3T3 fibroblasts transfected with a cDNA for mitochondrial aspartate aminotransferase express plasma membrane fatty acid-binding protein and saturable fatty acid uptake.

To explore the relationship between mitochondrial aspartate aminotransferase (mAspAT; EC 2.6.1.1) and plasma membrane fatty acid-binding protein (FABPpm) and their role in cellular fatty acid uptake, 3T3 fibroblasts were cotransfected with plasmid pMAAT2, containing a full-length mAspAT cDNA downstream of a Zn(2+)-inducible metallothionein promoter, and pFR400, which conveys methotrexate resistance. Transfectants were selected in methotrexate, cloned, and exposed to increasing methotrexate concentrations to induce gene amplification. Stably transfected clones were characterized by Southern blotting; those with highest copy numbers of pFR400 alone (pFR400) or pFR400 and pMAAT2 (pFR400/pMAAT2) were expanded for further study. [3H]Oleate uptake was measured in medium containing 500 microM bovine serum albumin and 125-1000 microM total oleate (unbound oleate, 18-420 nM) and consisted of saturable and nonsaturable components. pFR400/pMAAT2 cells exhibited no increase in the rate constant for nonsaturable oleate uptake or in the uptake rate of [14C]octanoate under any conditions. By contrast, Vmax (fmol/sec per 50,000 cells) of the saturable oleate uptake component increased 3.5-fold in pFR400/pMAAT2 cells compared to pFR400, with a further 3.2-fold increase in the presence of Zn2+. Zn2+ had no effect in pFR400 controls (P > 0.5). The overall increase in Vmax between pFR400 and pFR400/pMAAT2 in the presence of Zn2+ was 10.4-fold (P < 0.01) and was highly correlated (r = 0.99) with expression of FABPpm in plasma membranes as determined by Western blotting. Neither untransfected 3T3 nor pFR400 cells expressed cell surface FABPpm detectable by immunofluorescence. By contrast, plasma membrane immunofluorescence was detected in pFR400/pMAAT2 cells, especially if cultured in 100 microM Zn2+. The data support the dual hypotheses that mAspAT and FABPpm are identical and mediate saturable long-chain free fatty acid uptake.

Inhibition of in vitro VEGF expression and choroidal neovascularization by synthetic dendrimer peptide mediated delivery of a sense oligonucleotide

Ocular neovascularisation is the leading cause of blindness in developed countries and the most potent angiogenic factor associated with neovascularisation is vascular endothelial growth factor (VEGF). We have previously described a sense oligonucleotide (ODN-1) that possesses anti-human and rat VEGF activity. This paper describes the synthesis of lipid-lysine dendrimers and their subsequent ability to delivery ODN-1 to its target and mediate a reduction in VEGF concentration both in vitro and in vivo. Positively charged dendrimers were used to deliver ODN-1 into the nucleus of cultured D407 cells. The effects on VEGF mRNA transcription and protein expression were analysed using RT-PCR and ELISA, respectively. The most effective dendrimers in vitro were further investigated in vivo using an animal model of choroidal neovascularisation (CNV). All dendrimer/ODN-1 complexes mediated in a significant reduction in VEGF expression during an initial 24 hr period (40-60%). Several complexes maintained this level of VEGF reduction during a subsequent, second 24 hr period, which indicated protection of ODN-1 from the effects of endogenous nucleases. In addition, the transfection efficiency of dendrimers that possessed 8 positive charges (chi = 81(.)51%) was significantly better (P = 0(.)0036) than those that possessed 4 positive charges (chi = 56(.)8%). RT-PCR revealed a correlation between levels of VEGF protein mRNA. These results indicated that the most effective structural combination was three branched chains of intermediate length with 8 positive charges such as that found for dendrimer 4. Dendrimer 4 and 7/ODN-1 complexes were subsequently chosen for in vivo analysis. Fluorescein angiography demonstrated that both dendrimers significantly (P < 0(.)0001) reduced the severity of laser mediated CNV for up to two months post-injection. This study demonstrated that lipophilic, charged dendrimer mediated delivery of ODN-1 resulted in the down-regulation of in vitro VEGF expression. In addition, in vivo delivery of ODN-1 by two of the dendrimers resulted in significant inhibition of CNV in an inducible rat model. Time course studies showed that the dendrimer/ODN-1 complexes remained active for up to two months indicating the dendrimer compounds provided protection against the effects of nucleases. (C) 2004 Elsevier Ltd. All rights reserved.

