Permanent production and use of organic chemicals for many purposes has resulted in their introduction and accumulation in the environment. Depending on their physicochemical properties they can be transported by different ways from the source to very remote regions of the planet. Many organic chemicals are used in agriculture as pesticides for cultures protection or nutrient. Residues of these chemicals can always be found in fields, and under the effect of precipitations they leach and pass in streams and rivers. Pharmaceuticals and personal health care products and other house holding chemicals are continuously introduced in the environment through municipal wastewaters. These substances exhibit, in most of the cases, perturbation effects towards the living organisms, moreover the effect of many of them is not known yet. Despite their concentration in water is low, the exposure of organisms for long periods can lead to negative consequences, but these effects cannot be measured instantly. In order to reduce or avoid the pollution of water with chemicals many water treatment methods has been developed like adsorption of pollutants on adsorbents, membrane filtration, microbiological treatment, chemical oxidation with oxidizing agents and advanced oxidation processes. Most of the methods used in waste water treatment plants (WWTP) do not completely destroy the organic contaminants or they only separate the contaminants from water. Then they have to be deposed somewhere else remaining always a potential source of contamination. Advanced oxidation processes and in particular electrochemical advanced oxidation processes are methods developed later and are proven as more effective as they can completely oxidize the organic matter in water. The subject of this thesis is the use of electro-Fenton, an electrochemical advanced oxidation process for efficient destruction of organic pollutants in aqueous medium. In this method, organic pollutants are eliminated by H hydroxyl radicals (high oxidation power species) which are produced in situ through the Fenton's reagent (H2O2 + Fe2+) itself generated in the solution electrochemically and continuously. In this process, the electrode material is of fundamental importance in order to have an efficient process, so we have studied at large extent the influence of both cathode and anode material in this work. Firstly a systematic study on the oxidation capacity of the process of amoxicillin (AMX) as model pollutant with several anodes materials: BDD, Pt, DSA, PbO2 Carbon felt, Graphite and Carbon fibre was realised. In all cases a stainless steel electrode was used as cathode. The degradation of AMX was followed by HPLC analysis whereas the mineralization efficiency ot the process was measured by total organic carbon analyser (TOC). This revealed that BDD was the most efficient anode for AMX oxidation and DSA was the weakest one. Carbon felt showed a characteristic behaviour; it was very efficient on AMX oxidation but it could not transform AMX to CO2 and H2O. Afterwards four anodes were tested for their influence on electro-Fenton process efficiency namely Pt, BDD, DSA and Carbon felt, the cathode was always carbon felt. Sulfamethazine (SMT) was used as model pollutant. Apparent rate constants have given only moderate values of mineralization for currents lower than 100 mA. Here again the BDD anode was distinguished for its excellent mineralization capacity owing to the additional hydroxyl radicals and other oxidizing species introduced in the system. When electro-Fenton applied good degradation and mineralization results were obtained even with the DSA anode (...)

The global carbon cycle is subject to intense research, where sources and sinks for greenhouse gases, carbon dioxide in particular, are estimated for various systems and biomes. Lakes have previously been neglected in carbon balance estimations, but have recently been recognized to be significant net sources of CO2. This thesis estimates emission of carbon dioxide (CO2) from boreal lakes and factors regulating the CO2 saturation from field measurements of CO2 concentration along with a number of chemical, biological and physical parameters. Concentration of dissolved organic carbon (DOC) was found to be the most important factor for CO2 saturation in lake water, whereas climatic parameters such as precipitation, temperature and global radiation were less influential. All lakes were supersaturated with and, thus, sources of CO2. Sediment incubation experiments indicated that in-lake mineralization processes during summer stratification mainly occurred in the pelagial. Approximately 10% of the CO2 emitted from the lake surface was produced in epilimnetic sediments. The mineralization of DOC and emission of CO2 from freshwaters was calculated on a catchment basis for almost 80,000 lakes and 21 major catchments in Sweden, together with rates of sedimentation in lakes and export of organic carbon to the sea. The total export of terrestrial organic carbon to freshwaters could thereby be estimated and consequently also the importance of lakes for the withdrawal of organic carbon export from terrestrial sources to the sea. Lakes removed 30-80% of imported terrestrial organic carbon, and mineralization and CO2 emission were much more important than sedimentation of carbon. The carbon loss was closely related to water retention time, where catchments with short residence times (<1 year) had low carbon retentions, whereas in catchments with long residence times (>3 years) a majority of the imported TOC was removed in the lake systems. The Gulf of Bothnia was also studied in this thesis and found to be a net heterotrophic system, emitting large amounts of CO2 to the atmosphere on an annual basis. The rate of CO2 emission was depending on the balance between primary production and bacterial respiration, and the system was oscillating between being a source and a sink of CO2.

