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CAPES A mobilização miofascial vem sendo utilizada como intervenção para pacientes com fibromialgia (FM) por atuar sobre as vias nociceptivas ascendentes possivelmente envolvidas no processo da sensibilização central, modulando a experiência da dor. Entretanto, ainda existe uma lacuna da sua eficácia quando comparada com outra abordagem hands on, visto que a terapia manual possui efeitos inespecíficos, como o placebo. Desta forma, este trabalho teve como objetivo verificar o efeito da mobilização miofascial das cadeias fisiológicas do tronco sobre a dor, qualidade de vida e capacidade funcional de pacientes com FM comparado com alongamento muscular passivo e controle. Trata-se de um ensaio clínico controlado randomizado, cego para avaliadores e estatístico, composto por pacientes diagnosticados com FM que obedeceram aos critérios de elegibilidade. Foram realizadas 8 sessões (1x/semana) de mobilização miofascial da cadeia estática posterior e da cadeia de flexão do tronco no Grupo Mobilização (GM), 8 sessões (1x/semana) de alongamento muscular no Grupo Alongamento (GA) e 4 consultas de acompanhamento com reumatologista no Grupo Controle (GC). As avaliações foram realizadas em 4 momentos: baseline, após 4 e 8 semanas e no follow-up (1 mês), com os seguintes instrumentos: escala visual analógica da dor, índice de dor generalizada, questionário de impacto da FM, escala de percepção global de mudança e diário de medicamentos para dor. Para análise estatística foram utilizados os testes de Qui-Quadrado ou Exato de Fisher, ANOVA com Post Hoc de Tukey (distribuição normal) ou Kruskal Wallis com comparação múltipla de Dunn's (não normal) e modelo de regressão linear misto. No total, 38 pacientes com média de idade de 45,92 (± 9,01) foram randomizados, sendo 13 pertencentes ao GM, 13 ao GA, e 12 ao GC. Foi encontrada diferença significativa no escore total do impacto da FM entre o GM e GC, após 4 (Δ = -21,07; IC = -41,65; -0,49; p = 0,044) e 8 semanas (Δ = -33,91; IC = -52,87; -14,97; p < 0,001) e no follow-up (Δ = -20,53; IC = -40,7; -0,36; p = 0,045). Após 8 semanas, também foi encontrada diferença significativa na intensidade da dor entre os grupos GM e GC (Δ = -2,75; IC = -4,97; -0,53; p = 0,012) e entre o GM e GA (Δ = -2,47; IC = -4,75; -0,21; p = 0,030). Diante do exposto, a mobilização miofascial apresentou efeito significativo e clinicamente importante sobre a melhora da dor comparada ao alongamento e ao controle, assim como sobre a melhora da percepção do impacto da FM comparada ao controle. Myofascial release has been used as an intervention for patients with fibromyalgia (FM) because it acts on ascending nociceptive pathways possibly involved in the central sensitization process, modulating the pain experience. However, there is still a gap in its efficacy when compared to another hands-on approach, since manual therapy has nonspecific effects, such as placebo. The aim of this study was to verify the effect of the myofascial release of the trunk physiological chains on the pain, quality of life and functional capacity of FM patients compared to passive muscle stretching and control. It is a randomized controlled trial, blind for evaluators and statistician, composed of patients diagnosed with FM who met the eligibility criteria. Eight sessions (1x / week) of myofascial release of the posterior static chain and the trunk flexion chain were performed in the Mobilization Group (MG), 8 sessions (1x / week) of muscle stretching in the Stretching Group (SG) and 4 medical appointments with a rheumatologist in the Control Group (CG). The evaluations were performed in four moments: baseline, after 4 and 8 weeks and at the follow-up (1 month), with the following instruments: visual analog pain scale, generalized pain index, FM impact questionnaire, perception scale global change and diary drugs for pain. For statistical analysis, Chi-square or Fisher's exact tests were used for the categorical variables; and ANOVA with Post Hoc of Tukey (normal distribution) or Kruskal Wallis with multiple comparison of Dunn's (nonnormal) and mixed linear regression model for the quantitative variables. In total, 38 patients with a mean age of 45.92 (± 9.01) were randomized, 13 of whom were MG, 13 were SG, and 12 were CG. A significant difference was found in the total FM impact score between MG and CG, after 4 (Δ = -21.07, CI = -41.65, -0.49, p = 0.044) and 8 weeks (Δ = 33.91, CI = -52.87, -14.97, p