Skeletal muscle and nuclear hormone receptors: Implications for cardiovascular and metabolic disease

Skeletal muscle is a major mass peripheral tissue that accounts for similar to 40% of the total body mass and a major player in energy balance. It accounts for > 30% of energy expenditure, is the primary tissue of insulin stimulated glucose uptake, disposal, and storage. Furthermore, it influences metabolism via modulation of circulating and stored lipid (and cholesterol) flux. Lipid catabolism supplies up to 70% of the energy requirements for resting muscle. However, initial aerobic exercise utilizes stored muscle glycogen but as exercise continues, glucose and stored muscle triglycerides become important energy substrates. Endurance exercise increasingly depends on fatty acid oxidation (and lipid mobilization from other tissues). This underscores the importance of lipid and glucose utilization as an energy source in muscle. Consequently skeletal muscle has a significant role in insulin sensitivity, the blood lipid profile, and obesity. Moreover, caloric excess, obesity and physical inactivity lead to skeletal muscle insulin resistance, a risk factor for the development of type II diabetes. In this context skeletal muscle is an important therapeutic target in the battle against cardiovascular disease, the worlds most serious public health threat. Major risk factors for cardiovascular disease include dyslipidemia, hypertension, obesity, sedentary lifestyle, and diabetes. These risk factors are directly influenced by diet, metabolism and physical activity. Metabolism is largely regulated by nuclear hormone receptors which function as hormone regulated transcription factors that bind DNA and mediate the pathophysiological regulation of gene expression. Metabolism and activity, which directly influence cardiovascular disease risk factors, are primarily driven by skeletal muscle. Recently, many nuclear receptors expressed in skeletal muscle have been shown to improve glucose tolerance, insulin resistance, and dyslipidernia. Skeletal muscle and nuclear receptors are rapidly emerging as critical targets in the battle against cardiovascular disease risk factors. Understanding the function of nuclear receptors in skeletal muscle has enormous pharmacological utility for the treatment of cardiovascular disease. This review focuses on the molecular regulation of metabolism by nuclear receptors in skeletal muscle in the context of dyslipidemia and cardiovascular disease. (c) 2005 Published by Elsevier Ltd.

Capsule and fimbria interaction in Klebsiella pneumoniae

The capsular polysaccharide and type I fimbriae are two of the major surface-located virulence properties associated with the pathogenesis of Klebsiella pneumoniae. The capsule is an elaborate polysaccharide matrix that encases the entire cell surface and provides resistance against many host defense mechanisms. In contrast, type 1 fimbriae are thin adhesive thread-like surface organelles that can extend beyond the capsular matrix and mediate D-mannose-sensitive adhesion to host epithelial cells. These fimbriae are archetypical and consist of a major building block protein (FimA) that comprises the bulk of the organelle and a tip-located adhesin (FimH). It is assumed that the extended major-subunit protein structure permits the FimH adhesin to function independently of the presence of a capsule. In this study, we have employed a defined set of K. pneumoniae capsulated and noncapsulated strains to show that the function of type I fimbriae is actually impeded by the concomitant expression of a polysaccharide capsule. Capsule expression had significant effects on two parameters commonly used to define FimH function, namely, yeast cell agglutination and biofilm formation. Our data suggest that this effect is not due to transcriptional/translational changes in fimbrial gene/protein expression but rather the result of direct physical interference. This was further demonstrated by the fact that we could restore fimbrial function by inhibiting capsule synthesis. It remains to be determined whether the expression of these very different surface components occurs simply via random events of phase variation or in a coordinated manner in response to specific environmental cues.

