In vivo phosphorus spectroscopy in human subjects on a clinical 3T MRI system

Alors que l’Imagerie par résonance magnétique (IRM) permet d’obtenir un large éventail de données anatomiques et fonctionnelles, les scanneurs cliniques sont généralement restreints à l’utilisation du proton pour leurs images et leurs applications spectroscopiques. Le phosphore jouant un rôle prépondérant dans le métabolisme énergétique, l’utilisation de cet atome en spectroscopie RM présente un énorme avantage dans l’observation du corps humain. Cela représente un certain nombre de déEis techniques à relever dus à la faible concentration de phosphore et sa fréquence de résonance différente. L’objectif de ce projet a été de développer la capacité à réaliser des expériences de spectroscopie phosphore sur un scanneur IRM clinique de 3 Tesla. Nous présentons ici les différentes étapes nécessaires à la conception et la validation d’une antenne IRM syntonisée à la fréquence du phosphore. Nous présentons aussi l’information relative à réalisation de fantômes utilisés dans les tests de validation et la calibration. Finalement, nous présentons les résultats préliminaires d’acquisitions spectroscopiques sur un muscle humain permettant d’identiEier les différents métabolites phosphorylés à haute énergie. Ces résultats s’inscrivent dans un projet de plus grande envergure où les impacts des changements du métabolisme énergétique sont étudiés en relation avec l’âge et les pathologies.

Novel Strategies for Heterocyclic Constructions via 1, 4-Dipolar Intermediates

In the thesis entitled " Novel Strategies for Heterocyclic Constructions via 1 ,4-Dipolar Intermediates"Synthesis of a complex organic molecules essentially involves the formation of carbon-carbon and carbon-heteroatom bonds. Various synthetic methods are available for these processes involving ionic, pericyclic and radical reactions. Among the pericyclic reactions, dipolar cycloaddition reactions, introduced by Huisgen, have emerged as a very powerful tool for heterocyclic construction. Heterocyclic compounds remain an important class of organic molecules due to their natural abundance and remarkable biological activity, thus constituting an intergral part of pharmaceutical industry. In this respect, developing newer synthetic methodology for heterocyclic construction has been an area of immense interest. In recent years, 1,3-dipolar cycloaddition reactions proved to be efficient routes to a wide variety of five membered heterocycles, as attested by their application in the total synthesis of various complex organic molecules. However, the potential application of similar 1,4- dipolar cycloaddition reactions for the construction of six memebered heterocycles remained underexploited. In this context, a systematic investigation of the reactivity of 1,4-dipoles generated from nitrogen heterocycles (pyridine and its analogues) and dimethyl acetylenedicarboxy!ate (DMAD) towards various dipolarophiles has been carried out and the results are embodied.

Leguminous cover crops differentially affect maize yields in three contrasting soil types of Kakamega, Western Kenya

Maize production in smallholder farming systems in Kenya is largely limited by low soil fertility. As mineral fertilizer is expensive, green manuring using leguminous cover crops could be an alternative strategy for farmers to enhance farm productivity. However due to variability in soil type and crop management, the effects of green manure are likely to differ with farms. The objectives of this study were to evaluate Mucuna pruriens and Arachis pintoi on (i) biomass and nitrogen fixation (^15N natural abundance), (ii) soil carbon and nitrogen stocks and (iii) their effects on maize yields over two cropping seasons in Kakamega, Western Kenya. Mucuna at 6 weeks accumulated 1–1.3 Mg ha^{-1} of dry matter and 33–56 kg ha^{-1} nitrogen of which 70% was nitrogen derived from the atmosphere (Ndfa). Arachis after 12 months accumulated 2–2.7 Mg ha^{-1} of dry matter and 51–74 kg N ha^{-1} of which 52-63 % was from Ndfa. Soil carbon and nitrogen stocks at 0–15 cm depth were enhanced by 2-4 Mg C ha^{-1} and 0.3–1.0 Mg N ha^{-1} under Mucuna and Arachis fallow, irrespective of soil type. Maize yield increased by 0.5-2 Mg ha^{-1} in Mucuna and 0.5–3 Mg ha^{-1} in Arachis and the response was stronger on Nitisol than on Acrisol or Ferralsol. We concluded that leguminous cover crops seem promising in enhancing soil fertility and maize yields in Kenya, provided soil conditions and rainfall are suitable.

