Evidências sugerem que o fator de crescimento de fibroblasto 2 (FGF2), produzido por melanomas, possui importante papel no crescimento tumoral, angiogênese e metástase. Assim, o uso de anticorpo monoclonal (mAb) que reconhece e bloqueia a atividade de FGF2 é uma abordagem a ser considerada em oncologia. O propósito desse estudo foi avaliar a aplicação diagnóstica e terapêutica de um novo anticorpo anti-FGF2, 3F12E7 IgG1, em melanoma experimental B16-F10. Para isso, camundongos C57Bl/6 foram implantados subcutaneamente (ou intravenosamente, para ensaios de metástase) com células de melanoma murino B16-F10 (5x105 células/animal). Quando tumores alcançaram 3-4 mm de diâmetro (ou 24 h pós-inóculo de células B16-F10, no caso de ensaios de metástase), camundongos foram tratados com anti-FGF2 3F12E7 IgG. Animais controle receberam igual volume do veículo ou quantidade de anticorpo controle de isotipo. Grupos: animais tratados com (1) anti-FGF2 3F12E7 IgG1; (2) ligante de CEA IgG1 (controle de isotipo); e (3) veículo. O tratamento dos camundongos portadores de tumor com anti-FGF2 IgG resultou, comparado com os controles salina e de isotipo, em uma redução no número de focos metastáticos nos pulmões (ANOVA, p < 0,05), em ensaios de metástase experimental, bem como em uma menor taxa de crescimento de tumores subcutâneos (n=7/grupo). Esse resultado é acompanhado por uma redução na densidade vascular do tumor, conforme determinado por imunomarcação para CD34 ou CD31. A captação tumoral de anti-FGF2 3F12E7 IgG foi avaliada por métodos de medicina nuclear, usando esse anticorpo radiomarcado com tecnécio-99m. Estudos SPECT/CT in vivo e de biodistribuição ex vivo revelaram que 99mTc-anti-FGF2 3F12E7 IgG pode atingir eficientemente tumores subcutâneos e metastáticos de B16-F10. Assim, esses dados sugerem que anti-FGF2 3F12E7 IgG pode ser uma estratégia antitumoral promissora para melanoma, bem como uma potencial ferramenta de imagem a ser explorada, atuando como um possível traçador para rastrear tumores FGF2-positivos e mapear esse estímulo angiogênico no microambiente tumoral. Aprovado pelo comitê de ética (CAPPesq): número 0942/09 / Compelling evidence suggests that fibroblast growth factor 2 (FGF2), produced by melanomas, plays important role in tumor growth, angiogenesis and metastasis. Therefore, the use of a monoclonal antibody (mAb) that recognizes and blocks FGF2 activity is seen as an approach to be considered in oncology. The purpose of this study was to evaluate the diagnostic and therapeutic application of a new anti-FGF2 antibody, 3F12E7 IgG1, in experimental melanoma B16-F10. For this, C57Bl/6 mice were subcutaneously (or intravenously, for experimental metastasis assay) implanted with murine melanoma B16-F10 cells (5x105 cells/animal). When tumors reached 3-4 mm in diameter (or 24 h after B16-F10 cells injection, in the case of metastasis assay), mice started receiving anti-FGF2 3F12E7 IgG. Control mice received equal volume of vehicle or isotype control IgG amount. Groups: (1) anti-FGF2 3F12E7 IgG1-treated, (2) CEA-binding IgG1-treated (isotype control) and (3) vehicle-treated mice. The treatment of tumor-bearing mice with anti-FGF2 IgG, compared with saline and isotype controls, led to a reduction in the number of metastatic foci in the lungs (ANOVA test, p < 0.05), in experimental metastasis assays, as well as to a lower subcutaneous tumor growth rate (n=7 per group). This result is accompanied by a reduction in the tumor vascular density, as determined by CD34 or CD31 staining. The anti-FGF2 3F12E7 IgG tumor uptake was evaluated by nuclear medicine approaches, using this antibody radiolabeled with technetium-99m. In vivo SPECT/CT and ex vivo biodistribution studies reveled that 99mTc-anti-FGF2 IgG could efficiently achieved B16-F10 subcutaneous and metastatic tumors. Thus, these data suggest that the anti-FGF2 3F12E7 IgG may be a promising antitumor strategy for melanoma, as well as a potential imaging tool to be explored, working as a possible tracer to identify FGF2-positive tumors and map this angiogenic stimulus in the tumor microenvironment. Ethics committee (CAPPesq) approval number 0942/09

