A adoção da rotação de culturas com leguminosas em áreas de implantação e reforma de canaviais é uma estratégia promissora para recuperação e manutenção da fertilidade do solo, e tem potencial no aproveitamento do N para as soqueiras. A análise mais detalhada do comportamento do N no solo durante o ano agrícola e dos parâmetros de produção em locais que possuem solo e clima contrastantes, permitem aprimorar o manejo da adubação nitrogenada da cana-de-açúcar. As hipóteses deste trabalho são (1) a rotação de culturas irá aumentar a disponibilidade de N no solo, a qual suprirá parte da demanda do N das soqueiras subsequentes e (2) a rotação de culturas promoverá redução da resposta das soqueiras à adubação nitrogenada. Objetivou-se avaliar o efeito da rotação de culturas nas transformações do N no solo (N mineral, N-ISNT e N e C da biomassa microbiana) e na produtividade da primeira soqueira. Os experimentos foram instalados em quatro regiões representativas de cultivo de cana-de-açúcar na região Centro Sul do Brasil: Quatá/SP (Local A); Iracemápolis/SP (Local B); Chapadão do Céu/GO (Local C) e Quirinópolis/GO (Local D). Em cada local, foi instalado um experimento em esquema de parcelas subdivididas no delineamento blocos ao acaso com quatro repetições, em áreas submetidas (com rotação) ou não (sem rotação) ao plantio de leguminosa do gênero crotalária. Após a colheita da cana planta, foram implantados os seguintes tratamentos: 60, 120 e 180 kg ha-1 de N e um controle (sem N), aplicados superficialmente sobre a palhada. Foram realizadas três amostragens para determinação do conteúdo de N mineral, N-ISNT e C e N da biomassa microbiana. Entre 150 a 180 dias após a colheita da cana-planta, foram realizadas avaliações de perfilhamento e índice SPAD. Na colheita, avaliou-se matéria seca da parte aérea, produtividade e atributos tecnológicos. Os resultados demonstraram que a mineralização dos resíduos da crotalária influenciaram a dinâmica do C e N e os reservatórios de N no perfil do solo ao longo do ciclo da primeira soqueira, com destaque para o solo de textura média (Local A). Apesar da rotação de culturas ter modificado o conteúdo de N mineral, N-ISNT, e aumentado o perfilhamento e índice SPAD no local A, somente nos solos argilosos (Locais C e D), a rotação aumentou a produtividade da soqueira. A hipótese de que a rotação de culturas reduz a demanda de fertilizantes nitrogenados não foi aceita; houve resposta à adubação nitrogenada nos quatro locais, independentemente da rotação de culturas. Porém, a rotação demonstrou potencial em aumentar a produção de biomassa com manutenção da dose de N. Esta pesquisa demonstrou a viabilidade de rotação de culturas em solos argilosos com elevado conteúdo de C e N total, com potencial de aumentar a produtividade do canavial, porém sem reduzir a demanda de adubação nitrogenada. / The adoption of crop rotation with sugarcane is a promising strategy for the recovery and maintenance of soil fertility, and it shows potential in the use of N to the sugarcane ratoon. A more detailed analysis of the transformations N in soil during the agricultural year and production parameters in sites that have contrasting soil texture and climate, allows to improve management of N fertilization for sugarcane. The hypothesis of this work are (1) crop rotation will increase soil N availability, which will satisfy part of N requirements of sugarcane ratoon and (2) crop rotation will reduce the sugarcane ratoon response to N fertilization. The objective was to evaluate the effects of sunn hemp rotation in the transformations of N in soil (mineral N content, ISNT-N and microbial biomass C and N) and sugarcane ratoon yield. The field trials were installed in four representative regions of cultivation of sugarcane in Brazil: Quatá/SP (Site A); Iracemápolis/SP (Site B); Chapadão do Céu/GO (Site C) e Quirinópolis/GO (Site D). At each site, a field trial was installed in split plot in a randomized block experimental design with four replications, in areas sowed (rotation) and not sowed (no rotation) with sunn hemp. After the harvest of cane-plant, the following treatments were applied: 60, 120 and 180 kg ha-1 of N and a control (without N) over the straw. Three sampling were performed to determine the soil inorganic N content, ISNT-N, C and N of microbial biomass. Between 150 to 180 days after the harvest of the cane-plant, tillering was counted and SPAD index was measured. At harvest of the sugarcane ratoon, biometric parameters, yield and quality parameters were evaluated. The results showed that mineralization of residues of sunn hemp influenced the dynamics of C and N and N content in the soil profile throughout the cycle of the sugarcane ratoon, with emphasis on medium-textured soil (Site A). Despite crop rotation has modified mineral N stocks, ISNT-N, and increased tillering and SPAD index in site A, crop rotation only increased the sugarcane ratoon yield on clay soils (Sites C and D). The hypothesis that crop rotation reduces N-fertilizer requirement was not accepted; there was response to N fertilization in four sites, regardless of crop rotation. However, the rotation showed potential in increase biomass production with maintenance of N rate. This research demonstrated the viability of crop rotation in clay soils with high total C and N content, with potential to increase sugarcane yield, but without reducing N-fertilizer requirement.

