Le muscle strié squelettique est un tissu richement vascularisé. Au delà de l'apport en oxygène et en nutriments, de nouvelles fonctions des vaisseaux ont été récemment identifiées, par le biais des interactions établies entre les cellules du vaisseau (cellules endothéliales) et les cellules du muscle, en particulier les cellules souches musculaires (cellules satellites). Celles-ci interagissent étroitement avec les cellules endothéliales pour leur expansion et leur différenciation, puis avec les cellules péri-endothéliales pour leur auto-renouvellement et leur retour à la quiescence. Les vaisseaux participent ainsi au contrôle de l’homéostasie du muscle squelettique. Grâce à ces interactions, les cellules vasculaires jouent donc un rôle central dans le remodelage tissulaire après un phénomène destructif, survenant par exemple au cours d’un trauma ou d’une myopathie. Pour étudier, les mécanismes de la plasticité vasculaire au cours du remodelage tissulaire, deux situations paradigmatiques de muscle en régénération chez l’enfant : la dermatomyosite juvénile (DMJ) et la dystrophie musculaire de Duchenne (DMD) ont été étudiées. Il existe, dans ces deux pathologies une souffrance musculaire associée à des cycles de nécrose/régénération. Elles se différencient par leur plasticité vasculaire et par leur évolution. En effet, la DMJ, la myopathie inflammatoire la plus fréquente de l’enfant est caractérisée par une vasculopathie avec perte en capillaires. L’évolution peut être favorable avec restitution ad integrum du muscle. La DMD est une myopathie génétique conduisant à une dégradation progressive de la force musculaire associée à une néovascularisation compensatrice. Le volet clinique/histologique incluant une analyse multiparamétrique des critères évolutifs cliniques et de réponse thérapeutique couplée à une réévaluation des données histologiques de la DMJ (analyse morphométrique des muscles DMJ) a permis de montrer qu’il existait des sous groupes phénotypiques homogènes de sévérité différente dans la DMJ. Le degré de sévérité clinique est relié à la gravité de la vasculopathie musculaire Par ailleurs, des marqueurs cliniques et histologiques simples permettant de repérer au diagnostic les patients nécessitant une escalade thérapeutique rapide (CMAS>34, atteinte gastrointestinale, fibrose endomysiale musculaire au diagnostic) ont été identifiés. Le volet cellulaire a permis l’identification in vitro des interactions cellulaires spécifiques et différentielles des myoblastes issues de patients DMD et DMJ sur les cellules endothéliales normales par l’analyse de leur rôle sur la prolifération, migration et différenciation des cellules vasculaires. Dans la DMD, les myoblastes entrainent une réponse angiogénique importante mais non efficace (néovascularisation anarchique). Dans la DMJ, les myoblastes participent efficacement à la reconstruction vasculaire notamment via la sécrétion de facteurs proangiogéniques. Ces résultats ont été renforcés par analyse transcriptomique effectuée à partir de cellules endothéliales et satellites isolées de muscles de patients confirmant le rôle central de la vasculopathie associée à un contexte inflammatoire spécifique lié à l’interféron dans la physiopathologie de la DMJ et montrant dans la DMD une dérégulation de l’homéostasie normale des interactions vaisseau-muscle avec mise en jeu d’un remodelage tissulaire non efficace. Ces données permettent d'identifier de nouvelles fonctions des cellules vasculaires dans le remodelage du muscle strié squelettique au cours des pathologies musculaires de l'enfant, et devraient ouvrir la voie à de nouvelles approches thérapeutiques. / Skeletal muscle is highly vascularised. Beyond oxygen and nutriment supply, new functions for vessels have been recently identified, through the interactions that vessel cells (endothelial cells) establish with muscle cells, particularly with muscle stem cells (satellite cells). These latter closely interact with endothelial cells for their expansion and their differentiation, then with periendothelial cells for their self-renewal and return to quiescence. During skeletal muscle regeneration endothelial cells reciprocally interact with myogenic cells by direct contact or by releasing soluble factors to promote both myogenesis and angiogenesis processes. Skeletal muscle regeneration typically occurs as a result of a trauma or disease, such as congenital or myopathies. To better understand the role of vessel plasticity in tissue remodeling, we took advantage of two muscular disorders that could be considered as paradigmatic situations of regenerating skeletal muscle in the child: Juvenile Dermatomyositis (JDM), the most frequent inflammatory myopathy and Duchenne Muscular Dystrophy (DMD), the most common type of muscular dystrophy. Although these two muscular disorders share, at the tissue level, similar mechanisms of necrosis-inflammation, they differ regarding the vessel domain. In JDM patients, microvascular changes consist in a destruction of endothelial cells assessed by focal capillary loss. This capillary bed destruction is transient. The tissue remodeling is efficient and muscle may progressively recover its function. By contrast, in DMD, despite an increase of vessels density in an attempt to improve the muscle perfusion, the muscle function progressively alters with age. We identified clinical and pathological markers of severity and predictive factors for poor clinical outcome in JDM by computing a comprehensive initial and follow-up clinical data set with deltoid muscle biopsy alterations controlled by age-based analysis of the deltoid muscle capillarization. We demonstrated that JDM can be divided into two distinctive clinical subgroups. The severe clinical presentation and outcome are linked to vasculopathy. Furthermore, a set of simple predictors (CMAS<34, gastrointestinal involvement, muscle endomysial fibrosis at disease onset) allow early recognition of patients needing rapid therapeutic escalation with more potent drugs. We studied in vitro the specific cell interactions between myogenic cells issued from JDM and DMD patients and normal endothelial cells to explore whether myogenic cells participate to the vessel remodeling observed in the two pathologies. We demonstrated that MPCs possessed angiogenic properties depending on the pathological environment. In DMD, MPCs promoted the development of establishment of an anarchic, although strong, EC stimulation, leading to the formation of weakly functional vessels. In JDM, MPCs enhanced the vessel reconstruction via the secretion of proangiogenic factors. This functional analysis was supported by the transcriptomic analysis consistent with a central vasculopathy in JDM including a strong and specific response to an inflammatory environment. On the contrary, DMD cells presented an unbalanced homeostasis with deregulation of several processes including muscle and vessel development with attempts to recover neuromuscular system by MPCs. To summarize, our data should allow the definition of new functions of vessel cells in skeletal muscle remodelling during muscle pathologies of the child that will open the way to explore new therapeutic options and to gain further insights in the pathogenesis of these diseases.

