Covalent Protein Modification with ISG15 via a Conserved Cysteine in the Hinge Region

The ubiquitin-like protein ISG15 (interferon-stimulated gene of 15 kDa) is strongly induced by type I interferons and displays antiviral activity. As other ubiquitin-like proteins (Ubls), ISG15 is post-translationally conjugated to substrate proteins by an isopeptide bond between the C-terminal glycine of ISG15 and the side chains of lysine residues in the substrates (ISGylation). ISG15 consists of two ubiquitin-like domains that are separated by a hinge region. In many orthologs, this region contains a single highly reactive cysteine residue. Several hundred potential substrates for ISGylation have been identified but only a few of them have been rigorously verified. In order to investigate the modification of several ISG15 substrates, we have purified ISG15 conjugates from cell extracts by metal-chelate affinity purification and immunoprecipitations. We found that the levels of proteins modified by human ISG15 can be decreased by the addition of reducing agents. With the help of thiol blocking reagents, a mutational analysis and miRNA mediated knock-down of ISG15 expression, we revealed that this modification occurs in living cells via a disulphide bridge between the substrates and Cys78 in the hinge region of ISG15. While the ISG15 activating enzyme UBE1L is conjugated by ISG15 in the classical way, we show that the ubiquitin conjugating enzyme Ubc13 can either be classically conjugated by ISG15 or can form a disulphide bridge with ISG15 at the active site cysteine 87. The latter modification would interfere with its function as ubiquitin conjugating enzyme. However, we found no evidence for an ISG15 modification of the dynamin-like GTPases MxA and hGBP1. These findings indicate that the analysis of potential substrates for ISG15 conjugation must be performed with great care to distinguish between the two types of modification since many assays such as immunoprecipitation or metal-chelate affinity purification are performed with little or no reducing agent present

Higher Law: Can Christian Conservatives Transform Law Through Legal Education?

The allure of law schools as transformative institutions in the United States prompted Christian Right leaders to invest in legal education in the 1990s and early 2000s. The aspiration was to control the training of lawyers in order to challenge the secular legal monopoly on law, policy, and culture. In this article, we examine three leading Christian conservative law schools and one training program dedicated to transforming the law. We ask how each institution seeks to realize its transformative mission and analyze how they organize themselves to produce the kinds of capital (human, intellectual, social, cultural) needed to effectively change the law. To do so, we develop a typology of legal institutionbuilding strategies (infiltration, supplemental, and parallel alternative) to compare the relative advantages and disadvantages of institutional forms. We conclude by discussing implications of our findings for those looking to law schools as sites of broader transformation within the law

Schwingfestigkeit nitrierter Proben. Lebensdauervorhersage nitrierter bauteilaehnlicher Proben mit Hilfe normierter Woehlerstreubaender - Einfluss der Randschicht Abschlussbericht

Rotating bar fatigue tests have been performed yielding statistically verified Woehler lines of heat-treated or nitrided specimens of steel 31 CrMoV 9 (1.8519), the tests covering three different specimen diameters, two stress concentration factors, and five variants of surface layers, with comparable base materials properties. The Woehler line paramaters decisively depend on the place of incipient cracking (at or below the surface). The place of incipient cracking in turn is determined inter alia by the specimen geometry, depth of nitride-hardened phase, residual stress curve, and surface hardness. For unnotched or weakly notched specimens with incipient cracking below the surface, fatigue strength assessment can be done with the concept by Kogaev and Serensen, and the standardised Woehler scatter band can be applied for fatigue strength prediction for nitrided components irrespective of their surface layers or stress gradients. Accompanying tension-compression tests have shown that above (charge-dependent) overall strain amplitudes of 0.003 through 0.0055 mm/mm, early failure is observed induced by cracking emanating in the nitrided phase. The Woehler lines proved to be successful in predicting lifetimes of specimens tested by strain or stress-controlled, one-step or multistep tests. (orig.)Am vergueteten sowie am nitrierten Zustand des Stahls 31 CrMo V 9 (1.8519) wurden an drei Probendurchmessern, zwei Formzahlen und fuenf Randschichtvarianten bei vergleichbaren Grundwerstoffeigenschaften Umlaufbiegeversuche durchgefuehrt und statistisch abgesicherte Woehlerlinien ermittelt. Die Woehlerlinienparameter sind entscheidend abhaengig von der Lage des Anrissortes (an oder unter der Oberflaeche). Die Anrisslage wird u.a. durch die Probengeometrie, die Nitrierhaertetiefe, den Eigenspannungsverlauf und die Randhaerte bestimmt. Bei ungekerbten sowie schwach gekerbten Proben mit Anrissbildung unter der Oberflaeche kann die Abschaetzung der Dauerschwingfestigkeit nach dem Konzept von Kogaev und Serensen erfolgen und das normierte Woehlerstreuband zur Schwingfestigkeitsvorhersage unabhaengig vom Randschichtaufbau und vom Spannungsgradienten fuer nitrierte Bauteile verwendet werden. Die begleitenden Zugdruckuntersuchungen ergaben, dass oberhalb (chargenabhaengiger) Gesamtdehnungsamplituden von 0,003 bis 0,0055 mm/mm ein fruehzeitiges Versagen durch Anrissbildung in der Nitrierschicht ausgeloest wird. Mit PSWT-Woehlerlinien konnte die Lebensdauer dehnungs- und spannungsgeregelter Ein- sowie Mehrstufenversuche erfolgreich vorhergesagt werden. (orig.)SIGLEAvailable from TIB Hannover: RA 2383(197) / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Wirtschaft, Bonn (Germany); Arbeitsgemeinschaft Industrieller Forschungsvereinigungen e.V., Koeln (Germany)DEGerman

