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Impacto da umidade do solo sobre a estrutura das comunidades bacterianas e sobre as atividades enzimáticas em solos da Caatinga e da Mata Atlântica / Impact of the soil humidity on structure of the bacterial communities and on the enzymatic activities in soils of the Caatinga and Atlantic ForestLaura Bononi 04 February 2016 (has links)
As comunidades microbianas regulam a ciclagem de nutrientes no solo. O impacto das mudanças climáticas sobre estas poderia alterar a estrutura, a função e provocar um desequilíbrio dos nutrientes no ambiente. O principal objetivo desse trabalho foi estudar o impacto da umidade sobre a diversidade e atividades enzimáticas das comunidades bacterianas em microcosmos compostos por solos dos biomas Caatinga e Mata Atlântica. Amostras de solo foram coletadas nos seguintes estados brasileiros: Bahia, Pernambuco e São Paulo totalizando quatro pontos de coleta, com um ponto em cada estado e dois pontos no estado de São Paulo. As amostras de solo dos dois biomas, foram incubadas em microcosmos, com três ciclos de umidade (60 %- 30 %, umidade relativa) e para cada ciclo de umidade, foram construídas uma biblioteca do gene 16S rRNA. Nas análises de alfa diversidade, foram calculados os índices PD (diversidade filogenética baseado nos ramos da árvore filogenética) e Shannon. Para avaliar a beta diversidade foi utilizado o índice Bray-Curts (índice de similaridade baseado nos grupos dominantes) e a distância UniFrac. Simultaneamente, cada uma das amostras de cada tratamento foi analisada para as seguintes atividades enzimáticas: fosfatase ácida e alcalina, arilsulfatase, desidrogenase, celulase, amilase, urease e fitase. Os resultados aqui obtidos mostraram que os filos Acidobacteria, Proteobacteria e Chloroflexi foram mais abundantes nos solos da Mata Atlântica e Actinobacteria, Proteobacteria, Chloroflexi e Acidobacteria nos solos da Caatinga, considerando a característica em estudo do efeito adverso do estresse hídrico. As classes mais prevalentes na Mata Atlântica foram Acidobacteria, Alphaproteobacteria, Betaproteobacteria e Gammaproteobacteria. Enquanto que para a Caatinga, as classes prevalentes foram Ktedonobacteria, Actinobacteria-6 e Betaproteobacteria. No que diz respeito ao estresse hídrico sobre as atividades enzimáticas, os dados demonstraram efeito significativo para todos os solos amostrados, exceto para a enzima fitase. Por correlação entre as OTUs e as enzimas observou-se que grupos específicos foram correlacionados positivamente com a atividade das enzimas e com C e N total nos solos estudados. Estes resultados indicam que os ciclos de umidade afetam a distribuição taxonômica das comunidades bacterianas e as funções enzimáticas. / The microbial communities regulate the nutrient cycling in the soil. The impact of climate change on could alter the structure and function and cause an imbalance of nutrients in the environment. The main aim of this work was to study the impact of the soil humidity on diversity and enzymatic activities of bacterial communities in microcosms compounds soils of the Biosphere biomes of Caatinga and Atlantic Forest. Soil samples were collected in the following Brazilian states: Bahia, Pernambuco and Sao Paulo totaling four collection points, with a point in each state and two points in São Paulo. The soils samples of the two Biomes, were incubated in microcosms, with three humidity cycles (60% - 30% relative humidity) and for each humidity cycle, was constructed a 16S rRNA gene library. In the analyzes of alpha diversity, the PD indices were calculated (based on phylogenetic diversity branches of the phylogenetic tree) and Shannon and to test the beta diversity we used the Bray-Curts index (similarity index based on the dominant groups) and the distance UniFrac. Simultaneously, each soil sample of each treatment was analyzed for the following enzymatic activities: acid and alkaline phosphatase, arylsulfatase, dehydrogenase, cellulase, amylase, urease and phytase. The results herein obtained showed that the Phyla Acidobacteria, Proteobacteria and Chloroflexi were more abundant in the soils at Atlantic Forest, and Actinobacteria, Proteobacteria, Chloroflexi and Acidobacteria in the soils of Caatinga, considering the studied trait of the adverse effect of water stress. The most prevalent classes in the Atlantic Forest were Acidobacteria, Alphaproteobacteria, Betaproteobacteria and Gammaproteobacteria. While for the Caatinga, the prevalent classes were Ktedonobacteria, Actinobacteria-6 and Betaproteobacteria. In regard to the hydric stress on the enzymatic activities, the data showed significant effects for all sampling soils, except for the enzyme phytase. By correlations between OTUs and enzymes it was observed that specific groups were positively correlated with the activity of the enzymes and carbon and total nitrogen in the studied soils. These results indicate that humidity cycle affects the taxonomic distribution of bacterial communities and the enzymatic functions.
