Coupling Virus-Induced Gene Silencing to Exogenous Green Fluorescence Protein Expression Provides a Highly Efficient System for Functional Genomics in Arabidopsis and across All Stages of Tomato Fruit Development

Since the advent of the postgenomic era, efforts have focused on the development of rapid strategies for annotating plant genes of unknown function. Given its simplicity and rapidity, virus-induced gene silencing (VIGS) has become one of the preeminent approaches for functional analyses. However, several problems remain intrinsic to the use of such a strategy in the study of both metabolic and developmental processes. The most prominent of these is the commonly observed phenomenon of ""sectoring"" the tissue regions that are not effectively targeted by VIGS. To better discriminate these sectors, an effective marker system displaying minimal secondary effects is a prerequisite. Utilizing a VIGS system based on the tobacco rattle virus vector, we here studied the effect of silencing the endogenous phytoene desaturase gene (pds) and the expression and subsequent silencing of the exogenous green fluorescence protein (gfp) on the metabolism of Arabidopsis (Arabidopsis thaliana) leaves and tomato (Solanum lycopersicum) fruits. In leaves, we observed dramatic effects on primary carbon and pigment metabolism associated with the photobleached phenotype following the silencing of the endogenous pds gene. However, relatively few pleiotropic effects on carbon metabolism were observed in tomato fruits when pds expression was inhibited. VIGS coupled to gfp constitutive expression revealed no significant metabolic alterations after triggering of silencing in Arabidopsis leaves and a mild effect in mature green tomato fruits. By contrast, a wider impact on metabolism was observed in ripe fruits. Silencing experiments with an endogenous target gene of interest clearly demonstrated the feasibility of cosilencing in this system; however, carefully constructed control experiments are a prerequisite to prevent erroneous interpretation.

Complex rearrangements in patients with duplications of MECP2 can occur by fork stalling and template switching

Duplication at the Xq28 band including the MECP2 gene is one of the most common genomic rearrangements identified in neurodevelopmentally delayed males. Such duplications are non-recurrent and can be generated by a non-homologous end joining (NHEJ) mechanism. We investigated the potential mechanisms for MECP2 duplication and examined whether genomic architectural features may play a role in their origin using a custom designed 4-Mb tiling-path oligonucleotide array CGH assay. Each of the 30 patients analyzed showed a unique duplication varying in size from similar to 250 kb to similar to 2.6 Mb. Interestingly, in 77% of these non-recurrent duplications, the distal breakpoints grouped within a 215 kb genomic interval, located 47 kb telomeric to the MECP2 gene. The genomic architecture of this region contains both direct and inverted low-copy repeat (LCR) sequences; this same region undergoes polymorphic structural variation in the general population. Array CGH revealed complex rearrangements in eight patients; in six patients the duplication contained an embedded triplicated segment, and in the other two, stretches of non-duplicated sequences occurred within the duplicated region. Breakpoint junction sequencing was achieved in four duplications and identified an inversion in one patient, demonstrating further complexity. We propose that the presence of LCRs in the vicinity of the MECP2 gene may generate an unstable DNA structure that can induce DNA strand lesions, such as a collapsed fork, and facilitate a Fork Stalling and Template Switching event producing the complex rearrangements involving MECP2.

Genetic dissection of vitamin E biosynthesis in tomato

Vegetables are critical for human health as they are a source of multiple vitamins including vitamin E (VTE). In plants, the synthesis of VTE compounds, tocopherol and tocotrienol, derives from precursors of the shikimate and methylerythritol phosphate pathways. Quantitative trait loci (QTL) for alpha-tocopherol content in ripe fruit have previously been determined in an Solanum pennellii tomato introgression line population. In this work, variations of tocopherol isoforms (alpha, beta, gamma, and delta) in ripe fruits of these lines were studied. In parallel all tomato genes structurally associated with VTE biosynthesis were identified and mapped. Previously identified VTE QTL on chromosomes 6 and 9 were confirmed whilst novel ones were identified on chromosomes 7 and 8. Integrated analysis at the metabolic, genetic and genomic levels allowed us to propose 16 candidate loci putatively affecting tocopherol content in tomato. A comparative analysis revealed polymorphisms at nucleotide and amino acid levels between Solanum lycopersicum and S. pennellii candidate alleles. Moreover, evolutionary analyses showed the presence of codons evolving under both neutral and positive selection, which may explain the phenotypic differences between species. These data represent an important step in understanding the genetic determinants of VTE natural variation in tomato fruit and as such in the ability to improve the content of this important nutriceutical.