This thesis describes strategies for and examples ofcellulose fiber modification.The ability of an engineered biocatalyst, acellulose-binding module fused to theCandida antarcticalipase B, to catalyze ring-openingpolymerization of e-caprolactone in close proximity tocellulose fiber surfaces was explored. The water content in thesystem was found to regulate the polymer molecular weight,whereas the temperature primarily influenced the reaction rate.The hydrophobicity of the cellulose sample increased as aresult of the presence of surface-deposited polyester. A two-step enzymatic method was also investigated. Here,Candida antarctica lipase B catalyzed the acylation ofxyloglucan oligosaccharides.The modified carbohydrates werethen incorporated into longer xyloglucan molecules through theaction of a xyloglucan endotransglycosylase. The modifiedxyloglucan chains were finally deposited on a cellulosesubstrate. The action ofCandida antarcticalipase B was further investigated inthe copolymerization of e-caprolactone and D,L-lactide.Copolymerizations with different e-caprolactone-to-D,L-lactideratios were carried out. Initially, the polymerization wasslowed by the presence of D,L-lactide. During this stage,D,L-lactide was consumed more rapidly than ε-caprolactoneand the incorporation occurred dimer-wise with regard to thelactic acid units. Morphological studies on wood fibers were conducted using asol-gel mineralization method. The replicas produced werestudied, without additional sample preparation, by electronmicroscopy and nitrogen adsorption. Information concerning thestructure and accessibility of the porous fiber wall wasobtained. Studies of never-dried kraft pulp casts revealedmicro-cavities and cellulose fibrils with mean widths of 4.7(±2) and 3.6 (±1) nm, respectively. Finally, cationic catalysis by simple carboxylic acids wasstudied. L-Lactic acid was shown to catalyze the ring-openingpolymerization of ε-caprolactone in bulk at 120 °C.The reaction was initiated with methylß-D-glucopyranoside, sucrose or raffinose, which resultedin carbohydrate-functionalized polyesters. The regioselectivityof the acylation was well in agreement with the correspondinglipase-catalyzed reaction. The polymerization was alsoinitiated with a hexahydroxy-functional compound, whichresulted in a dendrimer-like star polymer. The L-lactic acidwas readily recycled, which made consecutive reactions usingthe same catalyst possible. Keywords:Candida antarcticalipase B, cationic catalysis,cellulose-binding module, dendrimer, enzymatic polymerization,fiber modification, silica-cast replica, sol-gelmineralization, organocatalysis, xyloglucanendotransglycosylase

Tissue Engineering is a scientific discipline that aims to regenerate tissues and organs that are diseased, lost or congenitally absent. It encompasses the use of suitable synthetic equivalents of native extracellular matrix that may or may not be supplemented with cells or relevant growth factors. Such scaffolds are designed to reside at the site of implantation for a variable period of time during which they induce the regeneration of native tissue. During this time, they also provide a template for new cells to attach, infiltrate, differentiate into appropriate phenotype and eventually restore function of the concerned tissue. Among the factors that affect the outcome are the composition of scaffold, methods of fabrication, bulk properties of the scaffold and topography and architecture at the cellular level. Bone is unique in the body in that it is one of the few tissues capable of complete regeneration even in adults, as seen during fracture healing. However, certain conditions (non-union of fractures, congenital and acquired bone deficiencies) exist in which the regenerative capacities of bone are exceeded and appropriate intervention becomes necessary. Current treatment options include autologous bone grafts harvested from iliac crest or de-cellularized allografts or synthetic substitutes made from metals, ceramics and polymers. However these options have serious limitations: while autografts are limited in supply, necessitate second surgery and show inadequate vascularization, allografts can transmit viral infections. Metals, ceramics and polymers are in essence structural replacements without performing any biological function. Other problems associated with these synthetic materials include adverse immune reactions, corrosion, stress-shielding and secondary fractures due to inadequate osseo-integration. Bone tissue engineering is a specialized field of research that provides an alternative strategy to repair bone defects by exploiting the advances in engineering and better understanding of bone biology. Scaffold-based tissue engineering approach is a promising field that involves implantation of a biomaterial that is specifically matched in terms of biological and material properties to the tissue it replaces. This study explores the feasibility of using electrospinning as a potential fabrication strategy for bone tissue engineering applications, more specifically intended for cleft palate repair. This model represents a congenital deformity that affects both hard and soft tissues and presents unique challenges and opportunities. Among the challenges are: the need for the implant allow growth of the most complex areas of the facial skeleton, integrate and grow with the patient through adolescence, the ability of the implant to not interfere with vital functions including breathing and feeding. Further the implant should provide a flexible matrix that can effectively support erupting teeth. In spite of these extreme demands, maxilla is a non load-bearing membranous bone, a favorable consideration from materials engineering perspective. The present study is organized into three independent sections. The first section investigates developing strategies intended to improve the material properties of electrospun bone scaffold. Bone is composed of a high volume