrinogen, serve as biomarkers associated with Bovine Mastitis. Specific metabolic changes and characteristics of the transitional period, blood parameters, overlapping the effect of drying employees and protocols, it is suggested that the systemic profile the periparto cow is associated with mastitis

Made available in DSpace on 2016-07-18T17:53:15Z (GMT). No. of bitstreams: 1 Guilherme Pepino Bastos.pdf: 4641063 bytes, checksum: b561cc3e6ce8ff140fb329e91d1cc713 (MD5) Previous issue date: 2015-09-25 / Background: The digital infrared imaging thermography is a non-invasive imaging exam of easy performance accurately to measure temperatures of the areas of the animals body. The objective was to study the physiological variations of the surface temperature of the mammary gland, vulva, pelvis, thorax and abdomen with digital infrared imaging thermography in negative dairy cattle with California mastitis test (CMT) in different seasons, and the influence of climatic factors in these temperatures. Materials, Methods & Results: 18 Holstein cows, pregnant and non pregnant were used, negative for the California Mastitis Test, grazing on Urochloa decumbens pasture, receiving 2 kg of corn / animal / day, mineral mix and ad libitum water. Infrared thermography was performed every 30 days, for five months: January, February, March, April and May with thermographic camera (E40®, FLIR, Sweden) in the areas of the body: vulva, mammary gland, pelvis, abdomen and thorax. The thermal images (thermograms) were processed using the Flir Tools 2.1® program. The climatic factors: room temperature and relative humidity were monitored with globe thermometer (ITitwtg 2000®, Instrutemp, Brazil). Data were analyzed by analysis of variance and the average compared by 5% Tukey test. For room temperature data, relative humidity, rectal temperature and for the areas of the body we used Pearson correlation to 5%. For climatic factors, among the samples, room temperature and relative humidity, there was a significant difference (P <0.05). To the skin surface temperatures of the body areas examined: vulva, pelvis, ischium, abdomen, thorax and mammary gland, among the samples, there were differences (P <0.05) with lower temperatures of the areas in March and May samples, compared to the months of January, February and April. The average temperatures of the surfaces of the examined areas vary for the vulva between 33 °C and 38 °C; ischium, pelvis and abdomen between 30 °C and 37 °C; teats between 28 °C and 37 °C; Cistern of mammary quarter between 32 °C and 38 °C; and thorax from 31 °C to 37 °C. There were significant correlations (P<0.01) between: rectal temperature x room temperature (0.49); Rectal temperature x relative humidity (- 0.37). To the surface temperatures of all areas of the body examined, there were significant correlations (P<0.01) with the room temperature between 0.73 and 0.85; and between - 0.57 and - 0.75 for the relative humidity. Discussion: The animals showed no behavioral change during the thermography examination. In dairy cattle it is recommended the use of infrared thermography as a routine test to measure the temperatures of the areas of the body. Climatic factors, room temperature and relative humidity influence the rectal temperatures and the body surfaces of the cows. Conclusions: The areas of the body examined by infrared thermography showed different temperatures, in the same data collection, showing physiological temperature variations that assist in the clinical evaluation of each of the areas examined. The thermographic images were saved and processed easily, quickly and in a practical way, recommending the use of thermal imaging by infrared for routine as imaging test complementary to the clinical examination of the mammary gland and of the body areas in dairy cattle. / Introdução: A termografia digital por infravermelho é um exame de imagem não invasive de fácil realização com precisão para mensurar temperaturas das áreas do corpo dos animais. Objetivou-se estudar as variações fisiológicas relevantes às variações de temperatura da superfície da glândula mamária, vulva, pelve, tórax e abdômen com termografia digital por infravermelho em vacas de leite negativas ao California mastitis test (CMT), em diferentes épocas do ano, e a influência de fatores climáticos nessas temperaturas. Materiais, Metodos & Resultados: Foram utilizadas 18 vacas da raça Holandesa, gestantes e não gestantes, negativas para a California Mastitis Test, mantidas em pastagem Urochloa decumbens, recebendo 2 kg de milho/animal/dia, sal mineral e água ad libitum. Termografia por infravermelho foi realizada a cada 30 dias, durante cinco meses; de janeiro a maio, com câmera termográfica (E40®, FLIR, Suécia) nas áreas do corpo: vulva, glândula mamária, pelve, abdome e tórax. As imagens térmicas (termogramas) foram processadas usando o programa Flir Tools 2.1®. Os fatores climáticos: temperatura ambiente e umidade relativa do ar foram monitoradas com termômetro de globo (ITitwtg 2000®, Instrutemp, Brasil). Os dados foram analisados pela análise de variância e as médias comparadas pelo teste de Tukey a 5%. Para os dados da temperatura