5,342 research outputs found
Structural properties of relaxin chimeras: NMR characterization of the R3/I5 relaxin peptide
No full textRelaxin-3 interacts with high potency with three relaxin family peptide receptors (RXFP1, RXFP3, and RXFP4). Therefore, the development of selective agonist and antagonist analogs is important for in vivo studies characterizing the biological significance of the different receptor–ligand systems and for future pharmaceutical applications. Recent reports demonstrated that a peptide selective for RXFP3 and RXFP4 over RXFP1 can be generated by the combination of the relaxin-3 B chain with the A chain from insulin-like peptide 5 (INSL5), creating an R3/I5 chimera. We have used NMR spectroscopy to determine the three-dimensional structure of this peptide to gain structural insights into the consequences of combining chains from two different relaxins. The R3/I5 structure reveals a similar backbone conformation for the relaxin-3 B chain compared to native relaxin-3, and the INSL5 A chain displays a relaxin/insulin-like fold with two parallel helices. The findings indicate that binding and activation of RXFP3 and RXFP4 mainly require the B chain and that the A chain functions as structural support. RXFP1, however, demonstrates a more complex binding mechanism, involving both the A chain and the B chain. The creation of chimeras is a promising strategy for generating new structure–activity data on relaxins
Structural insights into the function of relaxins
No full textThe relaxin peptide hormones are members of the insulin superfamily and share a structural fold that is characterized by two peptide chains which are cross-braced by three disulfide bonds. On this framework, various amino acid side chains are presented, allowing specific interactions with different receptors. The relaxin receptors belong to two unrelated classes of G-protein-coupled receptors, but interestingly they are not selective for a single relaxin peptide. Relaxin-3, which is considered to be an extreme example of the relaxin family, can activate receptors from both classes and in fact interacts to some degree with all four receptors identified to date. To deduce how changes in the primary sequence can fine-tune the overall structure and thus the ability to interact with the various receptors, we have studied a range of relaxin-like peptides using solution nuclear magnetic resonance analysis. Three-dimensional structures of relaxin-3, insulin-like peptide 3 (INSL3), and INSL5 were determined and revealed a number of interesting features. All peptides showed a significant amount of line-broadening in certain regions, in particular around the intra-A-chain disulfide bond, suggesting that despite the disulfide bonds the fold is rather dynamic. Although the peptides share a common structural core there are significant differences, particularly around the termini. The structural data in combination with mutational studies provide valuable insights into the structure–activity relationships of relaxins
Status prawny młodzieżowych rad powiatów
No full textSocial participation, which has its origin in the Constitution of the Republic of Poland (civil society principle), should also be seen on the self-government level. On the county level, citizens’ activity can be strengthened by a youth county council. The article first discusses the history of these bodies as well as their current popularity and condition. The author also tries to create their definition and outline the establishment procedure. While pointing to the legal basis of the establishment acts, the author gives several reasons why youth county councils are not legal in the current circumstances. Youth county councils and youth municipal councils are compared on the basis of their activity. In the final part of the article, the author points out why those bodies should have a legal basis on the national level.Partycypacja społeczna, wynikająca z konstytucyjnej zasady społeczeństwa obywatelskiego, powinna mieć silne oddziaływanie w ramach samorządu terytorialnego. Na poziomie powiatu jedną z form aktywności obywateli może być młodzieżowa rada powiatu. W artykule zarysowano historię tych podmiotów oraz przedstawiono ich obecne rozpowszechnienie. Podjęto próbę stworzenia definicji tych rad oraz omówiono procedury ich powstania. Wskazując na podstawy prawne aktów powołujących, przestawiono wątpliwości co do możliwości ich istnienia w obecnym stanie prawnym. Porównano także aktywność tych podmiotów i młodzieżowych rad gmin, posiadających umocowanie ustawowe. Na podstawie oceny działalności młodzieżowych rad powiatów zaprezentowano również powody, które przemawiają zdaniem autora za koniecznością nowelizacji ustawy o samorządzie powiatowym