Cytotoxic T-lymphocyte epitope vaccination protects against human metapneumovirus infection and disease in mice

Human metapneumovirus (hMPV) has emerged as an important human respiratory pathogen causing upper and lower respiratory tract infections in young children and older adults. In addition, hMPV infection is associated with asthma exacerbation in young children. Recent epidemiological evidence indicates that hMPV may cocircullate with human respiratory syncytial virus (hRSV) and mediate clinical disease similar to that seen with hRSV. Therefore, a vaccine for hMPV is highly desirable. In the present study, we used predictive bioinformatics, peptide immunization, and functional T-cell assays to define hMPV cytotoxic T-lymphocyte (CTL) epitopes recognized by mouse T cells restricted through several major histocompatibility complex class I alleles, including HILA-A*0201. We demonstrate that peptide immunization with hMPV CTL epitopes reduces viral load and immunopathollogy in the lungs of hMPV-challenged mice and enhances the expression of Th1-type cytokines (gamma interferon and interleukin-12 [IL-12]) in lungs and regional lymph nodes. In addition, we show that levels of Th2-type cytolkines (IL-10 and IL-4) are significantly lower in hMPV CTL epitope-vaccinated mice challenged with hMPV. These results demonstrate for the first time the efficacy of an hMPV CTL epitope vaccine in the control of hMPV infection in a murine model.

A study on uncertain dynamic disaster management tasks, knowledge sharing, and task performance

Each disaster presents itself with a unique set of characteristics that are hard to determine a priori. Thus disaster management tasks are inherently uncertain, requiring knowledge sharing and quick decision making that involves coordination across different levels and collaborators. While there has been an increasing interest among both researchers and practitioners in utilizing knowledge management to improve disaster management, little research has been reported about how to assess the dynamic nature of disaster management tasks, and what kinds of knowledge sharing are appropriate for different dimensions of task uncertainty characteristics. ^ Using combinations of qualitative and quantitative methods, this research study developed the dimensions and their corresponding measures of the uncertain dynamic characteristics of disaster management tasks and tested the relationships between the various dimensions of uncertain dynamic disaster management tasks and task performance through the moderating and mediating effects of knowledge sharing. ^ Furthermore, this research work conceptualized and assessed task uncertainty along three dimensions: novelty, unanalyzability, and significance; knowledge sharing along two dimensions: knowledge sharing purposes and knowledge sharing mechanisms; and task performance along two dimensions: task effectiveness and task efficiency. Analysis results of survey data collected from Miami-Dade County emergency managers suggested that knowledge sharing purposes and knowledge sharing mechanisms moderate and mediate uncertain dynamic disaster management task and task performance. Implications for research and practice as well directions for future research are discussed.^

