Étude in vivo de la relation entre la structure et la fonction de la boucle variable de l'ARN de transfert de la sélénocystéine d'E. coli

La sélénocystéine est le 21e acide aminé encodé génétiquement et on la retrouve à travers les trois domaines de la vie. Elle est synthétisée sur l'ARNtSec par un processus unique. L'ARNtSec se distingue également au niveau structural. La tige acceptrice possède 8 (procaryotes) et 9 (eucaryotes) paires de bases, contrairement aux ARNt canoniques qui ont invariablement 7 paires de bases dans la tige acceptrice. De plus, la tige D a 2 paires de bases additionnelles qui remplacent les interactions tertiaires universelles 8-14, 15-48 qui sont absentes chez l'ARNtSec. D'autre part, la longueur de la boucle variable de l'ARNtSec est plus longue que la majorité des ARNt de type II. Dans ce mémoire, on se concentre sur la région de la boucle variable de l'ARNtSec . La recherche consiste à distinguer les paires de bases de la boucle variable qui sont essentielles à la biosynthèse et l’insertion de la sélénocystéine. De plus, on regarde si la paire de base additionnelle de la tige acceptrice de l'ARNtSec (procaryote) est essentielle pour l'insertion de la sélénocystéine. Pour répondre à ces questions, on a utilisé l'approche expérimentale Évolution Instantanée qui consiste au criblage in vivo d'ARNtSec fonctionnels chez E. coli. Dans ce travail, on montre que l'insertion de la sélénocystéine ne nécessite pas une spécificité de la longueur ou de la séquence de l'ARNtSec. On montre aussi que ni la longueur de la tige acceptrice ou du domaine tige acceptrice/tige T n'est essentielle pour avoir un ARNtSec fonctionnel.

The mitochondrial genome of the stingless bee Melipona bicolor (Hymenoptera, Apidae, Meliponini): sequence, gene organization and a unique tRNA translocation event conserved across the tribe Meliponini

At present a complete mtDNA sequence has been reported for only two hymenopterans, the Old World honey bee, Apis mellifera and the sawfly Perga condei. Among the bee group, the tribe Meliponini (stingless bees) has some distinction due to its Pantropical distribution, great number of species and large importance as main pollinators in several ecosystems, including the Brazilian rain forest. However few molecular studies have been conducted on this group of bees and few sequence data from mitochondrial genomes have been described. In this project, we PCR amplified and sequenced 78% of the mitochondrial genome of the stingless bee Melipona bicolor (Apidae, Meliponini). The sequenced region contains all of the 13 mitochondrial protein-coding genes, 18 of 22 tRNA genes, and both rRNA genes (one of them was partially sequenced). We also report the genome organization (gene content and order), gene translation, genetic code, and other molecular features, such as base frequencies, codon usage, gene initiation and termination. We compare these characteristics of M. bicolor to those of the mitochondrial genome of A. mellifera and other insects. A highly biased A+T content is a typical characteristic of the A. mellifera mitochondrial genome and it was even more extreme in that of M. bicolor. Length and compositional differences between M. bicolor and A. mellifera genes were detected and the gene order was compared. Eleven tRNA gene translocations were observed between these two species. This latter finding was surprising, considering the taxonomic proximity of these two bee tribes. The tRNA Lys gene translocation was investigated within Meliponini and showed high conservation across the Pantropical range of the tribe.

Evolutionary History of Arabidopsis thaliana Aminoacyl-tRNA Synthetase Dual-Targeted Proteins

Aminoacyl-transfer RNA (tRNA) synthetases (aaRS) are key players in translation and act early in protein synthesis by mediating the attachment of amino acids to their cognate tRNA molecules. In plants, protein synthesis may occur in three subcellular compartments (cytosol, mitochondria, and chloroplasts), which requires multiple versions of the protein to be correctly delivered to its proper destination. The organellar aaRS are nuclear encoded and equipped with targeting information at the N-terminal sequence, which enables them to be specifically translocated to their final location. Most of the aaRS families present organellar proteins that are dual targeted to mitochondria and chloroplasts. Here, we examine the dual targeting behavior of aaRS from an evolutionary perspective. Our results show that Arabidopsis thaliana aaRS sequences are a result of a horizontal gene transfer event from bacteria. However, there is no evident bias indicating one single ancestor (Cyanobacteria or Proteobacteria). The dual-targeted aaRS phylogenetic relationship was characterized into two different categories (paralogs and homologs) depending on the state recovered for both dual-targeted and cytosolic proteins. Taken together, our results suggest that the dual-targeted condition is a gain-of-function derived from gene duplication. Selection may have maintained the original function in at least one of the copies as the additional copies diverged.

