971 resultados para LC-APCI-MS


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A Embrapa Solos, em parceria com a Secretaria de Estado de Desenvolvimento Agrário, da Produção, da Indústria, do Comércio e do Turismo - SEPROTUR, realizou o Zoneamento Agroecológico do Estado do Mato Grosso do Sul - Fase II - com objetivo de contribuir na indicação de áreas passíveis de exploração agrícola sustentável. No desenvolvimento desse trabalho foram considerados aspectos legais, restrições ambientais, potencial das culturas, aspectos do clima, de geomorfologia e dos solos, todos integrados em ambiente de sistema de informação geográfica com apoio de algebra de mapas, no intuito de avaliar a adequabilidade de uso das terras e apresentar uma proposição de planejamento de uso e ocupação das terras. Os resultados foram consolidados por município e dão origem a esse boletim de pesquisa. No município de Anastácio, as terras indicadas para o uso com lavouras somam 1.425 km², o que equivale a aproximadamente 50% da área total do município, enquanto que as recomendadas para pastagem equivalem a 26,5% e as áreas recomendadas para pastagem especial ou cultivo de arroz correspondem a cerca de 20% da área do município que corresponde a algo como 578 km². Nestas unidades é fundamental avaliar criteriosamente a utilização de pastagens nestas terras quando essas ainda se encontram sob cobertura vegetal, visto que, praticamente 20% destas terras ainda permanecem com vegetação natural em seus diversos graus de conservação. As terras recomendadas para conservação dos recursos naturais equivalem a menos de 80 km², as quais constituem áreas de alta fragilidade ambiental e/ ou apresentam restrições legais de uso como áreas de preservação permanente. As áreas identificadas como zonas recomendadas para recuperação ambiental equivalem a 80 km² e constituem áreas de moderada a alta fragilidade ambiental e/ou que apresentam restrições legais de uso e que já foram desmatadas para o uso com pastagens/agricultura. A área do município de Anastácio apresenta alto grau de ação antrópica das terras, onde mais de 75% das terras sendo utilizadas com pastagens e/ou com agricultura, enquanto que apenas 25% ainda apresentam certo grau de preservação.

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A Embrapa Solos, em parceria com a Secretaria de Estado de Desenvolvimento Agrário, da Produção, da Indústria, do Comércio e do Turismo - SEPROTUR, realizou o Zoneamento Agroecológico do Estado do Mato Grosso do Sul - Fase II - com objetivo de contribuir na indicação de áreas passíveis de exploração agrícola sustentável. No desenvolvimento desse trabalho foram considerados aspectos legais, restrições ambientais, potencial das culturas, aspectos do clima, de geomorfologia e dos solos, todos integrados em ambiente de sistema de informação geográfica com apoio de algebra de mapas, no intuito de avaliar a adequabilidade de uso das terras e apresentar uma proposição de planejamento de uso e ocupação das terras. As terras indicadas para o uso com lavouras somam cerca de 1.040 km², o que equivale a aproximadamente 16% da área total zoneada. As áreas recomendadas para pastagem equivalem a 47% e as áreas recomendadas para pastagem especial a menos de 1,5% da área do município que corresponde a algo como 80 km². Nestas unidades é fundamental avaliar criteriosamente a utilização de pastagens nestas terras quando essas ainda se encontram sob cobertura vegetal, visto que, praticamente 82% destas terras ainda permanecem com vegetação natural em seus diversos graus de conservação. As terras recomendadas para conservação dos recursos naturais e/ou recuperação ambiental equivalem a 210 km², as quais constituem áreas de alta fragilidade ambiental e/ou apresentam restrições legais de uso como áreas de preservação permanente. As áreas identificadas como zonas recomendadas para recuperação ambiental equivalem a 123 km² e constituem áreas de moderada a alta fragilidade ambiental e/ou que apresentam restrições legais de uso e que já foram desmatadas para o uso com pastagens/agricultura. O do município de Coxim apresenta um alto grau de ação antrópica das terras, onde cerca de 60% das terras são utilizadas com pastagens e/ou com agricultura, enquanto que apenas 40% ainda apresentam certo grau de preservação. O município de Coxim apresenta um bom potencial para o desenvolvimento da agropecuária, porém, devidos às características edafoclimáticas e de topografia, a recomendação preferencial é a utilização com pastagens.

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Jenkins, H., Beckmann, M., Draper, J., & Hardy, N. (2007). GC-MS Peak Labeling Under ArMet. Nikolau, B. J., & Wurtele, E. S. (Eds.), In: Concepts in Plant Metabolomics. (pp. 19-28). Dordrecht: Springer. Sponsorship: The authors gratefully acknowledge the United Kingdom Food Standards Agency (under the G02006 project) for support of their work in metabolomics.

