L’hyperplasie et l’hypertrophie contribuent à l'augmentation de la masse de muscle lisse bronchique observée dans le souffle. Les cellules musculaires lisses (CML) présentent deux phénotypes; prolifératif ou contractile. Le serum response factor (SRF), un facteur de transcription impliqué dans l’activation de nombreux gènes, contribuerait à cette modulation phénotypique. Notamment, lorsqu'associé au cofacteur Elk-1, un phénotype prolifératif serait observé, alors qu'en présence de la myocardine (MYOCD) il y aurait induction d'un profil contractile. Récemment, il a été démontré que SRF est surexprimé dans les voies périphériques chez les chevaux atteints du souffle suite à une exposition antigénique. Cette étude vise à caractériser l'expression protéique et génique de SRF, Elk-1 et MYOCD dans les CML des voies respiratoires centrales et périphériques chez des chevaux atteints du souffle et des chevaux contrôles. L'évaluation de l’expression protéique de SRF, Elk-1 et MYOCD s’est effectuée par immunodétection sur des tissus provenant de biopsies thoracoscopiques ou endobronchiques, et ce, avant, à 1 et 30 jours du défi antigénique. L'expression génique a été étudiée par qPCR sur du muscle lisse disséqué de la trachée, et des bronches, ainsi que sur des voies respiratoires intermédiaires et périphériques. Les expressions génique et protéique de MYOCD sont augmentées uniquement dans les voies périphériques. L’expression génique de SRF et Elk-1 varient dans les voies centrales alors que le taux de protéines demeure stable. En conclusion, SRF et MYOCD pourraient être impliquées dans l’hypertrophie des voies respiratoires périphériques dans le souffle alors que l’hyperplasie ne semble pas être activée par Elk-1. / Airway smooth muscle (ASM) cells hyperplasia and hypertrophy contribute to the increased airway smooth muscle mass present in heaves. ASM cells express either a synthetic proliferative or a contractile phenotype. Serum response factor (SRF) is a transcription factor that has been shown to regulate myocyte differentiation in vitro in vascular and intestinal smooth muscles. When SRF is associated with Elk-1, it promotes ASM proliferation while myocardin (MYOCD) promotes the expression of contractile elements. Recently, SRF was shown to be overexpressed in the peripheral airways of heaves affected horses following an antigenic challenge. The objective of this study was to characterize the protein and gene expression of SRF, Elk-1 and MYOCD in ASM cells from central and peripheral airways of heaves affected horses and controls. Protein expression of Elk-1 and MYOCD was evaluated using immunohistochemistry while immunofluorescence was used for SRF detection in pulmonary peripheral and endobronchial biopsies before and at 1 and 30 days of antigenic exposure. Gene expression was investigated in ASM cells dissected from trachea and bronchi as well as from intermediate and peripheral airways using qPCR. MYOCD gene and protein expressions are increased only in peripheral airways. SRF and Elk-1 gene expression varied in the central airways while the positive cell percentage remains stable. In conclusion, the pulmonary peripheral airways hypertrophy observed in heaves seems to implicate SRF and MYOCD while the hyperplasia doesn’t seem to be activated by Elk-1.

Introdução: O sedentarismo e obesidade têm sido descritos como fatores de risco relevantes para o desenvolvimento de hiperplasia prostática benigna (HBP). No presente trabalho investigamos o papel da atividade física nas vias proliferativas e de angiogênese da próstata e sua relação na prevenção da HPB. Métodos: Foram utilizados nesse experimento ratos machos Wistar com oito semanas de idade, divididos aleatoriamente em quatro grupos: 1. atividade física (AF) e dieta normal; 2. AF e dieta rica em gorduras; 3. Sedentários com dieta normal; 4. Sedentários e dieta rica em gorduras. Alimentos e água foram fornecidos ad libitum. Os ratos dos Grupos 1 e 2 foram submetidos a um protocolo de treinamento de natação por 10 semanas, executado cinco vezes por semana com duração de 60 minutos por dia. No final do protocolo, as glândulas prostáticas foram dissecadas, pesadas e armazenadas. Medimos os níveis de expressão gênica do fator de crescimento semelhante à insulina (IGF1), do genes relacionados ao eixo proliferativo IGF1/PI3K/Akt e dos genes relacionados à hipóxia e angiogênese através do método de reação em cadeia da polimerase quantitativa em tempo real (qRT-PCR) e do índice apoptótico tecidual por hibridização in-situ (TUNEL). Resultados: Após 10 semanas, os grupos 1 e 2 apresentaram menor gordura visceral quando comparados aos grupos 3 (29,4 vs 37,96 gramas, p < 0,05) e 4 (31,87 vs 41,96 gramas, p < 0,05), respectivamente. AF diminuiu o crescimento da próstata em ratos com dieta normal (grupo 1 vs grupo 3, p < 0,05), mas este achado não foi evidenciado em ratos alimentados com dieta rica em gorduras (grupo 2 vs grupo 4). Quando comparados os ratos com dieta normal (grupo 1 vs grupo 3), os genes relacionados ao IGF1, IRS1 e Akt foram subexpressos na próstata de ratos submetidos a AF (médias de 0.22, 0.04 e 0.27 respectivamente). Esses padrões de expressão também foram evidenciados quando comparamos ratos com dieta rica em gorduras (grupo 2 vs grupo 4), mas a subexpressão de IGF1 foi menos pronunciada (p < 0,001). AF aumentou a expressão de genes relacionados à angiogênese (HIF-1?, VEGF, VEGFR e mTOR) quando comparamos ratos submetidos a dieta rica em gorduras (grupo 2 vs grupo 4). O índice apoptótico (número de apoptoses/10 HPF) foi maior em ratos submetidos a AF (9,0 vs 2,0, grupo 1 vs grupo 3, p=0,07), (9,0 vs 1,43, grupo 1 vs grupo 4; p < 0,05). Conclusão: A atividade física inibe a expressão gênica do eixo proliferativo IGF1/Akt na próstata tanto em ratos normais quanto em ratos com dieta rica em gorduras, estimula a angiogênese em ratos com dieta rica em gorduras e estimula a apoptose prostática. Esses achados podem estar relacionados à prevenção de HPB / Introduction: Sedentarism and obesity have been reported as relevant risk factors for the development of benign prostatic hyperplasia (BPH). In the present work we investigated the role of physical activity in proliferative and angiogenic pathways on prostate and implications for BPH prevention. Methods: Male Wistar rats with eight weeks old were used in this experiment, randomly divided into four groups: 1. physical activity (PA) and normal diet; 2. PA and high fat diet; 3. sedentary (S) and normal diet; 4. S and high fat diet. Food and water were provided ad libitum. The rats in the Groups 1 and 2 were submitted to a swimming training protocol for 10 weeks, executed five times a week with duration of 60 minutes per day. At the end of the protocol, prostate glands were dissected, weighted and stored. We measured prostatic gene expression levels of insulin-like growth factor 1 (IGF1), IGF1/PI3K/Akt proliferative axis and genes related to angiogenesis through the quantitative real-time polymerase chain reaction (qRT-PCR) method, and apoptotic index (TUNEL). Results: After 10 weeks, groups 1 and 2 had less visceral fat when compared to the groups 3 (29,4 vs 37,96 grams; p < 0,05) and 4 (31,87 vs 41,96 grams; p < 0,05), respectively. According to prostate weight, PA decreased prostate growth in normal fed rats (group 1 vs group 3, p < 0,05), but this finding was not shown in high fat fed rats (group 2 vs group 4). When comparing normal fed rats (group 1 vs group 3), IGF1, IRS1 and Akt genes were less expressed in prostate of rats submitted to PA (means 0.22, 0.04 and 0.27 respectively). Those patterns of expressions were also shown when we compared high fat fed rats (group 2 vs group 4), but the IGF1 downregulation was less pronounced (p < 0,001). PA increased the expression of angiogenic related genes (HIF-1alpha, VEGF, VEGFR and mTOR) when comparing rats submitted to high fat diet. Apoptotic index (number of apoptosis per 10 HPF) was higher in rats submitted to PA (9,0 vs 2,0; group 1 vs group 3; p=0,07), (9,0 vs 1,43; group 1 vs group 4; p < 0,05). Conclusion: PA seems to inhibit IGF1/Akt proliferative axis on prostate in both normal and high fat fed rats, stimulates angiogenesis in high fed rats, and increase prostatic apoptosis. These findings can be related to BPH prevention