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Synthèse et caractérisation de nucléotides et oligonucléotides modifiés pour l'obtention de structures capables de mimer l'activité enzymatique des protéases à sérine / Nucleotides & oligonucleotides synthesis and caracterisation for the obtention of structures ables to mimics the enzymatic activity of serine proteasesAddamiano, Maria Claudia 25 October 2016 (has links)
Ce projet porte sur la synthèse d'oligonucléotides modifiés décorés par des groupements chimiques rappelant les chaines latérales des acides aminés impliqués dans la catalyse enzymatique des protéases à sérine, à savoir l'acide aspartique (Asp), la sérine (Ser) et l'histidine (His) afin d'en mimer l'activité protéolytique. L'approche synthétique de type phosphoramidite a été mise en oeuvre. De ce fait nous avons synthétisé des phosphoramidites fonctionnalisés avec une fonction acide carboxylique rappelant l'Asp, une fonction hydroxyle pour la Ser ou un imidazole pour l'His, correctement protégés pour être ensuite incorporés au sein de séquences oligonucléotidiques, et des phosphoramidites convertibles, qui portent une fonction chimique réactive permettant une conjugaison post synthèse supportée et automatisée de l'oligonucléotide. Ces deux types de phosphoramidites offrent la possibilité de les combiner et donc d'obtenir des séquences hautement fonctionnalisées. Les séquences oligonucléotidiques ont été choisies afin d'apporter un contrôle topologique structurant. Les structures secondaires envisagées sont de type bulge, épingle à cheveux et jonction trois voies car stables grâce aux appariements Watson et Crick entre les bases tout en ayant une flexibilité importante due à la présence de bases non appariées. Les travaux présentés dans ce manuscrit décrivent la synthèse des phosphoramidites modifiés ainsi que l'incorporation du nucléotide convertible de type alcyne au sein de séquences qui ont été conjuguées par CuAAC, et qui ont permi d'obtenir une jonction trois voies qui a été caractérisée par dichroïsme circulaire, gel de polyacrylamide et par dénaturation thermique. / We are interested in the synthesis of modified oligonucleotides decorated by chemical fonctions mimicking the amino acids side chains involved in enzymatic catalysis of serine proteases, acid aspartic (Asp), serine (Ser) and histidine (His), in order to mimic proteolytic activity. The phosphoramidite synthetic approach was choosen. We synthesised functionnalized phosphoramidites bearing either a carboxylic acid function reminding Asp, a hydroxyle function for Ser or an imidazole for His properly protected for their incorporation in oligonucleotides sequences, and convertible phosphoramidites, bearing a reactive function that permits a conjugation post automated oligonucleotides solid phase synthesis. Oligonucleotides sequences have been choosen in order to have a perfect structural control. The envisaged secondary structures are bulge, hairpin and three way junction because of their stability, thanks to Watson et Crick base pairing, and their flexibility thanks to the unpaired bases. The work presented here regards the synthesis of modified phosphoramidites and their incorporation of the convertible alkyne in sequences then conjugated by CuAAC reaction to form a three way junction whose stability and physico-chemical behavior have been evaluated.
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Characterisation of a novel soybean candidate glutathione peroxidase/thioredoxin-dependent peroxidase in soybean exposed to osmotic/drought stressPhillips, Kyle January 2012 (has links)
>Magister Scientiae - MSc / Drought stress is a major contributor to reduced soybean crop yield and quality, this can however be mitigated by the plant’s antioxidant defence mechanisms. One group of antioxidant enzymes that are active in these defence mechanisms are glutathione peroxidases (GPXs). GPXs are antioxidant proteins which are able to reduce H2O2, a toxic reactive oxygen species which accumulates under stress conditions. This study aims at isolating the protein encoded by Glyma01g42840 and determining if it has Phospholipid hydroperoxidase glutathione peroxidase (PHGPX) and/or Thioredoxin dependent peroxidase (TRX-PX) activity as well as assaying the effect of Drought stress on the expression of this putative GPX . This will be accomplished by molecular cloning, sequencing as well as the expression of the isolated protein to assay it enzymatic activity. It was found that the enzyme encoded by Glyma01g42840 is able to use glutathione and thioredoxin as electron donors for the detoxification peroxides, however enzymatic activity is more efficient when using glutathione as an electron donor. In conclusion it was found that glyma01g42840 encodes an enzyme which is able to utilise more than one electron donor and as glutathione produces the greatest amount of enzymatic activity it can be said that glyma01g42840 encodes a GPX.