Efficacy of condensed tannins against larval Hymenolepis diminuta (Cestoda) in vitro and in the intermediate host Tenebrio molitor (Coleoptera) in vivo

Natural anti-parasitic compounds in plants such as condensed tannins (CT) have anthelmintic properties against a range of gastrointestinal nematodes, but for other helminths such effects are unexplored. The aim of this study was to assess the effects of CT from three different plant extracts in a model system employing the rat tapeworm, Hymenolepis diminuta, in its intermediate host, Tenebrio molitor. An in vitro study examined infectivity of H. diminuta cysticercoids (excystation success) isolated from infected beetles exposed to different concentrations of CT extracts from pine bark (PB) (Pinus sps), hazelnut pericarp (HN) (Corylus avellana) or white clover flowers (WC) (Trifolium repens), in comparison with the anthelmintic drug praziquantel (positive control). In the in vitro study, praziquantel and CT from all three plant extracts had dose-dependent inhibitory effects on cysticercoid excystation. The HN extract was most effective at inhibiting excystation, followed by PB and WC. An in vivo study was carried out on infected beetles (measured as cysticercoid establishment) fed different doses of PB, HN and praziquantel. There was a highly significant inhibitory effect of HN on cysticercoid development (p = 0.0002). Overall, CT showed a promising anti-cestodal effect against the metacestode stage of H. diminuta.

Structural-electronic aspects related to the near-infrared light emission of Fe-doped silicon films

The need of efficient (fast and low consumption) optoelectronic devices has always been the driving force behind the investigation of materials with new or improved properties. To be commercially attractive, however, these materials should be compatible with our current micro-electronics industry and/or telecommunications system. Silicon-based compounds, with their matured processing technology and natural abundance, partially comply with such requirements-as long as they emit light. Motivated by these issues, this work reports on the optical properties of amorphous Si films doped with Fe. The films were prepared by sputtering a Si+Fe target and were investigated by different spectroscopic techniques. According to the experimental results, both the Fe concentration and the thermal annealing of the samples induce changes in their atomic structure and optical-electronic properties. In fact, after thermal annealing at similar to 750 degrees C, the samples partially crystallize with the development of Si and/or beta-FeSi(2) crystallites. In such a case, certain samples present light emission at similar to 1500 nm that depends on the presence of beta-FeSi(2) crystallites and is very sensitive to the annealing conditions. The most likely reasons for the light emission (or absence of it) in the considered Fe-doped Si samples are presented and discussed in view of their main structural-electronic characteristics. (C) 2011 Elsevier Ltd. All rights reserved.

Effect of root surface treatment with propolis and fluoride in delayed tooth replantation in rats

Replantation is an acceptable option for treatment of an avulsed permanent tooth. Nevertheless, an extended extraoral period damages the periodontal ligament and results in external root resorption. The purpose of this study was to assess by histologic and histometric analysis, the influence of propolis 15% (natural resinous substance collected by Apis mellifera bees from various plants) and the fluoride solution used as root surface treatment on the healing process after delayed tooth replantation. Thirty Wistar (Rattus norvegicus albinus) rats were submitted to extraction of their upper right incisor. The teeth were maintained in a dry environment for 60 min. After this, the pulp was extirpated and the papilla, enamel organ and periodontal ligament were removed with scalpel. The teeth were divided into three experimental groups: Group I - teeth immersed in 20 ml of physiologic saline; Group II - teeth immersed in 20 ml of 2% acidulated phosphate sodium fluoride; Group III - teeth immersed in 20 ml of 15% propolis. After 10 min of immersion in the solutions, the root canals were dried and filled with calcium hydroxide paste and the teeth were replanted. The animals were euthanized 60 days after replantation. The results showed that similar external root resorption was seen in the propolis and fluoride groups. Teeth treated with physiologic saline tended to have more inflammatory root resorption compared with those treated with fluoride or propolis. However, the comparative analysis did not reveal statistically significant differences (P > 0.05) between the treatment modalities when used for delayed tooth replantation.

