Centrin Is Necessary for the Formation of the Motile Apparatus in Spermatids of Marsilea

During spermiogenesis in the water fern, Marsilea vestita, basal bodies are synthesized de novo in cells that lack preexisting centrioles, in a particle known as a blepharoplast. We have focused on basal body assembly in this organism, asking what components are required for blepharoplast formation. Spermiogenesis is a rapid process that is activated by placing dry microspores into water. Dry microspores contain large quantities of stored protein and stored mRNA, and inhibitors reveal that certain proteins are translated from stored transcripts at specific times during development. Centrin translation accompanies blepharoplast appearance, while β-tubulin translation occurs later, during axonemal formation. In asking whether centrin is an essential component of the blepharoplast, we used antisense, sense, and double-stranded RNA probes made from the Marsilea centrin cDNA, MvCen1, to block centrin translation. We employed a novel method to introduce these RNAs directly into the cells. Antisense and sense both arrest spermiogenesis when blepharoplasts should appear, and dsRNA made from the same cDNA is an effective inhibitor at concentrations at least 10 times lower than either of the single-stranded RNA used in these experiments. Blepharoplasts are undetectable and basal bodies fail to form. Antisense, sense, and dsRNA probes made from Marsilea β-tubulin permitted normal development until axonemes form. In controls, antisense, sense, and dsRNA, made from a segment of HIV, had no effect on spermiogenesis. Immunoblots suggest that translational blocks induced by centrin-based RNA are gene specific and concentration dependent, since neither β-tubulin- nor HIV-derived RNAs affects centrin translation. The disruption of centrin translation affects microtubule distributions in spermatids, since centrin appears to control formation of the cytoskeleton and motile apparatus. These results show that centrin plays an essential role in the formation of a motile apparatus during spermiogenesis of M. vestita.

The Role of Gibberellin, Abscisic Acid, and Sucrose in the Regulation of Potato Tuber Formation in Vitro1

The effects of plant hormones and sucrose (Suc) on potato (Solanum tuberosum L.) tuberization were studied using in vitro cultured single-node cuttings. Tuber-inducing (high Suc) and -noninducing (low Suc or high Suc plus gibberellin [GA]) media were tested. Tuberization frequencies, tuber widths, and stolon lengths were measured during successive stages of development. Endogenous GAs and abscisic acid (ABA) were identified and quantified by high-performance liquid chromatography and gas chromatography-mass spectrometry. Exogenous GA4/7 promoted stolon elongation and inhibited tuber formation, whereas exogenous ABA stimulated tuberization and reduced stolon length. Indoleacetic acid-containing media severely inhibited elongation of stolons and smaller sessile tubers were formed. Exogenous cytokinins did not affect stolon elongation and tuber formation. Endogenous GA1 level was high during stolon elongation and decreased when stolon tips started to swell under inducing conditions, whereas it remained high under noninducing conditions. GA1 levels were negatively correlated with Suc concentration in the medium. We conclude that GA1 is likely to be the active GA during tuber formation. Endogenous ABA levels decreased during stolon and tuber development, and ABA levels were similar under inducing and noninducing conditions. Our results indicate that GA is a dominant regulator in tuber formation: ABA stimulates tuberization by counteracting GA, and Suc regulates tuber formation by influencing GA levels.

Formation of junctions involved in excitation-contraction coupling in skeletal and cardiac muscle.

During excitation-contraction (e-c) coupling of striated muscle, depolarization of the surface membrane is converted into Ca2+ release from internal stores. This process occurs at intracellular junctions characterized by a specialized composition and structural organization of membrane proteins. The coordinated arrangement of the two key junctional components--the dihydropyridine receptor (DHPR) in the surface membrane and the ryanodine receptor (RyR) in the sarcoplasmic reticulum--is essential for their normal, tissue-specific function in e-c coupling. The mechanisms involved in the formation of the junctions and a potential participation of DHPRs and RyRs in this process have been subject of intensive studies over the past 5 years. In this review we discuss recent advances in understanding the organization of these molecules in skeletal and cardiac muscle, as well as their concurrent and independent assembly during development of normal and mutant muscle. From this information we derive a model for the assembly of the junctions and the establishment of the precise structural relationship between DHPRs and RyRs that underlies their interaction in e-c coupling.