Neste trabalho, foi estudada uma rota alternativa para síntese de poliuretanos sem a utilização de isocianatos (NIPU) por um processo de fabricação seguro e utilizando CO2 como insumo. A produção destes poliuretanos \"verdes\" ocorreu através da reação entre ciclocarbonatos e diaminas. A caracterização da estrutura química e a análise dos produtos da reação de formação dos poliuretanos foi possível com a utilização de espectroscopia de infravermelho com transformada de Fourier (FTIR) e espectroscopia de ressonância magnética nuclear (RMN). Foram sintetizados também os ciclocarbonatos pela reação de cicloadição de CO2 ao grupo oxirano de uma resina epóxi e o co-catalisador (resinato de zinco) utilizado na obtenção dos carbonatos cíclicos. Para otimização da formação dos ciclocarbonatos desenvolveu-se um reator de síntese, o qual permitiu a análise do rendimento da reação em função dos parâmetros de síntese controlados: pressão de CO2, temperatura do meio reacional, tempo de síntese, gaseificação da resina epóxi com CO2 (por agitação mecânica ou sistema de refluxo de CO2) e fonte de aquecimento (convencional ou utilizando radiação de microondas). A estrutura química do carbonato obtido, o rendimento e os possíveis subprodutos da reação de cicloadição também foram analisados por FTIR e RMN. / Seeking to optimize the formation of ciclocarbonates, it was developed a reactor in a closed system (no atmospheric emissions of CO2) in which the parameters of synthesis could be controlled: CO2 pressure, temperature of the reaction medium, distribution and concentration of CO2 gas bubbles in epoxy resin. The development of the ciclocarbonate synthesis\'s reactor included new systems to improve the gas-liquid contact between the reactants (aerator and CO2 reflux system) and also a heating system using microwave radiation. It were synthesized in this research, the co-catalyst (zinc resinate), the ciclocarbonate under different conditions of: pressure, temperature, synthesis time, heating source, aeration and catalyst, and \"green\" polyurethanes by an alternative route instead of the use of isocyanates, which have potential in specific applications (control of properties through chemical structure of the polymer). The result of the carbonates formation reaction was analyzed by Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (RMN) to obtain the carbonates\'s chemical structure\'s, yield and the reaction\'s by-products. Non-isocyanate polyurethanes (NIPU) were obtained from a manufacturing process and secure using CO2 as an input. The characterization of the chemical structure and the yield and by-product\'s analysis of the polyurethanes formation reaction also happened by FTIR and RMN.