Introdução: A metástase linfonodal tem impacto significativo não somente no prognóstico do carcinoma espinocelular (CEC) de laringe, como também na sobrevivência. O índice de densidade linfonodal (IDL) combina a análise da extensão e a qualidade do esvaziamento cervical, com a habilidade na análise anatomopatológica. Objetivo: Avaliar o IDL como fator prognóstico no CEC de laringe. Métodos: Foi realizado um estudo retrospectivo com 186 pacientes submetidos a tratamento cirúrgico da laringe com esvaziamento cervical entre janeiro de 2009 a dezembro de 2016. Variáveis clínico-patológicas foram avaliadas, assim como o ponto de corte de IDL para determinar fatores prognósticos. Resultados: Foram calculados dois valores de IDL no estudo: considerando os pacientes N0 (IDL= 0,018) e considerando pN+ (IDL= 0,06). A curva de Kaplan-Meier (log-rank) em relação à sobrevivência acumulada mostrou que tanto os pacientes com IDL > 0,018 quanto considerando o IDL >= 0,06 tiveram maior taxa de mortalidade do que os pacientes com IDL <= 0,018 ou IDL < 0,06, mostrando uma forma mais agressiva da doença, com recidiva mais precoce. Entretanto, apenas o IDL > 0,06 mostrou impacto tanto no intervalo livre de doença quanto na sobrevivência global. Conclusão: O IDL mostrou ser um índice importante na avaliação prognóstica dos pacientes com CEC de laringe, tendo relação direta com recidiva da doença. Pacientes com IDL > 0,060 provavelmente têm doença mais agressiva e devem ser candidatos à terapia adjuvante / Introduction: Lymph node metastasis has a significant impact not only on the prognosis for laryngeal squamous cell carcinoma (SCC), but also on survival. The index of lymph node density (LND) combines the analysis of the extension and quality of the neck dissection with the skill in analyzing the pathology. Objective: To evaluate LND as a prognostic factor in patients with laryngeal SCC. Methods: A retrospective study was carried out with 186 patients submitted to laryngeal surgical treatment with neck dissection between January 2009 and December 2016. Clinical-pathological variables were assessed, as well as the cut-off point for LND, to determine prognostic factors. Results: Two LND values were calculated in the study: one considering N0 patients (LND=0.018) and the other considering pN+ (LND=0.060). The Kaplan-Meier curve (log-rank) related to cumulative survival demonstrated that not only patients with LND > 0.018 but also those with LND >= 0.060 had a higher mortality rate than those with LND <= 0.018 or LND < 0.060, presenting a more aggressive form of the disease, with earlier recurrence. However, only the LND >= 0.060 group had impact on both disease-free survival, and overall survival. Conclusion: The LND proved to be an important index in the prognostic evaluation of larynx SCC patients, having a direct relationship with disease recurrence. Patients with LND >= 0.060 are likely to have a more aggressive form of the disease and should be considered for adjuvant therapy

Le Lymphome à Cellules du Manteau (LCM) est une entité récemment identifiée qui se caractérise par la translocation génétique t(11 ;14)(q13 ;q32) et compte pour 2 à 10 % de tous les Lymphomes non-Hodgkiniens. Avec une survie médiane entre 3 et 5 ans après le diagnostic, le LCM est une pathologie agressive et malgré les récentes avancées thérapeutiques, peu d’informations sont disponibles concernant ses facteurs pronostiques. Certaines études ont analysé le rôle des caractéristiques clinicopathologiques et des nouvelles stratégies thérapeutiques, mais on connait peut le rôle des facteurs environnementaux et du mode de vie sur le pronostic des patients atteints de LCM. Entre 2008 et 2012, le groupe LYSA a mené en France deux essais cliniques prospectifs multicentriques : LM manteau 2010 SA "RiBVD" (NCI01457144) et Manteau 2007 SJ "LyMa" (NCT00921414). Après une comparaison de ces patients avec les patients de population générale, l’effet de facteurs socioéconomiques et des habitudes de vie sur la survie des patients a été étudié à l’aide d’un questionnaire qualitatif administré à tous les volontaires après le diagnostic. Nos résultats suggèrent qu’un faible niveau d’éducation, un indice de masse corporelle élevé et de la consommation d’alcool sont associés à un risque de décès plus élevé chez les patients atteints de LCM. Toutefois, l’étude de tels facteurs et de leur influence sur un sous-type de LNH aussi rare requiert des échantillons d’étude de taille plus importante. L’élargissement des critères d’inclusion des patients dans les essais cliniques permettrait de sélectionner davantage de patients mais aussi des patients mieux représentatifs de la population générale. Enfin, l’intégration systématique de ce type de questionnaire dans les protocoles d’essais cliniques serait aussi un atout majeur. / Mantle Cell Lymphoma (MCL) is a recently defined entity, typically characterised by the genetic translocation t(11 ;14)(q13 ;q32) and counting for 2 - 10% of all non-Hodgkin Lymphomas. With a median survival between 3 and 5 years after diagnosis, MCL is an agressive disease and despite the recent therapeutic advances little in know about its prognostic factors. Some studies had investigated clinicopathological features and new treatment strategies, but there is a lack of knowledge regarding the impact of lifestyle and environnemental factors on outcome of MCL patients. From 2008 to 2012, the LYSA Group conducted in France two prospective multi center clinical trials on MCL : LM manteau 2010 SA "RiBVD" (NCI01457144) and Manteau 2007 SJ "LyMa" (NCT00921414). After a comparison of these patients with population-based data, socioeconomic factors, lifestyle factors and their influence on survival had been investigated through a qualitative survey administrated to each volunteer after diagnosis. Our findings suggest that low educational attainment, low body body mass index and alcohol consumption are associated with a higher risk of death in MCL. However, to investigate lifestyle factors in this rare NHL subtype, larger studies should be carried out. Clinical trial inclusion criteria must be widen to select more patients and patients more representative of general population. Implementation of these epidemiological studies in clinical practice should be considered.