The value of idiosyncratic markers and changes to conserved tRNA sequences from the mitochondrial genome of hard ticks (Acari: Ixodida: Ixodidae) for phylogenetic inference

Idiosyncratic markers are features of genes and genomes that are so unusual that it is unlikely that they evolved more than once in a lineage of organisms. Here we explore further the potential of idiosyncratic markers and changes to typically conserved tRNA sequences for phylogenetic inference. Hard ticks were chosen as the model group because their phylogeny has been studied extensively. Fifty-eight candidate markers from hard ticks ( family Ixodidae) and 22 markers from the subfamily Rhipicephalinae sensu lato were mapped onto phylogenies of these groups. Two of the most interesting markers, features of the secondary structure of two different tRNAs, gave strong support to the hypothesis that species of the Prostriata ( Ixodes spp.) are monophyletic. Previous analyses of genes and morphology did not strongly support this relationship, instead suggesting that the Prostriata is paraphyletic with respect to the Metastriata ( the rest of the hard ticks). Parallel or convergent evolution was not found in the arrangements of mitochondrial genes in ticks nor were there any reversals to the ancestral arthropod character state. Many of the markers identified were phylogenetically informative, whereas others should be informative with study of additional taxa. Idiosyncratic markers and changes to typically conserved nucleotides in tRNAs that are phylogenetically informative were common in this data set, and thus these types of markers might be found in other organisms.

Os desafios da avaliação de desempenho na AP do sec. XXI

Comunicação apresentada no 5º Congresso Nacional de Administração Pública "Modernização, Desenvolvimento e Competitividade", em Lisboa, a 29 e 30 de Outubro de 2007.

Technologie de production du jambon sec en Espagne

L’élaboration du jambon sec est réalisée depuis des temps immémoriaux par un procédé de conservation dû à la salaison et au séchage. Le produit ainsi obtenu est conditionné, par la qualité de la matière première utilisée et par le processus technologique qu’il a suivi. Il existe différentes technologies pour produire du jambon sec, mais fondamentalement elles essayent toutes d’obtenir un produit qui d’une part peut être maintenu à température ambiante sans compromettre la santé, et d’autre part qui facilite le développement des caractéristiques sensorielles désirées. L’Espagne, l’Italie, l’Allemagne et la France sont les principaux producteurs de jambons secs et en 2000, la production dans l’Union européenne était de plus de 600 000 tonnes. L’objectif de cette présentation est de passer en revue les principaux aspects du processus d’élaboration, en comparant les paramètres liés à la matière première et ceux liés aux technologies qui affectent la qualité du jambon sec.

Evolució del paisatge de l'Horta de Lleida (1956/57-2003): transformació d'un paisatge de secà a regadiu

Els paisatges agraris al llarg del temps s’han transformat i han evolucionat segons les necessitats de la societat d’aquell moment. L’aigua ha estat el principal element transformador en l’agricultura del segle XX, i el cas de l’horta de Lleida, és un clar exemple de com l’aigua ha desenvolupat una agricultura que poc a poc s’ha caracteritzat per tenir majoritàriament cultius de regadiu, tot i ser una de les zones més àrides i amb menys precipitacions de Catalunya, a diferència de les zones de secà, que han anat desapareixen. Quantificar i analitzar els canvis que influeixen en la dinàmica del territori, han estat possibles a partir de l’estudi de fonts documentals històriques, tant escrites com gràfiques, com són els cadastres de rústica, les imatges aèries i els ortofotomapes. Mitjançant aquest valuós recurs històric s’ha pogut detectar i analitzar els principals canvis en els usos del sòl esdevinguts durant el període de 1956 a 2003 en determinades zones de l’horta de Lleida, a més ha servit per estimar les repercussions en el paisatge del consum d’aigua.