Solanum cernuum Vell. (Solanaceae), popularmente conhecida como \"panacéia\", é uma planta nativa do sul e sudeste do Brasil. Seu nome \"panacéia\" provavelmente é devido à grande amplitude de aplicações que possui na medicina popular: tratamento de úlceras, lesões hepáticas, infecções de pele, como antitumoral, diurético, depurativo, anti-hemorrágico, bem como para o tratamento da gonorreia e da hiperplasia benigna da prostática (HBP). Considerando os usos de S. cernuum na medicina popular brasileira e a escassez de estudos que os corroborrem, propomos neste trabalho, a partir da obtenção de diferentes extratos, frações e compostos isolados, testar as atividades biológicas gastroprotetora, citotóxica e antimicrobiana relatadas na medicina popular. Para tanto, folhas da espécie Solanum cernuum Vell. (Solanaceae) foram coletadas no município de Teresópolis/ RJ. O extrato hidroetanólico (ESC) e em acetato de etila foram obtidos a partir de folhas secas e trituradas. Frações em hexano (FHex), diclorometano (FDCM), acetato de etila (FAcOEt), n-butanol (FBuOH) e água (FAq) foram obtidas a partir do extrato ESC. A partir dos diversos fracionamentos do extrato ESC isolaram-se e identificaram-se quatro compostos: ácido trans-isoferulico, cernumidina, quercitrina e afzelina, sendo encontrados no extrato nas concentrações de 1,5%, 4,8%, 1,3% e 1,6%, respectivamente. Da fração em heptano, obtida da partição do extrato em acetato, isolaram-se ?-sitsterol e friedelina. No ensaio antimicrobiano, apenas as frações FHex, FDCM e FAq, oriundas da partição do extrato ESC, apresentaram atividade, em diferentes concentrações, frente a determinadas linhagens de bactérias e fungos testadas. Quanto aos compostos voláteis, na extração por hidrodestilação, não houve rendimento suficiente, mas a partir do hidrolato, por partição com diclorometano, foi possível determinar a composição dos voláteis por CG/EM. Os tricomas de S. cernuum são majoritariamente estrelados, do tipo tectores. No ensaio do XTT apenas a FDCM e o composto friedelina foram seletivos para células MCF-7 (carcinoma mamário) na concentração de 16,6 ?g/mL e 124,9 ?g/mL, respectivamente. O extrato ESC apresentou atividade gastroprotetora nos ensaios de indução de úlcera por etanol/HCl e por anti-inflamatório não esteroidal (AINE), bem como foi curativo no ensaio de úlcera crônica induzida por ácido acético na dose de 250 mg/kg. Os compostos isolados, exceto o alcaloide cernumidina, apresentaram atividade gastroprotetora pelo ensaio de indução de úlcera por etanol/HCl na dose de 30 mg/kg. Ainda sobre a gastroproteção, na avaliação de pH utilizando o ensaio de ligadura de piloro, todos os grupos de animais tratados com extrato ESC não apresentaram diminuição da acidez estomacal. Porém, houve diminuição da concentração de íons hidrogênio ([H+] mEq.g-1/L) na secreção gástrica nos grupos tratados. A produção de muco na mucosa gástrica também não foi aumentada nos grupos tratados com o extrato ESC. A avaliação da ação gastroprotetora por inibição da enzima iNOS e alquilação de grupos sulfidrilas evidenciou que a produção de óxido nítrico e de grupos sulfidrilas podem estar envolvidos na atividade gastroprotetora do ESC. Diferentes concentrações de ii extrato ESC e alcaloide cernumidina foram testadas nos tempos de 48, 72 e 96 h em cultura primária de células de músculo liso relacionadas a hiperplasia benigna da próstata (HPB), sendo possível observar inibição do crescimento celular de 60% na concentração de 3 mg/mL para o extrato e de 62% para o alcaloide na concentração de 1 mg/mL (3,3 x 10-6 M) no tempo de exposição de 96 h. Com o intuito de avaliar o potencial efeito indutor de apoptose, foi realizado o ensaio de anexina V Cy5 por citometria de fluxo. Tanto para o extrato ESC como para o acaloide cernumidina observou-se que a morte celular ocorreu mais por apoptose. Apenas para o alcaloide, na concentração de 1 mg/mL, houve significativa morte por necrose. Na atividade de inibição da angiogenese, avaliada em células HUVEC, a FBuOH (50 ?g/mL) e o alcaloide cernumidina (160 nM) inibiram a formação de vasos endoteliais em 62,75% e 77,73%. Na secagem do extrato por spray dryer o rendimento, a densidade aparente, a densidade compacta, o tamanho das partículas (D50) e o teor de cernumidina foram influenciados por alguns dos fatores avaliados: temperatura, vazão e porcentagem de adjuvante, linearmente ou na forma quadrática. Na otimização da secagem por spray dryer, utilizando a função desejabilidade, observou-se a proximidade dos valores preditos para rendimento do pó e teor de cernumidina com os valores reais, evidenciando a confiabilidade desse método matemático. Assim, os resultados obtidos corroboram alguns dos usos populares abrindo novas possibilidades para o desenvolvimento de fitoterápico a partir S. cernuum. / Solanum cernuum Vell. (Solanaceae), popularly known as \"panacéia\", is a native tree from Southern and Southeastern Brazil. Its traditional name is due to the wide range of uses in folk medicine including the treatment of ulcers, liver damage, skin infections, as diuretic, antitumor, depurative, anti-hemorrhagic, as well as to treat gonorrhoea and benign prostatic hyperplasia (BPH). Considering the importance of S. cernuum in folk medicine and the lack of research supporting its use, the aims of this project were to obtain different extracts, fractions and isolated compounds this plant to be tested for gastroprotective, cytotoxicity and antimicrobial activities. Leaves S. cernuum were collected in Teresópolis/RJ. The hydroethanolic (ESC) and ethyl acetate extracts were obtained from the dried and grinded leaves. Fractions in hexane (FHex), dichloromethane (FDCM), ethyl acetate (FAcOEt), n-butanol (FBuOH) and water (FAQ) were obtained from the ESC extract. The ESC extract was fractionated using different chromatographic techniques furnishing four isolated compounds: trans-isoferulic acid, cernumidine, quercitrin and afzelin. The content of these four compounds were quantified to be 1.5%, 4.8%, 1.3% and 1.6%, respectively, in the extract. ?-sitosterol and the friedelin were isolated by partitioning the ethyl acetate extract with heptane. In the antimicrobial assay, only FHex, FDCM and FAq fractions showed activity at different concentrations against the tested strains of bacteria and fungi. The hydrodistillation of the leaves of S. cernuum did not fusrnish essential oils, but the partition of the hydrolate with dichloromethane allowed the determination of its composition by GC/MS. The trichomes of S. cernuum are majority kind of stellate tector. Only FDCM fraction and friedelin compound showed activity in XTT assay against MCF-7 cells (breast carcinoma) at the concentration of 16.6 ?g/mL and 124.9 ?g/mL, respectively. The ESC extract showed gastroprotective effects in the acute gastric ulcer assay, which is induced by HCl/EtOH and nonsteroidal anti-inflammatory drugs (NSAIDs). Also, in the chronic ulcer assay, induced by acetic acid, the oral administration of ESC extract (250 mg/kg) reduced the ulcerated area, suggesting that it has a curative potential. The isolated compounds (30 mg/kg) exhibited gastroprotective activity in acute gastric ulcer assay, except cernumidine. The results indicated that the ESC extract was able to reduce the concentration of H+ ions in comparison with the control group. However, ESC