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Trávicí proteasy termitů / Digestive proteases of termitesČermáková, Markéta January 2011 (has links)
Digestive proteolysis in termites has not been studied yet. In this diploma thesis, proteolytic enzymes of the digestive tract of two significant pest species Reticulitermes santonensis and Coptotermes formosanus (Rhinotermitidae) were analyzed. Proteases were identified and quantified in gut compartments using a panel of specific substrates and inhibitors. Major proteases were localized in the midgut and were classified as endogenous serine proteases of trypsin type. Minor cysteine proteases were detected in the paunch and were most likely produced by symbionts. The trypsin protease from R. santonensis was chromatographically isolated and its N-terminal sequence was identified. The physiological importance of the digestive trypsin proteases was demonstrated using selective inhibitors tested in vivo with C. formosanus. Based on the analysis of proteases from additional 12 termite species, a general scheme of digestive proteolysis in the order Isoptera was proposed. (In Czech)
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Advances in Synthesis and Biophysical Analysis of Protein-Polymer BioconjugatesWright, Thaiesha Andrea 08 July 2020 (has links)
No description available.
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The use of different ecosystem components as indicators of ecosystem development during platinum mine tailings rehabilitation / Johanna Martina (Juanita) RossouwRossouw, Johanna Martina January 2005 (has links)
Platinum mining activities contribute substantially to South Africa's economy since it
exceeded gold as economical contributor in 2001. Mining activities contribute to large
amounts of waste production in the form of tailings and rock waste, deposited in the
surrounding environment of the mine premises. Mining companies are held
responsible for damages caused to the surrounding environment. These companies are
required to introduce the cost of ecological rehabilitation in their operation costs as
well as compile an environmental management plan. Numerous attempts to
rehabilitate mine waste have proven unsuccessful. New and improved rehabilitation
techniques are required to facilitate in the rehabilitation of these mine spoils.
Woodchip-vermicompost produced from platinum mining wastes (woodchips and
sewage sludge) was used as an alternative amendment to inorganic fertilisers during
the rehabilitation of platinum mine tailings. The effectiveness of the woodchip-vermicompost
as an alternative amendment during the platinum mine tailings
rehabilitation were monitored using different ecosystem components. A natural veldt
in the vicinity of the mine area was randomly selected to serve as a reference site.
These ecosystem components selected have previously been shown to be effective as
indicators of ecosystem quality. The components selected for this study includes the
use of microbial enzymatic activity, microbial community structure, nematode trophic
structures, and other mesofaunal groups such as micro-arthropods. The physical and
chemical properties of the platinum mine tailings and reference area as well as the
vegetation cover of the platinum mine tailings were determined. Statistical and
multivariate analyses were use to determine the correlation between the dependent
microbial components and dominate independent chemical properties. Nematode
trophic structure, Maturity Index, and Plant-Parasitic nematode Index were used to
compare the two rehabilitation techniques in terms of nematodes as indicators. Microarthropods
family structures were used to compare the two amendments in terms of
diversity and abundance. Enzymatic activity was positively affected by the addition of
woodchip-vermicompost, than in the sites treated with inorganic fertilisers. The
microbial community structure showed no statistically significant (p < 0.05)
differences between the two amendments. A higher abundance of nematodes
especially plant-parasitic nematodes and bacterivorous nematodes were observed in the woodchip-vermicompost sites than in the inorganic fertilised sites. According to
the Maturity Index, both amendments became more enriched during the study period,
while the Plant-Parasitic nematode Index showed that the carrying capacity for plantparasitic
nematodes on the woodchip-vermicompost sites increased while it decreased
in the inorganic fertilised sites, which can be related to the decrease in vegetation
cover on the inorganic fertilised sites. Both coloniser (Prostigmata) and persister
(Cryptostigmata and Mesostigmata) groups of the micro-arthropods, as well as a
higher diversity of micro-arthropods, were present on the woodchip-vermicompost
sites whereas the inorganic fertilised sites showed only the presence of colonisers,
with a decrease in diversity and abundance of micro-arthropods over the study. The
colonisation of micro-arthropods may have been affected by the addition of
woodchip-vermicompost and vegetation cover, which contribute to the establishment
of suitable microhabitats for these soil biota. By intercorrelating the results, it may be
concluded that the addition of woodchip-vermicompost may be an essential part of the
rehabilitation process, by contributing to soil organic material to the ecosystem
system, which may improve the recolonisation of soil biota and ecosystem processes.