De onde vem o endividamento feminino?: construção e validação de um modelo PLS-PM Where does the women debt come from?: construction and validation of a PLS-PM model

O consumo exacerbado pode levar muitos indivíduos a contraírem dívidas comprometendo uma parcela significativa de suas rendas e, em muitos casos, acabando por ser tornarem inadimplentes. A inadimplência trás consigo efeitos muitas vezes arrasadores tanto do ponto de vista macroeconômico, aumentando o risco das operações e produtos financeiros, como do ponto de vista do indivíduo, ao afetar suas relações sociais, seu estado psicológico e sua vida familiar. Por outro lado, a maior participação da mulher no mercado de trabalho trouxe uma maior independência financeira e consequentente maior poder na decisão de consumo e ao mesmo tempo, maiores responsabilidades sobre o gerenciamento financeiro e nas decisões de endividamento. Neste sentido, este estudo centrou-se na identificação e análise dos fatores que afetam na propensão ao endividamento, nas mulheres da Mesorregião Centro Ocidental Rio-grandense. Assim, este trabalho propõe um modelo estrutural para explorar as relações entre os fatores determinantes da propensão ao endividamento junto às mulheres da referida Mesorregião, considerando variáveis que compõem os construtos de STATUS SOCIAL, PREOCUPAÇÃO, ESTABILIDADE, PRAZER, PODER, ORÇAMENTO, ILUSÃO e MATERIALISMO. Para isso, foram aplicados 2.500 questionários espalhados estatisticamente entre os 31 municípios que compõem esta Mesorregião. Os dados foram analisados através da metodologia Partial Least Squares - Path Modeling (PLS-PM). Sumariamente, os resultados sugerem que o construto ENDIVIDAMENTO está associado aos construtos STATUS, PREOCUPAÇÃO e MATERIALISMO, corroborando com as teorias das Finanças Comportamentais, ao sugerir que as decisões que envolvem endividamento vão além da simples relação consumo e renda, ou seja, existem outras variáveis comportamentais que são importantes na hora do indivíduo contrair dívidas, tais como, o significado que o indivíduo atribui ao dinheiro e o nível de materialismo.<br>The exacerbate consumption may lead many individuals to contract debts which commite a significant portion of their income and, in many cases, eventually taking into default. The default often brings with it devastating effects in both macroeconomic point of view, increasing the risk of operations and financial products, such as from the individual standpoint, affecting his social, psychological state and family life. Moreover, the bigger women participation in labor market brought greater financial independence and empowerment in decision and consequently larger power of consumption decision and at the same time, more responsibility on financial management and borrowing decisions. Thus, this study focused on the identification and analysis of factors which affect the indebtedness propensity, among women in the Rio Grande do Sul western-central mesoregion. This study proposes a structural model to determine the relationships among the women indebtedness propensity determinants of that mesoregion, considering variables which compose the constructs of SOCIAL STATUS, CONCERN, STABILITY, PLEASURE, POWER, BUDGET, ILLUSION and MATERIALISM. For this, were applied 2,500 questionnaires statistically scattered among the 31 cities which compose the mesoregion. Data were analyzed by Partial Least Squares -Path Modeling (PLS-PM) methodology. In summary, the results suggest that the construct INDEBTEDNESS is associated with SOCIAL STATUS, CONCERN and MATERIALISM, corroborating with the Behavioral Finance theories, suggesting that decisions involving borrowing go beyond the simple consumption and income relationship, i.e., there are other behavioral variables which are important in the time that individual contract debts, such as the meaning that the individual attaches to money and the materialism level