extract did not modify the pH of the gastric juice acid and there was no significant increment in the amount of gastric adherent mucus in pretreated group. Further, it was evaluated the role of nitric oxide (NO) and sulfhydryl (SH) groups on the gastroprotective effect elicited by ESC extract. The results showed that gastroprotection induced by the ESC extract is correlated with the presence of endogenous NO and SH groups which were responsible for gastroprotective activity. In the anti-angiogenic activity assay evaluated in HUVEC cells, the FBuOH fraction (50 ?g/mL) and the cernumidine alkaloid (160 nM) inhibited the the formation of endothelial vessels by 62.75% and 77.73%, respectively. Different concentrations of the ESC extract and cernumidine alkaloid were further tested in primary culture of iv smooth muscle HPB for 48, 72 and 96 h. ESC extract (3 mg/mL) and the alkaloid (1 mg/mL; 3.3 x 10-6 M) treatment for 96 h inhibited the cell growth by 60% and 62%, respectively. In addition, it was assessed the potential effect of extract and its constituents to induce apoptosis by performing the annexin Cy5 assay by flow cytometry. Treatment with ESC extract and cernumidine alkaloid led to cell death by apoptosis. Only cernumidine at 1 mg/mL showed significant cells death by necrosis compared to control. Considering the spray dryer technique to dry the extract, the yield, bulk and tapped densities, particle size (D50) and the content of the cernumidine are influenced by some of the evaluated factors: temperature, flow and excipient percentage, linearly or quadratic form. To optimize the drying by spray dryer, it was used the desirability function. It was observed that the proximity of the predicted values of the yield and cernumidine content with the real values were found, showing the reliability of this mathematical method. Therefore, the obtained results corroborate the folk use and open new opportunities for the development of a phytotherapic from S. cernuum.

A hiperplasia do processo coronóide é uma afecção que causa uma limitada mobilidade mandibular e conseqüente limitação de abertura da boca. Não apresenta sintomatologia dolorosa, e tem progressão lenta, levando o paciente a procurar por tratamento somente se a limitação de abertura for severa a ponto de prejudicar as funções mastigatórias normais. Muitos casos levam clínicos a tratarem o paciente como portador de disfunção da articulação têmporo mandibular (DTM). Este trabalho se propõe a avaliar a tomografia computadorizada (TC) como método para auxiliar na elaboração do diagnóstico e no planejamento cirúrgico desta anomalia, estudando a presença da hiperplasia do processo coronóide pela observação de imagens obtidas por meio da Tomografia Computadorizada Helicoidal, considerando as imagens volumétricas multiplanar e 3D, e as suas associações. Foram utilizadas imagens de arquivo dos exames de tomografia computadorizada helicoidal de 152 pacientes que apresentavam sinais e sintomas de Disfunção Têmporo Mandibular (DTM) encaminhados para a Unidade de Diagnóstico Dento-Maxilo-Facial da Clínica Félix Boada, na cidade de Caracas - Venezuela, para exame tomográfico da ATM. Dos 152 pacientes, foram selecionados 20 casos, que eram de portadores de sinais e/ou sintomas de Disfunção Têmporo Mandibular, mas que, ao exame realizado, constatou-se a presença da hiperplasia do processo coronóide. Os dados dos 20 pacientes foram comparados, de maneira a identificar as possíveis diferenças entre as opiniões de 5 cirurgiões dentistas radiologistas e de 5 cirurgiões dentistas buco-maxilo-faciais. Utilizou-se a técnica da Análise de Médias (Analysis of Means - ANOM), que calcula uma média geral entre os 5 observadores de cada especialidade, e um intervalo que indica quais observadores tiveram opiniões semelhantes e quais tiveram opiniões diversas. Depois foi feita a comparação entre os dois grupos, utilizando o teste das proporções. Em conclusão, os observadores acharam as imagens MPR mais esclarecedoras que as reconstruções volumétricas 3D, na maioria dos casos de hiperplasia do processo coronóide, quando esses recursos foram avaliados separadamente. Contudo, acharam imprescindível a utilização de ambas as imagens, em associação ou não, indicando-as para o estudo da hiperplasia do processo coronóide. A hipomobilidade foi observada na articulação temporomandibular, no lado acometido pela hiperplasia do processo coronóide, em 55,2% dos casos / The Coronoid Process Hyperplasia is an affection that causes a limited mandibular mobility and a consequent limitation in mouth opening. It doesn’t present any painful symptomatology and has a slow progression, leading the patient to look for treatment only when the opening limitation is severe to the point of impairing regular masticatory functions. Many cases lead clinicians to treat the patient as he had a temporomandibular dysfunction. This work proposed an evaluation of the CT as an auxiliary method in the elaboration of the diagnosis and in the surgical planning of that anomaly by studying the presence of the Coronoid Process Hyperplasia through the observation of images obtained by means of the Helicoidal Computerized Tomography, considering the volumetric multiplane and 3D images and their associations. Filed images of Helicoidal Computerized Tomography were used, belonging to the examination of 152 patients who presented symptoms of temporomandibular dysfunction and were all directed to the Dento-Maxillo-Facial Diagnosis Unit of the Clinica Felix Boada in the city of Caracas – Venezuela for TMA tomographic examination. Of the 152 patients 20 were selected who showed signs or symptoms of TMA dysfunction and Coronoid Process Hyperplasia was detected. Data of the 20 patients were compared in order to identify possible differences among the opinions of 5 radiologist specialists and 5 buco-maxillo -facial specialists. The Analysis of Means – ANOM was used to calculate a general average among the 5 specialists in each specialty and a gap that opinions and which ones had different opinions. Afterwards the comparison between the 2 groups was made using the Proportion Test. The observers concluded that the MPR images were more clarifying than the 3D images in most of the cases of coronoid process hyperplasia, especially when those resources were evaluated separately. Although they found vital the utilization of both images in association or not and indicated them to the study of the Coronoid Process Hyperplasia. Hypo mobility was observed in the TMA in the affected side in 55,2% of the cases

INTRODUÇÃO: A HPB é a neoplasia benigna mais comum no homem. Embora amplamente estudada, a patogênese da HPB não foi totalmente elucidada. Fatores de crescimento são proteínas que regulam o crescimento, a diferenciação e morte celular programada. Estudos têm demonstrado que eles interagem com marcadores de inflamação e angiogênese na próstata e simulam a resposta que ocorre no processo de cicatrização de tecidos. Avaliamos o perfil de expressão de FC e marcadores de inflamação e angiogênese no tecido prostático de pacientes com próstata aumentada em relação ao grupo controle com próstata de tamanho normal. MÉTODO: Foram analisados tecidos prostáticos congelados de 62 pacientes submetidos à ressecção transuretral da próstata ou cirurgia aberta para tratamento da HPB. O grupo controle foi composto por amostras de tecido de 8 pacientes com próstata de pequeno volume (menor do que 40 gramas) obtidas por ressecção transuretral. Os níveis de expressão dos FC (EGF, FGF2, PDF, TGFbeta1, IGF1), de angiogênese (VEGF, CD105) e de inflamação (IL-2, IL-6, IL-8 e IL-17) foram avaliados por reação de cadeia da polimerase em tempo real quantitativo (qRT-PCR). A análise de subgrupo de pacientes com e sem uso de sonda vesical de demora antes da cirurgia bem como a correlação