Rad GTPase inhibits cardiac fibrosis through connective tissue growth factor
No full textAims Our previous studies documented that Rad (Ras associated with diabetes), a member of the RGK (Rad, Gem, and Kir) family of Ras-related small G protein, is significantly decreased in human failing hearts and plays an important role in attenuating cardiac hypertrophy. The goal of this study is to identify the effect of Rad on cardiac fibrosis and the underlying mechanisms. Methods and results Rad knockout (KO) mice showed more severe cardiac fibrosis compared with wild-type littermate controls as detected by Sirius Red staining. Western blot analyses demonstrated that the expression of connective tissue growth factor (CTGF), a key mediator of fibrosis, increased dramatically in Rad KO mice. Overexpression of Rad in cultured neonatal cardiomyocytes suppressed both basal and transforming growth factor-beta 1-induced CTGF expression. Elevated CTGF expression was observed in cardiomyocytes when Rad was reduced by RNA interference. Moreover, cardiac fibroblasts produced greater extracellular matrix (ECM) when stimulated with conditioned medium from Rad-knockdown cardiomyocytes. ECM production was completely abolished by adding a CTGF-neutralizing antibody into the medium. CCAAT/enhancer-binding protein delta(C/EBP-delta) was demonstrated to activate CTGF in cardiomyocytes. Chromatin immunoprecipitation assay and co-immunoprecipitation further demonstrated that Rad inhibited the binding of C/EBP-delta to the CTGF promoter via direct interaction with C/EBP-delta. Conclusion Our data reveal that Rad deficiency can lead to cardiac fibrosis. Rad inhibits CTGF expression through binding with C/EBP-delta, thus regulating ECM production in the heart. This study suggests a potential link between decreased Rad levels and increased cardiac fibrosis in human failing hearts.Cardiac & Cardiovascular SystemsSCI(E)PubMed13ARTICLE190-989
Platelet-derived growth factor induces Rad expression through Egr-1 in vascular smooth muscle cells.
No full textBACKGROUND: Ras associated with diabetes (Rad) inhibits vascular lesion formation by reducing the attachment and migration of vascular smooth muscle cells (VSMCs). However, the transcriptional regulation of Rad in VSMCs is unclear. METHODOLOGY AND PRINCIPAL FINDINGS: We found that Platelet-Derived Growth Factor (PDGF)induced Rad expression in a time- and dose-dependent manner in rat aortic smooth muscle cells (RASMCs) using quantitative real-time PCR. By serial deletion analysis of the Rad promoter, we identified that two GC-rich early growth response-1 (Egr-1) binding sites are essential for PDGF-induced Rad promoter activation. Overexpression of Egr-1 in RASMCs strongly stimulated Rad expression while the Egr-1 corepressor, NGFI-A binding protein 2 (NAB2), repressed PDGF-induced Rad up-regulation in a dose-dependent manner. Direct binding of Egr-1 to the Rad promoter region was further confirmed by chromatin immunoprecipitation assays. CONCLUSIONS: Our results demonstrate that Rad is regulated by PDGF through the transcriptional factor Egr-1 in RASMCs
Hochbegabte kinder - das unterdrückte genie -- was treibt Hans Giebenrath unter das rad?: eine neuere perspektive zu Herman Hesses Unterm Rad, in bezug auf die idee ,das lernen als strafe'
No full text2013 Fall.Includes bibliographical references.Text in German; title and abstract in English and German.The current discussion on Hermann Hesse's 1906 book, Unterm Rad, leaves many open-ended questions. Because the storyline so closely follows Hermann Hesse's personal biography, it obfuscates his authorial intentions and makes it difficult for scholars to differentiate between the two. Many critics also claim that the correlations between Unterm Rad's protagonist, Hans Giebenrath, and Hesse's personal life have actually stagnated later research about the book, as discussions always circle back to Hesse's personal struggles in the Prussian school system. This thesis, although acknowledging the similarities to Hesse's personal timeline, aims to frame the book in historical context in order to discuss its importance in a literary context. This thesis begins by analyzing the norm of educational methods in the early nineteenth century, and establishes that they are strongly based on a long tradition of child rearing by using force. This can be traced back to some accounts from 1752, and are based on a history of bourgeois childrearing. The headmaster and pastor's treatment of Hans in Unterm Rad clearly demonstrate the force