The alpha 5 beta 1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression.

Anchorage-dependent cells that are prevented from attaching to an extracellular matrix substrate stop proliferating and may undergo apoptosis. Cell adhesion to a substrate is mediated by the integrin family of cell surface receptors, which are known to elicit intracellular signals upon cell adhesion. We show here that Chinese hamster ovary cells expressing the alpha 5 beta 1 integrin, which is a fibronectin receptor, do not undergo apoptosis upon serum withdrawal when the cells are plated on fibronectin. However, the alpha v beta 1 integrin, which is also a fibronectin receptor and binds fibronectin on the same RGD motif as alpha 5 beta 1, did not prevent apoptosis on fibronectin of the same cells. The cytoplasmic domain of the integrin alpha 5 subunit was required for the alpha 5 beta 1-mediated cell survival on fibronectin. The fibronectin-mediated survival effect appeared to be independent of the level of tyrosine phosphorylation of the focal adhesion kinase, which is induced by integrin-mediated cell attachment. The expression of the Bcl-2 protein, which counteracts apoptosis, was elevated in cells attaching to fibronectin through alpha 5 beta 1; cells attaching through alpha v beta 1 survived only if exogenous Bcl-2 was provided. Thus, alpha 5 beta 1, but not the closely related alpha v beta 1 integrin, appears to suppress apoptotic cell death through the Bcl-2 pathway.

A mutation in the epithelial sodium channel causing Liddle disease increases channel activity in the Xenopus laevis oocyte expression system.

We have studied the functional consequences of a mutation in the epithelial Na+ channel that causes a heritable form of salt-sensitive hypertension, Liddle disease. This mutation, identified in the original kindred described by Liddle, introduces a premature stop codon in the channel beta subunit, resulting in a deletion of almost all of the C terminus of the encoded protein. Coexpression of the mutant beta subunit with wild-type alpha and gamma subunits in Xenopus laevis oocytes resulted in an approximately 3-fold increase in the macroscopic amiloride-sensitive Na+ current (INa) compared with the wild-type channel. This change in INa reflected an increase in the overall channel activity characterized by a higher number of active channels in membrane patches. The truncation mutation in the beta subunit of epithelial Na+ channel did not alter the biophysical and pharmacological properties of the channel--including unitary conductance, ion selectivity, or sensitivity to amiloride block. These results provide direct physiological evidence that Liddle disease is related to constitutive channel hyperactivity in the cell membrane. Deletions of the C-terminal end of the beta and gamma subunits of rat epithelial Na+ channel were functionally equivalent in increasing INa, suggesting that the cytoplasmic domain of the gamma subunit might be another molecular target for mutations responsible for salt-sensitive forms of hypertension.

Differential subcellular mRNA targeting: deletion of a single nucleotide prevents the transport to axons but not to dendrites of rat hypothalamic magnocellular neurons.

It has previously been shown that mRNA encoding the arginine vasopressin (AVP) precursor is targeted to axons of rat magnocellular neurons of the hypothalamo-neurohypophyseal tract. In the homozygous Brattle-boro rat, which has a G nucleotide deletion in the coding region of the AVP gene, no such targeting is observed although the gene is transcribed. RNase protection and heteroduplex analyses demonstrate that, in heterozygous animals, which express both alleles of the AVP gene, the wild-type but not the mutant transcript is subject to axonal compartmentation. In contrast, wild-type and mutant AVP mRNAs are present in dendrites. These data suggest the existence of different mechanisms for mRNA targeting to the two subcellular compartments. Axonal mRNA localization appears to take place after protein synthesis; the mutant transcript is not available for axonal targeting because it lacks a stop codon preventing its release from ribosomes. Dendritic compartmentation, on the other hand, is likely to precede translation and, thus, would be unable to discriminate between the two mRNAs.

Delivery of antisense oligodeoxyribonucleotides against the human epidermal growth factor receptor into cultured KB cells with liposomes conjugated to folate via polyethylene glycol.

