Transactivation in Drosophila of Human Enhancers by Human Transcription Factors Involved in Congenital Heart Diseases

Background: The human transcription factors (TFs) GATA4, NKX2.5 and TBX5 form part of the core network necessary to build a human heart and are involved in Congenital Heart Diseases (CHDs). The human natriuretic peptide precursor A (NPPA) and α-myosin heavy chain 6 (MYH6) genes are downstream effectors involved in cardiogenesis that have been demonstrated to be in vitro targets of such TFs. Results: To study the interactions between these human TFs and their target enhancers in vivo, we overexpressed them in the whole Drosophila cardiac tube using the UAS/GAL4 system. We observed that all three TFs up-regulate their natural target enhancers in Drosophila and cause developmental defects when overexpressed in eyes and wings. Conclusions: A strong potential of the present model might be the development of combinatorial and mutational assays to study the interactions between human TFs and their natural target promoters, which are not easily undertaken in tissue culture cells because of the variability in transfection efficiency, especially when multiple constructs are used. Thus, this novel system could be used to determine in vivo the genetic nature of the human mutant forms of these TFs, setting up a powerful tool to unravel the molecular genetic mechanisms that lead to CHDs. Developmental Dynamics 241:190–199, 2012. © 2011 Wiley Periodicals, Inc

The Hexapeptide and Linker Regions of the AbdA Hox Protein Regulate Its Activating and Repressive Functions

AbstractThe Hox family transcription factors control diversified morphogenesis during development and evolution. They function in concert with Pbc cofactor proteins. Pbc proteins bind the Hox hexapeptide (HX) motif and are thereby thought to confer DNA binding specificity. Here we report that mutation of the AbdA HX motif does not alter its binding site selection but does modify its transregulatory properties in a gene-specific manner in vivo. We also show that a short, evolutionarily conserved motif, PFER, in the homeodomain-HX linker region acts together with the HX to control an AbdA activation/repression switch. Our in vivo data thus reveal functions not previously anticipated from in vitro analyses for the hexapeptide motif in the regulation of Hox activity

Nano-architecture of gustatory chemosensory bristles and trachea in Drosophila wings

International audienceIn the Drosophila wing anterior margin, the dendrites of gustatory neurons occupy the interior of thin and long bristles that present tiny pores at their extremities. Many attempts to measure ligand-evoked currents in insect wing gustatory neurons have been unsuccessful for technical reasons. The functions of this gustatory activity therefore remain elusive and controversial. To advance our knowledge on this understudied tissue, we investigated the architecture of the wing chemosensory bristles and wing trachea using Raman spectroscopy and fluorescence microscopy. We hypothesized that the wing gustatory hair, an open-ended capillary tube, and the wing trachea constitute biological systems similar to nano-porous materials. We present evidence that argues in favour of the existence of a layer or a bubble of air beneath the pore inside the gustatory hair. We demonstrate that these hollow hairs and wing tracheal tubes fulfil conditions for which the physics of fluids applied to open-ended capillaries and porous materials are relevant. We also document that the wing gustatory hair and tracheal architectures are capable of trapping volatile molecules from the environment, which might increase the efficiency of their spatial detection by way of wing vibrations or during flight