da expressão com variáveis clínico-demográficas (volume prostático, PSA, presença de prostatite, idade e tempo de sintoma) foram realizadas. RESULTADOS: Encontramos superexpressão de todos os fatores de crescimento, marcadores de inflamação e angiogênese nos 62 pacientes com HPB em relação ao grupo controle. Verificamos na análise de subgrupo que a presença da sonda vesical parece induzir o aumento da expressão de FGF2, IGF1, TGFbeta1, IL-8 e VEGF. Estes dois últimos apresentaram-se ainda mais superexpressos em pacientes com prostatite associada a HPB. Pacientes com expressão de IGF1 acima da média também apresentaram expressão aumentada de outros fatores de crescimento (FGF2, TGFbeta1, CD105 e IL-2). E, também, o nível de expressão de IGF1 foi maior em pacientes com idade superior a 65 anos em relação aos mais jovens. VEGF apresentou maior superexpressão em pacientes com menos de 1 ano de sintomas do trato inferior e a IL-6 mostrou maiores níveis de expressão em pacientes com tempo de sintoma acima de 1 ano. A IL-2 mostrou-se ainda mais superexpressa no subgrupo de pacientes com sonda vesical e próstata maior do que 60g. CONCLUSÃO: Encontramos uma superexpressão geral de todos os FC, marcadores de angiogênese e inflamação nos pacientes com HPB e próstatas maiores do que 40g comparado ao grupo controle. Além disso demonstramos pioneiramente que o uso da sonda vesical pode aumentar a expressão dos FC (FGF2, IGF1 e TGFbeta1) de VEGF e também da IL-8 / INTRODUCTION: BPH is the most common benign tumor in men. Although widely studied, the pathogenesis of BPH has not been fully elucidated. Growth factors (GF) are proteins that regulate growth, differentiation, and programmed cell death. Studies have shown that they interact with markers of inflammation and angiogenesis in prostate and simulate the response that occurs in the tissue healing process. We evaluate the FC expression profile and inflammation and angiogenesis markers in prostate tissue from patients with enlarged prostate compared to the control group with normal sized prostate. METHOD: Frozen prostate tissues were analyzed from 62 patients who underwent transurethral resection of the prostate or open surgery for treating BPH. The control group was composed of tissue samples from 8 patients with small prostate volume (less than 40 grams) obtained by transurethral resection. The expression levels of GF (EGF, FGF2, PDF, TGFbeta1, IGF1), angiogenesis (VEGF, CD105) and inflammatory (IL-2, IL-6, IL-8 and IL-17) were evaluated by reaction Polymerase chain quantitative real-time (qRT-PCR). The subgroup analysis of patients with and without use of indwelling urinary catheter before surgery as well as the correlation of expression with clinical and demographic variables (prostate volume, PSA, presence of prostatitis, age and symptom of time) were performed. RESULTSs: We found overexpression of all growth factors, inflammation and angiogenesis markers in the 62 patients with BPH compared to the control group. We found in subgroup analysis that the presence of urinary catheter seems to induce increased FGF2, IGF1, TGF?1, IL-8 and VEGF expression. These latter had become even more overexpressed in patients with BPH associated with prostatitis. Patients with IGF1 expression above the average also showed increased expression of other GF (FGF2, TGFbeta1, IL-2 and CD105). And also, the level of IGF1 expression was higher in patients older than 65 years compared to younger patients. VEGF overexpression was higher in patients with less than 1 year of lower tract symptoms and IL-6 showed higher expression levels in patients with symptoms over 1 year of length. IL-2 was shown to be overexpressed in further subgroup of patients with bladder catheter and prostate greater than 60g size. CONCLUSION: We found a general overexpression of all GF, markers of angiogenesis and inflammation in patients with BPH and prostates larger than 40g compared to the control group. Also demonstrated pioneered that the use of indwelling catheter can increase the expression of GF (FGF2, IGF1 and TGFbeta1), VEGF and IL-8 as well

Die moderne Gefäßchirurgie bedient sich bei hohen Stadien der pAVK, spezieller Gefäßrekonstruktionen in Form von distalen End-zu-Seit-Gefäßanastomosen. Das langfristige Versagen der Gefäßanastomose hängt primär von der Entstehung einer subendothelialen Intimahyperplasie (IH) ab. Diese IH-Gebiete befinden sich je nach Anastomosengeometrie im Gebiet der Hauben- und Fersenzone sowie am Boden der Anastomose. // Mit Hilfe der Particle Image Velocimetry-Technik wird eine Taylor-Patch-, eine Miller-Cuff-Anastomose und eine femoro-crurale Patch-Prothese bezüglich ihrer Flussmuster sowie ihrer hämodynamischen Eigenschaften wie Geschwindigkeit, Scherstress und Rotation in z-Richtung (Vorticity) untersucht. // In einem hydrodynamischen Kreislaufmodell werden elastische, transparente Silikonmodelle der Anastomosen hergestellt und mit einem blutanalogen Newtonschen Fluid (Glycerol-Wasser-Gemisch) unter Simulation der femorocruralen Druckkurve, pulsatil bei Variation der Strömungsbedingung perfundiert. Der periphere Widerstand beträgt 0,5 mmHg/ml/min (PRU) und die Phasenverschiebung -12 Grad (zwischen Druck- und Flusskurve). // Die Flussmuster variieren zwischen den unterschiedlichen Ausstromverhältnissen erheblich. Bei den unterschiedlichen Flussstärken hingegen ähneln sich die Flussmuster. Alle drei Modelle zeigen ausgeprägte Flussseparationszonen im Hauben- und Fersengebiet sowie geometrieabhängig auch eine Stagnationszone am Boden. Diese Bereiche wiesen die geringsten Fluidgeschwindigkeiten, deutlich unter normalem Wandscherstressniveau liegende Scherstressverhältnisse sowie geringe Vorticitywerte auf. Im Bereich der Übergangszonen finden sich hohe Scherstress- sowie Vorticitywerte. Geschwindigkeitsunterschiede des Fluids zeigten sich im Bereich der Ausstromsegmente. Variable Stressverteilungen zeigen sich auch innerhalb der Separationszonen. Eine Erklärung für die unterschiedlich beschriebenen Offenheitsraten der drei Anastomosenformen wird durch diese Arbeit nicht gefunden. / Modern vascular surgery uses special termino-lateral anastomoses for treating high levels of peripheral arterial disease (PAD). Long term stenoses and occlusions of vascular anastomoses mostly depend on the development of subendothelial myointimal hyperplasia (MIH). There are characteristic areas within the anastomoses, where this process can be examined: The heel, the tow and the floor zone. // This examination observes local hemodynamics like velocity, shear stress and vorticity (rotation in z-direction) and flow patterns of a Taylor-Patch-, a Miller-Cuff-Anastomosis and a feroro-crural patch prothesis (FCPP) with the usage of a Particle Image Velocimetry. In a hydrodynamic circulation model various elastic, transparent silicon phantoms of termino-lateral anastomoses are perfused with a Newton fluid blood analogon (glycerol-water mixture) while simulating the femorocrural pressure curve in a pulsatile manner under variation of the flow conditions. The outflow resistance is 0.5 mmHg/ml/min (PRU, peripheral resistance units) and a phase shift of -12° between flow and pressure curve is simulated. // The flow patterns differed extremely in accordance of the various outflow ratios. Using different flow intensity, the flow patterns are very similar. // All three anastomoses show characteristic heel and toe separation zones. In the FCPP centre a stagnation zone on the floor can not be examined. Shear stress inside the flow separations was significantly lower than normal wall shear stress. High shear stress levels were found inside the transition zones between flow separation and high velocity mainstream. An explanation for the different stenoses and occlusions time of the three different anastomoses can not be found.