fraction (50%) of inorganic hydroxyapatite nanocrystals that is closely associated with collagen. The dispersal of brittle mineral is critical in not only strengthening the bone in compression but also contributes to the osteoconductivity of the matrix. Since loading of mineral in a bone scaffold is a serious limitation, we attempted to achieve improved loading of bone mineral by dual mineralization approach. We first incorporated nanocrystalline hydroxyapatite (nHA) directly into the scaffold by adding it to the electrospinning polymer solution. The second step involves inducing biomimetic mineralization of electrospun scaffolds by incubating them in simulated body fluid (SBF) for 2 weeks. The hypothesis was that the nanocrystalline hydroxyapatite seeded during electrospinning would act as sites for nucleation and further crystal growth when incubated in solution supersaturated with respect to calcium and phosphate ions. We tested this approach in two synthetic, biocompatible polymers-polydioxanone and poly (lactide: glycolide) and four formulations of SBF with differential loading of nHA (0-50% by wt. of polymer). A modified Alizarin Red S (ARS) staining that specifically binds to calcium was developed that allowed us to quantify the mineral content of 3D scaffold with great accuracy. Results indicated a unique combination of factors: PDO scaffolds containing 50% nHA incubated in 1x revised-SBF incubated under static conditions gave maximum mineralization over a period of two weeks. We then sought to exploit these findings to engineer a stiffer scaffold by stacking multiple layers together and cold welding them under high pressure. Electrospun scaffolds (1, 2 or 4 layered stacks) were either compressed before or after mineralizing treatment with SBF. After two weeks, scaffolds were analyzed for total mineral content and stiffness by uniaxial tensile testing. Results indicated while compression of multiple layers significantly increases the stiffness of scaffolds, it also had lower levels of mineralization partly due to increased density of fibers and loss of surface area due to fiber welding. However this can be offset to a reasonable degree by increasing the number of stacks and hence this strategy can be successfully adopted to improve the mechanical properties of electrospun scaffolds. The second section introduces a novel infrared imaging technique to quantify and characterize the biological activity of biomaterials, based on cell adhesion. Cells attach to the surface by the formation of focal contacts where multiple proteins including vinculin and talin assemble to signal critical processes like cell survival, migration, proliferation and differentiation. After allowing MG-63 osteoblasts to adhere to 2D biomaterial surface coated with extracellular matrix proteins (collagen, gelatin, fibronectin) cells were fixed and probed with antibodies for vinculin and talin. Secondary antibodies, tagged with infrared-sensitive fluorescent dyes, were used to quantify the molecules of interest. In addition, the kinetics of focal contact formation in these different substrates was followed. Successful quantification of focal contacts were made and further research revealed phosphorylation of vinculin at pY-822 as one potential mechanism for recruitment of vinculin to focal contacts. Hence it could represent a subset of vinculin and might serve as a specific molecular marker for focal contacts. As an extension, we evaluated the possibility of using such an assay to quantify 3D electrospun tissue engineering scaffolds. We fabricated scaffolds of graded biological activity by electrospinning blends of polydioxanone and collagen in different ratios. Vinculin and talin expressed by MG-63 cultured on these scaffolds for 24 hours were quantified in a similar manner. Results indicate that while talin does not show a significant difference in expression among different scaffolds, vinculin showed a positive correlation with increasing biological activity of scaffolds. In conclusion, we have identified vinculin as a reliable marker of focal contacts in 3D scaffolds while phosphovinculin (pY-822) was more specific to focal contacts in coated 2D substrates. In both instances, infrared imaging proved to be reliable in study of focal contacts. The third section aims to make the bone scaffolds osteoinductive- a property of a material to induce new bone formation even when implanted in subcutaneous and intramuscular heterotopic sites. Bone morphogenetic proteins (BMP) are potent cytokines that can induce migration, proliferation and differentiation of stem cells along osteoblastic lineage. The therapeutic efficacy of BMPs in the treatment of severe bone defects has been identified and is currently FDA approved for specific orthopedic applications. BMPs are clinically administered in a buffer form that not only makes the treatment expensive but less effective. Suitable delivery systems for BMP delivery have been an intense area of investigation. We rationalized electrospinning as a strategy to incorporate BMP within the scaffold and that would enable controlled release when implanted. One of the drawbacks of using electrospinning to deliver bioactive molecules is the potential denaturing effect and eventual loss of activity of BMPs. The final section of this dissertation tries to develop sensitive and relevant assays that could answer intriguing questions about solvent-protein interaction. We chose to use the BMP-2/7 heterodimer as the osteoinductive molecule of choice because of its superior potency compared to homodimer counterparts. We characterized the detection and quantification of BMP-2/7 using a slot blot technique. Further, we used a novel cell line (C2C12 BRA) to test the retention of activity of BMP-2/7 that has been exposed to organic solvents. Results indicate significant loss of activity when BMPs are exposed to organic solvents but complete recovery was possible by diluting the solvent with an aqueous buffer.

