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We assessed the effects of pharmacological spinal inhibition of voltage-gated calcium channels (VGCC) in mouse pruritus. The epidural administration of P/Q-type MVIIC or PhTx3.3, L-type verapamil, T-type NNC 55-0396 or R-type SNX-482 VGCC blockers failed to alter the scratching behavior caused by the PAR-2 activator trypsin, injected into the mouse nape skin. Otherwise, trypsin-elicited pruritus was markedly reduced by the spinal administration of preferential N-type VGCC inhibitors MVIIA and Phα1β. C. magus-obtained toxin MVIIA displayed significant effects when dosed from 1 to 4 h before trypsin, whereas the effects of P. nigriventer-derived Phα1β remained for up to 12 h. MVIIA or Phα1β also prevented the itching elicited by intradermal (i.d.) injection of SLIGRL-NH2, compound 48/80 or chloroquine, although they did not affect H2O2-induced itching. Furthermore, the co-administration of MVIIA or Phα1β markedly inhibited the pruritus caused by the spinal injection of gastrin-releasing peptide (GRP), but not morphine. Notably, spinal MVIIA or Phα1β greatly prevented chronic pruritus allied to dry skin. However, either toxin failed to alter the edema formation or neutrophil influx caused by trypsin. In addition, epidural MVIIA or Phα1β did not modify the expression of GRP receptor (GRP-R) in the spinal cord, whilst they brought c-Fos activation to control levels. Finally, the in vitro incubation of MVIIA or Phα1β prevented the calcium influx evoked by the synthetic PAR-2 agonist AC264613 in spinal cord synaptosomes. Data brings novel evidence on itching transmission mechanisms, pointing out the therapeutic relevance of N-type VGCC inhibitors to control refractory pruritus.

Foram avaliados os efeitos da inibi??o farmacol?gica dos canais de c?lcio voltagem-dependentes (CCVD) no prurido em camundongos. A administra??o intratecal (i.t.) dos bloqueadores CCVD do subtipo P/Q (MVIIA ou PhTx3.3), subtipo L (verapamil), subtipo T (NNC 55-0396) ou do subtipo R (SNX-482) n?o alterou o comportamento de co?ar induzido pela tripsina (agonista do receptor PAR-2), aplicada na regi?o dorsal do pesco?o de camundongos. Entretanto, o comportamento de co?ar induzido pela tripsina foi significativamente diminu?do pela administra??o i.t. dos bloqueadores de CCVD do subtipo N (MVIIA e Phα1β). A toxina MVIIA, derivada do C. magus, apresentou efeitos significativos quando administrada entre 1 a 4 horas antes da aplica??o de tripsina. Por outro lado, a toxina Phα1β derivada do veneno da aranha P. nigriventer demonstrou atividade, quando injetada at? 12 h antes da tripsina. O pr?-tratamento com MVIIA ou Phα1β foi efetivo em inibir o comportamento de co?ar induzido pela aplica??o intrad?rmica (i.d.) de SLIGRL-NH2, composto 48/80 ou cloroquina; entretanto, as…

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CAPES

The aim that this thesis was to evaluate the influence of differential nitrogen supply on metabolism, yield and quality of essential oil of Ocimum basilicum (basil). Two experiments were conducted in a greenhouse, in the first variety Sweet Italian Large Leaf subjected to contrasting doses of nitrate-N (50 and 300 kg N ha-1). In the second, the varieties Sweet Italian Large Leaf and Maria Bonita were cultured with doses of 50 or 100 kg N in three sources (nitrate, ammonium and ammonium nitrate). Analyses after harvesting of plants were: nitrate reductase activity, nitrate-N, ammonium-N, amino-N, total-N and soluble sugars. The essential oils from basil were extracted with steam distillation and essential oil components were analyzed using gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectroscopy (GC-MS) and 1HNMR. After the characterization of the essential oil tests were conducted to evaluate the allelopathic activity and antifungal against the fungi Rhizoctonia solani, Fusarium oxysporum, Geothricum spp, Candida albicans, C. guilliermondii, C. parapsilosis, C. tropicalis, C. krusei. The effect of oil on the morphology of fungi was performed by scanning electron microscopy. Gene expression was evaluated by real-time qPCR of 3-hydroxy 3-methyl glutaryl CoA reductase enzyme, 1-Deoxy-D-xylulose- 5-phosphate synthase, linalool synthase, phenylalanine ammonia lyase and eugenol synthase involved in the biosynthesis of essential oil. And anatomical analyzes evaluated the secretory trichomes. The supply of 50 Kg nitrate-N.ha-1 increased the yield (1.14%) and concentration of linalool (28.04%) in Sweet Italian Large Leaf. The sugars can be a strategy to ensure the availability of assimilates to mevalonate, methyl erythritol phosphate and shikimate pathways to produce secondary metabolites. At concentrations of 15 mg.mL-1 basil essential oil inhibited the germination of lettuce seeds. The basil essential oil showed fungistatic and according to the ultrastructural observations it is possible that its mechanism of action is by interaction with components of the cell membrane of fungi. The supply of 100 kg ha-1 of ammonium nitrate resulted in higher (1.46%) and linalool content (69.25%) in the essential oil of Maria Bonita variety, and Sweet Italian Large Leaf variety seems to be adapted conditions of nutrient scarcity with oil yield of 0.37% and 38.9% linalool. The differential N supply induced the expression of genes encoding key enzymes in the production of essential oil, and nitrogen is modulator of gene expression and thus the production of secondary…