Genomic Analysis of Wild Tomato Introgressions Determining Metabolism- and Yield-Associated Traits

With the aim of determining the genetic basis of metabolic regulation in tomato fruit, we constructed a detailed physical map of genomic regions spanning previously described metabolic quantitative trait loci of a Solanum pennellii introgression line population. Two genomic libraries from S. pennellii were screened with 104 colocated markers from five selected genomic regions, and a total of 614 bacterial artificial chromosome (BAC)/cosmids were identified as seed clones. Integration of sequence data with the genetic and physical maps of Solanum lycopersicum facilitated the anchoring of 374 of these BAC/cosmid clones. The analysis of this information resulted in a genome-wide map of a nondomesticated plant species and covers 10% of the physical distance of the selected regions corresponding to approximately 1% of the wild tomato genome. Comparative analyses revealed that S. pennellii and domesticated tomato genomes can be considered as largely colinear. A total of 1,238,705 bp from both BAC/cosmid ends and nine large insert clones were sequenced, annotated, and functionally categorized. The sequence data allowed the evaluation of the level of polymorphism between the wild and cultivated tomato species. An exhaustive microsynteny analysis allowed us to estimate the divergence date of S. pennellii and S. lycopersicum at 2.7 million years ago. The combined results serve as a reference for comparative studies both at the macrosyntenic and microsyntenic levels. They also provide a valuable tool for fine-mapping of quantitative trait loci in tomato. Furthermore, they will contribute to a deeper understanding of the regulatory factors underpinning metabolism and hence defining crop chemical composition.

Nyblivan föräldrars upplevelser av separation från sitt nyfödda barn efter förlossningen : en systematisk litteraturstudie

I olika sammanhang och i olika omfattning separeras mor och barn idag inom den perinatala vården. Då barnet behöver vård på neonatalavdelning leder det ofta till upprepad och långvarig separation mellan föräldrarna och barnet. Syftet med denna litteraturstudie är att beskriva föräldrars upplevelse av separation från sitt nyfödda barn efter förlossningen. Metod- studien är en systematisk litteraturstudie där 15 vetenskapliga artiklar ingår. Resultat- situationer som leder till separation mellan mamma och barn kan utgöra ett unikt tillfälle för pappan till nära kontakt och tidig anknytning mellan honom och barnet. Behovet att vara nära sitt barn är starkt och finns hos båda föräldrarna och separation leder till många negativa känslor och ökad stress hos föräldrarna samt utgör ett hinder för den tidiga anknytningen till barnet. Skillnader i behovet att vara nära barnet i den tidiga nyföddhetsperioden kan ses mellan mammor och pappor och mellan föräldrar till prematura barn och fullgångna. Nära hudkontakt och samvård mellan föräldrar och barn har visat sig ha många positiva effekter. Slutsatser- separation mellan föräldrar och barn har många negativa konsekvenser, såväl fysiska, psykiska som sociala. Kontinuerlig närhet efter förlossningen mellan mamman och det nyfödda barnet är viktigt och närhet mellan pappan och det nyfödda barnet är att föredra vid de tillfällen mor och barn måste separeras. Neonatal samvård skapar förutsättningar för en god anknytning mellan föräldrar och barn och ger möjlighet att påbörja föräldraskapet under optimala förutsättningar.

Förebyggande åtgärder vid ventilatorassocierad pneumoni : En systematisk litteraturstudie

Syftet med litteraturstudien var att beskriva förebyggande omvårdnadsåtgärder vid ventilatorassocierad pneumoni (VAP), vilket är en form av pneumoni som är sjukhusförärvad och relaterad till att patienten är intuberad, så kallad nosokomial infektion. Längre vårdtid, ökad kostnad samt mortalitet ses vid VAP. Resultatet som framkom var att personal ska ha god handhygien vilket var en av viktigaste åtgärden för att förebygga VAP. Andra förebyggande omvårdnadsåtgärder är att vårdpersonal ska hjälpa patienten som vårdas i respirator med munvården och använda hjälpmedel för detta. Personal ska även hjälpa patienten med lämplig höjning på huvudgärden då detta också minskar riken för VAP, även sugning och befuktning av luftvägar är en åtgärd som patienten behöver hjälp med.