En agriculture de conservation (AC), les résidus de cultures sous forme de paillis à la surface du sol associés à la suppression du travail du sol sont une composante intrinsèque des systèmes de culture. L'objectif principal de ce travail était de comprendre les effets de la nature et de la décomposition de mulch de résidus s sur les services d'approvisionnement et de régulation fournis par les sols dans les agrosystèmes en AC de régions tempérées (France) et tropicales (Madagascar et le Brésil). Des études expérimentales ont été réalisées pour obtenir des paramètres de décomposition pour une large gamme représentative de la qualité des résidus végétaux des agrosystèmes étudiés et pour tester et améliorer un modèle de décomposition des mulchs. Ensuite nous avons utilisé le modèle PASTIS_MULCH qui simule les biotransformations C et N et le transport de solutés dans les sols lors de la décomposition des mulchs.Pour la gamme de résidus de tiges testées, la rétention d'eau maximale a varié considérablement et ceci a été expliqué par les caractéristiques physiques du résidu. Avec les données obtenues par une série d'incubations de décomposition de résidus, nous avons proposé un ensemble de paramètres biologiques unique pour le module de décomposition CANTIS, simulant une large gamme de qualité des résidus de culture. L'étude expérimentale dans les colonnes de sol a montré que le mulch de maïs + dolique se décompose plus rapidement que mulch de blé + luzerne. Un régime de pluies fines et fréquentes augmente la décomposition du mulch par rapport à des pluies plus rares et plus fortes et cela est dû au maintien de l'humidité du mulch.Les simulations de scénarios avec PASTIS_MULCH ont montré que la pluie et les conditions d'évaporation classent les résidus de culture vis-à-vis de la décomposition lorsque ceux-ci sont placés en mulch à la surface des sols, tandis que les caractéristiques chimiques des résidus classent ceux-ci vis-à-vis de la décomposition lorsqu'ils sont incorporés. Aucune des situations étudiées pourrait être définie au vu des résultats de simulation comme étant adaptée à toutes les conditions pédo-climatiques et agricoles. / In conservation agriculture (CA), crop residues mulches are associated to reduction or suppression of soil tillage and are an intrinsic component of CA. The objective of this work was to understand the effects of nature and decomposition of crop residue mulches on the provisioning and regulating services of agrosystems provided by soils under temperate (France) and tropical (Madagascar and Brazil) conditions. Experimental studies were realized to get decomposition parameters for a large range of residue quality representative of the agrosystems studied and to test and improve a MULCH model. Then we used PASTIS_MULCH model which simulates the C and N biotransformations and solutes transport in soils during mulch decomposition.For a range of plant-stem residues tested, the maximal water retention varied greatly and was only explained by the physical features of the residue. With a series of decomposition incubations, we proposed a single set of biological parameters for CANTIS decomposition module, simulating a large range of crop residue quality. Experimental study in soil columns showed that maize+dolichos mulch decomposed faster than wheat+alfalfa mulch. Frequent and light rain enhanced mulch decomposition compared to infrequent and heavy rain and this was due to the mulch remaining wetter with frequent rain.The simulations of scenarios with PASTIS showed that rain and evaporation conditions ranked crop residues decomposition when placed as mulches while the residue chemical characteristics ranked crop residues decomposition when incorporated. None of the situations studied would be defined as suitable in all pedo-climatic and agricultural conditions.

A engenharia de tecido pulpar tem como objetivo substituir a polpa dentária inflamada ou necrosada por um tecido saudável e funcional, capaz de formar nova dentina para reparar a estrutura dentária perdida. Assim, os objetivos deste trabalho foram: avaliar a habilidade de diferenciação de célulastronco de dentes decíduos exfoliados humanos (SHED) em odontoblastos funcionais, demonstrando a formação de tecido mineralizado in vivo; e estudar o efeito de VEGF em SHED com relação à estimulação de vias de sinalização celular (STAT3, AKT e ERK), proliferação, migração, formação de estruturas tubulares e diferenciação em células endoteliais. O início do processo de mineralização de SHED tratadas com dexametasona, ácido ascórbico \'beta\' - glicerofosfato pôde ser detectado por meio da produção da enzima fosfatase alcalina a partir da segunda semana de cultura, mas a expressão de RNAm para DSPP só foi observada após 28 dias de indução. Utilizando-se o modelo de fatias de dentes e matrizes condutivas implantadas no dorso de camundongos imunodeprimidos, demonstrou-se a diferenciação de SHED em células semelhantes a odontoblastos, as quais tiveram imunomarcação positiva com o anticorpo DMP-1. A deposição de dentina, seguindo um ritmo centrípeto de crescimento, numa taxa de 14,1 µm por dia também foi demonstrada por meio da marcação com tetraciclina. O tratamento das SHED com VEGF estimulou a fosforilação de ERK e AKT e a diminuiu a fosforilação de STAT3 em um período de uma hora, provavelmente por meio de sua ligação com os receptores VEGFR-1 e NP-1 presentes nestas células. Além disso, VEGF intensificou a organização das SHED em estruturas tubulares, havendo diferença estatisticamente significativa entre os grupos tratado e não tratado a partir do 5o dia de tratamento. Entretanto, VEGF não estimulou a proliferação nem a migração destas células. Os resultados de RT-PCR mostraram que SHED cultivadas em fatias de dentes e matrizes condutivas expressaram VEGFR-2 já após o primeiro dia de estímulo com VEGF. Ademais, os quatro marcadores de células endoteliais (VEGFR-1, VEGFR-2, CD31 e