Antisense oligodeoxyribonucleotides targeted to the epidermal growth factor (EGF) receptor were encapsulated into liposomes linked to folate via a polyethylene glycol spacer (folate-PEG-liposomes) and efficiently delivered into cultured KB cells via folate receptor-mediated endocytosis. The oligonucleotides were a phosphodiester 15-mer antisense to the EGF receptor (EGFR) gene stop codon (AEGFR2), the same sequence with three phosphorothioate linkages at each terminus (AEGFR2S), a randomized 15-mer control of similar base composition to AEGFR2 (RC15), a 14-mer control derived from a symmetrized Escherichia coli lac operator (LACM), and the 5'-fluorescein-labeled homologs of several of the above. Cellular uptake of AEGFR2 encapsulated in folate-PEG-liposomes was nine times higher than AEGFR2 encapsulated in nontargeted liposomes and 16 times higher than unencapsulated AEGFR2. Treatment of KB cells with AEGFR2 in folate-PEG-liposomes resulted in growth inhibition and significant morphological changes. Curiously, AEGFR2 and AEGFR2S encapsulated in folate-PEG-liposomes exhibited virtually identical growth inhibitory effects, reducing KB cell proliferation by > 90% 48 hr after the cells were treated for 4 hr with 3 microM oligonucleotide. Free AEGFR2 caused almost no growth inhibition, whereas free AEGFR2S was only one-fifth as potent as the folate-PEG-liposome-encapsulated oligonucleotide. Growth inhibition of the oligonucleotide-treated cells was probably due to reduced EGFR expression because indirect immunofluorescence staining of the cells with a monoclonal antibody against the EGFR showed an almost quantitative reduction of the EGFR in cells treated with folate-PEG-liposome-entrapped AEGFR2. These results suggest that antisense oligonucleotide encapsulation in folate-PEG-liposomes promise efficient and tumor-specific delivery and that phosphorothioate oligonucleotides appear to offer no major advantage over native phosphodiester DNA when delivered by this route.

Mutações inativadoras no gene MKRN3 são causa de puberdade precoce central familial

A maioria dos casos de puberdade precoce central (PPC) em meninas permanece idiopática. A hipótese de uma causa genética vem se fortalecendo após a descoberta de alguns genes associados a este fenótipo, sobretudo aqueles implicados com o sistema kisspeptina (KISS1 e KISS1R). Entretanto, apenas casos isolados de PPC foram relacionados à mutação na kisspeptina ou em seu receptor. Até recentemente, a maioria dos estudos genéticos em PPC buscava genes candidatos selecionados com base em modelos animais, análise genética de pacientes com hipogonadismo hipogonadotrófico, ou ainda, nos estudos de associação ampla do genoma. Neste trabalho, foi utilizado o sequenciamento exômico global, uma metodologia mais moderna de sequenciamento, para identificar variantes associadas ao fenótipo de PPC. Trinta e seis indivíduos com a forma de PPC familial (19 famílias) e 213 casos aparentemente esporádicos foram inicialmente selecionados. A forma familial foi definida pela presença de mais de um membro afetado na família. DNA genômico foi extraído dos leucócitos do sangue periférico de todos os pacientes. O estudo de sequenciamento exômico global realizado pela técnica ILLUMINA, em 40 membros de 15 famílias com PPC, identificou mutações inativadoras em um único gene, MKRN3, em cinco dessas famílias. Pesquisa de mutação no MKRN3 realizada por sequenciamento direto em duas famílias adicionais (quatro pacientes) identificou duas novas variantes nesse gene. O MKRN3 é um gene de um único éxon, localizado no cromossomo 15 em uma região crítica para a síndrome de Prader Willi. O gene MKRN3 sofre imprinting materno, sendo expresso apenas pelo alelo paterno. A descoberta de mutações em pacientes com PPC familial despertou o interesse para a pesquisa de mutações nesse gene em 213 pacientes com PPC aparentemente esporádica por meio de reação em cadeia de polimerase seguida de purificação enzimática e sequenciamento automático direto (Sanger). Três novas mutações e duas já anteriormente identificadas, incluindo quatro frameshifts e uma variante missense, foram encontradas, em heterozigose, em seis meninas não relacionadas. Todas as novas variantes identificadas estavam ausentes nos bancos de dados (1000 Genomes e Exome Variant Server). O estudo de segregação familial em três dessas meninas com PPC aparentemente esporádica e mutação no MKRN3 confirmou o padrão de herança autossômica dominante com penetrância completa e transmissão exclusiva pelo alelo paterno, demonstrando que esses casos eram, na verdade, também familiares. A maioria das mutações encontradas no MKRN3 era do tipo frameshift ou nonsense, levando a stop códons prematuros e proteínas truncadas e, portanto, confirmando a associação com o fenótipo. As duas mutações missenses (p.Arg365Ser e p.Phe417Ile) identificadas estavam localizadas em regiões de dedo ou anel de zinco, importantes para a função da proteína. Além disso, os estudos in silico dessas duas variantes demonstraram patogenicidade. Todos os pacientes com mutação no MKRN3 apresentavam características clínicas e hormonais típicas de ativação prematura do eixo reprodutivo. A mediana de idade de início da puberdade foi de 6 anos nas meninas (variando de 3 a 6,5) e 8 anos nos meninos (variando de 5,9 a 8,5). Tendo em vista o fenômeno de imprinting, análise de metilação foi também realizada em um subgrupo de 52 pacientes com PPC pela técnica de MS-MLPA, mas não foram encontradas alterações no padrão de metilação. Em conclusão, este trabalho identificou um novo gene associado ao fenótipo de PPC. Atualmente, mutações inativadoras no MKRN3 representam a causa genética mais comum de PPC familial (33%). O MKRN3 é o primeiro gene imprintado associado a distúrbios puberais em humanos. O mecanismo preciso de ação desse gene na regulação da secreção de GnRH necessita de estudos adicionais