Specificity of dimer formation in tropomyosins: influence of alternatively spliced exons on homodimer and heterodimer assembly.

Tropomyosins consist of nearly 100% alpha-helix and assemble into parallel and in-register coiled-coil dimers. In vitro it has been established that nonmuscle as well as native muscle tropomyosins can form homodimers. However, a mixture of muscle alpha and beta tropomyosin subunits results in the formation of the thermodynamically more stable alpha/beta heterodimer. Although the assembly preference of the muscle tropomyosin heterodimer can be understood thermodynamically, the presence of multiple tropomyosin isoforms expressed in nonmuscle cells points toward a more complex principle for determining dimer formation. We have investigated the dimerization of rat tropomyosins in living cells by the use of epitope tagging with a 16-aa sequence of the influenza hemagglutinin. Employing transfection and immunoprecipitation techniques, we have analyzed the dimers formed by muscle and nonmuscle tropomyosins in rat fibroblasts. We demonstrate that the information for homo- versus heterodimerization is contained within the tropomyosin molecule itself and that the information for the selectivity is conferred by the alternatively spliced exons. These results have important implications for models of the regulation of cytoskeletal dynamics.

Suppression of synapsin II inhibits the formation and maintenance of synapses in hippocampal culture.

Numerous synaptic proteins, including several integral membrane proteins, have been assigned roles in synaptic vesicle fusion with or retrieval from the presynaptic plasma membrane. In contrast, the synapsins, neuron-specific phosphoproteins associated with the cytoplasmic surface of synaptic vesicles, appear to play a much broader role, being involved in the regulation of neurotransmitter release and in the organization of the nerve terminal. Here we have administered antisense synapsin II oligonucleotides to dissociated hippocampal neurons, either before the onset of synaptogenesis or 1 week after the onset of synaptogenesis. In both cases, synapsin II was no longer detectable within 24-48 h of treatment. After 5 days of treatment, cultures were analyzed for the presence of synapses by synapsin I and synaptophysin antibody labeling and by electron microscopy. Cultures in which synapsin II was suppressed after axon elongation, but before synapse formation, did not develop synapses. Cultures in which synapsin II was suppressed after the development of synapses lost most of their synapses. Remarkably, with the removal of the antisense oligonucleotides, neurons and their synaptic connections recovered. These studies lead us to conclude that synapsin II is involved in the formation and maintenance of synapses in hippocampal neurons.

Enteropathogenic Escherichia coli contains a putative type III secretion system necessary for the export of proteins involved in attaching and effacing lesion formation.

Enteropathogenic Escherichia coli (EPEC) causes a characteristic histopathology in intestinal epithelial cells called the attaching and effacing lesion. Although the histopathological lesion is well described the bacterial factors responsible for it are poorly characterized. We have identified four EPEC chromosomal genes whose predicted protein sequences are similar to components of a recently described secretory pathway (type III) responsible for exporting proteins lacking a typical signal sequence. We have designated the genes sepA, sepB, sepC, and sepD (sep, for secretion of E. coli proteins). The predicted Sep polypeptides are similar to the Lcr (low calcium response) and Ysc (yersinia secretion) proteins of Yersinia species and the Mxi (membrane expression of invasion plasmid antigens) and Spa (surface presentation of antigens) regions of Shigella flexneri. Culture supernatants of EPEC strain E2348/69 contain several polypeptides ranging in size from 110 kDa to 19 kDa. Proteins of comparable size were recognized by human convalescent serum from a volunteer experimentally infected with strain E2348/69. A sepB mutant of EPEC secreted only the 110-kDa polypeptide and was defective in the formation of attaching and effacing lesions and protein-tyrosine phosphorylation in tissue culture cells. These phenotypes were restored upon complementation with a plasmid carrying an intact sepB gene. These data suggest that the EPEC Sep proteins are components of a type III secretory apparatus necessary for the export of virulence determinants.