This investigation tested the hypothesis that iron, as a micronutrient, will affect proteins in Trichodesmium and therefore affect nitrogen fixation. Changes in proteins that are a result of iron enrichment were compared to naturally occuring diel changes. Alterations in the iron protein of nitrogenase were compared to nitrogen fixation rates using the acetylene reduction technique. The observed changes in proteins were compared in Trichodesmium colonies from the Caribbean Sea and the Sargasso Sea. Trichodesmium colonies were monitored for protein and iron content over a diel period on two cruises. The changes in protein and iron content in Trichodesmium colonies were variable but at times showed a cyclic diel pattern. Changes in protein bands on SDS-PAGE showed consistent changes in the banding pattern of a low molecular weight protein that responded to iron nutrition and time of day (Elardo and Rueter 1990). These changes were similar to changes m the iron protein of nitrogenase which also responded to changes associated with iron nutrition and time of day (Elardo 1991 ). Trichodesmium appear to alter certain proteins which appear as changes in banding patterns in response to environmental factors such as nutrients, temperature and light. My research shows that the pattern of modification of the iron protein of nitrogenase differs in colonies from the Caribbean Sea compared to those from the Sargasso Sea (Elardo 1991). The Caribbean Sea population in February had a clear pattern of active and inactive forms (day vs. night) of the enzyme. The Sargasso Sea population of Trichodesmium spp. had both forms of the enzyme at all times of the day during April and May when NO3 - is present in the euphotic zone due to recent mixing. These differences between the two populations may be due to different environmental conditions since the Caribbean Sea is permanently stratified, warmer and nutrient-depleted throughout the year. The Sargasso Sea undergoes seasonal breakdown of the thermocline during winter months, resulting in an injection of nitrate from deeper water, and minimum temperatures of 18oC.

No description available.

Stable isotope probing (SIP) is an established technique that can be applied to identify the metabolically active micro-organisms within a microbial population. The SIP method utilises an isotopically-labelled substrate and PCR techniques to discern the members of a microbial community that incorporate the isotope into their DNA or RNA. The current literature gap around using 15N isotopes with RNA-SIP offers real potential and advantages for targeting and identifying active members from mixed communities involved in global biogeochemical nitrogen cycling. This study specifically investigated whether nitrogen based compounds can be used as substrates in RNA-SIP methodologies and whether they can in turn be used to probe mixed community environments known to be actively fixing nitrogen. The nitrogen-limited systems targeted represented an ideal opportunity to assess the suitability of 15N-RNA-SIP approaches due to their known high nitrogen fixation rates. Identifying these nitrogen-fixing bacteria could provide a better representation analysis of the community, leading to an improved prediction on how to manage and optimise the treatment performance of target waste systems and to exploit the unique bioconversion properties of these types of organisms. Initially, the project undertook methodological proof of concept by using a soluble nitrogen source, 15NH4Cl, to label the RNA of Novosphingobium nitrogenifigens and a mixed microbial community. Successful separation of the 14N- (control) and 15N-RNA was achieved for both pure and mixed communities using isopycnic caesium trifluoroacetate (CsTFA) gradients in an ultracentrifuge. The usefulness of this technique to identify active diazotrophs in real environmental samples was tested using a nitrogen-fixing community from a pulp and paper wastewater treatment system. After growing the mixed culture with 15N2 as the sole nitrogen source, the labelled RNA was extracted and fractionated using isopycnic centrifugation in CsTFA gradients. The community composition of the active nitrogen-fixing community in the 15N2 enriched fraction was analysed by establishing a 16S rRNA gene clone library containing over 200 members. These were analysed by comparison with published sequences and by phylogenetic analysis. It was found that the more isotopic label substrate incorporated, the further the buoyant density (BD) separation between 15N- and 14N-RNA. Novosphingobium nitrogenifigens gave an average BD shift of 0.03 + 0.004 g ml-1 (95.0 atom % 15N) with 15NH4Cl. For mixed communities the average BD shift was 0.02 + 0.004 g ml-1 (80.0 atom % 15N) with 15NH4Cl and 0.013 + 0.002 g ml-1 (32.6 atom % 15N) when using 15N2. Clone library analysis of 16S rRNA genes present in the enriched 15N-RNA fraction of the mixed community was shown to consist of a diverse population of bacteria as indicated by a Shannon Weaver index value of gt;2.8. Three dominant genera (Aeromonas, Pseudomonas and Bacillus) were identified by comparison with published sequences and phylogenetic analysis. Many other groups not known as archetypal nitrogen-fixing bacteria were also identified, demonstrating that 15N2-RNA-SIP provides a useful tool for the identification of important and previously unknown contributors to nitrogen fixation in a range of environments. Overall, this project has established that nitrogen based RNA-SIP is a powerful tool that can be used successfully and reproducibly with both pure and complex mixed microbial communities to study active diazotrophs in environmental samples.