Nanoparticles can be used as carriers to transport biomolecules like proteins and synthetic molecules across the cell membrane because many molecules are not able to cross the cell membrane on their own. The uptake of nanoparticles together with their cargo typically occurs via endocytosis, raising concerns about the possible degradation of the cargo in the endolysosomal system. As the tracking of a dye-labelled protein during cellular uptake and processing is not indicative of the presence of the protein itself but only for the fluorescent label, a label-free tracking was performed with the red-fluorescing model protein R-phycoerythrin (R-PE). Four different eukaryotic cell lines were investigated: HeLa, HEK293T, MG-63, and MC3T3. Alone, the protein was not taken up by any cell line; only with the help of calcium phosphate nanoparticles, an efficient uptake occurred. After the uptake into HeLa cells, the protein was found in early endosomes (shown by the marker EEA1) and lysosomes (shown by the marker Lamp1). There, it was still intact and functional (i.e. properly folded) as its red fluorescence was detected. However, a few hours after the uptake, proteolysis started as indicated by the decreasing red fluorescence intensity in the case of HeLa and MC3T3 cells. 12 h after the uptake, the protein was almost completely degraded in HeLa cells and MC3T3 cells. In HEK293T cells and MG-63 cells, no degradation of the protein was observed. In the presence of Bafilomycin A1, an inhibitor of acidification and protein degradation in lysosomes, the fluorescence of R-PE remained intact over the whole observation period in the four cell lines. These results indicate that despite an efficient nanoparticle-mediated uptake of proteins by cells, a rapid endolysosomal degradation may prevent the desired (e.g. therapeutic) effect of a protein inside a cell. OA Förderung 2017

Pre-service teachers of all subject areas need learning opportunities in the field of German as a Second Language (GSL) in order to adequately facilitate second language learners in the content classroom. On the basis of a competency model including the sub-competencies Subject-specific registers, Multilinguaiism and Didactics, the BMBF-project DaZKom has conducted a validation study. This included developing a test instrument with 68 items and analyzing it with IRT-methods. The results of the validation study (N = 252) show the expected correlations between GSL competency and linguistic knowledge as well as pedagogical knowledge, yet that test measures a conceptually independent construct. Furthermore, it is found that pre-service teachers with a higher number of learning opportunities and semesters, and with German as a study subject, show a higher GSL competency.

Fachlehrkräfte benötigen in der Lehrerausbildung Gelegenheit zum Kompetenzerwerb in Deutsch als Zweitsprache (DaZ), um die spezifischen Lernvoraussetzungen von Schülerinnen und Schülern nicht-deutscher Herkunftssprache berücksichtigen zu können. Auf der Basis eines Kompetenzmodells mit den Dimensionen Fachregister, Mehrsprachigkeit und Didaktik wurde im BMBF-Projekt DaZKom derzeit ein Testinstrument entwickelt und mit IRT-Methoden ausgewertet. Die Ergebnisse der Validierungsstudie (N = 252) zeigen, dass Zusammenhänge zwischen DaZ-Kompetenz und linguistischem Wissen sowie pädagogischem Wissen bestehen, der Test aber konzeptuell ein eigenständiges Konstrukt misst. Darüber hinaus korrelieren eine höhere Semesterzahl, Deutsch als Studienfach und eine größere Anzahl an DaZ-Lerngelegenheiten mit einer höheren DaZ-Kompetenz. (DIPF/Orig.), Pre-service teachers of all subject areas need learning opportunities in the field of German as a Second Language (GSL) in order to adequately facilitate second language learners in the content classroom. On the basis of a competency model including the sub-competencies subject-specific registers, multilingualism and didactics, the BMBF-project DaZKom has conducted a validation study. This included developing a test instrument with 68 items and analyzing it with IRT-methods. The results of the validation study (N = 252) show the expected correlations between GSL competency and linguistic knowledge as well as pedagogical knowledge, yet that test measures a conceptually independent construct. Furthermore, it is found that pre-service teachers with a higher number of learning opportunities and semesters, and with German as a study subject, show a higher GSL competency. (DIPF/Orig.)

Der folgende Artikel fokussiert das im Rahmen der Studie DaZKom1 entwickelteund evaluierte Kompetenzmodell, das Fähigkeiten und Fertigkeiten von Lehrerinnen und Lehrern aller Unterrichtsfächer im Bereich Deutsch als Zweitsprache definiert und umschreibt. Begründung, Genese und empirische Überprüfung dieses Modells sollen im Folgenden dargelegt werden.