However further studies need to be conducted in order to determine the long-term
sustainability of the woodchip-vermicompost in providing organic material and
sustaining the ecosystem processes. The study also showed the necessity to integrate
various ecosystem components when evaluating ecosystem development due to the
unique role each component plays and the impact it may have on other components. / Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2006.
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Impact de l'utilisation d'un compost vert sur l'activité et la diversité de la microflore telluriqueDekaki, Anouar 16 December 2008 (has links)
Le compostage est une technique de valorisation des déchets organiques en un produit stable et riche en matières humiques. Certains composts « verts » se sont révélés être de 2 à 3 fois plus efficaces sur la croissance des plantes que les composts classiques. Notre étude réalisée sur un compost fabriqué à partir de déchets végétaux a permis de suivre l’évolution de la densité et de la diversité de la microflore (bactéries, champignons) au cours du processus de maturation puis de tester l’impact de ce compost sur la diversité et l’activité de la microflore tellurique. Cette analyse a été effectuée par des techniques complémentaires : biochimiques (dosages enzymatiques), microbiologiques (cultures in vitro) et de biologie moléculaire (PCR-DGGE, Séquençage). Les résultats montrent qu’au cours de sa maturation, le compost étudié présente une baisse significative de son taux d’humidité et une augmentation sensible de son pH. Sa microflore subit une complète restructuration avec apparition de souches bactériennes susceptibles de dégrader des composés polluants comme les plastiques, les pesticides et les hydrocarbures. L’ajout de ce compost à deux types de sol présentant des propriétés physico-chimiques différentes, n’a pas montré de modifications importantes et durables de la diversité microbienne et fonctionnelle de celui-ci. Les causes de l’effet remarquable de ce compost sur la croissance végétale sont discutées. / Composting is a technique of transformation organic waste in a stable product rich in organic materials. Some "green" compost proved to be from 2 to 3 times more benefit on the growth of the plants than traditional composts. The main of this study is to follow the evolution of density and diversity of the microflora (bacteria, fungi) during the process of maturation of green compost manufactured from vegetable wastes, and to investigate the impact of this compost on the diversity and the activity of the telluric microflora. This analysis was carried out by complementary techniques: biochemical (enzymatic activity), microbiological (in vitro cultures) and molecular biology (PCR-DGGE, DNA sequencing). The results show that during its maturation, the studied compost presents a significant decrease of its water content and an appreciable increase in its pH. The microflora undergoes a complete reorganization with appearance of bacterial strain suitable for degrade polluting compounds like the plastics, the pesticides and hydrocarbons. The addition of this compost with two types of soil presenting of the different physicochemical properties, did not show significant and durable modifications of the microbial and functional diversity of this one. The causes of the remarkable effect of this compost on the vegetable growth are discussed.