Axial rotation is regarded as an essential movement of the trunk that allows many individuals to participate in vocations, sports and activities of daily living. Unfortunately when the destabilising nature of rotation is combined with that of spinal flexion, the risk of injuring the spine can increase significantly. Few studies have investigated the potential benefits that maximizing trunk rotation has in certain vocation and sport-related arenas and none have looked at whether adopting certain spinal postures in the sagittal plane can maximise trunk rotation more than others. The aim of the study was to determine the effects of alterations of trunk inclination, spinal posture, pelvic fixation and turning direction on the active range of motion (ROM) of trunk rotation. Twenty healthy individuals participated in the main study. Retro-reflective markers were placed on key anatomical locations and used to track the movement of the thorax and pelvis during a series of repeated maximal trunk rotations in ten different spinal positions within the sagittal plane. Trunk kinematics and kinetics were recorded simultaneously using an optoelectronic motion analysis and force platform measuring system. A repeated-measures multiple analysis of variance (MANOVA) was used to test for the main effects of trunk inclination, spinal posture, fixation of pelvis and direction of turn on maximum active ROM of trunk rotation, maximum pelvic rotation and the anterior-posterior and lateral displacement of the centre of pressure (COP). To investigate test-retest reliability, ten participants were tested on two separate days. Repeatability for each outcome measure was investigated using interclass correlation coefficients (ICC) and Bland Altman graphs. The majority of subjects showed reasonable test-retest reliability for trunk rotation measures in each of the test positions, with ICC’s ranging between 0.562 – 0.731. Overall, trunk inclination (0°, 22.5°, 45°) forward in the sagittal plane had a significant effect on trunk and pelvic rotation (p<0.001) and lateral displacement of the COP (p<0.005) during trunk rotation. As trunk inclination increased from 0° to 45° there was an average increase in trunk rotation ROM of approximately 10 % (approximately 3.4°). Furthermore, increasing trunk inclination led to an increase in lateral displacement of the COP and a decrease in pelvic rotation. Spinal posture (neutral, flexed, extended) at a forward inclination of 45° had a significant effect on trunk rotation (p<0.01) and pelvic rotation (p<0.05), with a neutral spine averaging approximately 3 % (approximately 1.1°) more trunk rotation than a flexed or extended posture. The position and posture of the spine in the sagittal plane appears to have a significant influence on ranges of trunk rotation. The study suggests that rotating the trunk when adopting a neutral spine inclined to 45° will maximise range of trunk rotation and encourage a natural stabilisation of the lower body. This posture meets the unique set of biomechanical requirements for the sport of golf and may help to reduce the risk of injury in manual material handling tasks. Conversely, rotating the trunk whilst the thoracolumbar spine is flexed leads to a reduction in trunk rotation ROM, encourages greater pelvic and lower body rotation, reduces torque production of the trunk and may increase the risk of lower back injury. These findings have important implications in relation to the teaching of spinal position during vocations, sports and activities of daily living that seek to maximise trunk rotation.

Le problème de la gestion de la diversité est défini sur un ensemble partiellement ordonné d'élements possédant des demandes et des coûts unitaires de production. L'objectif est de produire un sous-ensemble de $k$ éléments références, $k$ étant un nombre donné, minimisant les coûts. Chaque élément non produit doit être remplacé par une référence qui lui est supérieure, ce qui implique un sur-coût. Après une étude théorique de complexité, nous modélisons ce problème grâce à un programme linéaire en nombres entiers, proche de ceux des problèmes de localisation $k$-médians. Pour résoudre ce programme, nous présentons un algorithme lagrangien, ainsi que de nombreux critères de fixation de variables permettant de réduire la taille du problème. Nous exploitons ensuite cet algorithme pour construire des solutions de bonne qualité. Nous développons enfin un algorithme exact de Séparation et Coupe. Nous étudions un certain type de coupes ainsi qu'une heuristique permettant de les générer. Nous concluons par des tests numériques effectués sur des instances réelles.