Pentoxifilina (PTF), um derivado da metilxantina, tem ação terapêutica como agente antifibrótico. In vitro, a PTF causa inibição na produção de colágeno, glicosaminoglicanos e fibronectina, bem como promove acentuada redução na proliferação dos próprios fibroblastos de quelóides. A PTF na concentração de 1.000 g/mL foi seletiva no controle da inibição da síntese protéica pelos fibroblastos. O objetivo deste estudo foi verificar o comportamento in vitro de fibroblastos oriundos de cicatrizes hipertróficas (HSHF) e de pele não-cicatricial (NHF) na presença e ausência de PTF (1.000 g/mL), quanto à: proliferação celular, produção de colágeno e capacidade dos fibroblastos gerarem contração em modelo experimental de matriz de colágeno. Para tanto, foram utilizados fibroblastos cultivados a partir de amostras de cicatrizes hipertróficas e pele não-cicatricial doadas, com finalidade de pesquisa, pelo banco de Tecidos do Instituto Central do Hospital das Clínicas da FMUSP. Culturas celulares expostas a PTF apresentaram diminuição na proliferação celular em HSHF (46,35%) e em NHF (37,73%) (p<0,0001). Na presença de PTF, foi observada seletividade de inibição na síntese de colágenos, havendo inibição mais expressiva de colágeno tipo III em HSHF e de colágeno tipo I em NHF (p<0,0001). O modelo experimental de matriz de colágeno povoada por fibroblastos de cicatriz hipertrófica apresentou contração menor (12%) na presença de PTF (p<0,0001), em relação à sua ausência / Fibroblasts are thought to be partially responsible for the persisting contractile forces that result in burn contractures. Using a monolayer and fibroblast populated collagen lattice (FPCL) three-dimensional (3D) model we subjected hypertrophic scar and non-cicatricial fibroblasts to the antifibrogenic agent pentoxifylline (PTF) 1000g/mL attempting to reduce proliferation, collagen type I and III synthesis and contraction in this 3D model. Fibroblasts were isolated from post burn hypertrophic scars (HSHF) and non-scarred skin (NHF). Cells were grown in monolayer or incorporated into FPCL\'s and exposed to PTF. In monolayer, cell number proliferation was reduced (46.35% in HSHF group and 37.73% in NHF group) p<0,0001. The PTF also demonstrated to be selective on collagen type I and III synthesis inhibition suggesting higher inhibition of collagen type III on HSHF group and more evident inhibition of type I on NHF group. FPCL\'s containing PTF had surface areas reduced in about 12% p<0,0001. PTF showed inhibition effects on cell proliferation and reduced contraction in both HSHF and NHF

Fluid pressures regulate endothelial cell (EC) tubulogenic activity involving fibroblast growth factor 2 (FGF-2) and its receptor, FGF receptor 2 (FGFR2). Our lab has recently shown that sustained 20 mmHg hydrostatic pressure (HP) upregulates EC sprout formation in a FGF2-dependent fashion. This upregulation of sprout formation may be due to enhanced FGF-2 / FGFR2 interactions in the presence of 20 mmHg HP. We hypothesize that exposure of ECs to 20 mmHg sustained HP enhances FGF-2 binding kinetics. We used a custom hydrostatic pressure system, immunofluorescence, and FACS to quantify FGF-2 binding by ECs in the absence or presence of a range of HPs for 30 minutes. Relative to cells maintained under control pressure, ECs exposed to 20, but neither 5 nor 40 mmHg, displayed a significant increase in binding affinity to FGF-2. EC binding of VEGF-A, another angiogenic growth factor, was unaffected by similar pressure stimuli. Additional studies showed that pressure-selective FGF-2 binding was independent of FGFR2 surface expression. These results implicate the FGF-2 axis in the pressure-sensitive, magnitude-dependent angiogenic processes which we have previously described. The present study provides novel insight regarding the involvement of FGF-2 signaling and interstitial pressure changes in various microvascular physiological and pathobiological processes.