Mitochondrial tRNA(Leu(UUR)) mutation m.3302A > G presenting as childhood-onset severe myopathy: threshold determination through segregation study.

Mitochondrial tRNA(Leu(UUR)) mutation m.3302A > G is associated with respiratory chain complex I deficiency and has been described as a rare cause of mostly adult-onset slowly progressive myopathy. Five families with 11 patients have been described so far; 5 of them died young due to cardiorespiratory failure. Here, we report on a segregation study in a family with an index patient who already presented at the age of 18 months with proximal muscular hypotonia, abnormal fatigability, and lactic acidosis. This early-onset myopathy was rapidly progressive. At 8 years, the patient is wheel-chair bound, requires nocturnal assisted ventilation, and suffers from recurrent respiratory infections. Severe complex I deficiency and nearly homoplasmy for m.3302A > G were found in muscle. We collected blood, hair, buccal swabs and muscle biopsies from asymptomatic adults in this pedigree and determined heteroplasmy levels in these tissues as well as OXPHOS activities in muscle. All participating asymptomatic adults had normal OXPHOS activities. In contrast to earlier reports, we found surprisingly little variation of heteroplasmy levels in different tissues of the same individual. Up to 45% mutation load in muscle and up to 38% mutation load in other tissues were found in non-affected adults. The phenotypic spectrum of tRNA(Leu(UUR)) m.3302A > G mutation seems to be wider than previously described. A threshold of more than 45% heteroplasmy in muscle seems to be necessary to alter complex I activity leading to clinical manifestation. The presented data may be helpful for prognostic considerations and counseling in affected families.

Effects of Trypanosoma brucei tryptophanyl-tRNA synthetases silencing by RNA interference

The kinetoplast genetic code deviates from the universal code in that 90% of mitochondrial tryptophans are specified by UGA instead of UGG codons. A single nucleus-encoded tRNA Trp(CCA) is used by both nuclear and mitochondria genes, since all kinetoplast tRNAs are imported into the mitochondria from the cytoplasm. To allow decoding of the mitochondrial UGA codons as tryptophan, the tRNA Trp(CCA) anticodon is changed to UCA by an editing event. Two tryptophanyl tRNA synthetases (TrpRSs) have been identified in Trypanosoma brucei: TbTrpRS1 and TbTrpRS2 which localize to the cytoplasm and mitochondria respectively. We used inducible RNA interference (RNAi) to assess the role of TbTrpRSs. Our data validates previous observations of TrpRS as potential drug design targets and investigates the RNAi effect on the mitochondria of the parasite.

Distinct subcellular localization of tRNA-derived fragments in the infective metacyclic forms of Trypanosoma cruzi

Small non-coding RNAs derived from transfer RNAs have been identified as a broadly conserved prokaryotic and eukaryotic response to stress. Their presence coincides with changes in developmental state associated with gene expression regulation. In the epimastigote form of Trypanosoma cruzi, tRNA fragments localize to posterior cytoplasmic granules. In the infective metacyclic form of the parasite, we found tRNA-derived fragments to be abundant and evenly distributed within the cytoplasm. The fragments were not associated with polysomes, suggesting that the tRNA-derived fragments may not be directly involved in translation control in metacyclics.

Technologies de procédé et de contrôle pour réduire la teneur en sel du jambon sec et des saucissons