Introdução: Os marcadores moleculares têm sido utilizados para identificar subgrupos de tumores com comportamento clínico distinto, inclusive quanto ao padrão de recidiva. Objetivo: Caracterizar a expressão imunoistoquímica de vimentina (VIM) e E-caderina (CDH1) em carcinomas ductais invasivos (CDI) de mama e sua associação com a expressão de citoqueratina 5 (CK5), e características clínico-patológicas. Métodos: Microarranjos teciduais (TMA) foram construídos a partir de 82 amostras de CDI de mama. Imunoistoquímica (IHQ) foi realizada para determinar os receptores hormonais (RH; receptores de estrógeno e progesterona) e receptor do fator de crescimento epidérmico humano 2 (HER2), VIM, CDH1, CK5 e Ki-67. Os tumores foram classificados como luminal A (RH+, HER2-), luminal B (RH+, HER2+ ou Ki-67 alto), HER2 hiperexpresso (RH-, HER2+) e triplo negativo (TN; RH-, HER2-). Resultados: O fenótipo VIM+/CDH1-/low não foi observado nos tumores luminal A, B e HER2 hiperexpresso, enquanto que este fenótipo estava presente em 61,9% dos TN (p= 0,0001). A mediana de Ki-67 em tumores VIM+/CDH1-/low foi 13,6 (variação, 17,8-45,4), em comparação com 9,8 (variação de 4,1-38,1) nos não- VIM+/CDH1-/low (p= 0,0007). O acometimento linfonodal foi menos freqüente em pacientes com VIM+/CDH1-/low do que nos não- VIM+/CDH1-/low (23% X 61%, teste de 2, p = 0,01). A sobrevida livre de doença em 5 anos foi de 61,5% e 83,7% (teste de log-rank, p= 0,02) e a sobrevida global em 5 anos foi de 51,2% e 83,5% (teste de log-rank, p= 0,03) em pacientes com tumores com fenótipo VIM+/CDH1-/low e não VIM+/CDH1-/low, respectivamente. Conclusão: A expressão de VIM e CDH1 identificaram um subconjunto de CDI de mama com fenótipo mesenquimal com alto grau histológico e alto índice mitótico. / Introduction: Molecular markers have been used to identify subgroups of tumors with distinct clinical behavior, including recurrence pattern. Objective: To characterize the immunohistochemical expression of vimentin (VIM) and E-cadherin (CDH1) in invasive ductal carcinoma (IDC) of the breast and its association with cytokeratin 5 (CK5) expression and clinicopathological features. Methods: A tissue microarray was constructed from 82 IDC breast cancer specimens. Immunohistochemistry (IHC) was used to determine hormone receptor (HR; estrogen and progesterone receptors) status and human epidermal growth factor receptor 2 (HER2), VIM, CDH1, CK5, and Ki-67 expression. Tumors were classified as luminal A (HR+, HER2-), luminal B (HR+, HER2+ or high Ki-67), HER2 enriched (HR-, HER2+), and triple negative (TNBC; HR-, HER2-). Results: The VIM+/CDH1-/low phenotype was not observed in luminal A, luminal B and HER2 enriched tumors, whereas this phenotype was present in 61.9% of triple negative (TNBC) tumors (p = 0.0001). The median Ki-67 index in VIM+/CDH1-/low tumors was 13.6 (range, 17.8-45.4) compared with 9.8 (range, 4.1-38.1) in non-VIM+/CDH1-/low tumors (p= 0.0007). The presence of lymph node metastasis was less frequent in patients with VIM+/CDH1-/low tumors than in those with non-VIM+/CDH1-/low tumors (23% vs. 61%; 2 test, p= 0.01). The 5-years disease-free survival was 61.5% and 83.7% (log-rank test; p= 0.02) and the 5-year overall survival was 51.2% and 83.5% (log-rank test; p= 0.03) in patients with VIM+/CDH1-/low phenotype and non-VIM+/CDH1-/low phenotype tumors, respectively. Conclusion: The expression of VIM and CDH1 identified a subset of IDC breast cancer of the mesenchymal phenotype with high histological grade and high mitotic index.