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Indicadores microbiológicos e químicos do solo sob diferentes sistemas de manejo agropecuário /Garcia, Martha Regina Lucizano. January 2007 (has links)
Orientador: Ely Nahas / Banca: Fábio Martins Mercante / Banca: Maria José Valarini / Banca: Antônio Carlos Monteiro / Banca: Roberto Alves de Oliveira / Resumo: Diversos estudos vêm buscando avaliar os diferentes impactos causados pelas várias formas de manejo do solo, procurando-se aliar a produção das culturas à preservação dos recursos naturais. O presente estudo objetivou avaliar a influência temporal (das estações do ano), do sistema de manejo do gado e das culturas nas características microbiológicas e químicas do solo em diferentes profundidades. O primeiro experimento foi conduzido no setor de bovinocultura da Estação Experimental da Faculdade de Ciências Agrárias e Veterinária, UNESP - Campus de Jaboticabal, localizada na região norte do Estado de São Paulo. Os sistemas de pastejo avaliados foram: pasto rotacionado, pasto contínuo e pasto controle (sem a presença de gado). As coletas das amostras de solo foram realizadas nas quatro estações do ano, no período de um ano (2005 - 2006). Cada amostra foi composta de 6 amostras simples, retiradas nas profundidades de 0 - 0,05, 0,05 - 0,10 e 0,10 - 0,20 m. O segundo experimento foi realizado nas fazendas Barracão e São João, no município de Guaíra, localizadas na região paulista da Alta Mogiana, Estado de São Paulo. Foram estudados quatro sistemas de manejo das culturas: plantio direto (PD), plantio direto integrado com pastagem (PD-I), cultivo convencional (CC) e Mata Natural (Mata). A coleta das amostras de solo foi feita somente na estação do verão, durante o período de janeiro e fevereiro do ano de 2006. Cada amostra foi composta de 10 amostras simples, retiradas nas profundidades de 0 - 0,10, 0,10 - 0,20 e 0,20 - 0,30 m. As variáveis microbiológicas, bioquímicas e químicas do solo avaliadas podem ser consideradas medidas sensíveis para determinar mudanças decorrentes dos efeitos das estações do ano, dos sistemas de manejo do gado e das culturas e da profundidade do solo. / Abstract: Several, studies come searching evaluated the different impacts caused by some forms of management of soil, to try to get the production of the cultures to the preservation of the natural resources. The objective of this study was determine the influence of the seasonal, of the cattle management system and crop and the depth of the soil in the microbiological and chemical variable of the soil. The first experiment was lead in the sector of cattle breeding of the Experimental Station of the College of Agrarian Sciences and Veterinary medicine, UNESP - Campus of Jaboticabal, located in the north of São Paulo State. The evaluated systems of pasture were: rotation pasture, continuous pasture and controlled pasture. The collections of soil samples were carried in four-year seasons during the period of one year (2005 - 2006). Each sample was composed of 6 simple samples, withdrawals with hoe, in the depths of 0 - 0,05, 0,05 - 0,10 and 0,10 - 0,20 m. The second experiment was carried in the Barracão and São João farms, in the Guaíra Cite, located in the region of the Alta Mogiana, in São Paulo State. In view of evaluation of the effect of the management system crop, four systems were studied: no-tillage (NT), pasture and crop integrated systems (PD-I), conventional tillage (CC) and forest. The collection of soil samples was made in summer, during the period of January and February in 2006. Each sample was composed of 10 simple samples withdrawals with hoe, in the depths of 0 - 0,10, 0,10 - 0,20 and 0,20 - 0,30 m. The evaluated microbiological, biochemical and chemical variables of soil can be considered sensitive measurements to determinate changes due to the effects of year seasons, the cattle management system and crop and the depth of the soil. / Doutor
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Avaliação comparativa de diferentes métodos de quantificação de formas teciduais de Trypanosoma cruzi na infecção experimental / Comparative evaluation of differents methodologies in tecidual types quantification of trypanosoma cruziAugusto, Mariana Bryan 02 August 2010 (has links)