Personalized Communications : A Cross Media tool for the future

Personalized communication is when the marketing message is adapted to each individual by using information from a databaseand utilizing it in the various, different media channels available today. That gives the marketer the possibility to create a campaign that cuts through today’s clutter of marketing messages and gets the recipients attention. PODi is a non-profit organization that was started with the aim of contributing knowledge in the field of digital printingtechnologies. They have created a database of case studies showing companies that have successfully implemented personalizedcommunication in their marketing campaigns. The purpose of the project was therefore to analyze PODi case studies with the main objective of finding out if/how successfully the PODi-cases have been and what made them so successful. To collect the data found in the PODi cases the authors did a content analysis with a sample size of 140 PODi cases from the year 2008 to 2010. The study was carried out by analyzing the cases' measurable ways of success: response rate, conversion rate, visited PURL (personalized URL:s) and ROI (Return On Investment). In order to find out if there were any relationships to be found between the measurable result and what type of industry, campaign objective and media vehicle that was used in the campaign, the authors put up different research uestions to explore that. After clustering and merging the collected data the results were found to be quite spread but shows that the averages of response rates, visited PURL and conversion rates were consistently very high. In the study the authors also collected and summarized what the companies themselves claim to be the reasons for success with their marketing campaigns. The resultshows that the creation of a personalized campaign is complex and dependent on many different variables. It is for instance ofgreat importance to have a well thought-out plan with the campaign and to have good data and insights about the customer in order to perform creative personalization. It is also important to make it easy for the recipient to reply, to use several media vehicles for multiple touch points and to have an attractive and clever design.

Vårdpersonals attityder till munvårdshandlingar hos äldre patienter

Syfte: Syftet med denna studie var att beskriva vårdpersonals attityder till utförandet av omvårdnadshandlingar i munhålan hos äldre patienter som var beroende av att få hjälp med munvårdshandlingar. Vidare var syftet att beskriva vilka åtgärder som gjorts för att skapa förändringar i personalens attityder, kunskaper och vårdhandlingar. Metod: Studien genomfördes som en litteraturstudie. Sökningarna gjordes i databaserna PubMed och CINAHL där både kvalitativa och kvantitativa studier söktes. Studierna granskades sedan med modifierade granskningsmallar för kvalitetsbedömning Resultat: De sökta studiernas resultat analyserades och utifrån detta skapades sex teman, Kunskap, Tidsbrist, Patientens motvillighet, Personalens motvilja, Utbildning och Rutiner. Dessa teman delades upp under de två frågeställningarna. Resultatet visade att bristande kunskap om munvårdshandlingar, brist på tid, motvilja från patienten och motvilja att utföra munvård var det som gjorde att munvården inte sköttes korrekt. Många patienter hade bristande munhygien i flertalet studier. Utbildning och nya rutiner var åtgärder som förbättrade munvården, personalens attityder och deras kunskap om munvård. Det visade sig även att patienternas munhygien förbättrades till stor del efter dessa interventioner. Slutsats: Munvård är ett ämne prioriteras ner i vårdarbetet och anses inte lika betydande som övrig vård. Interventioner som nya rutiner och utbildning påverkar personal att utföra munvårdshandlingar och därför borde utbildning prioriteras högt i förbättringsarbetet för god munvård. Samt att nya rutiner med munvårdskort och elektriska tandborstar är enkla medel för att skapa en bättre munhygien hos patienter.