Dysregulated wound healing is commonly associated with excessive fibrosis. Connective tissue growth factor (CTGF/CCN2) is characteristically overexpressed in fibrotic diseases and stimulated by transforming growth factor-β (TGF-β) in dermal fibroblasts. Reepithelialisation and epidermal wound coverage counteract excessive scar formation. We have previously shown that interleukin-1α (IL-1α) derived from keratinocytes conteracts TGF-β-stimulated CTGF-expression. The aim of this thesis was to further explore the effects of keratinocytes and IL-1α on gene and protein expression, as well as pathways, in TGF-β stimulated fibroblasts. Fibroblasts were studied in vitro by conventional two dimensional cell culture models and in a three dimensional keratinocyte-fibroblast organotypic skin culture model. The results showed that IL-1 suppresses basal and TGF-β-induced CTGF mRNA and protein, involving a possible TAK1 mechanism. Keratinocytes regulate the expression of fibroblast genes important for the turnover of the extracellular matrix. Most of the genes analysed (11/13) were regulated by TGF-β and counter regulated by keratinocytes. The overall results support a view that keratinocytes regulate fibroblasts to act catabolically (anti-fibrotic) on the extracellular matrix. Transcriptional microarray and gene set enrichment analysis showed that antagonizing effects of IL-1α on TGF-β were much more prominent than the synergistic effects. The most confident of these pathways was the interferon signaling, which were inhibited by TGF-β and activated by IL-1α. A proteomics study confirmed that IL-1α preferentially conteracts TGF-β effects. Six new fibroblast proteins involved in synthesis/ regulation were identified, being regulated by TGF-β and antagonized by IL-1α. Pathway analysis confirmed counter-regulation of interferon signaling by the two cytokines. These findings have implications for understanding the role of fibroblasts for inflammatory responses and development of fibrosis in the skin.

Abstract Chronic, non-healing wounds constitute a massive financial burden on health care system. The healing processes of these wounds and their underlying pathology are only partly understood. In this study, important biological functions performed by Retinoic acid with its regulatory protein cellular retinoic acid binding protein 2 (CRABP2) were discussed. Possibly, these biological func-tions might be linked with chronic wound therapeutic by inducing antiproliferative activity of cells which leads to reduction in migration and growth rate of fibroblast during skin regeneration pro-cess in chronic wound healing. The aim of this study was to comparatively analyze the expression pattern of CRABP2 and P16 cyclic dependent kinase inhibitor in dermal fibroblasts at mRNA levels along with their morphological pattern, migration and growth rate. Fibroblasts were cultured and their morphology were observed by phase-contrast imaging. Difference in viability, migratory capacity was examined by Cell titer blue and scratch assay respectively and expression were meas-ured by polymerase chain reaction. Interestingly, the date revealed that morphology was altered and growth rate and migration velocity was significantly lower in chronic wound fibroblasts and senescent fibroblasts when compared with their control. Expression pattern revealed that CRABP2 was highly up-regulated only by senescent cells but not in chronic wound fibroblasts which point novel function for this protein in term of replicative senescence. However, P16 was not signifi-cantly altered among all fibroblasts which demands supplementary studies to conform the role of CRABP2 in fibroblast dysfunction and cellular senescence in chronic wounds.