Endothelin-1 (ET-1) ist ein gefä?aktives Peptid, welches zusätzlich verschiedenste nicht kardiovaskuläre physiologische und pathophysiologische Effekte besitzt. So wurde z.B. beschrieben, dass ET-1 in der Brustdrüse während der Schwangerschaft und Stillzeit exprimiert wird. Zusätzlich zu den bekannten Nährstoffen und Wachstumsfaktoren konnte auch ET-1 in der Muttermilch nachgewiesen werden, was auf eine physiologische Rolle von ET-1 für die Laktation und den säugenden Nachwuchs hinweist. In der vorliegenden Arbeit sollte die Funktion von ET-1 in der Brustdrüsenentwicklung mit Hilfe von ET-1 transgenen Mäusen aufgeklärt werden. Die eingesetzten transgenen Tiere überexprimieren humanes ET-1 mit den entsprechenden 5''- und 3'' regulatorischen Sequenzen. Mit Hilfe dieser Strategie sollen die ET-1 spezifischen Funktionen während der Brustdrüsenentwicklung untersucht werden. Transgenes ET-1 wurde während der Tragzeit und Stillzeit in der Brustdrüse detektiert. Die Ergebnisse zeigten, dass säugende Neugeborene der ET-1 transgenen Mäuse eine geringere Gewichtszunahme und eine erhöhte Mortalität aufwiesen, welches auf einen Laktationsdefekt hinweist. Die histologische Untersuchung der Brustdrüse während der Tragzeit ergab eine reduzierte Milchkanalausbildung, kollabierte und nicht expandierende Alveoli, vermehrte Adipozytenausbildung und fortbestehende zytoplasmatische Lipidtropfen (CLDs). Zusätzlich war die Expression des Milchproteins WAP reprimiert. Interessanterweise wurde diese Repression nicht durch STAT5, einem beschriebenem Regulator der Milchproteinexpression und Alveolarexpansion, vermittelt, da dessen Aktivität unverändert war. Als Konsequenz dieses Laktationsdefekts konnte eine verfrühte Rückbildung der Brustdrüse festgestellt werden. Diese ging mit einer erhöhten Expression von STAT3 einher. Interessanterweise wies der bekannte Aktivator von STAT3, LIF, ebenfalls eine gesteigerte Aktivität auf, sowohl während der Tragzeit als auch während der Laktation. Zusätzlich zu den beschriebenen Defekten bei der Milchabgabe zeigten histologische Untersuchungen der Brustdrüse eine Laktationshyperplasie während der mittleren Laktationsphase. In diesem Zusammenhang wird darauf hingewiesen, dass ET-1 Rezeptoren, neben den klassischen Signalwegen dieser G Protein-gekoppelte Rezeptoren, auch mit Tyosinkinaserezeptoren wie z. B. dem EGFR interagieren können. Brustdrüsen von ET-1 transgenen Tieren zeigten eine erhöhte Aktivität sowohl von EGFR als auch von ERK1/2, welches im Zusammenhang mit dem hyperplastischen Phänotyp stehen könnte. Die mögliche tumorfördernde Wirkung von ET-1 wird ferner durch die erhöhte Expression von Amphiregulin, einem EGFR-Liganden, während der Tragzeit und der Laktation verstärkt. Zusammenfassend konnte festgestellt werden, dass ET-1 sowohl die Milchsekretion als auch den Milcheinschuss negativ beeinflusst, so dass eine ausreichende Versorgung säugender Jungtiere in der 1. Hälfte der Laktationsperiode nicht mehr gewährleistet ist. Zusätzlich verursachte ET-1 eine Laktationshyperplasie, welche auf die Induktion der EGFR-Achse zurückzuführen ist. Zusammenfassend kann somit festgestellt werden, dass die Ergebnisse auf eine wichtige Rolle von ET-1 in der Brüstphysiologie des Säugers hinweisen. / Endothelin-1 (ET-1) is a potent vasoactive peptide having wide physiological effects on vascular homeostasis and on a variety of pathophysiological processes unrelated to cardiovascular system. It has been noted that ET-1 is expressed in mammary glands during pregnancy and lactation periods. Furthermore, ET-1 is secreted into milk, suggesting additional physiological roles in the lactating mother and in the suckling neonate. Hence, the present study was proposed to elucidate the possible functional roles of ET-1 in mammary gland development employing ET-1 transgenic mice. ET-1 transgenic mice had been generated by using a human genomic ET-1 construct containing 5´ and 3´ regulatory sequences. This transgenic construction strategy grants to analyse the specific functions of ET-1 in normal mammary gland physiology. The transgene expression was found in mammary gland during pregnancy and lactation. ET-1 transgenic mice exhibited a lactational incompetence with reduced weight gain and increased mortality of their newborns, as a result of a secretory defect. In virtue of this defect, ET-1 transgenic mammary glands histologically revealed a reduced ductal outgrowth, collapsed alveoli with a reduced expansion capacity, increased adipocyte accumulation, and persistence of cystoplasmic lipid droplets (CLDs) during lactation. In addition, the expression of the milk protein, WAP, was found to be constantly suppressed in ET-1 mammary glands although the activity of STAT5, which is known to be a regulator of the expression of milk proteins and alveolar expansion, was found to be normal. Furthermore, as a consequence of the secretory defect, ET-1 transgenic mammary glands exhibited focal precocious involution during early stages of lactation along with an increased activity of STAT3. Consistently, the known activator of STAT3, LIF, was strongly upregulated during lactation and pregnancy. Besides the secretory defect of ET-1 transgenic mammary glands, histological analysis revealed a local lactational hyperplasia during the middle of lactation. Alternatively to the classical G protein-coupled receptors GPCR signalling pathways, endothelin receptors are able to communicate with tyrosine kinase receptors such as the epidermal growth factor receptor (EGFR) for which the term receptor transactivation was coined. Mammary glands of ET-1 transgenic animals exhibited an increased activity of the EGFR and ERK1/2, which could contribute to the observed hyperplastic phenotype. In support of the potential tumourigenicity of ET-1, one of the EGFR ligands, amphiregulin, was found significantly upregulated in ET-1 transgenic mammary glands, both during pregnancy and lactation periods. In summary, high levels of ET-1 affect the secretion and the milk let down process. Consequently the normal support of milk for the suckling neonates is severely impaired during the first half of the lactation period. In addition, ET-1 caused lactational hyperplasia in the mammary glands due to the induction of the EGFR axis. This suggests an important role for ET-1 in mammary gland physiology.