Knowledge and perceptions regarding sexual and reproductive health among high school students in Kathmandu, Nepal

Background: Nepal recently began teaching sexual education in the school system and has established youth friendly services in order to meet the need of increased sexual and reproductive knowledge among the youth. Objective: To examine the sexual and reproductive knowledge and perceptions among young people attending schools in Kathmandu. Method: A written questionnaire was distributed to 160 students, in a classroom environment, in four schools in Kathmandu. Results: Two thirds of the females and nearly 60% of the males knew that it was possible to get sexually transmitted infection (STI) during one sexual encounter and more than half of the students knew when in the menstrual cycle conception was more likely to occur . One third of the participants did not know that it was possible to become pregnant after having intercourse once. The males demonstrated less knowledge than the females regarding every aspect of sex and reproduction, with the exception of pregnancy prevention. Conclusion and clinical implications: For the youths in this study, it was more important to prevent unwanted pregnancies than to protect oneself from STIs. Establishment of a hotline on the internet, where personalized and confidential counselling can be offered may complement the comprehensive sexual education in schools.

The Potential Of Kohonen Method For Classifying Rivers Focused On Regional Environmental Flow Determination With ELOHA

This article highlights the potential benefits that the Kohonen method has for the classification of rivers with similar characteristics by determining regional ecological flows using the ELOHA (Ecological Limits of Hydrologic Alteration) methodology. Currently, there are many methodologies for the classification of rivers, however none of them include the characteristics found in Kohonen method such as (i) providing the number of groups that actually underlie the information presented, (ii) used to make variable importance analysis, (iii) which in any case can display two-dimensional classification process, and (iv) that regardless of the parameters used in the model the clustering structure remains. In order to evaluate the potential benefits of the Kohonen method, 174 flow stations distributed along the great river basin “Magdalena-Cauca” (Colombia) were analyzed. 73 variables were obtained for the classification process in each case. Six trials were done using different combinations of variables and the results were validated against reference classification obtained by Ingfocol in 2010, whose results were also framed using ELOHA guidelines. In the process of validation it was found that two of the tested models reproduced a level higher than 80% of the reference classification with the first trial, meaning that more than 80% of the flow stations analyzed in both models formed invariant groups of streams.

Análise da agenda do financiamento do SUS: diferentes contextos, mesmas escolhas?

O estudo aborda o tema do financiamento do SUS, procurando fazer uma reconstrução histórica evidenciando os embates intra Poder Executivo, entre as áreas de Saúde e Fazenda e procurando compreender quais foram os fatores que refletiram nas dificuldades de resolução da questão durante todo o período que se seguiu criação do SUS na Constituição Federal de 1988. Para isso, procuramos entender quais foram os constrangimentos e restrições impostas a essa política, especialmente nos governo de Fernando Henrique Cardoso e Lula. Procuramos entender em que medida os diferentes contextos sociais, políticos e econômicos desses governos levaram à opção pela não priorização do equacionamento da insuficiência de recursos. Nesse sentido, procuramos compreender se houve um caráter de continuidade entre esses governos. Destacamos todas as dificuldades de caráter financeiro enfrentadas por essa política universal na década de 90 até 2010, a luta pela ampliação de recursos e as soluções que se apresentaram, especialmente a CPMF, Emenda Constitucional n.º 29, bem como sua regulamentação. Ressaltamos também o papel do Executivo e do Legislativo na produção de legislação em saúde, bem como os conflitos entre as áreas de Saúde e Fazenda e o papel dos órgãos de fiscalização e controle (Tribunal de Contas da União e Ministério Público Federal). Como resultados, apresentamos as evidências de que houve continuidade na abordagem do tema do financiamento do SUS nos governos FHC e Lula e de que a agenda que prevaleceu foi a da área econômica. Apesar disso, a área de saúde conseguiu preservar-se como direito social universal e obteve muitos avanços.

Marketing de serviços públicos

Este trabalho aborda as possíveis aplicações do Marketing ao campo dos Serviços Públicos têm sido objeto de muita investigação e, ultimamente, diversos autores abordam esse tema em seus trabalhos. Esses estudos fazem parte de uma investigação mais ampla que indaga sobre a possibilidade de estender o campo do Marketing para as organizações não tipicamente comerciais ou para aquelas que não visam primordialmente o lucro. Foi iniciado portanto um longo debate, a nível teórico, o qual envolve uma discussão sobre a própria conceituação do Marketing, seu campo de aplicação e sua evolução recente. Inserindo nesse contexto mais amplo o objeto específico de nossa investigação: os serviços públicos. A partir desse modelo procuramos então fornecer alguns elementos para responder a questão básica sobre a possibilidade de se transferir ou adaptar o instrumental desenvolvido com base nos fenômenos de caráter comercial, micro, tangível para os serviços públicos.