Caderina-VE) foram observados após 21 dias sob estímulo de VEGF, resultado ainda mais evidente aos 28 dias. In vivo, observou-se que SHED transfectadas com o gene LacZ foram capazes de formar estruturas semelhantes a vasos sangüíneos quando implantadas em camundongos, mas a presença de sangue no seu interior não pôde ser observada após 21 dias de implante. Portanto, SHED podem ser estimuladas a se diferenciar em odontoblastos funcionais, capazes de produzir estrutura mineralizada semelhante à dentina. Ademais, VEGF interfere nas vias de sinalização STAT3, ERK e AKT e estimula a formação de estruturas tubulares e a diferenciação de SHED em células endoteliais, mas não a proliferação e migração de SHED. Acreditamos que tecnologia igual ou semelhante à empregada neste estudo poderá eventualmente fornecer ferramentas clínicas para tratamentos endodônticos que visem à regeneração de um tecido pulpar completo e formação de tecido dentinário num futuro não muito distante. / Dental pulp tissue engineering aims to replace the inflamed or necrotic pulp by a healthy and functionally competent tissue able to form new dentin in order to repair lost structure. The purposes of this work were: to evaluate the differentiation ability of stem cells from human exfoliated deciduous teeth (SHED) into functional odontoblasts, showing the formation of mineralized tissue in vivo; and to study the effect of VEGF on SHED with regards to the stimulation of cell signaling pathways (STAT3, AKT and ERK), the proliferation, migration, capillary sprouting, and the differentiation into endothelial cells. The beginning of the mineralization process of SHED treated with dexamethasone, ascorbic acid and beta-glycerophosphate could be detected through the production of alkaline phosphatase after the second week of culture, but the expression of DSPP mRNA was only observed after 28 days of induction. Using the tooth slice and scaffold model implanted in the dorsum of immunocompromised mice, the differentiation of SHED into odontoblast-like cells, which were immunostained with DMP-1 antibody, was demonstrated. Dentin deposition following a centripetal rhythm, in a rate of 14.1 µm per day, was also shown through the tetracycline labeling. VEGF treatment of SHED stimulated the ERK and AKT phosphorilation, and decreased the phosphorilation of STAT3 over 1 hour period, presumably due to its binding to VEGFR-1 and NP-1 receptors in these cells. In addition, VEGF enhanced SHED organization into tubular structures, with statistically significant difference between the treated group and the non-treated one after the 5th day of treatment. However, VEGF did not stimulate proliferation and migration of these cells. RT-PCR results demonstrated that SHED seeded in the tooth slices and scaffolds expressed VEGFR-2 after the first day of VEGF stimulation. Moreover, the four endothelial cell markers (VEGFR-1, VEGFR-2, CD31 and VE-Cadherin) were observed after 21 days of VEGF stimulation, and this result was even clearer after 28 days. In vivo, SHED transduced with LacZ gene were able to give rise to blood vessel-like structures when implanted in immunocompromised mice, but the presence of blood flow was not observed after 21 days of implantation. Therefore, SHED can be stimulated to differentiate into functional odontoblasts, which in turn are able to produce mineralized structure resembling dentin. Furthermore, VEGF interferes with the STAT3, ERK and AKT signaling pathways, and stimulates the formation of tubular structures and the differentiation of SHED into endothelial cells, but does not stimulate SHEDs proliferation and migration. We believe that the same technology employed in this study or a similar one can eventually provide clinical tools for the endodontic treatments aiming at regenerating a complete pulp tissue and forming a dentin tissue in a near future.

A associação de adubos verdes a fontes minerais de N para as culturas está tornando-se uma opção promissora de manejo da fertilização nitrogenada, com o objetivo de recuperar a fertilidade de solo, aumentar a matéria orgânica do solo, reduzir as perdas de N mineral de fontes prontamente disponíveis, e no caso das leguminosas incorporarem nitrogênio ao solo proveniente da fixação biológica. Os objetivos deste trabalho foram: (1) avaliar o aproveitamento do nitrogênio da crotalária, do milheto e da uréia pelo arroz e o efeito residual no feijoeiro cultivado em sucessão, (2) avaliar a contribuição do N das raízes e da parte aérea desses adubos verdes para o arroz e o feijoeiro, e (3) quantificar o N na planta proveniente dos adubos verdes em relação à época de aplicação da uréia (semeadura e cobertura). Foram desenvolvidos experimentos com Latossolo Vermelho distroférrico típico em casa de vegetação no CENA/USP em três fases: (a) marcação isotópica dos adubos verdes com 15N realizada com (NH4)2SO4 enriquecido em 10 e 5% de átomos de 15N para a crotalária e milheto, respectivamente, (b) cultivo de arroz em vasos de 4 kg de TFSA com delineamento experimental inteiramente casualizado com 12 tratamentos e três repetições, dispostos em esquema fatorial 3 x 4, correspondendo a combinação de crotalária, milheto e sem adição fonte orgânica de N com quatro doses de 15N-uréia (0; 28,6; 57,2; 85,8 mg kg-1 de N ), e (c) cultivo de feijão nos mesmos vasos após o cultivo de arroz. A produtividade de grãos de arroz foi influenciada positivamente pela utilização de crotalária quando comparada ao milheto e ao tratamento sem adubo verde, com maior efeito a medida que se aumentou a dose de N mineral. O aproveitamento do N da crotalária (18,9%) foi mais de duas vezes superior ao do milheto (7,8%). As doses de uréia, nos tratamentos com ou sem uso adubo verde, não influenciaram o aproveitamento de N do fertilizante pela planta de arroz (palha+grão), sendo a media geral de aproveitamento de N-uréia de 53,76%. O aproveitamento do N das raízes dos adubos verdes foi de 14,1%, enquanto