do ambiente, umidade do ar, temperatura retal e das áreas do corpo, utilizou-se correlação de Pearson a 5%. Para os fatores climáticos, entre colheitas, temperatura ambiente e umidade relativa do ar, houve diferença significativa (P<0,05). Para as temperaturas da superfície da pele das áreas do corpo examinadas: vulva, pelve, ísquio, abdomen, tórax e glândula mamária, entre colheitas, houve diferença (P<0,05) com menores temperaturas das áreas nas colheitas de março e maio, em relação aos meses de janeiro, fevereiro e abril. As temperaturas médias das superficies das áreas examinadas variaram para a vulva entre 33°C e 38°C; ísquio, pelve e abdomen entre 30°C e 37°C; tetos entre 28°C e 37°C; cisterna do quarto mamário entre 32°C e 38°C; e tórax entre 31°C a 37°C. Houve correlações significativas (P<0,01) entre: temperatura retal x temperatura ambiente (0,49); temperatura retal x umidade relativa do ar (- 0,37). Para as temperaturas da superfície de todas as áreas do corpo examinadas, houve correlações significativas (P<0,01) com a temperatura ambiente entre 0,73 e 0,85; e entre 0,57 e 0,75 para a umidade relativa do ar. Discussão: Os animais não apresentaram mudança de comportamento durante o exame de termografia. Em vacas leiteiras recomenda-se o uso da termografia por infravermelho como exame de rotina para mensurar as temperaturas das áreas do corpo. Os fatores climáticos temperatura ambiente e umidade relativa do ar influenciam nas temperaturas retal e das superficies do corpo das vacas. Conclusões: As áreas do corpo examinadas pela termografia por infravermelho apresentaram temperaturas distintas, em uma mesma colheita de dados, mostrando variações fisiológicas de temperatura que auxiliam na avaliação clínica de cada uma das áreas examinadas. A temperatura ambiente e a umidade relativa do ar influenciam na temperatura retal e de áreas da superfície do corpo. As imagens termográficas foram salvas e processadas com facilidade, rapidez e de forma prática, recomendando-se o uso da termografia por infravermelho de rotina como exame de imagem complementar ao exame clinico da glândula mamária e de áreas do corpo em vacas de leite.

L’épithélium mammaire est composé de cellules luminales et basales myoépithéliales. Il comprend des cellules souches et progénitrices qui gouvernent le développement de la glande. Leur dérégulation est probablement à l’origine de certains cancers du sein très agressifs, dits « triple-­‐négatifs » (négatifs pour les récepteurs hormonaux et HER2), pour lesquels il n’existe pas de thérapie ciblée. L’équipe a identifié un nouveau marqueur spécifique du compartiment basal comprenant les cellules souches, la Podoplanine (Pdpn), dont l’expression est concentrée aux contactsintercellulaires basal-­‐luminal. La Pdpn est une glycoprotéine transmembranaire connectée au cytosquelette, impliquée dans le développement denombreux tissus et la tumorigenèse. Son rôle dans la glande mammaire n’a jamais été étudié. A l’aide d’une délétion génique conditionnelle, nous avons trouvé que l’absence de Pdpn dans l’épithélium mammaire perturbe la fonction des cellules souches/progénitrices basales ainsi quel’expression de plusieurs composants du signalosome de la voie Wnt/b-­‐caténine. De plus, la perte de Pdpn dans un modèle murin de tumorigénèsemammaire induite par l’activation de la signalisation Wnt/b-­‐caténine réduit la fréquence des tumeurs mammaires triple-­‐négatives, limite l’expansiondes cellules initiatrices de tumeurs et favorise l’expression de marqueurs moléculaires associés à un programme de transition mésenchymo-­‐épithéliale. Au moyen d’expériences de gain de fonction dans une lignée de cellules basales mammaires, Nous avons montré que les phénotypes décrits précédemment reposent sur des mécanismes moléculaires impliquant la Pdpn dans le contrôle positif de l’activation de la voie Wnt/b-­‐caténine. Notre étude révèle un rôle pour la Pdpn dans le contrôle des cellules souches et de leur réponse à la signalisation Wnt/b-­‐caténine au cours du développement et de la tumorigénèse mammaire. / Stem cells (SC) drive mammary development, giving rise postnatally to an epithelial bilayer composed of luminal and basal myoepithelial cells. The molecular identity of SCs and the factors regulating their function remain poorly defined. We identified the transmembrane protein, Podoplanin (Pdpn), as a specific marker of the basal compartment, including multipotent SCs, and found Pdpn localized at the basal-luminal interface. Embryonic deletion of Pdpn targeted to basal cells diminished basal and luminal SC activity and affected expression of several Wnt/b-catenin (Wnt/b-cat) signaling components. Moreover, Pdpn deletion attenuated mammary tumor formation in a mouse model of b-cat-induced breast cancer, limiting tumor-initiating cell expansion and promoting molecular features associated with mesenchymal-to-epithelial cell transition. In line with the loss-of-function data, we demonstrated that mechanistically, Pdpn enhanced Wnt/b-cat signaling in mammary basal cells. Overall, our study reveals a role for Pdpn in mammary development and tumorigenesis through the control of Wnt/b-cat-responsive SCs.