Diagnóstico do Sistema de Tecnologia da Informação do Estado do Rio de Janeiro

A tecnologia de informação mudou nos últimos anos o modo de viver, de pensar e de se relacionar das pessoas. Influenciadas pelos avanços tecnológicos que se massificam cada vez mais rapidamente, as pessoas, que também são cidadãos dos Estados onde vivem, mudaram inclusive a forma de interagir com as instituições. Exemplos não faltam nos jornais sobre esse fenômeno. Os governos não podem ficar para traz nessa revolução, que já se compara em escala e efeito à revolução industrial do século XIX, sob pena de perderem a já combalida legitimidade. O desafio é grande, pois a velocidade da evolução técnica coloca mais uma pressão sobre os gestores públicos. Para acompanhar é preciso modernizar a forma de administrar os órgãos do governo e de relaciona-los com os cidadãos e a sociedade civil. Uma gestão da Tecnologia da Informação que parta dos primeiros escalões dos governos é condição inicial para mudar a forma de pensar e agir de toda burocracia. Neste estudo vamos avaliar como é a Governança de TI do Estado do Rio de Janeiro em face às estruturas dos demais estados, considerando também as visões orçamentária e de planejamento e ainda a experiência de profissionais integrantes do Sistema de Tecnologia da Informação do Estado do Rio de Janeiro. Pretendemos extrais lições sobre o que deve ser mantido, ampliado ou alterado no modelo de governança adotado.

Controle vetorial de velocidade de uma máquina de indução sem mancais trifásica com bobinado dividido utilizando estimação neural de fluxo

This work describes the study and the implementation of the vector speed control for a three-phase Bearingless induction machine with divided winding of 4 poles and 1,1 kW using the neural rotor flux estimation. The vector speed control operates together with the radial positioning controllers and with the winding currents controllers of the stator phases. For the radial positioning, the forces controlled by the internal machine magnetic fields are used. For the radial forces optimization , a special rotor winding with independent circuits which allows a low rotational torque influence was used. The neural flux estimation applied to the vector speed controls has the objective of compensating the parameter dependences of the conventional estimators in relation to the parameter machine s variations due to the temperature increases or due to the rotor magnetic saturation. The implemented control system allows a direct comparison between the respective responses of the speed and radial positioning controllers to the machine oriented by the neural rotor flux estimator in relation to the conventional flux estimator. All the system control is executed by a program developed in the ANSI C language. The DSP resources used by the system are: the Analog/Digital channels converters, the PWM outputs and the parallel and RS-232 serial interfaces, which are responsible, respectively, by the DSP programming and the data capture through the supervisory system

Viabilidade da Utilização de Máquinas de Indução Convencionais como Motores sem Mancais Mecânicos

Electrical Motors transform electrical energy into mechanic energy in a relatively easy way. In some specific applications, there is a need for electrical motors to function with noncontaminated fluids, in high speed systems, under inhospitable conditions, or yet, in local of difficult access and considerable depth. In these cases, the motors with mechanical bearings are not adequate as their wear give rise to maintenance. A possible solution for these problems stems from two different alternatives: motors with magnetic bearings, that increase the length of the machine (not convenient), and the bearingless motors that aggregate compactness. Induction motors have been used more and more in research, as they confer more robustness to bearingless motors compared to other types of machines building with others motors. The research that has already been carried out with bearingless induction motors utilized prototypes that had their structures of stator/rotor modified, that differ most of the times from the conventional induction motors. The goal of this work is to study the viability of the use of conventional induction Motors for the beringless motors applications, pointing out the types of Motors of this category that can be more useful. The study uses the Finite Elements Method (FEM). As a means of validation, a conventional induction motor with squirrel-cage rotor was successfully used for the beringless motor application of the divided winding type, confirming the proposed thesis. The controlling system was implemented in a Digital Signal Processor (DSP)