Development of novel therapeutics and in vitro models for spinal cord injury

Spinal cord injury (SCI) is a devastating condition, which results from trauma to the cord, resulting in a primary injury response which leads to a secondary injury cascade, causing damage to both glial and neuronal cells. Following trauma, the central nervous system (CNS) fails to regenerate due to a plethora of both intrinsic and extrinsic factors. Unfortunately, these events lead to loss of both motor and sensory function and lifelong disability and care for sufferers of SCI. There have been tremendous advancements made in our understanding of the mechanisms behind axonal regeneration and remyelination of the damaged cord. These have provided many promising therapeutic targets. However, very few have made it to clinical application, which could potentially be due to inadequate understanding of compound mechanism of action and reliance on poor SCI models. This thesis describes the use of an established neural cell co-culture model of SCI as a medium throughput screen for compounds with potential therapeutic properties. A number of compounds were screened which resulted in a family of compounds, modified heparins, being taken forward for more intense investigation. Modified heparins (mHeps) are made up of the core heparin disaccharide unit with variable sulphation groups on the iduronic acid and glucosamine residues; 2-O-sulphate (C2), 6-O-sulphate (C6) and N-sulphate (N). 2-O-sulphated (mHep6) and N-sulphated (mHep7) heparin isomers were shown to promote both neurite outgrowth and myelination in the SCI model. It was found that both mHeps decreased oligodendrocyte precursor cell (OPC) proliferation and increased oligodendrocyte (OL) number adjacent to the lesion. However, there is a difference in the direct effects on the OL from each of the mHeps; mHep6 increased myelin internode length and mHep7 increased the overall cell size. It was further elucidated that these isoforms interact with and mediate both Wnt and FGF signalling. In OPC monoculture experiments FGF2 treated OPCs displayed increased proliferation but this effect was removed when co-treated with the mHeps. Therefore, suggesting that the mHeps interact with the ligand and inhibit FGF2 signalling. Additionally, it was shown that both mHeps could be partially mediating their effects through the Wnt pathway. mHep effects on both myelination and neurite outgrowth were removed when co-treated with a Wnt signalling inhibitor, suggesting cell signalling mediation by ligand immobilisation and signalling activation as a mechanistic action for the mHeps. However, the initial methods employed in this thesis were not sufficient to provide a more detailed study into the effects the mHeps have on neurite outgrowth. This led to the design and development of a novel microfluidic device (MFD), which provides a platform to study of axonal injury. This novel device is a three chamber device with two chambers converging onto a central open access chamber. This design allows axons from two points of origin to enter a chamber which can be subjected to injury, thus providing a platform in which targeted axonal injury and the regenerative capacity of a compound study can be performed. In conclusion, this thesis contributes to and advances the study of SCI in two ways; 1) identification and investigation of a novel set of compounds with potential therapeutic potential i.e. desulphated modified heparins. These compounds have multiple therapeutic properties and could revolutionise both the understanding of the basic pathological mechanisms underlying SCI but also be a powered therapeutic option. 2) Development of a novel microfluidic device to study in greater detail axonal biology, specifically, targeted axonal injury and treatment, providing a more representative model of SCI than standard in vitro models. Therefore, the MFD could lead to advancements and the identification of factors and compounds relating to axonal regeneration.

A critical and interpretative investigation of the ethical dimension of policies about school digital technologies

This thesis examines digital technologies policies designed for Australian schools and the ways they are understood and interpreted by students, school staff, teachers, principals and policy writers. This study explores the ways these research participant groups interpret and understand the ‘ethical dimension’ of schools’ digital technologies policies for teaching and learning. In this thesis the ethical dimension is considered to be a dynamic concept which encompasses various elements including; decisions, actions, values, issues, debates, education, discourses, and notions of right and wrong, in relation to ethics and uses of digital technologies in schools. In this study policy is taken to mean not only written texts but discursive processes, policy documents including national declarations, strategic plans and ‘acceptable use’ policies to guide the use of digital technologies in schools. The research is situated in the context of changes that have occurred in Australia and internationally over the last decade that have seen a greater focus on the access to and use of digital technologies in schools. In Australian school education, the attention placed on digital technologies in schools has seen the release of policies at the national, state, territory, education office and school levels, to guide their use. Prominent among these policies has been the Digital Education Revolution policy, launched in 2007 and concluded in 2013. This research aims to answers the question: What does an investigation reveal about understandings of the ethical dimension of digital technologies policies and their implementation in school education? The objective of this research is to examine the ethical dimension of digital technologies policies and to interpret and understand the responses of the research participants to the issues, silences, discourses and language, which characterise this dimension. In doing so, it is intended that the research can allow the participants to have a voice that, may be different to the official discourses located in digital technologies policies. The thesis takes a critical and interpretative approach to policies and examines the role of digital technologies policies as discourse. Interpretative theory is utilised as it provides a conceptual lens from which to interpret different perspectives and the implications of these in the construction of meaning in relation to schools’ digital technologies policies. Critical theory is used in tandem with interpretative theory as it represents a conceptual basis from which to critique and question underlying assumptions and discourses that are associated with the ethical dimension of schools’ digital technologies policies. The research methods used are semi-structured interviews and policy document analysis. Policies from the national, state, territory, education office and school level were analysed and contribute to understanding the way the ethical dimension of digital technologies policies is represented as a discourse. Students, school staff, teachers, principals and policy writers participated in research interviews and their views and perspectives were canvassed in relation to the ethical use of digital technologies and the policies that are designed to regulate their use. The thesis presents an argument that the ethical dimension of schools’ digital technologies policies and use is an under-researched area, and there are gaps in understanding and knowledge in the literature which remain to be addressed. It is envisaged that the thesis can make a meaningful contribution to understand the ways in which schools’ digital technologies policies are understood in school contexts. It is also envisaged that the findings from the research can inform policy development by analysing the voices and views of those in schools. The findings of the policy analysis revealed that there is little attention given to the ethical dimension in digital technologies at the national level. A discourse of compliance and control pervades digital technologies policies from the state, education office and school levels, which reduces ethical considerations to technical, legal and regulatory requirements. The discourse is largely instrumentalist and neglects the educative dimension of digital technologies which has the capacity to engender their ethical use. The findings from the interview conversations revealed that students, school staff and teachers perceive digital technologies policies to be difficult to understand, and not relevant to their situation and needs. They also expressed a desire to have greater consultation and participation in the formation and enactment of digital technologies policies, and they believe they are marginalised from these processes in their schools. Arising from the analysis of the policies and interview conversations, an argument is presented that in the light of the prominent role played by digital technologies and their potential for enhancing all aspects of school education, more research is required to provide a more holistic and richer understanding of the policies that are constructed to control and mediate their use.