Monoamine oxidase inhibitory activities of indolylalkaloid toxins from the venom of the colonial spider Parawixia bistriata: Functional characterization of PwTX-I

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

The role of multiple partners in a digestive mutualism with a protocarnivorous plant

Background and aimsThe protocarnivorous plant Paepalanthus bromelioides (Eriocaulaceae) is similar to bromeliads in that this plant has a rosette-like structure that allows rainwater to accumulate in leaf axils (i.e. phytotelmata). Although the rosettes of P. bromelioides are commonly inhabited by predators (e.g. spiders), their roots are wrapped by a cylindrical termite mound that grows beneath the rosette. In this study it is predicted that these plants can derive nutrients from recycling processes carried out by termites and from predation events that take place inside the rosette. It is also predicted that bacteria living in phytotelmata can accelerate nutrient cycling derived from predators.MethodsThe predictions were tested by surveying plants and animals, and also by performing field experiments in rocky fields from Serra do Cipó, Brazil, using natural abundance and enriched isotopes of 15N. Laboratory bioassays were also conducted to test proteolytic activities of bacteria from P. bromelioides rosettes.Key ResultsAnalyses of 15N in natural nitrogen abundances showed that the isotopic signature of P. bromelioides is similar to that of carnivorous plants and higher than that of non-carnivorous plants in the study area. Linear mixing models showed that predatory activities on the rosettes (i.e. spider faeces and prey carcass) resulted in overall nitrogen contributions of 26·5 % (a top-down flux). Although nitrogen flux was not detected from termites to plants via decomposition of labelled cardboard, the data on 15N in natural nitrogen abundance indicated that 67 % of nitrogen from P. bromelioides is derived from termites (a bottom-up flux). Bacteria did not affect nutrient cycling or nitrogen uptake from prey carcasses and spider faeces.ConclusionsThe results suggest that P. bromelioides derive nitrogen from associated predators and termites, despite differences in nitrogen cycling velocities, which seem to have been higher in nitrogen derived from predators (leaves) than from termites (roots). This is the first study that demonstrates partitioning effects from multiple partners in a digestion-based mutualism. Despite most of the nitrogen being absorbed through their roots (via termites), P. bromelioides has all the attributes necessary to be considered as a carnivorous plant in the context of digestive mutualism. © 2012 The Author. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved.

Modulação das propriedade ópticas de semicondutores pela incorporação de íons terras raras

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A ictiofauna e a atividade pesqueira na Ilha das Onças, Barcarena - Pará