and suppression of new ideas, used as modes of teaching to ensure students conformed to societal norms: this is historically reconcilable. Hesse's fictional story is thus the ideal basis for an analysis of childrearing methods used during that time. In order to effectively introduce a new perspective on the discussion, this paper uses the New Historicism approach and begins with Roland Barthes's theory of authorial intention. It analyses the text within the constraints of The Death of the Author, and continues with Michel Foucault's What is an Author? The goal is to evaluate what Unterm Rad says about the child rearing at the turn of the century in southern Germany, particularly for gifted children, and how it can be applied to what is already known from a historical standpoint. This idea is then applied from Hans Giebenrath's point of view to German psychologist Katharina Rutschky's concept of "Schwarze Pädagogik" or "Black Pedagogy" and her theories of suppression. This idea is further supplemented by Alice Millers research on childrearing, in relation to Hans's experience at the school in Maulbronn
Role rad społecznych we współrządzeniu miastem
No full textUrban governance (UG) is a place-based network of cooperation between public and non-public actors where public authorities play a strategic role in deciding on city matters. A number of UG actors include advisory councils (ACs), characterized by a long tradition, universality, stability and objectives related to urban affairs. The article aims to identify AC roles in UG. In addition, the author hypothesizes a correlation between the number of official (statutory) and informal (not legally regulated) roles and differences between ACs in terms of the roles they play. The hypotheses were verified in qualitative research. 57 semi-structured interviews were conducted with members of the youth councils, senior citizen councils, sport councils, and councils for residents with disabilities in 16 Polish cities. The interview transcripts were uploaded to QDA Miner for encoding and analysis. It was found that all ACs, regardless of their official roles, play informal roles, and all these roles are related to UG. There is no correlation between the number of official and informal roles, which would make ACs very similar unless the mandatory character of the roles played by the council for residents with disabilities, which differs this AC from the other. The results of the study lead the author to recommend revisions to AC regulations in order to increase their participation in the UG.Współrządzenie miastem (WM) to związana z miejscem sieć współpracy między podmiotami publicznymi i niepublicznymi, w której władze publiczne odgrywają strategiczną rolę w podejmowaniu decyzji o sprawach miasta. Wśród aktorów WM są rady społeczne (RS), które charakteryzują się długą tradycją, powszechnością, stabilnością i celami związanymi z miastem. Artykuł ma na celu identyfikację ról RS w WM. Ponadto autorka stawia hipotezę o związku między liczbą ról oficjalnych (ustawowych) i nieformalnych (nieuregulowanych prawnie) oraz o różnicach między RS pod względem pełnionych ról. Hipotezy zweryfikowano w badaniach jakościowych. Przeprowadzono 57 wywiadów częściowo ustrukturyzowanych z członkami rad młodzieżowych, rad seniorów, rad sportu i rad ds. osób niepełnosprawnych w 16 polskich miastach. Transkrypcje wywiadów zostały wprowadzone do QDA Miner w celu zakodowania i analizy. Stwierdzono, że wszystkie RS, niezależnie od swoich ról oficjalnych, pełnią role nieformalne, a wszystkie te role są związane z WM. Nie ma związku między liczbą ról oficjalnych i nieformalnych, co sprawia, że RS wydałyby się bardzo podobne, gdyby nie role pełnione przez radę ds. osób niepełnosprawnych, które mają charakter obowiązku ustawowego, co istotnie różni tę radę od pozostałych. Wyniki badań skłoniły autorkę do zaproponowania zmian w przepisach dotyczących RS w kierunku zwiększenia ich udziału w UG
Rad na ulozi Sarah u predstavi "Ljubavnik"