Carcinoma de Células Renais (CCR) representa uma família de tumores distintos com evolução clínica imprevisível. Uma variedade de moléculas tem sido avaliada como marcadores prognósticos para CCR. Ciclina D1, uma proteína reguladora do ciclo celular, encontra-se superexpressa em vários tumores primários. Nosso objetivo é avaliar sua expressão como marcador prognóstico em CCR. Antes disso, traçamos um perfil clínico e histopatológico da amostra e verificamos sua relação com os fatores prognósticos considerados clássicos pela literatura. 109 espécimes de pacientes diagnosticados com CCR foram obtidos entre 2005 e 2010 no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto USP e submetidos à análise imunoistoquímica juntamente com 07 amostras de tecido renal normal. A maior parte das características epidemiológicas e clínicas de nossa amostra foi similar àquelas descritas na literatura mundial. Houve predomínio do gênero masculino, da raça branca, com idade próxima a 60 anos, frequência de pacientes assintomáticos em torno de 36% e grande prevalência do CCR de células claras (71,55%). A mortalidade específica da doença foi de 13,76%, sendo o CCR de células claras o tipo mais frequente entre os óbitos e casos metastáticos. Os casos que exibiram má evolução clínica, definida pela ocorrência de metástase e/ou óbito por CCR (22,01%), estiveram associados à presença de sintomas ao diagnóstico, maior tamanho tumoral, grupo de estágio alto (III ou IV), grau nuclear de Fuhrman alto (3 ou 4), presença de necrose e de diferenciação sarcomatóide no tumor, além de outros fatores histológicos desfavoráveis (p < 0,01). Isso indica que as variáveis utilizadas na avaliação de prognóstico em países desenvolvidos podem ser aplicadas aos nossos pacientes. Não houve expressão imunoistoquímica de Ciclina D1 nos casos de tecido renal normal. Observou-se heterogeneidade de marcação nuclear intratumoral no total de casos e menor expressão proteica entre os CCR papilífero e cromófobo. Pacientes com tumores com Ciclina D1baixa (até 30% de células positivas) apresentaram má evolução clínica (p = 0,03), maior tamanho tumoral (p = 0,01), presença de sintomas ao diagnóstico (p = 0,04), grau nuclear alto (p = 0,001), presença de necrose (p = 0,004) e de diferenciação sarcomatóide (p = 0,04) no tumor, além de menor sobrevida sem metástase e/ou óbito por CCR (p = 0,03). Após análise multivariada, a expressão de Ciclina D1 não apresentou valor prognóstico independente para má evolução clínica, embora tenha aumentado levemente a acurácia prognóstica do modelo adotado. Em todas as análises realizadas para o CCR de células claras isoladamente, observamos significância estatística semelhante à do total de casos (CCR). Nosso estudo demonstrou que: a proteína Ciclina D1 encontra-se superexpressa em CCR; os tipos de CCR parecem exibir diferentes padrões de marcação imunoistoquímica da Ciclina D1; alta marcação da proteína (acima de 30% de células positivas) esteve associada à boa evolução clínica e à maioria dos fatores prognósticos favoráveis bem estabelecidos na literatura. Novas investigações são necessárias para descobrir que mecanismos levam a seu acúmulo nas células neoplásicas e quais outros eventos podem estar contribuindo para a progressão da doença. / Renal Cell Carcinoma (RCC) is a family of distinct tumors with unpredictable clinical outcome. A variety of molecules have been evaluated as prognostic markers for RCC. Cyclin D1, a cell cycle regulatory protein, is overexpressed in several primary tumors. Our purpose is to evaluate its expression as a prognostic marker in RCC. Before that, we drew a clinical and histopathological profile of the sample and verified its relationship with prognostic factors regarded as classics in literature. 109 specimens from patients diagnosed with RCC were obtained between 2005 and 2010 at Hospital das Clínicas - Ribeirão Preto School of Medicine USP and submitted to immunohistochemical analysis, along with 07 normal kidney tissue samples. Most epidemiological and clinical characteristics of our sample were similar to those described in the literature. There was a predominance of male, Caucasian, aged about 60 years, the frequency of asymptomatic patients around 36%, and high prevalence of clear cell RCC (71.55%). The disease-specific mortality was 13.76%, being the clear cell RCC the most frequent type among deaths and metastatic cases. Cases that exhibited poor clinical outcome, defined by the occurrence of metastasis and/or death by RCC (22.01%), were related to the presence of symptoms at diagnosis, larger tumor size, high stage group (III or IV), high Fuhrman nuclear grade (3 or 4), presence of necrosis and sarcomatoid differentiation in the tumor and other unfavorable histological factors (p < 0.01). This indicates that the variables used in the assessment of prognosis in developed countries can be applied to our patients. There was no immunohistochemical expression of Cyclin D1 in cases of normal kidney tissue. There was intratumoral heterogeneity in nuclear staining in all cases and lower protein expression among papillary and chromophobe RCC. Patients with Cyclin D1low tumors (up to 30% positive cells) showed poor clinical outcome (p = 0.03), larger tumor size (p = 0.01), presence of symptoms at diagnosis (p = 0.04), high nuclear grade (p = 0.001), presence of necrosis (p = 0.004) and sarcomatoid differentiation (p = 0.04) in the tumor and lower survival without metastasis and/or death by RCC (p = 0.03). After multivariate analysis, the expression of Cyclin D1 showed no independent prognostic value for poor clinical outcome, although it has slightly increased the prognostic accuracy of the model adopted. In all analyzes performed for clear cell RCC alone, we observed statistical significance similar to that of the total cases (RCC). Our study showed that: Cyclin D1 protein is overexpressed in RCC; RCC types seem to exhibit different patterns of immunohistochemical staining for Cyclin D1; high protein expression (over 30% positive cells) was related to good clinical outcome and to most favorable prognostic factors well established in the literature. Further investigations are necessary to reveal which mechanisms lead to its accumulation in neoplastic cells and what other events might be contributing to the progression of the disease.