Nature of reading difficulties in Brazilian children with development dyslexia

According to an information processing approach, different reading strategies prevail at different stages and depending on print characteristics.The first stage, logographic, is marked by the prevalence of the logographic strategy in which recognition is limited to a few familiar words and plagued with paralexias. The second stage, alphabetical, is marked by the prevalence of a phonological strategy, based on grapheme-phoneme decoding, which permits reading new words, provided their spelling is regular. The third stage, orthographic, is marked by the prevalence of a lexical strategy, based on visual recognition, which permits reading irregularly-spelled words, provided they are familiar. Reading Competence Test (RCT) analyzes systematically the ability of using each strategy. RTC is made of seven types of print-picture pairs, i.e., a print (either a word or a nonword) associated with a picture. The task is to circle correct print-picture pairs and to cross out incorrect ones. The former consist of either regularly-spelled words or irregularly-spelled words associated with their corresponding pictures. The latter consist of either words associated with unrelated pictures, or nonwords nonwords that look similar to words, nonwords that sound similar to words, nonwords that neither sound nor look similar to words. Using RCT, this study analyzed reading strategies used by 13 dyslexic children and compared them to those used by 2196 1st-7th grade normolexic students. Results revealed that the difficulty presented by dyslexic children is eminently phonological: In word reading, dyslexic children performed equivalent to normolexic children, whereas in nonword reading, they performed significantly worse, especially for nonwords that look or sound similar to words. The study corroborated the notion that phonological processing is a major component of alphabetical reading and spelling, and that phonological processing disorders are mainly responsible for the specific reading difficulties that characterize developmental dyslexia.Segundo uma abordagem de processamento de informação, diferentes estratégias de leitura prevalecem em diferentes estágios, dependendo das características da escrita. No primeiro estágio, logográfico, prevalece a estratégia logográfica, em que o reconhecimento é limitado a umas poucas palavras familiares e dificultado por paralexias. No segundo, alfabético, prevalece a estratégia fonológica de decodificação grafemafonema, que permite a leitura de palavras novas, desde que sejam regulares grafofonemicamente irregulares, desde que sejam familiares. O Teste de Competência de Leitura de Palavras e Pseudopalavras (TCLPP) analisa, sistematicamente, a habilidade de usar cada estratégia. Compõe-se de sete tipos de pares figura-escrita, i.e., uma figura associada a uma palavra ou pseudopalavra. A tarefa é circular os pares corretos e cruzar os incorretos. Os dois primeiros consistem de palavras grafofonemicamente regulares ou irregulares associadas a suas correspondentes figuras. Os cinco últimos consistem de palavras associadas a figuras diferentes, ou a pseudopalavras de quatro tipos: pseudopalavras com pronúncia idêntica à de palavras; pseudopalavras com aspecto similar ao de palavras; pseudopalavras com pronúncia similar em relação à de palavras; e pseudopalavras sem semelhança visual ou fonológica com palavras. Usando TCLPP, o estudo analisou estratégias de leitura de 13 disléxicos, comparando-as às de 2196 normoléxicos de 1ª-7ª série. Resultados mostraram que as dificuldades são basicamente fonológicas. Disléxicos foram tão bem quanto normoléxicos na leitura de palavras, mas significativamente pior na de pseudopalavras, especialmente naquelas com aspecto similar ao de palavras e naquelas com pronúncia similar à de palavras. O estudo corroborou a noção de que o processamento fonológico é o principal quesito de leitura e escrita alfabéticas, e que distúrbios naquele processamento são responsáveis pelas dificuldades específicas de leitura que caracterizam a dislexia do desenvolvimento

Comércio justo como geração de renda para assentados da Reforma agrária: feira permanente de produtos da Agricultura familiar.

Trabalho apresentado no 31º SEURS - Seminário de Extensão Universitária da Região Sul, realizado em Florianópolis, SC, no período de 04 a 07 de agosto de 2013 - Universidade Federal de Santa Catarina.Os produtores associados à Cooperativa dos Agricultores Familiares do Vale do Ivaí - Cooperivaí, tinham a necessidade de comercializarem seus produtos de forma justa, sem a presença de atravessadores. Diante disso surgiu a iniciativa de implantação da Feira Permanente da Reforma Agrária – Cooperivaí, que é espaço físico fixo destinado para comercialização de produtos da reforma agrária e da agricultura familiar. Neste local são encontrados frutas, verduras, legumes, pães, bolos, doces, geléias, mel, queijos e derivados do leite, grãos (arroz, feijão, ervilha, milho), mandioca, temperos e conservas, erva mate, ervas medicinais, ovos, polpa de frutas, sorvetes, congelados, produtos de limpeza, artesanatos, entre outros; tudo produzido pelos agricultores familiares e assentados rurais. Este empreendimento é uma organização sem fins lucrativos, conta com 30 produtores que abastecem a loja assiduamente com seus produtos. Os agricultores recebem individualmente por aquilo que é vendido e pequena parte das vendas (18%) é destinada à manutenção do espaço. Com isso, 30 famílias estão sendo diretamente beneficiadas pela geração de renda fomentada pela feira, além da população local, que tem acesso a produtos frescos, de qualidade e com preços acessíveis, movimentando assim, a economia local. Atualmente o faturamento da loja gira em torno de 10 mil reais mensais, sendo este valor gradualmente crescente desde a abertura da mesma. As frutas e hortaliças são os produtos mais vendidos, responsáveis por mais de 70% do valor total das vendas. Nestes 13 meses de abertura e funcionamento do espaço de comercialização foi observado que este modelo é uma alternativa viável e rentável para os pequenos agricultores familiares se inserirem em um modelo de comércio justo