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Rothwell, W., Femina (Trinity College, Cambridge MS B.14.40); edited with an Introduction and Notes (The Anglo-Norman On-Line Hub, 2005) RAE2008

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Traditional methods for phenotyping skeletal muscle (e.g., immunohistochemistry) are labor-intensive and ill-suited to multixplex analysis, i.e., assays must be performed in a series. Addressing these concerns represents a largely unmet research need but more comprehensive parallel analysis of myofibrillar proteins could advance knowledge regarding age- and activity-dependent changes in human muscle. We report a label-free, semi-automated and time efficient LC-MS proteomic workflow for phenotyping the myofibrillar proteome. Application of this workflow in old and young as well as trained and untrained human skeletal muscle yielded several novel observations that were subsequently verified by multiple reaction monitoring (MRM).We report novel data demonstrating that human ageing is associated with lesser myosin light chain 1 content and greater myosin light chain 3 content, consistent with an age-related reduction in type II muscle fibers. We also disambiguate conflicting data regarding myosin regulatory light chain, revealing that age-related changes in this protein more closely reflect physical activity status than ageing per se. This finding reinforces the need to control for physical activity levels when investigating the natural process of ageing. Taken together, our data confirm and extend knowledge regarding age- and activity-related phenotypes. In addition, the MRM transitions described here provide a methodological platform that can be fine-tuned to suite multiple research needs and thus advance myofibrillar phenotyping.

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De acuerdo a la normativa de TFEs el repositorio no puede dar acceso a este trabajo. Para consultarlo póngase en contacto con el tutor del trabajo. Puede acceder al resumen del mismo pinchando en el pdf adjunto

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15 fotografías, nueve a color y seis en tono de grises.

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A simple procedure for the isolation of caffeine from energy drinks by solid phase extraction on a C18 cartridge. Quantitative analysis of the amount of caffeine by LC/MS is determined by referencing a standard curve.

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In this study, marine sponges collected in Irish waters were analysed for their associated microbiota. Of the approximately 240 bacterial isolates obtained from two sponges several showed antimicrobial activity; among them members of genera which have rarely been shown to produce antimicrobial compounds. Differences observed from the sponge-derived groups of isolates in terms of bioactivity suggests that S. carnosus isolates may be a better source of antibacterial compounds, while Leucosolenia sp. isolates appear to be a better source of antifungal compounds. More than 60% of fungal isolates obtained from 12 sponge samples proved to be bioactive. One of the isolates, which was closely related to Fusarium oxysporum and showed activity against bacteria and fungi, was investigated for its secondary metabolite genes. At least 5 different NRPS genes, with a sequence similarity as low as 50 % to known genes, were identified highlighting the likelihood that this isolate may be capable of producing novel secondary metabolites. A Micromonospora sp. was isolated from a Haliclona simulans sample collected in Irish waters. The isolate inhibited the growth of Gram positive bacterial test strains in three different antimicrobial assays. Employing preparative layer chromatography the compound responsible for the bioactivity could be isolated. According to LC-MS andNMR data the bioactive compound could indeed be novel. Finally, two deep water sponges were shown to host a remarkably different bacterial and archaeal diversity by application of 454 Pyrosequencing. The L. diversichela –proteobacterial community was dominated by a single ƴ-proteobacterial bacterium whereas the S. normani sample hosted a largely sponge specific microbial community, even more diverse than has been previously reported for shallow water sponges. Organisms potentially involved in nitrification, sulphate reduction and secondary metabolite production were found to be spatially distributed in the sponge. Furthermore, a deep sea specific population was implied.

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The standard early markers for identifying and grading HIE severity, are not sufficient to ensure all children who would benefit from treatment are identified in a timely fashion. The aim of this thesis was to explore potential early biomarkers of HIE. Methods: To achieve this a cohort of infants with perinatal depression was prospectively recruited. All infants had cord blood samples drawn and biobanked, and were assessed with standardised neurological examination, and early continuous multi-channel EEG. Cord samples from a control cohort of healthy infants were used for comparison. Biomarkers studied included; multiple inflammatory proteins using multiplex assay; the metabolomics profile using LC/MS; and the miRNA profile using microarray. Results: Eighty five infants with perinatal depression were recruited. Analysis of inflammatory proteins consisted of exploratory analysis of 37 analytes conducted in a sub-population, followed by validation of all significantly altered analytes in the remaining population. IL-6 and IL-6 differed significantly in infants with a moderate/severely abnormal vs. a normal-mildly abnormal EEG in both cohorts (Exploratory: p=0.016, p=0.005: Validation: p=0.024, p=0.039; respectively). Metabolomic analysis demonstrated a perturbation in 29 metabolites. A Cross- validated Partial Least Square Discriminant Analysis model was developed, which accurately predicted HIE with an AUC of 0.92 (95% CI: 0.84-0.97). Analysis of the miRNA profile found 70 miRNA significantly altered between moderate/severely encephalopathic infants and controls. miRNA target prediction databases identified potential targets for the altered miRNA in pathways involved in cellular metabolism, cell cycle and apoptosis, cell signaling, and the inflammatory cascade. Conclusion: This thesis has demonstrated that the recruitment of a large cohortof asphyxiated infants, with cord blood carefully biobanked, and detailed early neurophysiological and clinical assessment recorded, is feasible. Additionally the results described, provide potential alternate and novel blood based biomarkers for the identification and assessment of HIE.