Atmosphere : a framework for contextual awareness

This article enhances existing approaches to present-day asynchronous awareness concepts by providing the means to explicitly represent and mediate contextual information. The resulting concept of contextual awareness takes different notions of the term context into account. Following a human-centered approach, the proposed methods serve as mediators for context between persons rather than automatically detecting context. Based on this variant of awareness, the atmosphere framework is introduced to provide mechanisms to deal with the problem of workload in tandem with contextual information. Atmosphere provides a highly tailorable structure and interface to deal with a wide variance of user and organizational requirements. The article closes with the description of a partial implementation of the framework and its evaluation.

Discovery of an archetypal protein transport system in bacterial outer membranes

Bacteria have mechanisms to export proteins for diverse purposes, including colonization of hosts and pathogenesis. A small number of archetypal bacterial secretion machines have been found in several groups of bacteria and mediate a fundamentally distinct secretion process. Perhaps erroneously, proteins called 'autotransporters' have long been thought to be one of these protein secretion systems. Mounting evidence suggests that autotransporters might be substrates to be secreted, not an autonomous transporter system. We have discovered a new translocation and assembly module (TAM) that promotes efficient secretion of autotransporters in proteobacteria. Functional analysis of the TAM in Citrobacter rodentium, Salmonella enterica and Escherichia coli showed that it consists of an Omp85-family protein, TamA, in the outer membrane and TamB in the inner membrane of diverse bacterial species. The discovery of the TAM provides a new target for the development of therapies to inhibit colonization by bacterial pathogens.

UpaG, a new member of the Trimeric Autotransporter family of Adhesins in Uropathogenic Escherichia coli

The ability of Escherichia coli to colonize both intestinal and extraintestinal sites is driven by the presence of specific virulence factors, among which are the autotransporter (AT) proteins. Members of the trimeric AT adhesin family are important virulence factors for several gram-negative pathogens and mediate adherence to eukaryotic cells and extracellular matrix (ECM) proteins. In this study, we characterized a new trimeric AT adhesin (UpaG) from uropathogenic E. coli (UPEC). Molecular analysis of UpaG revealed that it is translocated to the cell surface and adopts a multimeric conformation. We demonstrated that UpaG is able to promote cell aggregation and biofilm formation on abiotic surfaces in CFT073 and various UPEC strains. In addition, UpaG expression resulted in the adhesion of CFT073 to human bladder epithelial cells, with specific affinity to fibronectin and laminin. Prevalence analysis revealed that upaG is strongly associated with E. coli strains from the B2 and D phylogenetic groups, while deletion of upaG had no significant effect on the ability of CFT073 to colonize the mouse urinary tract. Thus, UpaG is a novel trimeric AT adhesin from E. coli that mediates aggregation, biofilm formation, and adhesion to various ECM proteins.

