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.

Development of new nuclear markers and characterization of single nucleotide polymorphisms in kelp gull (Larus dominicanus)

The present study seeks to develop nuclear markers for the kelp gull (Larus dominicanus). We hereby report the characterization of 12 independent nuclear introns, where 104 single nucleotide polymorphisms (SNPs) in 8138 sequenced base pairs were observed. These SNP markers are the first to be designed for genotyping a gull species. The markers will provide useful tools for understanding which processes act or acted upon kelp gulls to cause their low genetic variability in mitochondrial DNA. In addition, these markers open a new opportunity for population genetic and evolutionary studies in the Laridae group.

Effects of Plasmodium gallinaceum on hemolymph physiology of Aedes aegypti during parasite development

Insect disease vectors show diminished fecundity when infected with Plasmodium. This phenomenon has already been demonstrated in laboratory models such as Aedes aegypti, Anopheles gambiae and Anopheles stephensi. This study demonstrates several changes in physiological processes of A. aegypti occurring upon infection with Plasmodium gallinaceum, such as reduced ecdysteroid levels in hemolymph as well as altered expression patterns for genes involved in vitellogenesis, lipid transport and immune response. Furthermore, we could show that P. gallinaceum infected A. aegypti presented a reduction in reproductive fitness, accompanied by an activated innate immune response and increase in lipophorin expression, with the latter possibly representing a nutritional resource for Plasmodium sporozoites. (C) 2010 Elsevier Ltd. All rights reserved.

Metallopeptidase inhibitors arrest vital biological processes in the fungal pathogen Scedosporium apiospermum

P>Scedosporium apiospermum is an emerging agent of opportunistic mycoses in humans. Previously, we showed that mycelia of S. apiospermum secreted metallopeptidases which were directly linked to the destruction of key host proteins. In this study, we analysed the effect of metallopeptidase inhibitors on S. apiospermum development. As germination of inhaled conidia is a crucial event in the infectious process of S. apiospermum, we studied the morphological transformation induced by the incubation of conidia in Sabouraud-dextrose medium at 37 degrees C. After 6 h, some conidia presented a small projection resembling a germ-tube. A significant increase, around sixfold, in the germ-tube length was found after 12 h, and hyphae were exclusively observed after 24 h. Three distinct metallopeptidase inhibitors were able to arrest the transformation of conidia into hyphae in different ways; for instance, 1,10-phenanthroline (PHEN) completely blocked this process at 10 mu mol l-1, while ethylenediamine tetraacetic acid (EDTA) and ethylene glycol-bis (beta-aminoethyl ether; EGTA) only partially inhibited the differentiation at up to 10 mmol l-1. EGTA did not promote any significant reduction in the conidial growth, while PHEN and EDTA, both at 10 mmol l-1, inhibited the proliferation around 100% and 65%, respectively. The secretion of polypeptides into the extracellular environment and the metallopeptidase activity secreted by mycelia were completely inhibited by PHEN. These findings suggest that metallo-type enzymes could be potential targets for future therapeutic interventions against S. apiospermum.

Phosphoproteomics Profiling Suggests a Role for Nuclear beta IPKC in Transcription Processes of Undifferentiated Murine Embryonic Stem Cells

Protein kinase C (PKC) plays a key role in embryonic stem cell (ESC) proliferation, self-renewal and differentiation However, the function of specific PKC Isoenzymes have yet to be determined Of the PKCs expressed in undifferentiated ESCs, beta IPKC was the only isoenzyme abundantly expressed in the nuclei To investigate the role of beta IPKC in these cells, we employed a phosphoproteomics strategy and used two classical (cPKC) peptide modulators and one beta IPKC-specific inhibitor peptide We identified 13 nuclear proteins that are direct or indirect beta IPKC substrates in undifferentiated ESCs These proteins are known to be involved in regulating transcription, splicing, and chromatin remodeling during proliferation and differentiation Inhibiting beta IPKC had no effect on DNA synthesis in undifferentiated ESCs However, upon differentiation many cells seized to express beta IPKC and beta IPKC was frequently found in the cytoplasm Taken together, our results suggest that beta IPKC takes part in the processes that maintain ESCs in their undifferentiated state