L'ATP synthase ou ATPase de type F, ancrée aux membranes internes des mitochondries, est un complexe macromoléculaire qui utilise le gradient électrochimique généré par l'oxydation de petites molécules (NADH2, FADH2) dans les différents complexes de la chaîne respiratoire pour former l'ATP, vecteur énergétique universel. Le gradient électrochimique ou pmf est transformé en une énergie mécanique qui se traduit par le mouvement du rotor de l'ATP synthase dans un sens horaire vu depuis la membrane. La rotation de la sous-unité déforme successivement les trois sites catalytiques et permet ainsi la synthèse d'ATP. Dans certains cas, comme ceux de l'anoxie ou de l'hypoxie, le gradient électrochimique peut s'effondrer et l'ATP synthase hydrolyse alors l'ATP. Pour éviter cette hydrolyse futile, un petit peptide nommé IF1, régulateur spécifique des ATP synthases mitochondriales, vient s'insérer entre les sous-unités d'une interface catalytique et bloque instantanément le fonctionnement de l'ATPase. Cette inhibition est réversible puisque le peptide se décroche lorsque la membrane interne mitochondriale se réénergise. Dans ce travail de thèse, nous nous sommes intéressés à caractériser le mécanisme d'inhibition de l'ATPase de S.cerevisiae par son peptide endogène IF1 en s'appuyant essentiellement sur les quelques données structurales qui ont été publiées sur le peptide et sur le complexe inhibé IF1-F1-ATPase de B.taurus. Constitué de 63 acides aminés chez S.cerevisiae et 84 acides aminés chez B.taurus, IF1 est majoritairement structuré en hélice α. Les études menées par Elena Cabezón ont montré qu'IF1 possédait différentes formes dont la prédominance et l'activité dépendait essentiellement du pH. Chez B.taurus, il existe une forme inhibitrice dimérique prédominante à pH inférieurs à 6,5 et une forme tétramérique dont nous connaissons la structure 3D qui est non inhibitrice et prépondérante à pH supérieurs à 6,5. Chez S.cerevisiae, il existe une forme monomérique inhibitrice prépondérante à pH supérieur à 6,5 et une forme dimérique prédominante à pH inférieurs à 6,5 et dont le caractère inhibiteur ou non n'a pas encore été déterminé. Sur la base de la structure 3D de l'IF1 bovin, nous avons voulu identifier les régions de dimérisation du peptide de levure en utilisant la technique de marquage de spin couplée à de la spectroscopie RPE. En plaçant des marqueurs de spin (MTSL) en partie médiane (E33C) ou en C-terminale (L54C), nous avons pu favoriser l'interface de dimérisation plutôt en partie médiane du peptide. Ce travail est encore au stade embryonnaire et ne nous permet pas, à ce jour, d'identifier la zone exacte de dimérisation. Dans un deuxième volet, nous avons voulu caractériser le mécanisme d'inhibition d'un point de vue dynamique et nous avons pu en préciser les différentes étapes : reconnaissance, verrouillage et stabilisation. Pour cela, nous avons associé la mutagenèse sur le peptide et sur l'enzyme aux cinétiques d'inhibition. Nous avons tout d'abord évalué le rôle de plusieurs résidus situés en Cterminal de la sous-unité β, dans la région de l'interface α/ β qui se referme sur le peptide IF1, dans la reconnaissance moléculaire spécifique d'IF1 par l'ATPase mitochondriale. Nous avons ensuite montré que la partie N-terminale d'IF1 joue un rôle mineur dans la reconnaissance moléculaire mais son enroulement autour de la sous-unité constitue un loquet important dans la stabilisation du complexe inhibé. Enfin, la fermeture de l'interface catalytique sur IF1 crée une zone de contact entre la "bosse" de la sous-unité γ et la partie C-terminale de la sous-unité α qui constitue la dernière clef de blocage du peptide au sein de la F1-ATPase. Ce dernier point de fermeture est le seul qui n'implique aucun résidu du peptide IF1.