da parte aérea foi de 16,8%. Com o uso de crotalária, observou-se maior aproveitamento de N-uréia aplicado na semeadura (61,7%) em relação à cobertura (47,1%), mas esse efeito não foi observado para o milheto e sem adição de adubo verde. O aproveitamento do N residual dos adubos verdes pelo feijão foi de 3,56% para o N da crotalária e de 3,43% para o N de milheto, e superior ao do N do fertilizante (2,63%). / The association of green manures and mineral N sources is becoming a promising option for nitrogen fertilizer management, with the objective of recovering the soil fertility, increase of soil organic matter, reduce the loss of mineral N from the readily available sources, and in the case of legume incorporate biologically fixed N to the soil. The objectives of this research were to: (1) evaluate the utilization of N by the rice crop from crotalaria (sunnhemp), millet and urea and its residual effect on bean crop (Phaseolus vulgaris L.) grown in succession; (2) evaluate the contribution of N from the roots and above ground part of these green manures to rice and bean crops, and (3) quantity the N in the plant derived from the green manures in relation to the urea application time (at seeding and topdressing). Experiments were carried out with typic dystrophic Red Latosol (oxisol) in green house at CENA/ USP, in three phases: (a) green manures labeling with 15N, using (NH4)2SO4 enriched with 10 and 5% 15N, for crotalaria and millet, respectively; (b) rice growth in pots containing 4 kg of air dried soil, in completely randomized design with 12 treatments and three replicates, arranged in a 3 x 4 factorial scheme, corresponding to combination of crotalaria, millet and without addition of organic N, with four rates of urea-15N (0; 28.6; 57.2; 85.8 mg N Kg-1), and (c) bean plant growth after rice cultivations. The rice grain productivity was affected positively by the crotalaria when compared to millet and treatment without green manure, with greater effect with increasing mineral N rate. The crotalaria N utilization (18.9%) by the rice plants was more than two times higher than from the millet (7.8%). The urea rates, in the treatments with or without green manure, did not affect the utilization by the rice plants (grains + husk) of N form the fertilizer (53.76% in average). The utilization of green manure roots N by the rice plants was 14.1%, while of above ground parts was 16.8%. With crotalaria, higher utilization of N occurred from urea applied at seeding (61.7%) compared to the top dressed application (47.1%), but this effect was not observed for millet and without addition of green manure. The utilization of green manures residual N by the bean crop was only 3.56% from crotalaria and 3.43% from millet, which were higher than from fertilizer N (2.63%).

[pt] O objetivo deste trabalho foi o desenvolvimento de um método analítico para determinação simultânea de elementos na faixa de concentração de traços em óleos através das técnicas de espectrometria de massas (ICPMS) e espectrometria de emissão ótica (ICPOES), ambas com plasma indutivamente acoplado, após procedimento de digestão em equipamento de mineralização que trabalha sob altas temperaturas (300 °C) e pressões (100 bar). A digestão dos petróleos foi realizada em frascos fechados de quartzo de volume de 90 mL utilizando-se uma mistura de ácido nítrico e peróxido de hidrogênio concentrado, de elevada pureza. Foram avaliadas as melhores condições de digestão dos petróleos, possíveis interferentes resultantes das matrizes formadas durante o processo de digestão e as condições operacionais para a determinação simultânea dos elementos de interesse. O teor de carbono residual foi determinado para avaliar a extensão do processo de digestão e possíveis interferências em ambas às técnicas utilizadas. O efeito da concentração residual de ácido nítrico na intensidade dos sinais analíticos foi estudado, bem como o desempenho de diferentes padrões internos para corrigi- lo. Curvas analíticas com lineariadades r2 > 0,999 foram obtidas para a maioria do elementos determinados e limites de detecção (LDM-3(sigma)): < 1 (mi) g kg-1 para Sb, Be, Cr, Co, Cu, Fe, Li, Mg, Mn, Hg, Ni, Ag, Na, Sr, Ti, V, Zn, ETR, EGP, e na faixa de 1 a 10 (mi)g kg-1, para Al, Ba, Cd, Pb, K, Se. O método foi validado através da quantificação de elementos certificados nas amostras de referência NIST 1634c e NIST 1084a, obtendo-se repetitividades e exatidões melhores que 5% para a maioria deles. / [en] An analytical procedure was developed for the determination of trace elements in crude oil by inductively coupled plasma mass spectrometry (ICPMS) and inductively coupled plasma optical emission spectrometry (ICPOES) after sample digestion in a high pressure asher system operating at 100 bar and a temperature of 300 °C. Wet digestion was performed in closed quartz vessels of 90 mL volume using high purity nitric acid and hydrogen peroxide. Best conditions for the digestion of petroleum samples were studied and possible interferences caused by the resulting matrix were evaluated, as well as best measuring conditions for the analytes of interest. Residual carbon content was determined in order to assess the completeness of digestion and interferences caused by this element in both techniques. The effect of residual acid concentration on the analytical signals were studied, as well as the performance of different internal standards for its correction. Linear analytical curves (r2 >0.999) were obtained and method detection limits (MDL - 3(sigma)) of 1 (mi)g kg-1 for Sb, Be, Cr, Co, Cu, Fe, Li, Mg, Mn, Hg, Ni, Ag, Na, Sr, Ti, V, Zn, ETR, EGP, and in the range of 1 to 10 (mi)g kg-1 for Al, Ba, Cd, Pb, K and Se were obtained. Method validation was performed by quantification of certified elements in two standard reference materials (NIST: 1634c; 1084a) resulting in repeatabilities and accuracies of better than 5 % for most of the elements studied.