Development of an Analytical Methodology for Quantification of Strong Acid in Diesel Oil

Many factors can affect the quality of diesel oil, in particular the degradation processes that are directly related to some organosulfur compounds. During the degradation process, these compounds are oxidized into their corresponding sulfonic acids, generating a strong acid content during the process. p-Toluene sulfonic acid analysis was performed using the linear sweep voltammetry technique with a platinum ultramicroelectrode in aqueous solution containing 3 mol L(-1) potassium chloride. An extraction step was introduced prior to the voltammetric detection in order to avoid the adsorption of organic molecules, which inhibit the electrochemical response. The extraction step promoted the transference of sulfonic acid from the diesel oil to an aqueous phase. The method was accurate and reproducible, with detection and quantification limits of 5 ppm and 15 ppm, respectively. Recovery of sulfonic acid was around 90%.

Photovoltaic Process Development and innovative Techniques

Photovoltaic processing is one of the processes that have significance in semiconductor process line. It is complicated due to the no. of elements involved that directly or indirectly affect the processing and final yield. So mathematically or empirically we can’t say assertively about the results specially related with diffusion, antireflective coating and impurity poisoning. Here I have experimented and collected data on the mono-crystal silicon wafers with varying properties and outputs. Then by using neural network with available experimental data output required can be estimated which is further tested by the test data for authenticity. One can say that it’s a kind of process simulation with varying input of raw wafers to get desired yield of photovoltaic mono-crystal cells.

Processes of feelings in a society with a violent past : A qualitative study of the communication for Societal healing in the Truth Commissions in East Timor, Sri Lanka and Ghana between 2002-2011

The research investigates in what extent and how communication for meeting feelings is provided in Truth Commission work. It examines if and in what way feelings are addressed in the communication officially published by the Truth Commissions in East Timor, Ghana and Sri Lanka, occurring between 2002-2011. The research is also looking at the healing processes in a time perspective to find out if there is a communication for Societal healing to be continued in a longer term. My conclusion is that two cases of three in my research, the TRCs in Ghana and East Timor, have communication clearly directed to meet feelings caused by the war. One of the three cases (East Timor) has a communication with a clear ambition to heal over a longer period, to continue after the existence of the Truth Commission. The research suggests that communication with a clear ambition to reach out widely in the society, a communication directed to meet and process feelings over a longer period, can make Societal healing more effective. It also concludes that, in the future, Societal healing, as a field in conflict resolution, will be more based on representational media than today, provided through web communication.

Development, technological change and innovation, Schumpeter and neo-Schumpeterians

This review essay is devoted to a discussion of some central aspects of the Schumpeterian and neo-Schumpeterian approaches to the dynamic processes of development, technological change and innovation. This essay is organised in two parts. In the first, Schumpeter's insightful distinction between circular flow and development is discussed. In the second, some central elements of the neo-Schumpeterian interpretation and extension of Schumpeter's views are critically outlined, special emphasis being placed on some recent attempts to formalize several of his insights on the cyclical dynamics of the processes of technological change and innovation. I should stress that due to space constraints I will focus primarily upon macrotheoretic issues, thus paying only secondary attention to the neo-Schumpeterian literature on the microeconomics of technological change and to the burgeoning empirical developments along those lines.

Social entrepreneurship, microcredit and development challenges: a case study analysis of Banco Pérola