No full textDjelo Harolda Pintera Ljubavnik zaživjelo je kao diplomska predstava Gordana Marijanovića i Matee Bublić pod mentorstvom doc. art. Jasmina Novljakovića na Akademiji za umjetnost i kulturu u Osijeku na odsjeku za kazališnu umjetnost, smjer gluma i lutkarstvo. U ovom diplomskom radu Matea opisuje proces rada na diplomskoj predstavi koji uključuje rad na tekstu, rad u prostoru te rad na ulozi. Na početku ukratko iznosi činjenice o autoru i djelu nakon čega kronološki navodi faze rada unutar kojih opisuje proces svojeg glumačkog istraživanja i spoznajne činjenice o utvrđenom znanju. Kroz sva poglavlja čitatelja upoznaje s režijskim konceptom predstave, opisuje mu scenografiju, kostimografiju i rekvizite te mu približava karakter predstave. Svoj diplomski rad Matea zaokružuje spoznajom i iznošenjem činjenica o stečenom znanju kroz petogodišnje studiranje te svojom idejom o karakteristikama koje čine kvalitetnog glumca.Drama „Lover“ by Harold Pinter was set on the scene as a final exam of Gordan Marijanović and Matea Bublić, mentored by assist. prof. art. Jasmin Novljaković and produced by Academy for art and culture in Osijek under the Department of Theatre Arts, study of acting and puppetry. In this written thesis Matea is describing the working process on the final exam which is consisted of chapters which are talking about: working on the text, working on the scene and creating a role. On the beginning she is writing about author and drama, chronologically describes the working phases and analyse process of acting reaserch. Through all chapters Matea introduces the reader to the concept of the play, describes the scenography, costumography and tries to bring the character of the play closer to the reader. She is concluding her written thesis by bringing the facts about acquired knowledge through the study and sharing her opinion about characteristics of a good actor
Engineering of chimeric peptides as antagonists for the G protein-coupled receptor, RXFP4
No full textInsulin-like peptide 5 (INSL5) is a very important pharma target for treating human conditions such as anorexia and diabetes. However, INSL5 with two chains and three disulfide bridges is an extremely difficult peptide to assemble by chemical or recombinant means. In a recent study, we were able to engineer a simplified INSL5 analogue 13 which is a relaxin family peptide receptor 4 (RXFP4)-specific agonist. To date, however, no RXFP4-specific antagonist (peptide or small molecule) has been reported in the literature. The focus of this study was to utilize the non-specific RXFP3/RXFP4 antagonist ΔR3/I5 as a template to rationally design an RXFP4 specific antagonist. Unexpectedly, we demonstrated that ΔR3/I5 exhibited partial agonism at RXFP4 when expressed in CHO cells which is associated with only partial antagonism of INSL5 analogue activation. In an attempt to improve RXFP4 specificity and antagonist activity we designed and chemically synthesized a series of analogues of ΔR3/I5. While all the chimeric analogues still demonstrated partial agonism at RXFP4, one peptide (Analogue 17) exhibited significantly improved RXFP4 specificity. Importantly, analogue 17 has a simplified structure which is more amenable to chemical synthesis. Therefore, analogue 17 is an ideal template for further development into a specific high affinity RXFP4 antagonist which will be an important tool to probe the physiological role of RXFP4/INSL5 axis
Differential expression of mesotocin receptors in the uterus and ovary of the pregnant tammar wallaby
No full textMesotocin, an oxytocin-like peptide, is released in highest concentrations during parturition in macropodid marsupials. In late pregnant wallabies, uterine sensitivity to mesotocin increases markedly in the myometrium of the gravid uterus. This coincides with a significant increase in myometrial mesotocin receptor concentrations 3-4 days before term. To date, there is no information on mesotocin receptor gene expression in female wallaby reproductive tissues. This study aimed to examine mesotocin receptor gene expression in the uterus and ovaries of pregnant tammar wallabies, and to localise mesotocin receptors within the uterus. An RT-PCR strategy produced a consensus nucleotide sequence of 834 bp, which encoded 278 amino acids of transmembrane domains I to VI. This protein sequence has approximately 80% homology with the bovine and rat oxytocin receptor exon 2 region. Only one mesotocin receptor was detected in the tammar genome. The myometrium and mammary gland both expressed a 4.1 kb mesotocin receptor gene transcript. Myometrial mesotocin receptor gene expression increased on day 22 of the 26-day gestation and was significantly higher in the gravid than the non-gravid uterus in late pregnancy. This pattern of mesotocin receptor gene expression paralleled mesotocin receptor concentrations. Mesotocin binding sites were localised only to the myometrium, the highest densities being observed in the gravid uterus. Finally, this study showed high expression of mesotocin receptors in the corpus luteum. The pattern of luteal mesotocin receptor expression differed from the myometrium, with a decrease in mesotocin receptors occurring on the day of expected births
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