Durch die intraoperative Anwendung der extrakorporalen Bypassflow-Messung wurde zum ersten Mal die Messung und Berechnung der hydraulischen Impedanz möglich. Von den in der vorgelegten Untersuchung berücksichtigten Variablen des hydraulischen Impedanzkonzeptes zeigte sich, dass die Phasenverschiebung zwischen Druck- und Flusskurven, gemessen als erster Impedanzphasenwinkel, die größte klinische Bedeutung hatte. Mit Hilfe der Phasenverschiebung wurde die Langzeitprognose von femoro-poplitealen und insbesondere cruralen Bypässen eindeutig vorhersagbar. Sämtliche Bypässe zu Ausstromgebieten, die einen hohen Wellenwiderstand von < -40° aufwiesen, waren innerhalb von 16 Monaten nach der Operation verschlossen. Das Verhalten der Impedanzparameter nach Prostaglandinapplikation bestätigte die bisherigen tierexperimentellen Erfahrungen. Durch die induzierte Vasodilatation kam es neben der Senkung des linearen Strömungswiderstandes auch zu einem deutlichen Abfall des pulsatilen Wellenwiderstandes. Es zeigte sich, dass die isolierte Prostaglandinreaktion ein weiterer Prognosefaktor von cruralen Bypässen war. Sämtliche Rekonstruktionen, die zu Anschlussgefässen geführt wurden, welche keine Prostaglandinreaktion mehr zeigten, waren innerhalb von 9 Monaten postoperativ verschlossen. Hierdurch konnte die Genauigkeit der Vorhersagbarkeit von Bypassverschlüssen noch verbessert werden. Es zeigte sich außerdem in der vorgelegten Studie, dass konventionelle Angiographien weder mit der Prognose der peripheren Rekonstruktionen noch mit den intraoperativ gemessenen linearen oder pulsatilen Widerständen korrelierten. Dieses ist um so mehr von Bedeutung, da die Operationsplanung in der Regel anhand der Angiographien erfolgt. In kritischen Fällen, in denen der periphere Abstrom aufgrund einer schwachen Kontrastierung der Anschlussgefässe nicht sicher zu bestimmen ist, sollten aus diesem Grund zusätzliche dynamische Untersuchungen, wie die farbkodierte Duplexsonographie oder Magnetresonanz-Angiographie erfolgen. Der hohe lineare und pulsatile Widerstand ist zusammen mit der Anastomosengeometrie weiterhin dafür verantwortlich, dass es innerhalb der Anastomosen zu ausgeprägten Flussseparationen kommt. Wie mit Hilfe der Particle Image Velocimetry-Untersuchung gezeigt wurde, finden diese sich innerhalb der klassischen End-zu-Seit-Anastomosen im Bereich der Haube und der Fersenregionen. Darüber hinaus kommt es in Abhängigkeit von der Geometrie zu einer Stagnationszone auf dem Boden der Anastomose. Innerhalb dieser Flussablösungszonen fanden sich Scherstressmuster, die deutlich unterhalb des normalen Wandscherstressniveaus lagen. Zwischen dem Zentralstrom und den Separationszonen ließen sich darüber hinaus Übergangszonen mit deutlich höherem Scherstress nachweisen. Diese Befunde unterstützen die sogenannte low-shear Theorie, welche zur Erklärung der subendothelialen Intimahyperplasie herangezogen wird. Die räumliche Korrelation der typischen Prädilektionsstellen der Intimahyperplasie mit den durch PIV nachgewiesenen Flussablösungen zeigte eine sehr hohe Übereinstimmung. Neben den Hauben- und Fersenseparationen entwickelte sich eine Stagnationszone am Boden des Empfängergefässes. Ein weiterer wichtiger Aspekt in diesem Zusammenhang war der erstmals erhobene Befund, dass in der Übergangszone zwischen Zentralstrom und Separationen normale Wandscherbedingungen vorlagen. Hierbei könnte es sich um das erforderliche Stopsignal für die Endothelzellen handeln, die Stimulation der subendothelialen Myozyten einzustellen. / The intraoperative use of the extracorporeal bypass flow method allowed for the first time measurement and calculation of hydraulic impedance. The phase shift between flow and pressure waves, as represented by the first phase angle, showed a strong correlation to graft patency. All grafts presenting with high impedance values of < -40°, were occluded within 16 month after the operation. The reaction of the impedance parameters following application of prostaglandin E1 correlated with the previously known experimental results. The induced vasodilation led to a significant decrease of the first phase angle. The prostaglandin reaction itself also proved to be a valuable prognostic factor. All reconstructions to a vasculature with a negative prostaglandin reaction were occluded within 9 months after the operation. The prostaglandin response therefore provided an enhancement of the prognostic tools associated with graft patency. In addition it was shown, that the conventional angiographies did not correlate with graft prognosis. This is particularly important, since angiographies are used for the preoperative planning of the operation. In critical cases, additional assessment of runoff by means of duplex sonography or magnetic resonance angiography is necessary. The high pulsatile resistance is responsible for flow separation inside distal anastomoses of peripheral bypasses. Using Particle Image Velocimetry it was shown, that significant separation areas were present at the heel, the hood and the floor of the anastomoses. The shear stress inside these areas was significantly lower than normal wall shear stresses. At the transition between separation and mainstream very high shear stresses were present, which could provide the stop signal for the signal cascades, leading to subendothelial hyperplasia, which leads to graft stenoses.