Defining Satisfactory Methods of Treatment in Rare Diseases When Evaluating Significant Benefit-The EU Regulator's Perspective

Since the implementation of the EU Orphan Regulation in 2000, the Committee for Orphan Medicinal Products at the European Medicines Agency has been evaluating the benefits of proposed orphan medicines vs. satisfactory treatment methods. This type of evaluation is foreseen in the Orphan Regulation as the orphan designation criterion called the "significant benefit." In this article, based on 20 years of experience, we provide a commentary explaining what is considered a satisfactory method of treatment in the context of the EU Orphan Regulation and for the purpose of the assessment of significant benefit. We discuss the challenges posed by continuously changing clinical practise, which is associated with the increasing number of treatment options, evolving nature of medicinal therapeutic indications and our understanding of them

Towards sustainability in European agricultural firms

Agricultural activity plays an important role in all 28 Member States (MS) of the European Union (EU) in terms of: -Economics, -Environmental, -Social, and -Political activity. The sector also provides: -agricultural goods and services to support food security; -exports and imports at European and World level; -creation of direct and indirect jobs; and -the maintenance of population living in rural and regional areas.N/

Reduction of Fmr1 mRNA Levels Rescues Pathological Features in Cortical Neurons in a Model of FXTAS

Fragile X-associated tremor ataxia syndrome (FXTAS) is a rare disorder associated to the presence of the fragile X premutation, a 55–200 CGG repeat expansion in the 5′ UTR of the FMR1 gene. Two main neurological phenotypes have been described in carriers of the CGG premutation: (1) neurodevelopmental disorders characterized by anxiety, attention deficit hyperactivity disorder (ADHD), social deficits, or autism spectrum disorder (ASD); and (2) after 50 years old, the FXTAS phenotype. This neurodegenerative disorder is characterized by ataxia and a form of parkinsonism. The molecular pathology of this disorder is characterized by the presence of elevated levels of Fragile X Mental Retardation 1 (FMR1) mRNA, presence of a repeat-associated non-AUG (RAN) translated peptide, and FMR1 mRNA-containing nuclear inclusions. Whereas in the past FXTAS was mainly considered as a late-onset disorder, some phenotypes of patients and altered learning and memory behavior of a mouse model of FXTAS suggested that this disorder involves neurodevelopment. To better understand the physiopathological role of the increased levels of Fmr1 mRNA during neuronal differentiation, we used a small interfering RNA (siRNA) ap

Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1

BACKGROUND: Anosmin-1, the protein implicated in the X-linked Kallmann's syndrome, plays a role in axon outgrowth and branching but also in epithelial morphogenesis. The molecular mechanism of its action is, however, widely unknown. Anosmin-1 is an extracellular protein which contains a cysteine-rich region, a whey acidic protein (WAP) domain homologous to some serine protease inhibitors, and four fibronectin-like type III (FnIII) repeats. Drosophila melanogaster Kal-1 (DmKal-1) has the same protein structure with minor differences, the most important of which is the presence of only two FnIII repeats and a C-terminal region showing a low similarity with the third and the fourth human FnIII repeats. We present a structure-function analysis of the different DmKal-1 domains, including a predicted heparan-sulfate binding site. RESULTS: This study was performed overexpressing wild type DmKal-1 and a series of deletion and point mutation proteins in two different tissues: the cephalopharyngeal skeleton of the embryo and the wing disc. The overexpression of DmKal-1 in the cephalopharyngeal skeleton induced dosage-sensitive structural defects, and we used these phenotypes to perform a structure-function dissection of the protein domains. The reproduction of two deletions found in Kallmann's Syndrome patients determined a complete loss of function, whereas point mutations induced only minor alterations in the activity of the protein. Overexpression of the mutant proteins in the wing disc reveals that the functional relevance of the different DmKal-1 domains is dependent on the extracellular context. CONCLUSION: We suggest that the role played by the various protein domains differs in different extracellular contexts. This might explain why the same mutation analyzed in different tissues or in different cell culture lines often gives opposite phenotypes. These analyses also suggest that the FnIII repeats have a main and specific role, while the WAP domain might have only a modulator role, strictly connected to that of the fibronectins