La morphogenèse crâniofaciale chez les vertébrés est un phénomène important, strictement régulé dans l’espace et dans le temps. Elle est basée sur une série complexe d'événements moléculaires et morphogénétiques qui implique un réseau interactionnel de gènes et de facteurs de transcriptions, tels les homéoboîtes. La crête neurale (CN) est au cœur de ce processus. Cette dernière fournit la principale source du mésenchyme crâniofacial. Cette population de cellules embryonnaires transitoires va subir une transition épithélio-mésenchymateuse et migrer en plusieurs vagues vers des sites prédéfinis puis se différencier en divers types cellulaires. La CN est à l’origine de plusieurs structures : une grande partie du squelette facial dont le maxillaire, la mandibule, l’os alvéolaire qui entoure les dents ainsi qu’une partie des tissus conjonctifs crâniofaciaux. Les cellules issues des CN sont pluripotentes et offrent un espoir en régénération osseuse et cartilagineuse. Ces caractéristiques ont généré un intérêt particulier des chercheurs pour les utiliser en thérapie cellulaire afin de réparer les défauts osseux des mâchoires. Parmi les tissus crâniofaciaux, nous avons choisi d’étudier plus avant la gencive et les cellules gingivales car leur accès est le plus facile et leurs capacités de différenciation autorisent l’observation d’autres phénotypes cellulaires.La gencive est un tissu kératinisé qui entoure les dents et recouvre l’os alvéolaire. Ce tissu est composé principalement de fibroblastes gingivaux (GFs). Parmi ces cellules, se trouvent des cellules souches gingivales (GSCs) caractérisées par leur auto-renouvellement et leur multipotence. Les GSCs sont facilement recueillies chez les patients adultes, elles montrent une plasticité importante et une activité immunomodulatrice qui en font un outil de choix pour la thérapie cellulaire. De plus, la biopsie se fait sans douleur et n’entraîne ni cicatrice ni problème fonctionnel.La première partie de mon travail de doctorat avait pour objectif d’évaluer le rôle de Msx1 dans la morphogenèse crâniofaciale et par la suite d’analyser l’os alvéolaire après une extraction dentaire afin d’analyser les mécanismes associés à ce processus et l’impact de Msx1 sur la cicatrisation osseuse.La deuxième partie de mon travail est axé sur la gencive et avait pour objectif de mettre en évidence l’origine embryologique des cellules souches orales, dont les GSCs, et de déterminer si elles proviennent des crêtes neurales, du mésoderme ou d’une mosaïque des deux. Enfin, pour appliquer nos connaissances sur l’origine embryologique des cellules souches gingivales, nous avons étudié le profil immunitaire des cellules dérivées des CN. Pour cela, nous avons déterminé la capacité phagocytaire des cellules souches gingivales murines dérivées des CN et comparé à des cellules de CN d’autres espèces vertébrées. / Craniofacial morphogenesis in vertebrates is an important phenomenon, strictly regulated in space and in time. It is based on a complex series of molecular and morphogenetic events involving an interactional network of genes and transcription factors, such as the homeobox. Neural crest (NC) is at the heart of this process. The latter provides the main source of craniofacial mesenchyme. This transient population of embryonic cells will undergo epithelial-mesenchymal transition and migrate in waves to predefined sites and to differentiate into various cell types. NC is the source of several structures: a large part of the facial skeleton including the maxillary, mandibular alveolar bone around the teeth as well as connective tissue in craniofacial portion. Cells from NC are pluripotent and offer hope for bone and cartilage regeneration. These characteristics have generated particular interest to researchers for use in cell therapy to repair bone defects of the jaw. Among the craniofacial tissues, we decided to further investigate the gums and gingival cells because their access is easier and differentiation capabilities allow observation of other cellular phenotypes.The gum is a keratinized tissue around the teeth and covers the alveolar bone. This tissue is composed mainly of populations of gingival fibroblasts (GFs). Among these populations, there are gingival stem cells (GSCs) characterized by their self-renewal and pluripotency. The GSCs are easily collected in adult patients, they show significant plasticity and immunomodulatory activity that make it a tool of choice for cell therapy. In addition, the biopsy is painless and involves neither scar nor functional problem.The first part of my PhD work was to evaluate the Msx1 role in craniofacial morphogenesis and subsequently analyse the alveolar bone after tooth extraction to analyse the mechanisms involved in this process and the impact of Msx1 on bone healing.The second part of my work focuses on the gingiva and was intended to highlight the embryological origin of oral stem cells, including GSCs and determine if they come from the neural crest, mesodermal or mosaic two. Finally, to apply our knowledge of the embryological origin of gum stem cells, we studied the immune profile derived NC cells. For this, we determined the phagocytic capacity gingival murine stem cells derived from CN and compared to cells of CN other vertebrate species.