La aplicación de stop-motion en arquitectura: materia y luz

Se inicia un análisis de los procesos de trabajo de stop-motion porque ayudan a comprender las diferentes escalas en arquitectura donde las maquetas se convierten en futuros prototipos de infraestructuras de edificios o de paisaje. Stop motion es una técnica de animación fotograma a fotograma de objetos estáticos mediante la manipulación de figuras de plastilina en entornos fijos con cambios de luz, color y sonido. Igual que dicha técnica reúne lo mejor del rodaje tradicional -story board, escenografía, fotografía, personajes, iluminación- la animación de maquetas de interiores sintetiza micro-procesos de mayor repercusión -habitaciones con cambios de humedad, de temperatura, de ventilación y de iluminación- incorporando efectos especiales que son procesados digitalmente en post-producción. Se construyen varios prototipos de habitación con parámetros fijos como el tamaño y la posición de la cámara y otros variables como los materiales, los personajes y la iluminación. Representan un mundo en miniatura que intenta aportar un acercamiento sensorial y atmosférico analizando la magia y la fantasía que Junichirô Tanizaki describe en la penumbra de las construcciones tradicionales japonesas y estudiando las imperfecciones de los escenarios que Tim Burton manipula en su películas de animación con una textura que las tecnologías digitales no pueden igualar. El objetivo es utilizar una escala micro para realizar unos modelos interiores donde las condiciones atmosféricas están controladas y reducidas, y tomar datos que se podrían aplicar a un proceso de modelado a escala intermedia para testar prototipos de edificios como el túnel de viento; o, finalmente, a una escala macro con maquetas de un sector de la costa o de un río donde los fenómenos meteorológicos son los protagonistas para simular inundaciones y diseñar futuras medidas de prevención y seguridad.

Please Stop Rubbing Your Relationship In My Face(book): An Investigation of Online Romantic Social Comparison

It is well-documented that social networking sites such as Facebook set the stage for social comparison. Such comparison has been linked to a number of negative outcomes including envy, negative moods, and lower self-esteem. The present research aims to extend current understanding of online social comparison by investigating how it pertains to romantic relationships. I hypothesized that for individuals high in attachment anxiety (compared to those low in this construct), online romantic social comparison might be related to negative consequences—which, in the current project, was operationalized as lower mood/affect and state self-esteem. Further, I hypothesized that there would be an interaction between attachment anxiety and relationship insecurities on these negative outcomes, such that the expected difference of attachment anxiety would be more pronounced under conditions priming relationship insecurities, relative to a control condition. Two experiments were conducted, one of which focused on single individuals, and the second focusing on individuals who were themselves in dating relationships. The paradigms of each entailed experimental manipulation of a key relationship-related variable (for single individuals, pessimism for future relationships; for dating individuals, the presence or absence of rejection threat), subsequent exposure to romantic content from Facebook, and finally, measures of affect and state self-esteem. I discovered partial support for the hypothesis that some single individuals—particularly those with higher, rather than lower, attachment anxiety—do indeed report feeling more negative moods and lower state self-esteem following exposure to romantic online content, in contrast to single individuals who had instead viewed neutral online content. The association between attachment anxiety and negative outcome was especially pertinent if individuals had been primed to believe that their own future romantic prospects were grim, or if attention had been drawn to their singleness. Among dating individuals, less support for hypotheses was found; however, exploratory post-hoc analyses revealed a promising (albeit weak) trend indicating that reinvestigation of the current hypotheses would be prudent.