A Ilha das Onças é uma das numerosas ilhas que se encontram na margem esquerda da Baía do Guajará, sendo conhecida principalmente pela presença da palmeira que produz o açaí como fruto (Euterpe oleracea,) e pela importância que seus canais representam para a navegação regional. Entre os principais canais que compõem a ilha, destaca-se o Canal do Piramanha, que atravessa a Ilha das Onças. A composição e a distribuição da ictiofauna presente na Ilha é pouco conhecida, com base nisso, esse trabalho teve como objetivo descrever as espécies da região, bem como do seu uso pelo ambiente, e a importância que possuem para a população local. Foram feitas entrevistas com os pescadores onde foram obtidas informações básicas sobre a pesca no local. Foram realizadas pescarias trimestrais de acordo com os períodos climáticos com rede de tapagem no igarapé e de emalhe no canal. No momento da despesca foram registrados in situ os dados abióticos (pH, salinidade, temperatura), que não apresentaram diferenças entre os períodos. Foram capturados 1200 indivíduos de 20 famílias de maioria Aspredinidae, Sciaenidae e Pimelodidae, distribuídos em 39 espécies identificadas (27 no canal e 26 no igarapé). As espécies foram em sua maioria constantes (FO>75%), e a diversidade foi similar ao longo do tempo, não havendo espécies dominantes em nenhum período de coleta. A estatística W mostra ambientes perturbados, nos meses mais chuvosos de março e dezembro, onde também foram registrados as maiores abundâncias (CPUEn e CPUEb), indicando a análise pode não ser adequada a ambientes onde há dominância de indivíduos de pequeno porte, fazendo com que o método confunda a existência natural de grande quantidade de indivíduos pequenos com uma troca de dominância. As análises multivariadas mostraram que os ambientes são significativamente diferentes, principalmente pela grande abundância numérica de A. aspredo no canal, e da P. squamosissimus no igarapé, que juntamente com o L. dorsalis, o H. marginatus, o A. aff. ucayalensis formam as principais espécies capturadas na Ilha. No canal houve maioria de espécies estuarinas, que se alimentam principalmente de zoobentos e peixes, e no igarapé, houve dominância de espécies dulcícolas, cujas preferências alimentares mudam ao longo do ano, aparentemente de acordo com o regime de chuvas. De maneira geral as espécies são de pequeno e médio porte (CT < 30 cm), imaturas (maioria em estádio A) e alimentando-se em pelo menos um período do ano. Apesar do ambiente favorável a pesca, 90% dos entrevistados tem no açaí sua principal fonte de renda, sendo a pesca colocada como fonte secundária, baseada nos períodos de safra e entre safra do açaí. A principal arte de pesca é o matapi que tem como alvo o camarão da Amazônia (Macrobachium amazonicum), seguida das malhadeiras e tapagens, cujas espécies alvo são na sua maioria as mesmas capturadas em abundancia nesse estudo. Pelo pequeno porte e baixo valor econômico, a pesca na ilha é realizada principalmente como fonte suplementar de proteínas, visto que a intensa atividade pesqueira e a perturbação causada pelas embarcações diminuem ainda mais esse número.

Boron uptake and translocation in some cotton cultivars

Boron (B) is the most deficient micronutrient in cotton (Gossypium hirsutum L.). It is generally accepted that B is immobile in cotton phloem, but some cultivars could remobilize the nutrient. In order to further understand B uptake and mobility in various cotton cultivars two experiments were conducted.In experiment-1, cotton cultivars were grown in B-10 enriched or natural abundance nutrient solutions for 4 weeks and transferred to nutrient solutions ranging from deficient to sufficient in B. In experiment-2 B-10 enriched boric acid was applied to cotton leaves and B mobilization was determined.In deficient plants, B previously supplied to roots was remobilized from older to younger plant tissues, but the amount was insufficient to maintain growth. Boron deficiency symptoms appeared and progressed with time. Boron applied to leaves was taken up and remobilized within 24 h. Boron mobilization was higher to plant parts above the treated region.Boron uptake and mobilization was similar among cotton cultivars. Boron applied to cotton leaves shows a preferential translocation to younger tissues. Foliar sprays of B to cotton may be used to cope with a temporary deficiency, but to achieve full growth and development B must be available to cotton throughout the plant cycle.

Evaluation of Fe uptake and translocation in transgenic and non-transgenic soybean plants using enriched stable Fe-57 as a tracer

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

Isotope determination of sulfur by mass spectrometry in soil samples

Sulphur plays an essential role in plants and is one of the main nutrients in several metabolic processes. It has four stable isotopes (32S, 33S, 34S, and 36S) with a natural abundance of 95.00, 0.76, 4.22, and 0.014 in atom %, respectively. A method for isotopic determination of S by isotope-ratio mass spectrometry (IRMS) in soil samples is proposed. The procedure involves the oxidation of organic S to sulphate (S-SO4(2-)), which was determined by dry combustion with alkaline oxidizing agents. The total S-SO4(2-) concentration was determined by turbidimetry and the results showed that the conversion process was adequate. To produce gaseous SO2 gas, BaSO4 was thermally decomposed in a vacuum system at 900 ºC in the presence of NaPO3. The isotope determination of S (atom % 34S atoms) was carried out by isotope ratio mass spectrometry (IRMS). In this work, the labeled material (K2(34)SO4) was used to validate the method of isotopic determination of S; the results were precise and accurate, showing the viability of the proposed method.