Uma premissa da Biologia Molecular atual estabelece a ativação de programas de transcrição voltados a processos biológicos específicos. Neste trabalho o objetivo é descrever genes regulados que, uma vez agrupados, são capazes de indicar os programas disparados na linhagem corticoadrenal murina Y1 quando tratada com o fator de crescimento de fibroblasto 2 (FGF2) ou pelo hormônio adrenocorticotrópico (ACTH). O papel de ACTH em células Y1 ainda não está bem estabelecido quanto ao seu potencial mitogênico, uma vez que este hormônio é capaz de agir por diferentes vias de sinalização, apresentando um comportamento dual. Ao traçar o perfil de genes regulados por este hormônio, comparando com o padrão observado por tratamentos de indutores clássicos, espera-se determinar o papel de ACTH frente ao ciclo celular. FGF2 é classicamente conhecido por sua atividade mitogênica, de forma que, em células Y1, é capaz de induzir a passagem G0?G1?S do ciclo celular. Recentemente foi descrita uma nova e surpreendente ação de FGF2, como um indutor de morte seletivo, agindo apenas em células potencialmente tumorigênicas (Costa e Armelin, dados não publicados). Foram feitos estudos de expressão gênica com ensaios de microarray, observando-se o padrão de transcrição da linhagem Y1, bem como de sub-linhagens de Y1 resistentes a morte desencadeada por FGF2, submetidas a tratamentos de FGF2, ACTH e soro. Os resultados indicam que a) o padrão de expressão gênica observado quando a linhagem Y1 é submetida a tratamentos de ACTH é diferente daqueles desencadeados por mitógenos clássicos; b) o tratamento de FGF2 regula genes envolvidos na via de MAPK, controle do ciclo celular e genes relacionados a processos de adesão, comunicação intercelular e sinalização a partir da matriz extracelular (ECM); c) o comportamento de morte induzida por FGF2 está relacionado a alterações na estrutura da célula, envolvendo mecanismos de adesão, remodelagem de citoesqueleto e transdução de sinais a partir da ECM. / Nowadays, a molecular biology premise establishes the activation of transcription programs related to specific biological processes. At this work the objective is to describe regulated genes that, once clustered, are able to indicate the running programs when murine adrenocortical lineage Y1 is treated with fibroblast growth factor (FGF2) or by adrenocorticotropin hormone (ACTH). The mitogenic potential of ACTH treatments in Y1 cells is not well established since this hormone acts by different signaling pathways, presenting a dual behavior. By tracing the profile of genes regulated by this hormone and comparing it to the transcription patterns observed in response to classic mitogens it is expected to determine the ACTH role in the cell cycle. FGF2 is known by its mitogenic activity, inducting the G0? G1?S cell cycle transitions in Y1 cells. Recently it has been described a new and surprising feat of FGF2, acting as a selective death inductor, only in potentially tumorigenic cells (Costa and Armelin, unpublished data). Microarray assays were used to determine the transcription patterns observed in Y1 lineage, as well as in FGF2 death-resistant Y1 sub-lineages, submitted to FGF2, ACTH and serum treatments. The results indicate that a) the gene expression profile displayed when Y1 cells are under ACTH treatments is different from the patterns observed when this lineage is submitted to classic mitogens; b) FGF2 treatment regulates genes involved in the MAPK pathway, cell cycle control and genes related to adhesion processes, intercellular communication and signaling from the extra cellularmatrix (ECM); c) the death behavior initiated by FGF2 is related to structural alterations in the cell, involving adhesion mechanisms, citoskeleton remodeling and signal transduction from the ECM.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23062号 / 医博第4689号 / 新制||医||1048(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 武藤 学, 教授 妹尾 浩, 教授 小川 誠司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Orientador: Sergio A.R. de Paiva / Resumo: A deficiência/insuficiência de vitamina D tem aumentado nos últimos anos e tornou-se problema mundial de saúde pública. Além do raquitismo, a deficiência de vitamina D também está associada com maior risco de desenvolver câncer, doenças imunológicas e, inclusive, doenças cardiovasculares. Estes fatores têm incentivado o uso indiscriminado da suplementação de vitamina D na população saudável. Suplementação de vitamina D, em altas doses, promove estresse oxidativo, inflamação, apoptose, altera o metabolismo energético, morfologia