Caracterização do perfil audiológico de escolares de escola pública do município de Bauru/SP

Muitos dos problemas auditivos não são notados por pais e professores. Este fato prejudica a aprendizagem da criança principalmente no ambiente escolar. Por isso, programas de triagem auditiva podem ser utilizados com o intuito de detectar e, posteriormente, diagnosticar escolares a fim de que se possa prevenir ou minimizar o impacto a que possíveis sequelas auditivas venham prejudicar o rendimento escolar da criança. Hoje em dia podemos contar com programas que permitem o melhor acompanhamento de populações que necessitam de cuidados preventivos e curativos, e a audição é um aspecto muito importante que pode ser avaliado quando estes programas são colocados em prática. O Programa Nacional de Reorientação da Formação Profissional em Saúde (Pró-Saúde), que visou reorientar a formação profissional, teve como objetivo integrar ensino-serviço e promover atenção básica por meio da abordagem integral do processo saúde-doença. Ambientes externos podem ser utilizados por alunos e professores universitários para que possam colocadas em prática ações que possibilitem a humanização das práticas de atenção a saúde e a integralidade das mesmas, por meio da articulação de ações e serviços de saúde, preventivos e curativos, individuais e coletivos. A escola é considerada um dos ambientes que este trabalho pode ser realizado. O Programa Saúde na Escola (PSE) abre o ambiente escolar com a finalidade de contribuir para a formação integral dos estudantes da rede pública de educação básica por meio de ações de prevenção, promoção e atenção à saúde. Sendo um estudo do tipo retrospectivo transversal, como objetivo principal caracterizar o perfil audiológico de escolares de escola pública do município de Bauru SP, contando com a integração de profissionais da área da saúde e educação no ambiente escolar, o que teve como base os programas citados acima. A triagem auditiva foi realizada com a aplicação dos seguintes procedimentos: imitanciometria, inspeção visual do meato acústico externo, emissões otoacústicas por produto de distorção e audiometria tonal liminar. Observou-se que do total de 652 estudantes, a grande maioria (97,1%) dos participantes com faixa etária entre 10 e 18 anos, apresentaram audição normal. Em 2,9% desta população foi encontrada alguma alteração auditiva temporária. Com a exceção de um único participante, portador de perda auditiva sensorioneural. Apesar de encontrarmos muitas crianças e adolescentes com audição normal, o que mais ressalta a importância deste trabalho é a necessidade da triagem auditiva em ambientes escolares e, essencialmente, o acompanhamento das mesmas nesta faixa etária, já que são escassos os estudos referentes a ela. Apesar das poucas alterações auditivas encontradas serem passageiras, são exatamente estas que interferem no bom rendimento escolar e outros fatores.

Sociología y drogodependencia: la mirada ténica en la intervención social. Fundamentos para la constitución de una sociología de las drogodependencias

Almost thirty years have passed since the City Council of Madrid approved the first Municipal Plan against Drugs, thus laying the foundations of the current commitment to offer assistance to drug addicts. A Service that has been continuously growing (in funding), maturing (in organizations) and diversifying (in actions) during these decades, in the same manner as the scenario in which these actions are deployed has been evolving. But, what can be said today about the status of drugaddiction intervention? This study adopts a sociological approach that starts with and is focused on what has been defined as the hegemonic vision on drug addiction, with the aim of studying (precisely and progressively) the different and most widely accepted theoretical-practical developments and to understand how certain conceptions or positions determine the form of this social fact. Therefore, the main objective of this study is to understand and evaluate the performative effects of certain practices, techniques and professionals on defining and addressing a reality that is in question and/or in conflict, as a social problem. In addition, this study is focused on identifying a series of fundamental elements to address what is understood as the state of the issue of drug addiction in depth (discourse of drugs Vs. discourse on drugs) in order to offer a series of sociological questions and insights to focus attention onto this reality. For this purpose, once its intentions have been defined and due to the social nature of the object under study, the present research deploys qualitative methodology as a tool for approximation, delimiting the scope that professional technicians assume regarding the issue in question as well as their own professional practices within their organizations. Thus, we have contacted certain public as well as non-governmental organizations with long-term experience in drug addiction, so as to find out first hand, in addition to their writings, the different meanings they find in the performance of their specific practice (micro) as well as the framework within which their practice is defined and materialised (macro) on a population defined or characterised by (drug) addiction...