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Described here is a mass spectrometry-based screening assay for the detection of protein-ligand binding interactions in multicomponent protein mixtures. The assay utilizes an oxidation labeling protocol that involves using hydrogen peroxide to selectively oxidize methionine residues in proteins in order to probe the solvent accessibility of these residues as a function of temperature. The extent to which methionine residues in a protein are oxidized after specified reaction times at a range of temperatures is determined in a MALDI analysis of the intact proteins and/or an LC-MS analysis of tryptic peptide fragments generated after the oxidation reaction is quenched. Ultimately, the mass spectral data is used to construct thermal denaturation curves for the detected proteins. In this proof-of-principle work, the protocol is applied to a four-protein model mixture comprised of ubiquitin, ribonuclease A (RNaseA), cyclophilin A (CypA), and bovine carbonic anhydrase II (BCAII). The new protocol's ability to detect protein-ligand binding interactions by comparing thermal denaturation data obtained in the absence and in the presence of ligand is demonstrated using cyclosporin A (CsA) as a test ligand. The known binding interaction between CsA and CypA was detected using both the MALDI- and LC-MS-based readouts described here.

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Mitochondria are responsible for producing the vast majority of cellular ATP, and are therefore critical to organismal health [1]. They contain thir own genomes (mtDNA) which encode 13 proteins that are all subunits of the mitochondrial respiratory chain (MRC) and are essential for oxidative phosphorylation [2]. mtDNA is present in multiple copies per cell, usually between 103 and 104 , though this number is reduced during certain developmental stages [3, 4]. The health of the mitochondrial genome is also important to the health of the organism, as mutations in mtDNA lead to human diseases that collectively affect approximately 1 in 4000 people [5, 6]. mtDNA is more susceptible than nuclear DNA (nucDNA) to damage by many environmental pollutants, for reasons including the absence of Nucleotide Excision Repair (NER) in the mitochondria [7]. NER is a highly functionally conserved DNA repair pathway that removes bulky, helix distorting lesions such as those caused by ultraviolet C (UVC) radiation and also many environmental toxicants, including benzo[a]pyrene (BaP) [8]. While these lesions cannot be repaired, they are slowly removed through a process that involves mitochondrial dynamics and autophagy [9, 10]. However, when present during development in C. elegans, this damage reduces mtDNA copy number and ATP levels [11]. We hypothesize that this damage, when present during development, will result in mitochondrial dysfunction and increase the potential for adverse outcomes later in life.

To test this hypothesis, 1st larval stage (L1) C. elegans are exposed to 3 doses of 7.5J/m2 ultraviolet C radiation 24 hours apart, leading to the accumulation of mtDNA damage [9, 11]. After exposure, many mitochondrial endpoints are assessed at multiple time points later in life. mtDNA and nucDNA damage levels and genome copy numbers are measured via QPCR and real-time PCR , respectively, every 2 day for 10 days. Steady state ATP levels are measured via luciferase expressing reporter strains and traditional ATP extraction methods. Oxygen consumption is measured using a Seahorse XFe24 extra cellular flux analyzer. Gene expression changes are measured via real time PCR and targeted metabolomics via LC-MS are used to investigate changes in organic acid, amino acid and acyl-carnitine levels. Lastly, nematode developmental delay is assessed as growth, and measured via imaging and COPAS biosort.

I have found that despite being removed, UVC induced mtDNA damage during development leads to persistent deficits in energy production later in life. mtDNA copy number is permanently reduced, as are ATP levels, though oxygen consumption is increased, indicating inefficient or uncoupled respiration. Metabolomic data and mutant sensitivity indicate a role for NADPH and oxidative stress in these results, and exposed nematodes are more sensitive to the mitochondrial poison rotenone later in life. These results fit with the developmental origin of health and disease hypothesis, and show the potential for environmental exposures to have lasting effects on mitochondrial function.