L’hyperplasie bilatérale macronodulaire des surrénales (HBMS) conduit au développement de nodules corticosurrénaliens bilatéraux entraînant un syndrome de Cushing diagnostiqué souvent au cours de la cinquième décennie. L’étiologie de cette maladie n’était que partiellement connue mais le caractère bilatéral de l’atteinte surrénalienne et l’observation de cas familiaux suggéraient, au début de ce travail, une origine génétique. L’analyse de l’ADN tumoral et leucocytaire d’une série de 33 patients opérés a permis de mettre en évidence chez 25% des patients une perte d’hétérozygotie neutre en nombres de copies (LOH) de tout le bras court du chromosome 16 (16p) au niveau du tissu surrénalien. Un séquençage complet du génome de 5 patients (ADN germinal et tumoral) a permis de mettre en évidence une mutation du gène ARMC5 localisé en 16p pour 4 patients. Le séquençage direct des parties codantes d’ARMC5 à partir de l’ADN somatique et germinal de l’ensemble des patients opérés de la cohorte a montré qu’au total, 55% des patients avaient une mutation d’ARMC5. L’inactivation du gène se fait selon la théorie de Knudson (un événement germinal inactivant le premier allèle associé à un autre événement somatique inactivant le second allèle) ce qui laisse supposer qu’ARMC5 est un gène suppresseur de tumeur. L’analyse de la cohorte d’HBMS de nos collaborateurs américains au National Institute Health (laboratoire de CA. Stratakis, Bethesda, USA) a permis de confirmer que les mutations d’ARMC5 étaient un événement fréquent. Des variants de ce gène sont aussi associés à l’hypertension à rénine basse chez les patients noirs-américains. Afin de déterminer des corrélations génotype-phénotype, notre cohorte initiale a été élargie pour constituer une série consécutive de 98 cas index de patients présentant des formes légères à sévères de la maladie, opérées ou non. Vingt-quatre patients (25%) présentaient une altération d’ARMC5. Par ailleurs, 31 nodules surrénaliens de 19 patients ont été analysés en somatique. Le second événement était une mutation dans 68% des cas et une LOH du locus pour les 32% restant. Chez un même patient, le second événement était différent dans chaque nodule présenté. Les patients mutés avaient un syndrome de Cushing plus sévère cliniquement et biologiquement par rapport aux patients non mutés. La taille de leurs surrénales étaient plus grandes avec un plus grand nombre de nodules. Les patients mutés étaient aussi plus jeunes au diagnostic et plus souvent hypertendus. Ces patients étaient ainsi plus souvent opérés. La fonction de la protéine ARMC5 n’était pas connue lors de son identification comme gène de l’HBMS. In vitro, la surexpression du gène sauvage induit l’apoptose. La surexpression des mutants faux-sens et du mutant p.F700del retrouvés chez les patients entraîne moins d’apoptose qu’ARMC5 sauvage. La protéine ARMC5 contient des domaines Armadillo et BTB, connus pour être impliqués dans l’interaction protéine-protéine. En physiologie, l’ACTH stimule la production d’AMPc et la voie de la protéine kinase A (PKA) est impliquée dans différentes pathologies corticosurrénaliennes. Nous avons pu montrer qu’ARMC5 interagissait avec la sous-unité catalytique alpha de la PKA. L’invalidation d’ARMC5 conduit in vitro à une diminution de l’expression d’enzymes de la stéroïdogénèse, de la production de cortisol et une diminution de l’activité PKA. Ainsi, l’hypothèse pour expliquer les HBMS liées à une inactivation d’ARMC5 est que la perte d’apoptose conduit à une hyperplasie nodulaire du tissu corticosurrénalien et que, même si la production de cortisol est diminuée à l’échelle unicellulaire, l’effet de masse global conduit au total à un hypercortisolisme. Nos travaux ont donc identifié et caractérisé un premier gène causal, ARMC5, fréquemment impliqué dans l’HBMS et associé à des formes plus sévères de la maladie. Cette découverte ouvre des perspectives pour le diagnostic familial et la prise en charge des patients. (...) / Primary bilateral macronodular adrenal hyperplasia (PBMAH) is a rare cause of adrenal Cushing’s syndrome and bilateral adrenal tumors. We suspected a genetic origin of the disease on the basis of the report of some familial cases and the involvement of both adrenal glands. The aim of this study was to find a genetic cause of non syndromic PBMAH. To look at chromosomal abnormalities, we use single-nucleotide polymorphism (SNP) arrays and microsatellite markers analysis in a first series of 33 patients all operated for PBMAH. We realize whole genome sequencing of 5 patients (blood and tumor DNAs matched). Then we genotyped by Sanger sequencing the gene Armadillo Repeat Containing 5 (ARMC5) in this first series and 66 additional patients. Clinical data were collected to establish genotype-phenotype correlation. In addition, the cohort of patients of our collaborators at the National Institute Health (Dr. Stratakis, Bethesda, USA) was studied. The effects of ARMC5 inactivation and overexpression and the partners of the protein were sought in cell-culture models. The most frequent somatic alteration was a loss of heterozygosity at 16p observed in tumors of 25% of the patients. The gene ARMC5, located at 16p11.2, was the most frequently mutated by whole genome sequencing: a mutation was found in 4/5 patients. 55% of the patients of the first cohort (33 patients treated by adrenalectomy for PBMAH) had ARMC5 alteration. One patient presented with germline microdeletion of the locus identified by SNP array. Every patient had two events: either a mutation or a deletion at the germline level, either a second mutation or a LOH at the somatic level. We showed that the two events were present on different alleles suggesting that ARMC5 is a tumor suppressor gene. In addition, we showed for several patients that the second hit was different in each adrenal nodules of a same patient. This first cohort included only operated patients with serious forms of the disease. The study of the American cohort and the analysis of the total cohort of our lab including non-operated patients and milder forms showed an alteration of ARMC5 in about 25% of the patients. Genotype-phenotype correlation showed that ARMC5 defects are associated with younger age at the diagnosis, higher hypercortisolism, bigger adrenals and higher number of nodules. In addition, a mutation of ARMC5 was shown in a patient with a PBMAH secreting both aldosterone and cortisol. Analysis of a series of patient affected by primary hyperaldosteronism suggested that ARMC5 may be associated with hypertension especially in African-American subjet. Overexpression of ARMC5 leads in vitro to cell apoptosis. We showed that this apoptosis was reduced when transfecting vector harboring missense mutations or single amino-acid deletion found in our cohort. Invalidation of ARMC5 leads to a decreased steroidogenic enzymes expression, cortisol production and reduced protein kinase A (PKA) activity. We showed that ARMC5 interacts with the calaytic subunit alpha of the PKA dissociated from the cAMP-bound regulatory subunits. More than one quarter of sporadic PBMAH patients present a pathogenic germline ARMC5 defect and these index cases present a more severe disease. Systematic genotyping of ARMC5 may help for early diagnosis of PBMAH, familial counseling, and patients’ management. ARMC5 appears to be a new regulator of PKA and might represent a new target for the development of pharmacological agents controlling PKA function and cortisol production.