Simultaneous implementation of the Bachelor, Masters and PhD degrees in nursing in the Universidad Jaume I. Castellón de la Plana, Spain

We present an experience in Nursing Education, accredited and implemented under the Spanish University System Reform in a Public University (Jaume I, Castellón) which had no previous nursing studies. The academics offered included all three educational levels (Bachelor, Master's and Doctorate), with an integrated theoretical-practical-clinical teaching methodology for the Bachelor Degree, competence acquisition in research in the Master's degree, and a doctorate formed by lines of research in the field of Nursing. Studies are accredited by the National Agency for Quality Assessment, which were authorized by the Spanish Ministry of Education and implanted between 2009 and 2011.

Satisfaction of clinical nurses with the formation of a university tutorial program

Degree in nursing from the Universitat Jaume I (UJI) maintains the continuity of learning with an integrated learning methodology (theory, simulated practice and clinical practice). The objective of this methodology is to achieve consistency between the knowledge, abilities and skills acquired in the classroom, laboratory and clinic to ensure skills related. Reference Nurse is a key figure in this process, you receive accredited training on Educational Methods, assessment of competence, and Evidence-Based Practice that plays the role of evaluating in conjunction with the subjects. It does not perceive economic remuneration. The main objective of this study is to determine the level of satisfaction of clinical nurses on the Nurses Training Program Reference in UJI (Castellon- Spain). A cross sectional study was performed and conducted on 150 nurses. 112 questionnaires were completed, collected and analysed at the end of training. The survey consists of 12 items measured with the Likert scale with 5 levels of response and two open questions regarding the positive and negative aspects of the course and to add in this formation. The training is always performed by the same faculty and it's used four sessions of 2012. We perform a quantitative analysis of the variables under study using measures of central tendency. The completion rate of the survey is 95.53% (n=107). Anonymity rate of 54,14% The overall satisfaction level of training was 3.65 (SD = 0.89) on 5 points. 54.2% (n = 58) of the reference nurses made a contribution in the open questions described in the overall results. The overall satisfaction level can be considered acceptable. It is considered necessary to elaborate a specific survey to detect areas of improvement of nurse training program reference and future recruitment strategies. The main objective of the present work is the selection and integration of different methodologies among those applicable within the framework of the European Higher Education Area to combine teaching methods with high implication from both lecturers and students.

Spanish Nursing Students' Practical Experience of Computer Applications in Nursing

This study aimed to determine the level of computer practical experience in a sample of Spanish nursing students. Each student was given a Spanish language questionnaire, modified from an original used previously with medical students at the Medical School of North Carolina University (USA) and also at the Education Unit of Hospital General Universitario del Mar (Spain). The 10-item self-report questionnaire probed for information about practical experience with computers. A total of 126 students made up the sample. The majority were female (80.2%; n=101). The results showed that just over half (57.1%, n=72) of the students had used a computer game (three or more times before), and that only one third (37.3%, n=47) had the experience of using a word processing package. Moreover, other applications and IT-based facilities (e.g. statistical packages, e-mail, databases, CD-ROM searches, programming languages and computer-assisted learning) had never been used by the majority of students. The student nurses' practical experience was less than that reported for medical students in previous studies.

Construction of Structures I in Moodle: a new teaching methodology in the Degree of Building Engineer