Croatia and the structural funds – Doing without an Ottoman Empire bypass bridge. CEPS Commentary, 29 July 2013

In his latest commentary, Associate Senior Fellow Michael Emerson hails Croatia’s accession to the EU as a fine step forward towards the official goal of integration of the whole of the Balkans into the EU. At the same time, he warns of the folly underway of the serious attention being paid by the European Commission to the newest member state’s petition to construct a multi-million euro bridge over a 9-km stretch of Bosnian land (the ‘Neum corridor’) that inconveniently but temporarily interrupts a non-stop drive along Croatia’s Dalmatian coast.

Falling short of expectations? Stress-testing the European banking system. CEPS Policy Brief No. 315, 15 January 2014

Before the ECB takes over responsibility for overseeing Europe’s largest banks, as foreseen in the establishment of a eurozone banking union, it plans to conduct an Asset Quality Review (AQR) throughout the coming year, which will identify the capital shortfalls of these banks. This study finds that a comprehensive and decisive AQR will most likely reveal a substantial lack of capital in many peripheral and core European banks. The authors provide estimates of the capital shortfalls of banks that will be stress-tested under the AQR using publicly available data and a series of shortfall measures. Their analysis identifies which banks will most likely need capital, where a public back stop is likely to be needed and, since many countries are already highly leveraged, where an EU-wide backstop might be necessary.

The EU's Use of 'Targeted' Sanctions: Evaluating effectiveness. CEPS Working Document No. 391, March 2014

This study analyses the use by the European Union of the novel concept of ‘targeted sanctions’ in the framework of its Common Foreign and Security Policy. It examines two sets of sanctions regimes featuring different degrees of efficacy: in Myanmar and Zimbabwe, the EU wielded measures in support of human rights and democracy objectives in the absence of a United Nations mandate, while it supplemented UN sanctions to stop nuclear proliferation in Iran and North Korea. The study highlights a number of facilitators of, or hindrances to, the efficacy of sanctions, such as the degree of support by regional powers or the presence of UN legitimation. It concludes that the EU sanctions regimes could be optimised by using more robust measures, designing them on the basis of ex ante assessments, enabling faster upgrades, monitoring their impact and adjusting them regularly and improving outreach efforts.

The future of human rights protection in the new Libya: prospects and challenges. Jean Monnet Occasional Paper No. 6/2014

There is no doubt that demand for the respect of human rights was one of the factors behind the Arab Spring and Libya is no exception. Four decades of absolute dictatorship headed by Muammar Gaddafi had been further tainted with gross violations of human rights of Libyan citizens and restrictions on their basic freedoms. Before the revolution, Libya was a country where no political parties were allowed. Freedom of expression and the press were extremely restricted. Reports about the country’s human rights violations published by a number of international organizations documented large scale human rights abuses at the hands of the Gaddafi regime. The 17 February 2011 revolution in Libya led to a turning point in the country’s history. The regime of Muammar Gaddafi which had dominated the country since 1969 eventually collapsed, leading to the beginning of the painful task of reconciliation and state building. Nonetheless it is estimated that more than 7000 prisoners are held captive by various militias and armed groups without due process. This in addition to thousands of internally displaced persons. State building involves the consolidation of a democratic state based on a democratic constitution. In 2011, a constitutional declaration was adopted to replace the one that had been in effect since 1969. This was intended as a stop-gap solution to allow the new political forces unleashed in the country time to write a new democratic constitution. To help consolidate the democratic state, three elements are required: that human rights be placed at its core; that these rights are truly implemented and applied; and lastly that the independence of the judiciary is safeguarded. For all this to happen it is also essential to strengthen education on human rights by encouraging non-governmental organizations to take a stronger role in promoting human rights. Libyan citizens can only avail themselves of these rights and strengthen their implementation if they know what they are and how they can benefit from their implementation

Rosyjski rewizjonizm wobec zachodu. Prace OSW 33/2009. = Russia's revisionist policy towards the west. OSW Study 33/2009

Russia’s actions so far have led to a kind of deadlock. Moscow has managed to stop NATO enlargement into the CIS area, persuade the USA not to deploy the missile shield in Poland and the Czech Republic, and avoid major consequences after the war with Georgia; nevertheless, the full implementation of its objectives remains unlikely.