Em grande parte das vezes, a excisão cirúrgica apropriada de mastocitomas em cães requer a realização de técnicas de reconstrução para o fechamento da ferida resultante, e o seu conhecimento se torna essencial para todo o cirurgião. Alcançar margens livres tem uma influência significativa sobre o tempo de sobrevida, e deve ser o objetivo para a maioria dos pacientes. A avaliação da evolução de retalhos cutâneos e a mensuração da aderência e neovascularização do tecido no local intencionado é extremamente importante para diagnóstico precoce de falhas na implantação do tecido. Não há evidências em literatura de trabalhos relacionados a avaliação da perfusão tecidual de retalhos cutâneos em animais de companhia, com o uso de técnicas de termografia. Neste trabalho, foram obtidas imagens, do tipo padrão e do tipo termográficas, de 63 mastocitomas, provenientes de 60 cães. A classificação histológica determinada neste estudo foi significativa na influência à sobrevida (p&lt;0,001). A pontuação prognóstica (0-13) aqui proposta, adaptada de Melo e colaboradores (2015) teve forte associação com sobrevida (p&lt;0,001). A presença de metástase foi observada em 17% dos casos, e de recidiva em 14%. Em ambas as ocorrências 90% dos animais acometidos vieram a óbito. Por meio de analises estatísticas comprovamos a associação entre esses dois fatores (metástase e recidiva) com o tempo de sobrevida (em ambas p&lt;0,001). Dentro do nosso estudo foi observado que, em tumores com a presença de AIM (agrupamentos independentes de mastócitos) há um risco de mortalidade 8,57 vezes maior do que animais com margens livres ou mesmo comprometidas. Esse risco é inclusive maior do que o risco de óbito em animais com recidiva (HR 5,13). VEGF-A se mostrou de significância estatística perante sobrevida; confirmando estudo anteriores e sugerido mais uma vez a inclusão desse marcador no perfil prognóstico do mastocitomas. Todas as formações apresentadas neste estudo tiveram análise termográfica concluída e documentada. Nota-se que mesmo quando considerado ponto central ou quando considerada toda área tumoral, 65 e 67% respectivamente, dos mastocitomas eram mais quentes que a pele sadia circundante. A causa destas mudanças de temperatura não é totalmente compreendida, mas sugere-se que esteja associada a neoangiogênese e inflamação local (XIE et al., 2004). Por meio das análises estatísticas é possível afirmar que as regiões tumoral e não tumoral são significativamente diferentes, tanto na avaliação de ponto central (SpT e SPNT) como na avaliação da área (AT e ANT) do tumor e pele circundante sadia (p &lt; 0,001). Ainda por meio de termografia, pudemos estabelecer que retalhos cutâneos pediculados apresentaram chance de deiscência 5,57 vezes maior do que o uso de outras técnicas de anaplastia e deslizamento de tecidos. Por fim, conseguimos estabelecer uma curva térmica de evolução das feridas da nossa população do estudo, bem como diferenciar o comportamento térmico quando há ou não deiscência. Isso pode ser útil a estudos futuros ou mesmo à prática clínico-cirúrgica, de modo a comparar a evolução de pacientes com as curvas previamente aqui estabelecidas, sendo factível assim prever a cicatrização da ferida. Acreditamos, por meio deste estudo, que a análise termográfica da evolução de retalhos cutâneos pode ser usada como correspondente a perfusão tecidual, conforme indicado em literatura e ser usada como ferramenta de reconhecimento precoces de deiscência da ferida cirúrgica, conforme proposto por diversos autores (SALMI et al., 1995; EICHHORN et al., 2009; WEERD et al., 2009; WEERD et al., 2011). / In most cases, proper surgical excision of mast cell tumors in dogs requires reconstructive techniques for the closure of the resulting wound, and it knowledge becomes essential for the surgeon. Achieving free margins has a significant influence on survival time, and should be the goal for most patients. The evaluation of the evolution of cutaneous flaps and the measurement of tissue adherence and vascularization at the intended site is extremely important for early diagnosis of defects in tissue implantation. There is no evidence in literature of studies related to evaluate tissue perfusion of skin flaps in companion animals, using thermography techniques. In this study, standard images and thermographic ones were obtained from 63 mast cell tumors from 60 dogs. The histological classification of the tumors in this study was significant in survival time (p &lt;0.001). The prognostic score (0-13) proposed here, adapted from Melo et al. (2015), had also strong association with survival time (p &lt;0.001). The presence of metastasis was observed in 17% of cases, and relapse in 14%. In both cases, 90% of the affected animals died. By means of statistical analyzes, we verified the association between these two factors (metastasis and relapse) in survival time (for both p &lt;0.001). Within our study it was observed that in tumors with the presence of AIM (independent mast cell groups) there is a mortality risk 8.57 times higher than animals with free or even compromised margins. This risk is even greater than the risk of death in animals with relapse (HR 5,13). Also, VEGF-A was shown to be statistically significant at survival time; confirming previous studys and leading us to suggets once again the inclusion of this marker in the prognostic profile of mast cell tumors. All the tumors presented in this study had a thermographic analysis completed and documented. It is noted that even when considered central point or tumor area, 65 and 67% respectively, of the mast cell tumors were warmer than the surrounding healthy tissue. The cause of these temperature changes is not fully understood, but it is suggested to be associated with neoangiogenesis and local inflammation (XIE et al., 2004). By means of the statistical analyzes it is possible to affirm that the tumoral and non-tumoral regions are significantly different, as well as in the evaluation of the central point (SpT and SPNT) and in the evaluation of the tumoral area (TA and ANT) im comparison with healthy surrounding skin (p &lt;0.001 ). Also through thermography, we could establish that skin flaps had a chance of dehiscence 5.57 times greater than the use of other techniques of reconstructive surgery. Finally, we were able to establish a thermal curve of evolution of the wounds of our study population, as well as to differentiate the thermal behavior when there is or not dehiscence. This may be useful for future studies or even clinical-surgical practice, in order to compare the evolution of patients with the curves previously established here, and it is feasible to predict wound healing. We believe, by means of this study, that the thermographic analysis of the evolution of cutaneous flaps can be used as corresponding to tissue perfusion, as indicated in the literature and be used as an early recognition tool for surgical wound dehiscence, as proposed by several authors (SALMI et al., 1995; EICHHORN et al., 2009; WEERD et al., 2009; WEERD et al., 2011).