FHOD1 gehört zur Unterfamilie der Diaphanous-verwandten Formine, deren Aktivität durch die Rho-GTPasen vermittelte Aufhebung intramolekularer Wechselwirkungen reguliert wird. Eine konstitutiv aktive Mutante von FHOD1 ruft in Zellen die Generierung dicker Stressfaserbündel hervor. Jedoch sind die zelluläre Relevanz und die molekularen Mechanismen der FHOD1-vermittelten Aktin-Reorganisation nur sehr bruchstückhaft aufgeklärt. Aus diesem Grund sollte in dieser Arbeit die molekulare Rolle von FHOD1 in der Regulation des Aktinzytoskeletts sowie in komplexen Aktin-basierten Prozessen charakterisiert werden. Durch RNAi-vermittelte Depletion des endogenen FHOD1 in U2OS-Zellen sowie die Expression konstitutiv aktiver FHOD1-Mutanten konnten in dieser Arbeit distinkte Funktionen von FHOD1 bei der Generierung unterschiedlicher Stressfasertypen beschrieben werden. Die Depletion des endogenen FHOD1 führte zu starken Defekten in der Aktinzytoskelett-Organisation, die häufig im vollständigen Kollaps des Stressfasersystems resultierte. Diese massiven Störungen gingen mit der Inhibierung von dynamischen Protrusionen an der Zellfront sowie Defekten in der Maturierung und Auflösung fokaler Adhäsionen einher und führten schlussendlich zu einer signifikant verringerten Zellmigration. Detaillierte Studien der Stressfaserdynamik zeigten, dass aktives FHOD1 die Entstehung kontraktiler ventraler Stressfasern über die verstärkte Generierung transversaler Aktinbögen fördert. Im Gegensatz dazu inhibiert das Formin das Wachstum nicht-kontraktiler dorsaler Stressfasern. In Lokalisationsanalysen konnte die distinkte Assoziation von aktivem FHOD1 entlang der induzierten kontraktilen Fasern beobachtet werden, was auf eine direkte Rolle des Formins in der Generierung dieser Strukturen hinweist. Dabei deutet insbesondere die partielle Kolokalisation mit dem Motorprotein Myosin und der Ausschluss beider Proteine entlang nicht-kontraktiler dorsaler Stressfasern darauf hin, dass FHOD1 zusammen mit Myosin transversale Aktinbögen in einem kooperativen Prozess induziert. Auf molekularer Ebene deutet die Verstärkung der transversalen Aktinbögen durch FHOD1, in der primär kurze Aktinfilamente verknüpft und gebündelt werden, und die Inhibierung dorsaler Fasern auf eine duale Funktion des Formins als ein Capping- bzw. Aktinfilament-bündelndes Protein. Zusammenfassend konnten in dieser Arbeit grundlegende Erkenntnisse hinsichtlich der zellulären Funktion von FHOD1 in der Regulation des Aktinzytoskeletts und Aktin-basierten dynamischen Prozessen gewonnen werden. Die vorgestellten Befunde stellen die Grundlage für weiterführende Studien zum Verständnis der Formin-vermittelten molekularen Mechanismen der Aktinzytoskelett-Regulation dar. FHOD1 is a member of the subfamily of Diaphanous-related formins, which are activated by Rho GTPase mediated release of their intramolecular autoinhibition. The expression of an activated FHOD1 mutant induces the formation of thick stress fiber bundles in cells. However, the cellular relevance of this effect as well as molecular mechanisms leading to FHOD1 mediated stress fiber formation are not well understood. The aim of the present study was to characterize the molecular function of FHOD1 in the dynamic reorganization of the actin cytoskeleton and the role of formin mediated actin dynamics in actin-based cellular processes. RNAi-mediated depletion of endogenous FHOD1 as well as expression of constitutively active FHOD1 mutants revealed distinct functions of the formin during the generation of different stress fiber types in U2OS cells. Overall, depletion of FHOD1 lead to massive defects in the formation of actin stress fibers in the leading edge which often resulted in the entire collapse of the actin filament network. These striking defects were accompanied by decreased protrusion dynamics at the cell front as well as perturbed turn over of focal adhesions, ultimately leading to significantly decreased cell migration. Detailed studies of stress fiber dynamics showed that active FHOD1 leads to the accumulation of contractile ventral stress fibers due to enhanced formation of transversal arcs. In contrast, growth of non-contractile dorsal fibers was inhibited in cells expressing active FHOD1. Localization studies revealed distinct localization patterns of FHOD1 along the induced contractile stress fibers suggesting a direct function of the formin in the generation of these structures. Additionally active FHOD1 colocalized partially with Myosin at contractile stress fibers, whereas both proteins were absent along non-contractile dorsal fibers. These findings indicate a possible interplay of both proteins in the process of transversal arc generation. The FHOD1 mediated enhanced formation of transversal arcs, which are primarily formed by fusion and bundling of short actin filaments, along with the inhibited growth of dorsal stress fibers indicate a potential dual molecular function of the formin as a capping and actin bundling protein. In summary, the present work provides fundamental molecular knowledge of FHOD1 function and upstream regulatory mechanisms in the organization of the actin cytoskeleton. Thus, the presented findings offer a critical basis for future work to deepen our understanding of formin mediated actin-based dynamic processes.