Shaping (reflexive) professional identities across an undergraduate degree programme: A longitudinal case study

In our complex and incongruous professional worlds, where there is no blueprint for dealing with unpredictable people and events, it is imperative that individuals develop reflexive approaches to professional identity building. Notwithstanding the importance of disciplinary knowledge and skills, higher education has a crucial role to play in guiding students to examine and mediate self in relation to context for effective decision-making and action. This paper reports on a small-scale longitudinal project that investigated the ways in which ten undergraduate students over the course of a three-year Radiation Therapy degree shaped their professional identities. Theories of reflexivity and methods of discourse analysis are utilised to understand the ways in which individuals accounted for their professional identity projects at university. The findings suggest that, across time, the participants negotiated professional ‘becoming’ through four distinct kinds of reflexive modalities. These findings have implications for teaching strategies and curriculum design in undergraduate programs.

The mast cell as a regulator of atherosclerotic plaque stability

Atherosclerosis is an inflammatory disease progressing over years via the accumulation of cholesterol in arterial intima with subsequent formation of atherosclerotic plaques. The stability of a plaque is determined by the size of its cholesterol-rich necrotic lipid core and the thickness of the fibrous cap covering it. The strength and thickness of the cap are maintained by smooth muscle cells and the extracellular matrix produced by them. A plaque with a large lipid core and a thin cap is vulnerable to rupture that may lead to acute atherothrombotic events, such as myocardial infarction and stroke. In addition, endothelial erosion, possibly induced by apoptosis of endothelial cells, may lead to such clinical events. One of the major causes of plaque destabilization is inflammation induced by accumulated and modified lipoproteins, and exacerbated by local aberrant shear stress conditions. Macrophages, T-lymphocytes and mast cells infiltrate particularly into the plaque’s shoulder regions prone to atherothrombotic events, and they are present at the actual sites of plaque rupture and erosion. Two major mechanisms of plaque destabilization induced by inflammation are extracellular matrix remodeling and apoptosis. Mast cells are bone marrow-derived inflammatory cells that as progenitors upon chemotactic stimuli infiltrate the target tissues, such as the arterial wall, differentiate in the target tissues and mediate their effects via the release of various mediators, typically in a process called degranulation. The released preformed mast cell granules contain proteases such as tryptase, chymase and cathepsin G bound to heparin and chondroitin sulfate proteoglycans. In addition, various soluble mediators such as histamine and TNF-alpha are released. Mast cells also synthesize many mediators such as cytokines and lipid mediators upon activation. Mast cells are capable of increasing the level of LDL cholesterol in the arterial intima by increasing accumulation and retention of LDL and by decreasing removal of cholesterol by HDL in vitro. In addition, by secreting proinflammatory mediators and proteases, mast cells may induce plaque destabilization by inducing apoptosis of smooth muscle and endothelial cells. Also in vivo data from apoE-/- and ldlr-/- mice suggest a role for mast cells in the progression of atherosclerosis. Furthermore, mast cell-deficient mice have become powerful tools to study the effects of mast cells in vivo. In this study, evidence suggesting a role for mast cells in the regulation of plaque stability is presented. In a mouse model genetically susceptible to atherosclerosis, mast cell deficiency (ldlr-/-/KitW-sh/W-sh mice) was associated with a less atherogenic lipid profile, a decreased level of lipid accumulation in the aortic arterial wall and a decreased level of vascular inflammation as compared to mast-cell competent littermates. In vitro, mast cell chymase-induced smooth muscle cell apoptosis was mediated by inhibition of NF-kappaB activity, followed by downregulation of bcl-2, release of cytochrome c, and activation of caspase-8, -9 and -3. Mast cell-induced endothelial cell apoptosis was mediated by chymase and TNF-alpha, and involved chymase-mediated degradation of fibronectin and vitronectin, and inactivation of FAK- and Akt-mediated survival signaling. Subsequently, mast cells induced inhibition of NF-kappaB activity and activation of caspase-8 and -9. In addition, possible mast cell protease-mediated mechanisms of endothelial erosion may include degradation of fibronectin and VE-cadherin. Thus, the present results suggest a role for mast cells in destabilization of atherosclerotic plaques.