Completando 100 anos da descoberta da molestia, a Doenca de Chagas, causada pelo agente etiologico Trypanosoma cruzi, ainda e considerada nas Americas Central e do Sul um problema de saude publica, atingindo mais de 8 milhoes de individuos. Sua transmissao ocorre de distintas maneiras, como por vetores, transfusao sanguinea, transplante de orgaos, acidentes em laboratorios, via oral. O parasita e conhecido por sua heterogeneidade no genotipo e fenotipo, baseado em mutacoes cumulativas em diferentes sub-populacoes do parasita. A cepa CL Brener e uma cepa padrao para pesquisa com diversas peculiaridades interessantes como baixa infectividade em animais, resposta aos tratamentos existentes e seus marcadores geneticos serem estaveis. O clone CL B5 de T. cruzi e originado a partir da cepa CL Brener modificada geneticamente. A cepa possui um gene reporter, o LacZ de Escherichia coli que sintetiza uma enzima, a À-galactose que pode catalisar uma reacao colorimetrica com o substrato vermelho de clorofenol À-galactopiranosideo (CPRG). A atividade enzimatica e diretamente proporcional ao numero de parasita. No presente trabalho propomos a introducao de uma tecnica colorimetrica para caracterizacao de parametros para quantificacao do parasitismo tecidual e comparacao com a tecnica histologica e Real Time-PCR. Camundongos Balb/C machos foram divididos em grupos controle (nao infectado) e experimental variando a via de inoculacao, a saber, intraperitoneal (IP), subcutanea (SC) e oral (OR). As curvas parasitemicas foram realizadas por meio da coleta de sangue da cauda do animal. Apos morte dos mesmos, foram estudados os tecidos cardiaco, hepatico, esplenico, enterico e muscular (musculo esqueletico). Os orgaos foram retirados, divididos em tres porcoes equitativas para analise histologica, enzimatica e molecular. Na atividade enzimatica, comparando os tecidos em suas vias de inoculacao, o figado apresentou resultado significativo no grupo inoculado via IP. Tecido cardiaco apresentou diferenca estatistica (P<0,05) entre as vias de inoculacao OR e SC, apresentando maior atividade enzimatica na primeira, o que esta relacionado a uma maior quantidade de parasita tecidual. Nos cortes histologicos o grupo SC mostrou-se mais infectado com grande numero de ninhos de forma amastigota em praticamente todos os tecidos, enquanto nos demais grupos foram observados ninhos de amastigota em sua maioria no coracao e viii musculo esqueletico. A presenca do parasita nos demais tecidos foi confirmada pela eletroforese em gel de agarose 1,5% apos tecnica de PCR convencional. A Real Time-PCR nao se mostrou satisfatoria no trabalho devido a dificuldade na sua padronizacao. Concluindo, a metodologia enzimatica se mostrou favoravel e adequada na quantificacao parasitaria tecidual, podendo ser aperfeicoada para corroborar sua eficacia. / One hundred years after the discovery of Chagas Disease, an illness caused by blood born protozoan Trypanosoma cruzi, is still consider a public health problem. It is a major cause of morbidity and mortality in Central and South Americas where more than 8 million individuals are already infected. This illness can be contracted by different forms as vector transmission, blood transfusion, tissue transplant, laboratory accidents and oral route. The parasite has a heterogeneous genotype and phenotype due to accumulating mutation in their sub-populations. CL Brener is considered as a pattern strain due to some intrinsic characteristics as low infectivity, good response to therapy drugs and a stable genetic heritage. The CL B5 clone of T. cruzi was originated by a genetic modification of the CL Brener strain which has a reporter gene LacZ of Escherichia coli, which induces the syntesis of â-galactosidase which is able to catalyze a colorimetric reaction using the substrat Chlorophenol red-â-D-galactopyranoside (CPRG). The enzymatic activity is directly related with the number of the parasites. In this study we proposed a new colorimetric assay to quantify T. cruzi load in different animal tissues by comparing with other methodologies such histological and molecular assay (Real Time-PCR). Male Balb/C mice were separated into control and experimental groups according to the route of infection (IP-intraperitoneal, SC-subcutaneous, OR-oral). Parasitemic curves were made by collecting blood samples from the tail of experimental animals. Fifteen days after, animals were euthanized and their tissues removed (heart, liver, spleen, intestine and skeletal muscles) and divided in three portions for the colorimetric analyses, histopathology and molecular assay. The enzymatic activity was performed comparing the number of parasites in different tissues according to the route of infection. Comparing them, it was observed that heart and liver displayed the highest number of parasites as compared with the other studied tissues. In the liver, IP route triggered the highest number of parasites while in the heart OR and SC routes displayed enhanced parasites. The histopathology analysis revealed that SC group presented the highest number of amastigote nests. For IP and OR groups, nests were mostly observed in heart and skeletal muscles. T. cruzi DNA were detected using electrophoreses 1,5 % agarose gel after conventional PCR technique, displaying characteristic bands of DNA. The Real Time-PCR was not a satisfactory x assay for this study due to difficulties on its patronization. To conclude, the enzymatic methodology was advantageous and appropriated on quantifying tissue parasites. Some further experiments will be needed to improve methodology and its efficacy.