La diversité des organismes est essentielle pour leur capacité à évoluer et s'adapter aux variations environnementales. De ce fait, déterminer les facteurs responsables de l'origine de cette diversité ainsi que de la maintenance de la variabilité génétique observée reste un objectif fondamental en recherche. L'objectif de cette thèse était de comprendre les facteurs évolutifs maintenant le polymorphisme neutre. L'influence des processus évolutifs tels que la sélection naturelle et la dérive génétique étant complexes, nous avons combiné des approches complémentaires expérimentale et théorique. Le système expérimental utilisé, la bactérie Escherichia coli et l'amibe sociale Dictyostelium discoideum nous a permis d'étudier dans un premier temps la variabilité naturelle des interactions existant entre les deux espèces. Dans une seconde partie, nous avons étudié les traits bactériens impliqués dans cette variabilité. Nous avons montré que les bactéries portant des facteurs de virulence sont plus résistantes à la digestion des amibes, ce qui est en accord avec l'hypothèse de coïncidence évolutive des facteurs de virulence. Le deuxième volet de cette thèse concerne les aspects de génétique des populations de ce système. La troisième partie de notre expérimentation était de suivre les variations temporelles des fréquences alléliques de populations bactériennes comportant un marqueur neutre, durant 300 générations et sous quatre conditions environnementales : avec ou sans structuration spatiale et avec ou sans facteur biotique. Nous avons observé que les variations des fréquences alléliques observées étaient compatibles avec la dérive génétique. L'objectif du modèle théorique a été dans un premier temps d'étudier les effets de la stochasticité démographique sur les probabilités de fixation d'un nouvel allèle neutre arrivant dans une population résidente ainsi que sur le temps de fixation. La probabilité de fixation ainsi que le temps de fixation sont modifiés par les effets stochastiques lorsque l'on compare nos modèles à taille de population fluctuantes à un modèle à taille de population constante tel que le modèle de Moran.

<p>Discoveries in recent years have disclosed the importance of marine cyano-bacteria in the context of primary production and global nitrogen cycling. It is hypothesized here that microbial mats in tropical coastal habitats harbour a rich diversity of previously uncharacterized cyanobacteria and that benthic marine nitrogen fixation in coastal zones is substantial.</p><p>A polyphasic approach was used to investigate cyanobacterial diversity in three tropical benthic marine habitats of different characters; an intertidal sand flat and a mangrove forest floor in the Indian Ocean, and a beach rock in the Pacific Ocean. In addition, nitrogenase activity was measured over diel cycles at all sites. The results revealed high cyanobacterial diversity, both morphologically and genetically. Substantial nitrogenase activity was observed, with highest rates at daytime where heterocystous species were present. However, the three habitats were dominated by non-heterocystous and unicellular genera such as <i>Microcoleus</i>, <i>Lyngbya</i>, <i>Cyanothece</i> and a large group of thin filamentous species, identified as members of the Pseudanabaenaceae family. In these consortia nocturnal nitrogenase activities were highest and <i>nifH</i> sequencing also revealed presence of non-cyanobacterial potential diazotrophs. A conclusive phylogenetic analysis of partial nifH sequences from the three sites and sequences from geographi-cally distant microbial mats revealed new clusters of benthic potentially ni-trogen-fixing cyanobacteria. Further, the non-heterocystous cyanobacterium <i>Lyngbya majuscula</i> was subjected to a physiological characterization to gain insights into regulatory aspects of its nitrogen fixation. The data demon-strated that nitrogenase activity is restricted to darkness, which called upon a re-evaluation of its diazotrophic behaviour.</p>