Vozes e caminhos do decrescimento: consequências humanas

O objetivo deste trabalho é apresentar o contemporâneo movimento sociopolítico do Decrescimento, partindo das ideias inaugurais de Serge Latouche, economista e filósofo francês responsável pela criação deste termo, sua historicidade, premissas de análise, e novas formas de subjetivação que propõe, cuja argumentação se sustenta e deriva também, dentre outros autores - como Castoriadis, Gorz e Mészáros - da tese de David Harvey que estabelece que desenvolvimento não é o mesmo que crescimento, e que é possível promover desenvolvimento nas dimensões das relações pessoais, sociais do cotidiano e das relações com o meio ambiente sem as orientações que favoreçam o capital e sua máxima de acumulação. Trata-se, segundo os autores, de um movimento anticapitalista que busca denunciar questões críticas contemporâneas e contradições e crises do capitalismo, apontando um conjunto de passos a serem dados para o lado de fora desta lógica que valoriza e incentiva o desejo e a necessidade do excesso. Em diálogo pleno com a Ética e seus mais recentes estudos, nos quais desponta a autora Victoria Camps, o Decrescimento se preocupa com as escolhas do indivíduo, com o que as mobiliza e com as maneiras pelas quais devemos viver. Segundo esta autora o reconhecimento em torno de nossa não autossuficiência faz com que nos percebamos vulneráveis e o espírito do que a maioria dos autores do Decrescimento chama de espírito do Dom do Decrescimento e de a Economia da Felicidade dialoga diretamente com a busca do homem por governar estas vulnerabilidades, compreender suas virtudes, desenvolvê-las e praticar o bem e seu senso coletivo. Sua contribuição por sua vez para a construção contínua e transformadora de uma Psicologia Social Crítica deve-se à busca por um novo sujeito, e cuja subjetividade possa ser ressignificada e emancipada, neste exercício alguns conceitos nos foram muito caros, como por exemplo, o conceito de liberdade. Discutiu-se o Decrescimento destacando suas viabilidades práticas, importância, dimensões planetárias e aquilo que particularmente mais me chamou atenção em relação ao movimento a partir de minha própria experiência na 3 Conferência Internacional do Decrescimento que ocorreu em Veneza em setembro de 2012. A espinha dorsal deste trabalho foi o próprio discurso do Decrescimento e o discurso construído em torno dele, e suas vozes que foram dispostas e organizadas em análise nesta pesquisa em forma de interlocução e diálogos abertos, interdisciplinares, teóricos e práticos. A interlocução foi exercício metodológico capaz de reunir as vozes do Decrescimento e transportá-las para dentro do texto desta pesquisa, sendo a própria pesquisa em si, pelo dar a conhecer deste sujeito e que pode ser outro e mediar sua existência com o mundo de outras maneiras, e pelo dar a conhecer em torno do próprio movimento, e sobre o qual não se pretende conclusões, pois não se trata de encerrar a discussão trata-se de valorizá-la por ela mesma e potencializar possibilidades dialéticas, críticas e reflexivas, ficando, portanto ao leitor o convite a problematizar-se diante das questões aqui destacadas, em busca de si, para si e pelas alternativas existenciais mais diversas, dentre elas, o Decrescimento.

RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination

Modification of proteins by ubiquitination plays important roles in various cellular processes. During this process, the target specificity is determined by ubiquitin ligases. Here we identify RNF220 (RING finger protein 220) as a novel ubiquitin ligase for Sin3B. As a conserved RING protein, RNF220 can bind E2 and mediate auto-ubiquitination of itself. Through a yeast two-hybrid screen, we isolated Sin3B as one of its targets, which is a scaffold protein of the Sin3/HDAC (histone deacetylase) corepressor complex. RNF220 specifically interacts with Sin3B both in vitro and in vivo. Sin3B can be regulated by the ubiquitin-proteasome system. Co-expression of RNF220 promotes the ubiquitination and proteasomal degradation of Sin3B. Taken together, these results reveal a new mechanism for regulating the Sin3/HDAC complex. (C) 2010 Elsevier Inc. All rights reserved.