Objetivos: Determinar a expressão imunohistoquímica do fator indutor de hipóxia 1-alfa (HIF-1-alfa) e suas variáveis associadas em pacientes com câncer de mama localmente avançado. Pacientes e método: Vinte e sete mulheres foram biopsiadas para diagnóstico histopatológico do carcinoma mamário e submetidas a tratamento quimioterápico pré-cirúrgico. Analisou-se a associação do HIF-1-alfa com idade, tamanho tumoral, grau histológico, estadio clínico, status hormonal e axilar, resposta clínica e patológica após tratamento quimioterápico, expressão do receptor de estrogênio, progesterona e cerbB2. Resultados: A expressão de HIF-1-alfa foi presente em 66,7% das pacientes. O único fator associado à sua presença foi o status axilar positivo (p=0,02), tendo permanecido durante a análise univariada. As demais variáveis não apresentaram associação estatisticamente significante. Conclusão: Existe uma associação estatisticamente significante entre o acometimento linfonodal e a presença de HIF-1-alfa em pacientes com câncer de mama localmente avançado. / Objectives: To assess the expression of HIF-1 and its associated variables with locally advanced breast cancer (LABC) patients. Methods: Twenty-seven women were submitted to incisional biopsy for histopathological diagnosis of breast carcinoma and undertaken to neoadjuvant chemotherapy (NACT). It was studied the association of HIF-1 with age, tumoral size, histological grade, clinical stage, hormonal and axillary status, clinical and pathological response after NACT, expression of estrogen and progesterone receptors, as well as the presence of cerbB2 antigen. Results: HIF-1-alpha expression was found in 66.7% of patients. Only axillary status was the associated factor with its presence (p=0.02), and remained after univariate analysis. The others did not present any significant statistically difference. Conclusion: There is a significant statistically association between axillary status and HIF-1-alpha expression in LABC patients.

Zielsetzung: Das Ovarialkarzinom ist die häufigste Krebstodesursache innerhalb der gynäkologischen Malignome. Im Rahmen dieser retrospektiven Arbeit werden die klinischen Ergebnissen von 84 Patientinnen dargestellt, die an Ovarialkarzinom erkrankten und im Krankenhaus im Friedrichshain in Berlin behandelt wurden. Das Ziel dieser Studie war die Analyse des Einflusses verschiedener Prognosefaktoren auf das Gesamtüberleben, Identifikation unabhängiger Prognosefaktoren und Erarbeitung von Prognoseregeln in Bezug auf das Gesamtüberleben sowie Beurteilung des Therapieerfolges und kritische Diskussion des second-look Verfahrens. Methodik: Zur Bestimmung der kumulierten Überlebensraten wurde die Kaplan-Meier-Methode verwendet, zur Identifikation unabhängiger Prognosefaktoren die multivariate Analyse. Als statistisch signifikant galten Ergebnisse mit einer Irrtumswahrscheinlichkeit von p < 0,05. Ergebnisse: Das mediane Alter zum Zeitpunkt der Erstdiagnose betrug 61 Jahre. Die kumulierte Fünfjahresüberlebensrate betrug 53 %. Die kumulierten Fünfjahresüberlebensraten lagen für die Stadien FIGO I, II, III bzw. IV bei 93 %, 83 %, 20 % bzw. 11 %. Seitens der Histologie handelte es sich überwiegend um serös-papilläre Karzinome. Die Rezidivrate lag bei 36 %. Der Median bis zum Auftreten eines Rezidivs betrug 11,5 Monate. Schlussfolgerungen: Eine gute Prognose ergibt sich bei geringem Alter bei Erstdiagnose, gutem Allgemeinzustand, normwertigem präoperativem Tumormarker CA 125, fehlendem Aszites bzw. möglichst geringer Aszitesmenge, hochdifferenzierten Tumoren, möglichst vollständiger Tumorentfernung und Lymphonodektomie. Als unabhängige Prognosefaktoren haben sich nur das FIGO-Stadium, Alter und Grading erwiesen. Anhand der unabhängigen Prognosefaktoren lässt sich die prognostizierte Überlebenswahrscheinlichkeit angeben. / Objective: Ovarian carcinoma is the most frequent reason of the cancer death among malignant gynaecological tumours. The clinical outcome of 84 patients, who had ovarian carcinoma and underwent the treatment in "The Hospital in Friedrichshain" in Berlin, is presented in this retrospective study. The objective of this study was to analyse the influence of different prognostic factors related to overall survival, to identify the independent prognostic factors, to set up prognostic rules for patients with ovarian carcinoma related to overall survival, to assess the benefit of the therapy and the critical discussion of the second-look operation. Methods: The Kaplan-Meier method was applied in order to estimate overall survival rates and multivariate analysis in order to evaluate the independent prognostic factors. The results with p < 0,05 were of statistic significance. Results: The median age at diagnosis was 61 years. The cumulated 5-year survival rate was 53 %. The cumulated 5-year survival rates for the stages FIGO I, II, III and IV were 93 %, 83 %, 20 % and 11 %, respectively. Concerning histology there were mostly serous-papillar carcinomas. The recurrence rate was 36 %. The median until the recurrence occurred was 11,5 months. Conclusions: A good prognosis is associated with lower patient age at diagnosis, good performance status, normal preoperative serum CA-125 level, absence or minimal presence of ascites, well differentiated tumours, minimal size of the residual disease after primary cytoreductive surgery and lymphadenectomy. However, on multivariate analysis, only tumour stage, patient age and tumour grade retained prognostic significance as independent prognostic factors. Due to independent prognostic factors the survival probability can be predicted.