Heteronuclear recoupling methods in solid-state NMR

This thesis is focused on the development of heteronuclear correlation methods in solid-state NMR spectroscopy, where the spatial dependence of the dipolar coupling is exploited to obtain structural and dynamical information in solids. Quantitative results on dipolar coupling constants are extracted by means of spinning sideband analysis in the indirect dimension of the two-dimensional experiments. The principles of sideband analysis were established and are currently widely used in the group of Prof. Spiess for the special case of homonuclear 1H double-quantum spectroscopy. The generalization of these principles to the heteronuclear case is presented, with special emphasis on naturally abundant 13C-1H systems. For proton spectroscopy in the solid state, line-narrowing is of particular importance, and is here achieved by very-fast sample rotation at the magic angle (MAS), with frequencies up to 35 kHz. Therefore, the heteronuclear dipolar couplings are suppressed and have to be recoupled in order to achieve an efficient excitation of the observed multiple-quantum modes. Heteronuclear recoupling is most straightforwardly accomplished by performing the known REDOR experiment, where pi-pulses are applied every half rotor period. This experiment was modified by the insertion of an additional spectroscopic dimension, such that heteronuclear multiple-quantum experiments can be carried out, which, as shown experimentally and theoretically, closely resemble homonuclear double-quantum experiments. Variants are presented which are well-suited for the recording of high-resolution 13C-1H shift correlation and spinning-sideband spectra, by means of which spatial proximities and quantitative dipolar coupling constants, respectively, of heteronuclear spin pairs can be determined. Spectral editing of 13C spectra is shown to be feasible with these techniques. Moreover, order phenomena and dynamics in columnar mesophases with 13C in natural abundance were investigated. Two further modifications of the REDOR concept allow the correlation of 13C with quadrupolar nuclei, such as 2H. The spectroscopic handling of these nuclei is challenging in that they cover large frequency ranges, and with the new experiments it is shown how the excitation problem can be tackled or circumvented altogether, respectively. As an example, one of the techniques is used for the identification of a yet unknown motional process of the H-bonded protons in the crystalline parts of poly(vinyl alcohol).

Analyse von Biotransformationen und Naturstoffsynthesen in pflanzlichen Zellkulturen unter Anwendung der in vivo NMR-Spektroskopie ohne Markierung

Die Aufklärung von Biosynthesewegen erfolgt häufig mit Hilfe von Fütterungsexperimenten mit radioaktiven oder stabilen Isotopen markierten Präkusoren oder auf der Basis der Enzymreinigung mit anschließender molekularbiologischer Charakterisierung. Die erstgenannte Methode verlangt die Isolierung der Produkte. Jedoch besteht bei Aufarbeitung und Extraktion immer die Gefahr, daß sich der Metabolit teilweise oder vollständig chemisch verändert. Ein weiterer Nachteil der genannten Methoden ist, daß diese generell mühsam und zeitaufwendig sind. Mit Hilfe der in vivo NMR-Spektroskopie können diese Nachteile umgangen werden. In der vorliegenden Arbeit wurden Biotransformationen und Biosynthesesequenzen des Ajmalin-Biosyntheseweges mit Hilfe der in vivo NMR-Spektroskopie in Pflanzenzellkulturen von Rauvolfia serpentina und Rauvolfia serpentina x Rhazya stricta anhand der natürlichen 13C-Häufigkeit untersucht. Dafür wurden ein 700 MHz, 800 MHz und ein 500 MHz CryoProbe Spektrometer eingesetzt, um die Biotransformationen von Isatin-3-oxim und Isatin sowie die Metabolisierungen der Alkaloide Vellosimin, Vinorin, Vomilenin, Ajmalin, Nß-Methyl-dihydrochano-ajmalin und Perakin mit der 1H-13C invers korrelierten NMR-Spektroskopie zu verfolgen.