The end of an overlooked European currency war. CEPS Commentary, 21 January 2015

On January 15th the Swiss National Bank (SNB) abandoned the efforts it had taken since September 2011 to ensure that the Swiss franc/euro exchange rate would not fall below 1.2 Swiss francs per euro. The Swiss franc appreciated immediately by almost 20% (after a temporary overshot of an even larger amount).The justification was that speculative capital flows induced by the euro crisis were driving the Swiss franc above its equilibrium value. Daniel Gros draws some important general lessons in this Commentary from the Swiss case and finds that the move by the SNB to stop its interventions will have an important impact on the euro-area economy.

Will this time be different for Greece? How to assess its ability to deliver on the reform agenda. CEPS Commentary, 13 March 2015

Taking its inspiration from the ongoing debate on whether this time will be different for Greece and whether Syriza will deliver on its reform promises to the European partners, this Commentary expresses bemusement that the public debate on such an important issue as well as internal discussions among senior policy-makers frequently resort to ‘gut feelings’ or simple stereotypes. To counteract this tendency, the author presents a simple analytical framework that can be used to assess the likelihood that a government will deliver on its reform agenda. Its purpose is not to allow for a precise probabilistic calculation, but to enable better structuring of the knowledge we have. It emphasises that the change depends NOT only on the capacity of the state to design and deliver policies, but even more crucially on state autonomy from both illegitimate and legitimate interests and cognitive models used by policy-makers to make sense of the world.

Renewables: The great uncertainty of the EU energy strategy. Egmont Paper No. 71, November 2014

Summary. For more than two decades, the development of renewable energy sources (RES) has been an important aim of EU energy policy. It accelerated with the adoption of a 1997 White Paper and the setting a decade later of a 20% renewable energy target, to be reached by 2020. The EU counts on renewable energy for multiple purposes: to diversify its energy supply; to increase its security of supply; and to create new industries, jobs, economic growth and export opportunities, while at the same time reducing greenhouse gas (GHG) emissions. Many expectations rest on its development. Fossil fuels have been critical to the development of industrial nations, including EU Member States, which are now deeply reliant upon coal, oil and gas for nearly every aspect of their existence. Faced with some hard truths, however, the Member States have begun to shelve fossil fuel. These hard truths are as follows: firstly, fossil fuels are a finite resource, sometimes difficult to extract. This means that, at some point, fossil fuels are going to be more difficult to access in Europe or too expensive to use.1 The problem is that you cannot just stop using fossil fuels when they become too expensive; the existing infrastructure is profoundly reliant on fossil fuels. It is thus almost normal that a fierce resistance to change exists. Secondly, fossil fuels contribute to climate change. They emit GHG, which contribute greatly to climate change. As a consequence, their use needs to be drastically reduced. Thirdly, Member States are currently suffering a decline in their own fossil fuel production. This increases their dependence on increasingly costly fossil fuel imports from increasingly unstable countries. This problem is compounded by global developments: the growing share of emerging economies in global energy demand (in particular China and India but also the Middle East) and the development of unconventional oil and gas production in the United States. All these elements endanger the competitiveness of Member States’ economies and their security of supply. Therefore, new indigenous sources of energy and a diversification of energy suppliers and routes to convey energy need to be found. To solve all these challenges, in 2008 the EU put in place a strategy based on three objectives: sustainability (reduction of GHG), competitiveness and security of supply. The adoption of a renewable energy policy was considered essential for reaching these three strategic objectives. The adoption of the 20% renewable energy target has undeniably had a positive effect in the EU on the growth in renewables, with the result that renewable energy sources are steadily increasing their presence in the EU energy mix. They are now, it can be said, an integral part of the EU energy system. However, the necessity of reaching this 20% renewable energy target in 2020, combined with other circumstances, has also engendered in many Member States a certain number of difficulties, creating uncertainties for investors and postponing benefits for consumers. The electricity sector is the clearest example of this downside. Subsidies have become extremely abundant and vary from one Member State to another, compromising both fair competition and single market. Networks encountered many difficulties to develop and adapt. With technological progress these subsidies have also become quite excessive. The growing impact of renewable electricity fluctuations has made some traditional power plants unprofitable and created disincentives for new investments. The EU does clearly need to reassess its strategy. If it repeats the 2008 measures it will risk to provoke increased instability and costs.