Le muscle strié squelettique est un tissu richement vascularisé. Au delà de l'apport en oxygène et en nutriments, de nouvelles fonctions des vaisseaux ont été récemment identifiées, par le biais des interactions établies entre les cellules du vaisseau (cellules endothéliales) et les cellules du muscle, en particulier les cellules souches musculaires (cellules satellites). Celles-ci interagissent étroitement avec les cellules endothéliales pour leur expansion et leur différenciation, puis avec les cellules péri-endothéliales pour leur auto-renouvellement et leur retour à la quiescence. Les vaisseaux participent ainsi au contrôle de l’homéostasie du muscle squelettique. Grâce à ces interactions, les cellules vasculaires jouent donc un rôle central dans le remodelage tissulaire après un phénomène destructif, survenant par exemple au cours d’un trauma ou d’une myopathie. Pour étudier, les mécanismes de la plasticité vasculaire au cours du remodelage tissulaire, deux situations paradigmatiques de muscle en régénération chez l’enfant : la dermatomyosite juvénile (DMJ) et la dystrophie musculaire de Duchenne (DMD) ont été étudiées. Il existe, dans ces deux pathologies une souffrance musculaire associée à des cycles de nécrose/régénération. Elles se différencient par leur plasticité vasculaire et par leur évolution. En effet, la DMJ, la myopathie inflammatoire la plus fréquente de l’enfant est caractérisée par une vasculopathie avec perte en capillaires. L’évolution peut être favorable avec restitution ad integrum du muscle. La DMD est une myopathie génétique conduisant à une dégradation progressive de la force musculaire associée à une néovascularisation compensatrice. Le volet clinique/histologique incluant une analyse multiparamétrique des critères évolutifs cliniques et de réponse thérapeutique couplée à une réévaluation des données histologiques de la DMJ (analyse morphométrique des muscles DMJ) a permis de montrer qu’il existait des sous groupes phénotypiques homogènes de sévérité différente dans la DMJ. Le degré de sévérité clinique est relié à la gravité de la vasculopathie musculaire Par ailleurs, des marqueurs cliniques et histologiques simples permettant de repérer au diagnostic les patients nécessitant une escalade thérapeutique rapide (CMAS>34, atteinte gastrointestinale, fibrose endomysiale musculaire au diagnostic) ont été identifiés. Le volet cellulaire a permis l’identification in vitro des interactions cellulaires spécifiques et différentielles des myoblastes issues de patients DMD et DMJ sur les cellules endothéliales normales par l’analyse de leur rôle sur la prolifération, migration et différenciation des cellules vasculaires. Dans la DMD, les myoblastes entrainent une réponse angiogénique importante mais non efficace (néovascularisation anarchique). Dans la DMJ, les myoblastes participent efficacement à la reconstruction vasculaire notamment via la sécrétion de facteurs proangiogéniques. Ces résultats ont été renforcés par analyse transcriptomique effectuée à partir de cellules endothéliales et satellites isolées de muscles de patients confirmant le rôle central de la vasculopathie associée à un contexte inflammatoire spécifique lié à l’interféron dans la physiopathologie de la DMJ et montrant dans la DMD une dérégulation de l’homéostasie normale des interactions vaisseau-muscle avec mise en jeu d’un remodelage tissulaire non efficace. Ces données permettent d'identifier de nouvelles fonctions des cellules vasculaires dans le remodelage du muscle strié squelettique au cours des pathologies musculaires de l'enfant, et devraient ouvrir la voie à de nouvelles approches thérapeutiques. / Skeletal muscle is highly vascularised. Beyond oxygen and nutriment supply, new functions for vessels have been recently identified, through the interactions that vessel cells (endothelial cells) establish with muscle cells, particularly with muscle stem cells (satellite cells). These latter closely interact with endothelial cells for their expansion and their differentiation, then with periendothelial cells for their self-renewal and return to quiescence. During skeletal muscle regeneration endothelial cells reciprocally interact with myogenic cells by direct contact or by releasing soluble factors to promote both myogenesis and angiogenesis processes. Skeletal muscle regeneration typically occurs as a result of a trauma or disease, such as congenital or myopathies. To better understand the role of vessel plasticity in tissue remodeling, we took advantage of two muscular disorders that could be considered as paradigmatic situations of regenerating skeletal muscle in the child: Juvenile Dermatomyositis (JDM), the most frequent inflammatory myopathy and Duchenne Muscular Dystrophy (DMD), the most common type of muscular dystrophy. Although these two muscular disorders share, at the tissue level, similar mechanisms of necrosis-inflammation, they differ regarding the vessel domain. In JDM patients, microvascular changes consist in a destruction of endothelial cells assessed by focal capillary loss. This capillary bed destruction is transient. The tissue