The subject of Construction of Structures I studies, from a constructive point of view and taking into account current legislation, reinforced concrete structures used in buildings, through the acquisition of knowledge and construction criteria required in the profession of a Technical Architect. The contents acquired in this course are essential for further professional development of technicians and are closely related to many of the subjects taught in the same or other courses of the Degree in Technical Architecture at the University of Alicante. The aim of this paper is to present, analyze and discuss the development of a new methodology proposed in the mentioned subject, as it supposed an important change in the traditional way of teaching Construction and Structures I. In order to incorporate new teaching tools in 2013-2014, the course has been implemented by using a Moodle software tool to promote blended learning with online exercises. Our Moodle community allows collaborative work within an open-source platform where teachers and students share a new and personalized learning environment. Students are easily used to the interface and the platform, value the constant connection with teachers or other fellows and completely agree with the possibility of making questions or share documents 24 hours a day. The proposed methodology consists of lectures and practical classes. In the lectures, the basics of each topic are discussed; class attendance, daily study and conducting scheduled exercises are indispensable. Practical classes allow to consolidate the knowledge gained in theory classes by solving professional exercises and actual construction problems related to structures, that shall be compulsorily delivered online. So, after the correction of the teacher and the subsequent feedback of students, practical exercises ensure lifelong learning of the student, who can download any kind of material at any time (constructive details, practical exercises and even corrected exams). Regarding the general evaluation system, goals achievement is assessed on an ongoing basis (65% of the final mark) along the course through written and graphic evidences in person and online, as well as a individual development of a workbook. In all cases, the acquisition of skills, the ability to synthesize, the capacity of logical and critical thinking are assessed. The other 35 % of the mark is evaluated by a complementary graphic exam. Participation in the computing platform is essential and the student is required to do and present, at least 90% of the practices proposed. Those who do not comply with the practices in each specific date could not be assessed continuously and may only choose the final exam. In conclusion, the subject of Construction of Structures I is essential in the development of the regulated profession of Technical Architect as they are considered, among other professional profiles, as specialists in construction of building structures. The use of a new communication platform and online teaching allows the acquisition of knowledge and constructive approaches in a continuous way, with a more direct and personal monitoring by the teacher that has been highly appreciated by almost 100% of the students. Ultimately, it is important to say that the use of Moodle in this subject is a very interesting tool, which was really well welcome by students in one of the densest and important subjects of the Degree of Technical Architecture.

Framing Banking Union in the Euro Area: Some empirical evidence. CEPS Working Document No. 389, 3 February 2014

Evidence shows that financial integration in the euro area is retrenching at a quicker pace than outside the union. Home bias persists: Governments compete on funding costs by supporting ‘their’ banks with massive state aids, which distorts the playing field and feeds the risk-aversion loop. This situation intensifies friction in credit markets, thus hampering the transmission of monetary policies and, potentially, economic growth. This paper discusses the theoretical foundations of a banking union in a common currency area and the legal and economic aspects of EU responses. As a result, two remedies are proposed to deal with moral hazard in a common currency area: a common (unlimited) financial backstop to a privately funded recapitalisation/resolution fund and a blanket prohibition on state aids.

Belarus-Russia: Whither Integration? OSW Point of View (unspecified), May 2003

Of the re-integration processes currently taking place in the former Soviet Union, the formation of a Russian-Belarusian so-called 'Union State' is one of the most advanced. A customs union was formally announced between the two countries as early as 1995 and the process of constructing the Union State itself was launched in December 1999. However, both events were largely driven by the perceived need to match societal demands, without much concrete action and the Union State remained largely 'virtual'. Only in the last few years has the Russian initiative allowed for moving from symbolic gestures to political action and since late 2002 debate and policy have intensified on specific issues of economic and political co-operation. However, despite such advances in the integration process, its objectives remain vague and there is little or no agreement on the principles that should govern the process. Furthermore, current bilateral relations questions still dominate the dialogue. The project seems at present to be driven mainly by the political interests of both countries' presidents and also, to a lesser extent, by the interests of business, political, military and security elites, each apparently motivated by self- and group-interest in the emerging dialogue of integration. In contrast to EU integration, the societies of the two countries involved appear to have had little or no say in the process. Thus, several questions naturally arise. What is the real nature of such integration? What motivates the parties involved? What stage has the process reached? What likely future course will it take? What might be the consequences of it for Belarusian independence? Answers to these questions should ultimately determine the stance and policies of the enlarged EU in this area.

Innovations of the European Central Bank in the Context of Financial and Monetary Integration: A Chinese Assessment. ZEI Discussion Paper C229, 2015

In the 15 years since the introduction of the Euro, the integration process within the European Economic and Monetary Union has seen rapid development in terms of both breadth and depth. Exclusively responsible for the monetary policy of the Eurozone, the European Central Bank has continued to adjust to meet the challenges brought about by these changes. The paper explores financial and monetary integration in the Eurozone and reviews the reasons, specific performance and impact of changes in the European Central Bank’s decision-making mechanisms. The purpose of which is to deepen and expand understanding in academic circles of the European economy and the European Economic and Monetary Union, as well as their development trends.