Das Ziel der Studien, die im Rahmen dieser Arbeit vorgestellt werden, war es neue Informationen über die elementare Zusammensetzung des mineralisierten Knochens zu gewinnen. In einer ersten Studie wurden zwei Parameter verglichen, die beide eng mit der Knochenmineralisierung verknüpft sind. So zeigte die Gegenüberstellung des mineral/matrix Raman-Wertes und der Kalziumkonzentration gute Übereinstimmung mit theoretischen Überlegungen. Diese Methoden wurden auch verwendet um Knochengewebe von Mäusen zu charakterisieren bei denen ein genetischer Defekt zu einem Mangel von Sclerostin führte. So war es möglich nachzuweisen, dass eine hierdurch verstärkte Knochenneubildung zu einer veränderten Mineralisationskinetik des Knochens führen kann. Nachdem zukünftig Sclerostinantikörper für die Behandlung von Knochenkrankheiten eingesetzt werden sollen, haben diese Erkenntnisse große medizinische Bedeutung. Es wurde auch die Mineraldichteverteilung eines Mausmodells mit fragilem Knochen (Osteogenesis Imperfecta, OI) untersucht. Die Mäuse wurden mit Sclerostinantikörpern behandelt. Es zeige sich ein signifikanter Knochenzuwachs doch die Mineraldichteverteilung veränderte sich gleichermaßen für gesunde und für OI Mäuse. In einer Studie am humanen Knochen konnten der Zusammenhang zwischen Osteozytennetzwerk und Knochenzusammensetzung untersucht werden. Elemente wie Na, Mg und S wiesen typische Konzentrationsverteilungen auf. Die Routinen wurden auch verwendet um Mineralisationsfronten zu charakterisieren. Es zeigte sich, dass die Konzentrationen von K, Mg, Na und Cl abhängig von dem analysierten anatomischen Ort, stark voneinander abweichen. Abschließend kann gesagt werden, dass durch die Entwicklung neuer Routinen zusätzliche Erkenntnisse über die Knochenmineralisierung und Zusammensetzung gewonnen werden konnten. Die Resultate sind von medizinischer und biologischer Bedeutung und tragen zu aktuellen Debatten über die Knochenentwicklung bei. / The purpose of the presented work was to gain new insight into the elemental composition of mineralized bone matrix at different sites of human bone tissue, and in mouse models linked to human genetic diseases. Using novel tools and routines, human (femur cross sections from healthy adults and children) and murine samples (femur long-and cross sections of two mouse models) were analyzed with focus on the elemental composition. In a methodological study the consistency of matrix mineralization measured by Raman microspectroscopy (e.g. the mineral/matrix ratio) and the Calcium content (wt%Ca) as measured by qBEI was proved. Both methods were applied to a mouse model exhibiting induced bone overgrowth due to a genetic defect causing a lack of Sclerostin, which is a negative regulator for bone formation. We found changes in the mineralization kinetics depending on the anatomical site. This result is of clinical importance since Sclerostin antibodies are suggested for future treatment of diseases characterized by fragile bone. Hence, also a mouse model of a brittle bone disease (Osteogenesis Imperfecta) was analyzed with and without Sclerostin antibody treatment. A significant increase in bone mass was documented while the mineralization pattern revealed no interaction between genotype and treatment. The correlation between OLCN and the composition of the mineralized matrix was examined in the same regions of human compact bone. Characteristic distributions of the minor elements (Mg, Na, S) were found. The developed tools were also used to investigate mineralization fronts, reflecting a critical stage of bone development. Differences in the Ca/P ratio and in the concentrations of K, Mg, Na and Cl depending on the anatomical site were revealed. In conclusion, using newly developed measurement routines, it was possible to gain novel information of bone mineralization and composition. The results contribute to actively debated issues of biological and medical importance.