The formin FHOD1 (formin homology 2 domain containing protein 1) can act as a capping and bundling protein in vitro. In cells, active FHOD1 stimulates the formation of ventral stress fibers. However, the cellular mechanisms by which this phenotype is produced and the physiological relevance of FHOD1 function are not currently understood. Here, we first show that FHOD1 controls the formation of two distinct stress fiber precursors differentially. On the one hand, it inhibits dorsal fiber growth, which requires the polymerization of parallel bundles of long actin filaments. On the other hand, it stimulates transverse arcs that are formed by the fusion of short antiparallel actin filaments. This combined action is crucial for the maturation of stress fibers and their spatio-temporal organization, and a lack of FHOD1 function perturbs dynamic cell behavior during cell migration. Furthermore, we show that the GTPase-binding and formin homology 3 domains (GBD and FH3) are responsible for stress fiber association and colocalization with myosin. Surprisingly, a version of FHOD1 that lacks these domains nevertheless retains its full capacity to stimulate arc and ventral stress fiber formation. Based on our findings, we propose a mechanism in which FHOD1 promotes the formation of short actin filaments and transiently associates with transverse arcs, thus providing tight temporal and spatial control of the formation and turnover of transverse arcs into mature ventral stress fibers during dynamic cell behavior.

Duisburg, Essen, Univ., Diss., 2013 FHOD1 gehört zur Unterfamilie der Diaphanous-verwandten Formine, deren Aktivität durch die Rho-GTPasen vermittelte Aufhebung intramolekularer Wechselwirkungen reguliert wird. Eine konstitutiv aktive Mutante von FHOD1 ruft in Zellen die Generierung dicker Stressfaserbündel hervor. Jedoch sind die zelluläre Relevanz und die molekularen Mechanismen der FHOD1-vermittelten Aktin-Reorganisation nur sehr bruchstückhaft aufgeklärt. Aus diesem Grund sollte in dieser Arbeit die molekulare Rolle von FHOD1 in der Regulation des Aktinzytoskeletts sowie in komplexen Aktin-basierten Prozessen charakterisiert werden. Durch RNAi-vermittelte Depletion des endogenen FHOD1 in U2OS-Zellen sowie die Expression konstitutiv aktiver FHOD1-Mutanten konnten in dieser Arbeit distinkte Funktionen von FHOD1 bei der Generierung unterschiedlicher Stressfasertypen beschrieben werden. Die Depletion des endogenen FHOD1 führte zu starken Defekten in der Aktinzytoskelett-Organisation, die häufig im vollständigen Kollaps des Stressfasersystems resultierte. Diese massiven Störungen gingen mit der Inhibierung von dynamischen Protrusionen an der Zellfront sowie Defekten in der Maturierung und Auflösung fokaler Adhäsionen einher und führten schlussendlich zu einer signifikant verringerten Zellmigration. Detaillierte Studien der Stressfaserdynamik zeigten, dass aktives FHOD1 die Entstehung kontraktiler ventraler Stressfasern über die verstärkte Generierung transversaler Aktinbögen fördert. Im Gegensatz dazu inhibiert das Formin das Wachstum nicht-kontraktiler dorsaler Stressfasern. In Lokalisationsanalysen konnte die distinkte Assoziation von aktivem FHOD1 entlang der induzierten kontraktilen Fasern beobachtet werden, was auf eine direkte Rolle des Formins in der Generierung dieser Strukturen hinweist. Dabei deutet insbesondere die partielle Kolokalisation mit dem Motorprotein Myosin und der Ausschluss beider Proteine entlang nicht-kontraktiler dorsaler Stressfasern darauf hin, dass FHOD1 zusammen mit Myosin transversale Aktinbögen in einem kooperativen Prozess induziert. Auf molekularer Ebene deutet die Verstärkung der transversalen Aktinbögen durch FHOD1, in der primär kurze Aktinfilamente verknüpft und gebündelt werden, und die Inhibierung dorsaler Fasern auf eine duale Funktion des Formins als ein Capping- bzw. Aktinfilament-bündelndes Protein. Zusammenfassend konnten in dieser Arbeit grundlegende Erkenntnisse hinsichtlich der zellulären Funktion von FHOD1 in der Regulation des Aktinzytoskeletts und Aktin-basierten dynamischen Prozessen gewonnen werden. Die vorgestellten Befunde stellen die Grundlage für weiterführende Studien zum Verständnis der Formin-vermittelten molekularen Mechanismen der Aktinzytoskelett-Regulation dar.