Dans certains pays européens, les produits carnés élaborés peuvent représenter près de 20% de la consommation journalière de sodium. De ce fait, les industries de la viande tentent de réduire la teneur en sel dans les produits carnés pour répondre, d’une part aux attentes des consommateurs et d’autre part aux demandes des autorités sanitaires. Le système Quick‐Dry‐Slice process (QDS®), couplé avec l’utilisation de sels substituant le chlorure de sodium (NaCl), a permis de fabriquer, avec succès, des saucisses fermentées à basse teneur en sel en réduisant le cycle de fabrication et sans ajout de NaCl supplémentaire. Les technologies de mesure en ligne non destructives, comme les rayons X et l’induction électromagnétique, permettent de classifier les jambons frais suivant leur teneur en gras, un paramètre crucial pour adapter la durée de l’étape de salaison. La technologie des rayons X peut aussi être utilisée pour estimer la quantité de sel incorporée pendant la salaison. L’information relative aux teneurs en sel et en gras est importante pour optimiser le processus d’élaboration du jambon sec en réduisant la variabilité de la teneur en sel entre les lots et dans un même lot, mais aussi pour réduire la teneur en sel du produit final. D’autres technologies comme la spectroscopie en proche infrarouge (NIRS) ou spectroscopie microondes sont aussi utiles pour contrôler le processus d’élaboration et pour caractériser et classifier les produits carnés élaborés, selon leur teneur en sel. La plupart de ces technologies peuvent être facilement appliquées en ligne dans l’industrie afin de contrôler le processus de fabrication et d’obtenir ainsi des produits carnés présentant les caractéristiques recherchées.

Cabo Verde Uma Experiência Colonial Acelerada (Sec.XVI-XVII)

Não vamos aqui trazer matéria nova relativamente ao que está escrito nos dois volumes já publicados da História Geral de Cabo Verde e nos vários artigos preparatórios e/ou resultantes desse labor, quase contínuo, de historiadores caboverdianos e portugueses. Propomo-nos antes observar o resultado do nosso trabalho, procurando uma perspectiva distanciada que permita encontrar algumas linhas de força na média duração de cerca de dois séculos. Poderíamos começar pela verificação de que, aqui se aprontou o primeiro cadinho de interacção de povos e culturas luso euro/africanos, na África ao Sul do Saara. Mas sabemos como os avanços e retrocessos, as continuidades e rupturas eliminam, e invertem até, prioridades de zonas geográficas. A História está cheia desses exemplos. Por isso, o primeiro indicador que se impõe, na perspectiva da média duração, não está tanto na antecipação e precocidade dos fenómenos sociais e culturais, resultantes da colonização europeia na África, mas no ritmo com que a nova sociedade os vive, apropria, integra e reinterpreta.

A evolução da sociedade foguense: Através de um estudo prosopográfico da elite da ilha (sec. XVI-XVIII)

No seu admirável romance, “Ilhéu da Contenda”, Teixeira de Sousa, relata o pensamento de Chiquinho, um jovem mestiço foguense que enquanto matuta sobre a sua posição social, como filho ilegítimo de um branco da terra, expõe-nos sucintamente os seus pensamentos sobre a história da sua família e da sua ilha: “...De que lhe valia ser filho de branco se não usava o nome do pai? /.../ Preferia não receber coisa alguma do pai se em troca apenas lhe desse o seu apelido. Não queria nada mais de Nhô Eusébio. Francisco Medina da Veiga seria nome mais bonito do que só Francisco de Pina. Francisco de Pina. Nome de negro. Ao passo que os Medinas e os Veigas foram sempre gentes graúdas desde o povoamento da ilha. O primeiro Veiga foi capitão-mor do Fogo, e tão rico que os descendentes se mantiveram abastados até os que ainda viviam. Ouvia contar a Nha Caela que esse Afonso Sanches Veiga capitão-mor da ilha, foi o tronco da família do marido. Pois Nho Pedro Simplício da Veiga descendia de uma linha directa desse grande homem que se celebrizou pela sua riqueza e também pela sua crueldade. Os escravos não brincavam com ele. Quando pisavam o risco, mandava-os amarrar a calabaceira de Ilhéu de 2 Contenda e ele próprio os ia castigar com varas de marmeleiro entrançadas em três. Depois mandava botar sal e vinagre nos lanhos sangrentos. Era violento como tudo, esse Sanches da Veiga, tetravô de Pedro Simplício” (pp.61-62)

Albereda del Riu Sec

Projecte presentat per al: Concurs de projectes per a la redacció de l’avantprojecte general d’ordenació de l’area d’equipaments culturals, educatius i esportius anomenada “Can Xarau”, així com el projecte d’execució i la direcció d’obra de la urbanització del parc, i dels vials d’accés, a Cerdanyola del Vallés