Die subendotheliale myointimale Hyperplasie (MIH) stellt eine der Hauptursachen für die Ausbildung von Stenosen und Verschlüssen im Bereich von Anastomosen dar. Besondere Bedeutung hat die MIH in termino-lateralen Anastomosen. An der Entstehung der MIH sind hämodynamische Faktoren entscheidend beteiligt. Ausgehend von der Annahme, daß eine Veränderung der Anastomosenform die Hämodynamik beeinflussen kann, wurde in der vorliegenden Arbeit eine neue Anastomosenform, die femorocrurale Patchprothese (FCPP), entwickelt mit dem Ziel einer Optimierung der Strömungsverhältnisse im Anastomosenbereich. In einem hydrodynamischen Kreislaufmodell wurden elastische, transparente Silikonmodelle von termino-lateralen Anastomosen mit einem blutanalogen Newtonschen Fluid (Glycerol-Wasser-Gemisch) unter Simulation der femorocruralen Druckkurve pulsatil bei Variation der Strömungsbedingungen perfundiert. Die konventionellen und klinisch erprobten Anastomosenformen (termino-laterale Anastomose, Composite Bypass, Linton Patch, Miller Collar, Taylor Patch) wurden in vitro mit der FCPP-Anastomose und zwei Modifikation dieser Anastomosenform verglichen. Die Visualisierung des Strömungsfeldes wurde mit drei verschiedenen Methoden erreicht. Bei der farboptischen Methode wurde die Verteilung und Bewegung von Farbteilchen im Anastomosenbereich nach Injektion in das strömende Fluid mittels Video aufgezeichnet. Für die Ultraschalluntersuchung mittels hochauflösender farbcodierter Dopplersonographie (FKDS) wurde das Fluid mit Sephadex-Partikeln dotiert. Sämtliche Ultraschalluntersuchungen wurden ebenfalls mittels Video aufgezeichnet. Die Dopplerspektren korrespondierender Punkte in den Randzonen der Anastomosen wurden off-line der Fast Fourier Analyse (FFT) unterzogen und diese dreidimensional dargestellt. Die semiquantitative Analyse anhand der farboptischen Methode zeigte eine signifikante Verringerung bis Elimination MIH-assoziierter Strömungsphänomene in der FCPP-Anastomose und deren Modifikationen. Die Ausprägung der einzelnen Strömungsphänomene war abhängig von der Reynolds-Zahl und von dem Verhältnis von proximalem und distalem Stromzeitvolumen. Als Vorteil der FCPP erwies sich die Möglichkeit der Anpassung ihrer Form an die entsprechende Flußsituation durch Modifikation der Gabelform mit annähernd laminarer Strömung im gesamten Anastomosenbereich. Die FKDS bestätigte die Ergebnisse der farboptischen Methode. In der Peak-Systole und am Beginn der Diastole waren Rezirkulationszonen und Zonen niedriger Strömungsgeschwindigkeit in der FCPP am geringsten ausgeprägt oder nicht mehr nachweisbar (modifizierte FCPP). Der Vergleich der 3D-Darstellungen der FFT der Dopplerspektren zeigte niedrige Frequenzen und damit Flußgeschwindigkeiten und partiell eine Strömungsumkehr nahezu konstant in allen Anastomosenformen außer der FCPP und deren Modifikationen. In einer prospektiven Studie zum klinischen Einsatz der FCPP Anastomose wurden im Zeitraum von 6 / 1992 bis 7 / 1998 135 PTFE-Prothesenbypass-Rekonstruktionen mit distaler FCPP Anastomose bei 129 Patienten im klinischen Stadium III und IV der paVK analysiert. Die kumulativen primären und sekundären 1-, 2-, 3-, 4-, und 5-Jahres-Offenheitsraten nach Kaplan / Meier betrugen jeweils 63,0%, 44,9%, 35,7%, 33,1% und 27,6% bzw. 74,5%, 55,2%, 44,8%, 43,0% und 37,6%. Die kumulativen 1-, 3-, und 5-Jahres-Wahrscheinlichkeiten für den Erhalt der Extremität betrugen jeweils 86,8%, 79,2% und 77,5%. Eine Optimierung des Strömungsverhaltens innerhalb der Anastomosenregion ist in der FCPP derart möglich, daß der Anteil der für die Pathogenese der MIH ursächlichen Strömungsmuster minimiert wird. Diese Anastomose ist klinisch anwendbar und führt bei ausschließlicher Verwendung von ePTFE als Bypassmaterial im femorodistalen Bereich zu akzeptablen Langzeitergebnissen, die denen bei Anwendung von alternativ möglichen Venenpatchplastiken nicht nur vergleichbar, sondern teilweise überlegen sind. / The subendothelial myointimal hyperplasia (MIH) represents one of the main etiological factors in the formation of stenoses and occlusions of vascular anastomoses. MIH plays a role especially in termino-lateral anastomoses. Hemodynamic factors have a decisive impact on the development of MIH. Assuming that changes in the morphology of the anastomoses influence the hemodynamics, a novel anastomosis form, the femorocrural patch prosthesis (FCPP), was developed with the goal of optimizing the blood flow-dynamics within the anastomotic site. In a hydrodynamic circulation model, various elastic, transparent silicon phantoms of termino-lateral anastomoses were perfused with a Newton fluid blood analog (glycerol-water mixture) while simulating the femorocrural pressure curve in a pulsatile manner under variation of the flow conditions. The conventional and clinically tested anastomosis forms (termino-lateral anastomosis, composite bypass, Linton patch, Millar collar, Taylor patch) were compared with the FCPP-anastomosis and two modifications of the FCPP in vitro. The visualization of the flow velocity field was achieved using three different methods. By means of the color-optic method, the distribution and motion of color elements in the anastomotic area were video-recorded following injection in the flowing liquid. For the ultrasound examination with high resolution, color-coded Doppler sonography, the fluid was marked with Sephadex particles. All ultrasound examinations were also recorded on video. The corresponding doppler spektrum points in the marginal zones of the anastomosis were subjected to the Fast Fourier Transform (FFT) analysis off-line and then displayed three-dimensionally. The semiquantitative analysis using the color-optic method showed a significant decrease or elimination of MIH-associated current phenomena in the FCPP-anastomosis and its corresponding modifications. The intensity of singular flow phenomena was dependent upon the Reynolds-number and upon the relation of proximal to distal flow volume over time. The possibility to adapt the FCPP to the flow phenomena by modifying the bifurcated form and thereby achieving almost laminar flow in the complete anastomotic area proved to be advantageous. The color-coded Doppler sonography confirmed the results of the color-optic method. During peak systole and at the beginning of the diastole, recirculation zones and zones with low flow velocity were least intense in the FCPP or not detectable at all (in the modified FCPPs). The comparison between the FFT 3D-reconstructions from the Doppler spektrum showed low frequencies and thus flow velocities and a partial flow reversal almost constantly in all anastomosis forms except the FCPP and its modifications. In a prospective study on the clinical application of the FCPP anastomosis during the period of June 1992 to July 1998, 135 ePTFE prosthetic bypass reconstructions with distal FCPP anastomosis were analyzed in 129 patients suffering from peripheral arterial occlusive disease stages III and IV. The cumulative primary and secondary 1-, 2-, 3-, 4-, and 5-year patency rates calculated with the Kaplan / Meier method were 63.0%, 44.9%, 35.7%, 33.1%, and 27.6%, and 75.5%, 55.2%, 44.8%, 43.0% and 37.6%, respectively. The cumulative 1-, 3-, and 5-year probabilities for limb salvage were 86.8%, 79.2% and 77.5%, respectively. As a result of anastomotic engineering, FCPP optimizes the flow properties within the anastomosis region to such an extent that the pathogenetic role of flow disturbances is minimized. This anastomosis is clinically employable when using ePTFE as bypass material in femoro-distal vascular reconstruction and leads to acceptable long-term results that are not only comparable but sometimes superior to the results obtained when using alternative vein cuff techniques.