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No. of bitstreams: 1 Carolina Gonçalves Leandro 2017 Dissertação.pdf: 10712768 bytes, checksum: ee554a9308600c58ba9930f1d2d302ae (MD5) Previous issue date: 2017 / A Mina Cerro Rico localiza-se aproximadamente a 7 km a leste da cidade de Lavras do Sul, na região central do estado do Rio Grande do Sul e está sob licença de pesquisa mineral da empresa Amarillo Mineração do Brasil Ltda. A área de pesquisa possui 15,758 km², delimitada pelas longitudes 53º 47’ 42,7” W - 53º 50’ 56,4” W e pelas latitudes 30º 48’ 5,7” S - 30º 49’ 46” S. A ocorrência mineral de Au, Ag e Cu desta área, está relacionada a presença de zonas de alteração hidrotermal caracterizadas por filões de quartzo de direções NW e EW, alojados nas rochas vulcânicas da Formação Hilário (~ 580 Ma). Os dados aerogeofísicos pertencem ao levantamento gamaespectrométrico – Projeto Lavras do Sul – Rio Grande do Sul, o qual foi executado no ano de 2007 pela empresa Prospectors Aerolevantamentos e Sistemas Ltda. A partir de setores identificados pela caracterização de anomalias geofísicas oriundas do processamento dos dados aerogamaespectrométricos onde foi definida uma área para levantamentos geofísicos terrestres em escala de detalhe, aproximadamente de 1: 3.000, com os métodos geoelétricos de Eletrorresistividade e de Polarização Induzida, utilizando as técnicas de Caminhamento Elétrico e de Sondagem Elétrica Vertical. Os dados aerogamaespectrométricos permitiram a partir da interpretação do mapa Ternário dos radioelementos (K (%), eU (ppm), eTh (ppm)), a identificação de cinco domínios radiométricos que sugerem a presença de diferentes litologias, em termos de possíveis variações na textura e na mineralogia das rochas e solos presentes na área. Os perfis gamaespectrométricos integrados com os dados geoelétricos possibilitaram a identificação de possíveis zonas de fraturas e/ou falhas, com provável percolação de fluído hidrotermal e/ou presença de mineralizações metálicas. O modelo geofísico-geológico 2D proposto para a área da Mina Cerro Rico, sugere que o topo do embasamento andesítico se encontra entre 20 a 30 metros de profundidade, caracterizado pela presença de um relevo irregular, afetado por fraturas e/ou falhas. Conforme a integração dos resultados oriundos dos dados de resistividade e da cargabilidade, definiu-se que patamares anômalos com valor de resistividade em torno de 200 Ohm.m e de cargabilidade em torno de 8 mV/V a 12 mV/V, estão associados a presença de zonas de fraturas com possível mineralização metálica. Os setores da área com presença de fraturas preenchidas por veios de quartzo e com provável mineralização, resultam em potenciais atrativos, para dar continuidade e orientar futuras pesquisas relacionadas a prospecção aurífera na Mina Cerro Rico. / The Cerro Rico Mine is located approximately 7 km east of the Lavras do Sul city, in the central region of the state of Rio Grande do Sul and is under mineral exploration license of the Amarillo Mineração do Brasil company. The research area is 15,758 km², delimited by longitudes 53º 47 '42.7 "W - 53º 50' 56.4" W and latitudes 30º 48 '5,7 "S - 30º 49' 46" S. The occurrence of gold, silver and cooper minerals in this area are related to the presence of hydrothermal alteration zones characterized by quartz filaments through NW and EW directions, intruded in the volcanic rocks of the Hilário Formation (~ 580 Ma). The aerogeophysical data belongs to the Lavras do Sul Gammaspectrometric Survey Project, which was executed by Prospectors Aerial Surveys and Systems in 2007. The sectors was identified by the characterization of geophysical anomalies through the processing of aerogamaespectrometric data, in this area the geophysical surveying was made on a scale of approximately 1: 3.000 and defined with the geoelectric methods of Eletroresistivity and Induced Polarization using the techniques of 2D Horizontal Profiling and Vertical Electrical Sounding. The aerogamaespectrometric data allowed the identification of five radiometric domains that suggests the presence of differents lithologies in terms of possible variations in the radioelements (K (%), eU (ppm), eTh (ppm)), texture and mineralogy of the rocks and soils present in the area. The gamma spectrometric profiles integrated with the geoelectric data allowed the identification of possible zones of fractures and/or faults, with probable percolation of hydrothermal fluid and/or presence of metallic mineralizations. The 2D geophysical-geological model proposed for the Cerro Rico Mine area, suggests that the depth of the top of the andesitic basement is between 20 and 30 meters, characterized by the presence of irregular relief, affected by fractures and/or faults. According to the integration of the results from the resistivity and chargeability data, the anomalous levels with a resistivity value around 200 Ohm.m and chargeability of 8 mV/V to 12 mV/V are associated with the presence of fracture zones with possible metallic mineralization. The sectors of the area with presence of fractures filled by quartz veins and probable mineralization, results in attractives potentials, to give continuity and guide future research related to gold prospecting in the Cerro Rico Mine.