remodeling is efficient and muscle may progressively recover its function. By contrast, in DMD, despite an increase of vessels density in an attempt to improve the muscle perfusion, the muscle function progressively alters with age. We identified clinical and pathological markers of severity and predictive factors for poor clinical outcome in JDM by computing a comprehensive initial and follow-up clinical data set with deltoid muscle biopsy alterations controlled by age-based analysis of the deltoid muscle capillarization. We demonstrated that JDM can be divided into two distinctive clinical subgroups. The severe clinical presentation and outcome are linked to vasculopathy. Furthermore, a set of simple predictors (CMAS<34, gastrointestinal involvement, muscle endomysial fibrosis at disease onset) allow early recognition of patients needing rapid therapeutic escalation with more potent drugs. We studied in vitro the specific cell interactions between myogenic cells issued from JDM and DMD patients and normal endothelial cells to explore whether myogenic cells participate to the vessel remodeling observed in the two pathologies. We demonstrated that MPCs possessed angiogenic properties depending on the pathological environment. In DMD, MPCs promoted the development of establishment of an anarchic, although strong, EC stimulation, leading to the formation of weakly functional vessels. In JDM, MPCs enhanced the vessel reconstruction via the secretion of proangiogenic factors. This functional analysis was supported by the transcriptomic analysis consistent with a central vasculopathy in JDM including a strong and specific response to an inflammatory environment. On the contrary, DMD cells presented an unbalanced homeostasis with deregulation of several processes including muscle and vessel development with attempts to recover neuromuscular system by MPCs. To summarize, our data should allow the definition of new functions of vessel cells in skeletal muscle remodelling during muscle pathologies of the child that will open the way to explore new therapeutic options and to gain further insights in the pathogenesis of these diseases.

Zur Abschätzung der Prognose von Tumorerkrankungen und zur Therapieplanung können neben konventionellen klinischen Parametern auch molekulare Prognosemarker im Tumorgewebe bestimmt werden. In der vorliegenden Studie haben wir vier verschiedene potentielle molekulare Prognosefaktoren im Ovarialkarzinom und teilweise auch im Mammakarzinom untersucht: die Cyclooxygenase-2 (COX-2), das humane ELAV-ähnliche Protein HuR, das Oberflächenantigen CD24 und die Mitogen-aktivierte Protein Kinase Phosphatase-1 (MKP-1). Dabei lag der Schwerpunkt auf der Untersuchung der Cyclooxygenase-2 (COX-2), die sowohl in der Entzündungsreaktion als auch bei der Entstehung und Progression maligner Tumoren eine wichtige Rolle spielt. Wir konnten zeigen, dass eine erhöhte Expression der COX-2 beim Ovarialkarzinom und beim Mammakarzinom signifikant mit einer schlechteren Prognose assoziiert ist. In Zellkulturmodellen haben wir verschiedene Strategien zur Inhibition der COX-2 angewendet, nämlich die pharmakologische Inhibition durch NS-398 sowie die spezifische Inhibition durch RNA Interferenz. Dabei ergab sich, dass COX-2 Inhibitoren neben der Wirkung auf die COX-2 auch über anderen Zielproteine die Proliferation von Ovarialkarzinomzellen hemmen und zu einem Zellzyklusarrest führen. Bei weiteren Untersuchungen zur Regulation der COX-2 konnten wir zeigen, dass das RNA-stabilisierende Protein HuR mit der COX-2 Expression korreliert und ebenfalls ein Prognosefaktor für das Ovarialkarziom ist. Unsere Ergebnisse bilden eine Grundlage für klinische Studien zur Untersuchung des möglichen Effektes von COX-Inhibitoren in der Therapie maligner Tumoren. / Molecular prognostic markers can be determined in tumor tissue and can be used - in addition to conventional clinicopathological parameters - to estimate patient prognosis and to plan the therapy of malignant tumors. In this study we have investigated the expression of four different molecular prognostic factors in ovarian carcinoma and partially in breast carcinoma: cyclooxygenase-2 (COX-2), the human ELAV-like protein HuR, the surface antigen CD24, as well as the mitogen-activated protein kinase phosphatase-1 (MKP-1). For further evaluation, we have focused on COX-2, which plays an important role in tumor biology and inflammation. Increased expression of COX-2 in tumor tissue was associated with poor prognosis in ovarian carcinoma and breast carcinoma. In cell culture models, we have used two different strategies for inhibition of COX-2: pharmacological inhibition and RNA interference. We found that COX-2 inhibitors act on other cellular targets in addition to COX-2 and inhibit proliferation of ovarian carcinoma cells by induction of cell cycle arrest. In further studies we could show that the RNA-stabilizing protein HuR is associated with increased COX-2 expression and is an prognostic factor in ovarian carcinoma, as well. These results provide a basis for further evaluation of COX-inhibitors in tumor therapy.