The term “social entrepreneurship” has been attracting growing interest from different sectors in the past years, driven by the possibility of employing business techniques to tackle recurrent social and environmental issues. At the forefront of this global phenomenon is microcredit, seen by many as an effective anti-poverty tool and having the Grameen Bank as its flagship program. While the prospects of social entrepreneurship seem promising, the newness of the concept and its somewhat confusing definition make conditions difficult to analyze this contemporary phenomenon. Therefore, the objective of this study was to discuss the challenges faced by social entrepreneurs and alternatives of development for social businesses through a case study on a Brazilian microcredit institution and inclusive business, Banco Pérola. The case addresses a growing need for case studies designed for teaching in the field of social entrepreneurship. It was focused mainly on understanding the development challenges within Banco Pérola, and built based on interviews carried out with top management, credit officer and clients of the institution, as well as on secondary data collected. An analysis of the case study was performed under a Teaching Notes. As illustrated by the Banco Pérola case, the main difficulties encountered by social entrepreneurs relate to the systematization of processes and creation of operational routines, including for performance evaluation (impact assessment tools); to the capture and management of both financial and human capital; to scaling up the business model and to the need of forging closer and more personal relationships with customers as against in traditional banking practices. In spite of certain limitations, such as the fact that the case might soon become outdated due to the fast-changing environment surrounding Banco Pérola, or the fact that not all relevant stakeholders (e.g. partners) were selected for interviews, the research objective has been achieved and the study can be seen as a contribution to spreading the concept of social entrepreneurship.

Effect of density on population development in the Amazon River prawn Macrobrachium amazonicum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development of mint (Mentha piperita L.) grown on biosolids: evaluation of productivity and essential oil content

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Development of amorphous carbon protective coatings on poly(vinyl)chloride

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Phantom development for radiographic image optimization of chest, skull and pelvis examination for nonstandard patient

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A new vision of the origin and the oocyte development in the Ostariophysi applied to Gymnotus sylvius (Teleostei: Gymnotiformes)

Tendo por base os novos conhecimentos oriundos de recentes estudos com Perciformes marinho, a origem e o desenvolvimento dos oócitos no Ostariophysi Gymnotus sylvius são aqui descritos. da mesma maneira que ocorre nos Perciformes, em Gymnotus sylvius as oogônias são encontradas no epitélio germinativo que margeia as lamelas ovígeras. No início da foliculogênese, a proliferação das oogônias e sua entrada em meiose dão origem a ninhos de células germinativas que se projetam em direção ao estroma ovariano, a partir do epitélio germinativo. Os ninhos e o epitélio germinativo são suportados pela mesma membrana basal que os separa do estroma. Coincidindo com a paralisação da meiose os oócitos, presentes nos ninhos, são separados uns dos outros por processos citoplasmáticos das células pré-foliculares. As células pré-foliculares derivam do epitélio germinativo sendo, portanto, inicialmente células epiteliais. Durante a foliculogênese, ao mesmo tempo em que envolvem os oócitos individualizando-os, as células pré-foliculares sintetizam a membrana basal ao seu redor. Os oócitos entram em crescimento primário ainda dentro dos ninhos. Ao término da foliculogênese, o oócito e as células foliculares que compõem o folículo são circundados pela membrana basal. O folículo permanece conectado ao epitélio germinativo uma vez que ambos compartilham uma porção comum da membrana basal. Células oriundas do estroma circundam o folículo ovariano exceto na região de compartilhamento da membrana basal formando a teca. O folículo, a membrana basal e a teca formam o complexo folicular. O desenvolvimento do oócito ocorre dentro do complexo folicular e compreende os estágios de crescimento primário e secundário, maturação e ovulação. Os alvéolos corticais surgem no ooplasma momentos antes do início do crescimento secundário ou estágio vitelogênico que tem início com a deposição de vitelo, progride até o oócito esteja completamente desenvolvido e o ooplasma preenchido pelos glóbulos de vitelo. A maturação é caracterizada pela migração do núcleo ou vesícula germinativa, pela quebra da vesícula germinativa, ou seja, pela fragmentação do envoltório nuclear e, retomada da meiose. Na ovulação o ovo é liberado do complexo folicular para o lúmen ovariano. em comparação com os Perciformes marinhos com ovos pelágicos, o desenvolvimento oocitário em Gymnotus sylvius tem menos etapas dentro dos estágios de desenvolvimento, sendo as duas mais notáveis delas as ausências da formação das gotas de lipídio durante os crescimentos primário e secundário (e a consequente fusão das gotas para formar um único glóbulo de lipídio durante a maturação) e, a hidrólise do vitelo antecedendo a ovulação.