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Sistema antioxidante de cana-de-açúcar em resposta à seca / Antioxidant system of sugarcane to drought responseCamargo, Isabela Amaral de 05 August 2013 (has links)
Cultivares de cana-de-açúcar com tolerância à seca e alta produtividade são desejáveis para a rápida expansão do cultivo da cana-de-açúcar em regiões caracterizadas por um período de déficit hídrico prolongado. Nas plantas, o estresse hídrico induz o dano oxidativo devido ao aumento da produção de espécies reativas de oxigênio (ROS) sem o seu consequente controle. A tolerância à seca pode incluir mecanismos distintos que permitam que as plantas mantenham o metabolismo em níveis normais, exigindo um sistema antioxidante robusto para inativar ROS, o qual consiste de vias enzimáticas, incluindo superóxido dismutase (SOD), catalase (CAT), ascorbato peroxidase (APX), glutationa redutase (GR), glutationa-S-transferase (GST) e glutationa peroxidase (GPX). A elucidação dos mecanismos bioquímicos e moleculares adotados pela cana-de-açúcar em resposta à deficiência hídrica seria relevante para o desenvolvimento de genótipos tolerantes, contribuindo para uma redução na necessidade de água para irrigação. Duas cultivares selecionadas com base em seu comportamento contrastante na resposta à seca foram avaliadas, \'IACSP94-2094\' considerada tolerante e \'IACSP97-7065\' considerada sensível à seca. O déficit hídrico foi imposto via suspensão da irrigação 4 meses após o plantio em casa de vegetação. A desidratação indicada tanto pelo potencial hídrico da folha quanto pelo conteúdo relativo de água (CRA) corrobora com a redução na assimilação de CO2 (A), condutância estomática (gS), transpiração (E) em ambas as cultivares. Também foram detectadas, quedas no redimento quântico da fotoquímica do fotossistema II, que comprovam o estabelecimento do déficit hídrico para a cultivar \'IACSP94-2094\', corroborado pelas análises de peroxidação lipídica, indicadora da ocorrência do estresse oxidativo. Com relação à atividade específica e expressão gênica das enzimas do sistema antioxidante, foi demonstrado que a superóxido dismutase (SOD), ascorbato peroxidase (APX) e glutationa redutase (GR) são as enzimas chave para o controle do estresse oxidativo desencadeado pela seca na cultivar tolerante. O perfil das isoformas da SOD revelou que a cultivar tolerante (\'IACSP94-2094\') possui um maior número de isoformas do que a cultivar sensível (\'IACSP97-7065\'), sendo a Fe-SOD a mais representativa. Teores de prolina e compostos fenólicos em folhas não mostraram relação direta com resposta ao déficit hídrico para a \'IACSP94-2094\', e somente sob máximo déficit hídrico a \'IACSP97-7065\' apresentou incrementos no teor de fenólicos, sugerindo que as cultivares podem adotar mecanismos alternativos para superar os efeitos prejudiciais da seca / Sugarcane cultivars with drought tolerance and high yield are highly desirable to the rapid expansion of sugarcane plantings to regions characterized by a period of prolonged water deficit. Water deficit stress in plants induces oxidative damage due to the increase of the production of reactive oxygen species (ROS) without their subsequent control. Drought tolerance may derive from distinct mechanisms that allow plants to maintain metabolism at normal levels, requiring a robust antioxidant system to inactivate ROS, which consists of enzymatic pathways including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), glutathione S-transferase (GST) and glutathione peroxidase (GPX). The elucidation of the biochemical and molecular mechanisms adopted by sugarcane in response to water deficit would be relevant to the development of tolerant cultivars, contributing to the reduction in the requirement for irrigation. Two cultivars selected based on their contrasting behavior in response to drought were evaluated, \'IACSP94-2094\' selected as tolerant, and \'IACSP97-7065\' as more sensitive to drought. Water deficit was imposed by withholding water in plants 4 months after planting in the greenhouse. Water stress was indicated by both leaf water potential and the leaf relative water content (CRA), together by the reduction in CO2 assimilation (A), stomatal conductance (gS), transpiration (E) in both cultivars. A decrease in quantum yield of photochemistry from photosystem II corroborated the occurrence of water deficit for \'IACSP94-2094\', together with results from lipid peroxidation analysis, indicator of oxidative stress. In relation of the specific activity and gene expression of antioxidant enzymes system, it was demonstrated that superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) were the key enzymes controlling oxidative stress triggered by drought in the tolerant cultivar. SOD isoform activity profiles showed that the tolerant cultivar (\'IACSP94-2094\') displayed more isoforms than the sensitive cultivar (\'IACSP97-7065\'), with the Fe-SOD being the most representative. Proline and phenolic compound contents in leaves did not show direct relationship with response to drought for \'IACSP94-2094\', and only under maximum water deficit \'IACSP97-7065\' presented increments in the phenolics content, suggesting that cultivars can adopt alternative mechanisms to overcome the harmful effects of dry
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