Lastly, we are currently working to investigate the potential for irreparable mtDNA lesions to drive mutagenesis in mtDNA. Mutations in mtDNA lead to a wide range of diseases, yet we currently do not understand the environmental component of what causes them. In vitro evidence suggests that UVC induced thymine dimers can be mutagenic [12]. We are using duplex sequencing of C. elegans mtDNA to determine mutation rates in nematodes exposed to our serial UVC protocol. Furthermore, by including mutant strains deficient in mitochondrial fission and mitophagy, we hope to determine if deficiencies in these processes will further increase mtDNA mutation rates, as they are implicated in human diseases.

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Los métodos tradicionales usados por pequeños agricultores de Corrientes ocasionaron la degradación de los suelos. Se hipotetizó que la siembra directa (SD) y la avena negra (Av) como cultivo de cobertura incrementan la densidad aparente (Da) y la resistencia mecánica (RM) sin reducir el rendimiento (Rto) del maíz y algodón, que aumentan la estabilidad de agregados (EA), la cobertura de suelo (Cob) y los rastrojos en superficie (BM), mejorando la infiltración (Ib), la porosidad total (PT) y el espacio aéreo a capacidad de campo (EaCC). El objetivo fue establecer el sistema de labranza y secuencia de cultivos que mejoran las propiedades físicas de un Argiudol hipertérmico, y evaluar los cambios en el tiempo. Se instaló un ensayo con diseño en parcelas completamente aleatorizadas con cuatro repeticiones y arreglo factorial 3x4: sistema de labranza (SL), con tres niveles: labranza convencional (LC), reducida (LR) y SD; y secuencia de cultivos, con maíz (M), algodón (A), descanso (D) y Av, con cuatro niveles: M-Av-A-Av, M-D-A-D, A-Av-M-Av y A-D-M-D. Se realizó un ANOVA en la situación inicial, a la siembra y cosecha de los cultivos durante 2007-10 y un ANOVA de medidas repetidas en el tiempo. Los efectos de los SL en las propiedades físicas fueron más evidentes que los ocasionados por las secuencias y Av. La SD mantuvo los rendimientos afectando poco las condiciones superficiales. Durante tres ciclos la SD mejoró la protección de la superficie del suelo (BM y Cob), produjo mayor EA que LC y LR, y mejoró la Ib. Un incremento progresivo de la Da y mayor RM se dio bajo SD, afectando la PT y el EaCC pero sin comprometer el rendimiento de los cultivos. Los resultados sugieren que la SD puede ser usada como alternativa al manejo tradicional por pequeños agricultores de Corrientes con suelos de régimen hipertérmico

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La intervención de un extensionista - ingeniero agrónomo de la Agencia de Extensión Rural Orán, del Instituto Nacional de Tecnología Agropecuaria/INTA- en el territorio de una Comunidad Tupí Guaraní del norte salteño, exige una profunda revisión de antecedentes dando lugar a reflexiones, interrogantes y propuestas relacionadas con distintos aspectos del trabajo de campo que trascienden a esta situación en particular. Esto está aquí directamente relacionado a la evolución y al desarrollo de esta zona de históricos conflictos de intereses, desigualdad social creciente y fuertes procesos de aculturación. Incluso existen contradicciones dentro de las instancias del Estado relacionadas con el desarrollo rural, en donde el INTA no constituye una excepción, generándose arduas discusiones internas. En este escenario, la praxis destaca la importancia de las técnicas participativas como la mejor metodología para sustentar procesos de desarrollo local, ya que es en terreno se debe trabajar dentro de un proceso de pérdida de la percepción social -pero más importante aún, de la propia percepción- en la que los pueblos originarios se consideren como un actor más. Entonces es desde la revalorización de las propias capacidades y posibilidades de la comunidad que se analiza una experiencia que refleja cómo, a partir del único agente del estado presente en terreno, se intenta avanzar en la trasformación de la realidad en donde se ve naturalizada y no problematizada la diferencia en la distribución de los ingresos y la pobreza estructural. El comienzo de la intervención, por lo tanto, se inició con la identificación clara de una amenaza -el avance de la erosión del río sobre la seguridad y posesiones de los campesinos/aborígenes- lo cual creó la necesidad ineludible de trabajar comunitariamente. Esta coyuntura generó la necesidad de organizarse para llevar a cabo el proyecto necesario para superar la emergencia, que gracias a la forma participativa de encarar el trabajo, así como el trabajo inter-institucional de los técnicos del INAI (Instituto Nacional de Asuntos Indígenas) y del INTA, confluyeron en crear un capital social entre productores y técnicos/instituciones. La articulación interinstitucional lograda hizo posible la formulación y ejecución en forma participativa de un proyecto evaluado satisfactoriamente -social e institucionalmente-, ya que posibilitó controlar la erosión fluvial evitando nuevas pérdidas de tierras, fortaleciendo a la vez a la organización comunitaria y los vínculos con distintos actores públicos, revalorizando sus propias capacidades.