e função cardíaca. Adicionalmente, esta suplementação aumenta a concentração sérica de fósforo, que pode estimular a liberação do fator de crescimento de fibroblasto-23 (FGF-23). Esta maior concentração sérica de FGF-23 pode estar associada à remodelação cardíaca. Outros autores sugerem que a ação do FGF-23 no coração ocorre por meio da via de sinalização calcineurina/fator nuclear das células T ativadas (NFAT). Desta maneira, a menor fosfatemia, promovida pelo uso de sevelamer, poderia atenuar as alterações cardíacas provocadas pela suplementação de vitamina D. Assim, o objetivo deste trabalho é verificar se o tratamento com sevelamer diminui a concentração de FGF-23 e, consequentemente, atenua a remodelação cardíaca, decorrente da suplementação de vitamina colecalciferol, em altas doses. Foram utilizados 169 ratos machos da raça Wistar alocados em seis grupos: 1) Grupo controle alimentado com ração padrão (C, n=27); 2) Grupo controle + 3% de sevelame... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Vitamin D deficiency/insufficiency has increased in recent years and has become a worldwide public health problem. In addition to rickets, vitamin D deficiency is also associated with increased risk of developing cancer, immune diseases and even cardiovascular diseases. These factors have encouraged the indiscriminate use of vitamin D supplementation in the healthy population. High-dose of vitamin D supplementation promotes oxidative stress, inflammation, apoptosis, changes in energy metabolism, morphology and cardiac function. In addition, this supplementation increases the serum phosphorus concentration which can stimulate the release of fibroblast growth factor-23 (FGF-23). Higher serum concentration of FGF-23 can be associated with cardiac remodeling. Other authors suggest that the action of FGF-23 in heart occurs through the signaling pathway calcineurin/nuclear factor of activated T cells (NFAT). Thus, decrease in phosphatemia, promoted by the use of sevelamer, can attenuate the cardiac changes promoted by vitamin D supplementation. The aim of this study is to verify whether treatment with sevelamer decreases the concentration of FGF-23 and, consequently, attenuates cardiac remodeling, due to supplementation of cholecalciferol vitamin, in high doses. 169 male Wistar rats were allocated in six groups: 1) Control group fed standard chow (C, n=27); 2) Control group + 3% sevelamer (C+S, n=26); 3) Group supplemented with 3,000 IU cholecalciferol/kg of chow (VitD 3, n=29); 4)... (Complete abstract click electronic access below) / Doutor

Im Rahmen der vorliegenden Dissertation wurde die prognostische Relevanz von Angiopoietin-2 (Ang-2) und Fibroblast Growth Factor 23 (FGF-23) als Prädiktor der 30-Tages sowie Ein-Jahres Mortalität bei Patienten im infarktbedingten kardiogenen Schock (CS) untersucht und dargestellt. Das betrachtete Patientenkollektiv dieser Substudie rekrutierte sich dabei aus den 218, im Rahmen der IABP- SHOCK II Studie in Leipzig eingeschlossenen Patienten. Nach Hospitalisierung und Randomisierung in den jeweiligen Therapiearm (IABP, Nicht-IABP) erfolgte eine prospektiv geplante Blutentnahme an den Tagen eins bis drei. Zur laborchemischen Bestimmung der Serum-, beziehungsweise Plasma Proteinkonzentration mittels ELISA von Ang-2 standen 189, für das Phosphathormon FGF-23 182 Blutproben zur Verfügung. Die 30-Tages Mortalität der untersuchten Substudien-Kohorte betrug 40%, die Ein-Jahres Mortalität 57%. Die vorliegende Arbeit bestätigt Ang-2 als einen starken und unabhängigen negativen Prädiktor des Kurz- und Langzeitverlaufs bei Patienten im CS auf Grund eines akuten Myokardinfarks. Weiterhin zeigt die Auswertung, dass die prognostische Relevanz des betrachteten Biomarkers im zeitlichen Verlauf signifikant zunimmt und verschiedene klinische Parameter wie beispielsweise eine akut eingeschränkte Nierenfunktion oder Blutungskomplikationen unabhängig mit erhöhten Ang-2 Konzentrationen assoziiert sind. Somit sind im infarktbedingten CS hohe Werte von Ang-2 unabhängig mit einem schlechteren klinischen Verlauf des Patienten, sowie dem Reperfusionserfolg und weiteren Komplikationen assoziiert. Patienten der Substudien-Kohorte im infarktbedingten CS waren durch signifikant erhöhte Konzentrationen von FGF-23 charakterisiert. Weiterhin zeigte sich, dass diese signifikant erhöhten Werte unabhängig mit einer deutlich schlechteren Prognose der 30-Tages und Ein-Jahres Mortalität verbunden sind. Diese Assoziation konnte jedoch ausschließlich bei Patienten mit eingeschränkter Nierenfunktion nachgewiesen werden.