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Gliomas are the most common and devastating tumors of the central nervous system (CNS). Among the gliomas group, the GBM is the most prevalent, aggressive and deadly malignant. Many pieces of evidence point out the relevance of natural compounds for cancer therapy and prevention, including chalcones. Chalcones are group of natural precursors of flavonoid and display a wide variety of biological and pharmacological proprieties that include anti-proliferative and anti-cancer activities. This study aimed at evaluating the in vitro anti-proliferative activity and cell viability inhibition of nine quinoxaline derived chalcones, structurally based on the selective PI3Ky inhibitor AS605240. These synthetic compounds were tested at different time-periods of incubation (24, 48 e 72 h) and concentrations (0,1; 0,1; 1; 5 e 10 μg/mL) in glioma cell lines from human and rat origin (U-138 MG and C6, respectively). The results showed by MTT assay and cell couting revealed that four chalcones (compounds N2, N9, N10 and N12), displayed higher efficacies and potencies, being able to inhibit either cell proliferation or viability, in a time- and concentration dependent manner. These four compounds which present methoxy groups at A-ring and their efficacy was greater than that seen for the positive control compound AS605240. Flow cytometry analysis demonstrated that incubation of C6 cells with chalcone N9 led to G1 phase arrest, likely indicating an interference with apoptosis. Furthermore, chalcone N9 was able to visibly inhibit AKT activation, allied to the stimulation of ERK 1/2 MAP-kinase. The chalcones tested herein, especially those displaying a methoxy substituent at A-ring, might well represent promising molecules for the treatment of gliomas.

Os gliomas s?o os tumores do SNC mais comuns e devastadores. Dentre os gliomas, o GBM ? o mais prevalente, agressivo, maligno e apresenta um mau progn?stico. A relev?ncia dos compostos naturais, incluindo as chalconas, no tratamento e na preven??o do c?ncer est? sendo muito evidenciada. As chalconas formam um grupo de compostos naturais derivados dos flavon?ides que apresentam diferentes propriedades biol?gicas e farmacol?gicas, incluindo as atividades anti-proliferativas e anti-tumorais. O objetivo deste estudo foi avaliar a a??o anti-proliferativa e a capacidade de inibi??o da viabilidade celular in vitro de nove chalconas derivadas da quinoxalina, baseadas estruturalmente no inibidor seletivo de PI3Ky, o AS605240. Estes compostos sint?ticos foram testados em diferentes tempos de incuba??o (24, 48 e 72 h) e concentra??es (0,1; 0,1; 1; 5 e 10 μg/mL) em linhagens de glioma humano e de rato (U-138 MG e C6, respectivamente). Os resultados observados nos experimentos de MTT e na contagem celular revelaram que quatros chalconas (compostos N2, N9, N10 e N12), apresentaram grande efic?cia e…

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS ([email protected]) on 2016-05-17T11:43:35Z No. of bitstreams: 1 DIS_FERNANDA_FERNANDES_CRUZ_COMPLETO.pdf: 1282066 bytes, checksum: 5b0d649de7173c518fd09040b6012a0b (MD5)

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Zebrafish (Danio rerio) is a bony fish originated in Pakistan and India with anatomical and physiological characteristics and its genome is homologous to the more developed organisms, which suggests that many genes involved in human diseases can be coincident to the zebrafish genome. Copper is a heavy metal involved in formation of ROS, responsible for the lipid peroxidation in the membrane, direct protein oxidation and cleavage of DNA and RNA molecules. Purines (ATP, ADP and adenosine) and pyrimidine (UTP and UDP) can act as extracellular signaling molecules, exerting different physiological effects when binding to purninergic receptors. There are two families of purinergic receptors: adenosine receptors or P1 (P1R), and P2 receptor (P2R). The P1R are a family of G protein-coupled receptors and are divided inA1, A2A, A2B, and A3 subtypes. Adenosine is the major ligand for P1R, and is involved in important physiological functions such as modulation of neurotransmitter release, synaptic plasticity, neuroprotection to ischemic events, hypoxia and oxidative stress. Studies suggest that drugs that interfere in adenosine signaling may weaken the protective mechanisms and lead to an exacerbation of the inflammatory response. Caffeine is the most widely consumed psychoactive substance in the world and is a nonselective antagonist of adenosine receptors. This study aimed to evaluate the effect of caffeine in copper-induced inflammation in zebrafish larvae. The animals were treated with copper at a concentration of 10 ?M and three different concentrations of caffeine (100 ?M, 500 ?M e 1 mM) to determine the survival rate and to define timepoints and concentrations for the following experiments. It was observed that high concentrations of caffeine combined with copper caused a significant decrease of larvae survival after 24 h of treatment. For the following experiments, we chose 4 and 24 h of exposure, 500 ?M caffeine and 10 ?M copper concentrations. After 4 h of treatment, caffeine was able to change the behavior parameters. It was shown that both copper and caffeine might have an anxiogenic effect on zebrafish larvae after 24 h exposure. For the evaluation of inflammatory profile, inflammation markers PGE2, COX-2, TNF, IL-6 and IL-10 were chosen. In 4 hours of exposure, PGE2 levels are increased in groups treated with copper and caffeine. Caffeine showed an additive effect on the PGE2 release. The gene expression of TNF, as well as COX-2 was increased by copper treatment, however, in treatments with caffeine there was a reduction of COX-2 gene expression. In 24 h of treatment,…

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Superparamagnetic iron oxide nanoparticles (SPIONs) are of great interest in nanomedicine due to their capability to act simultaneously as a contrast agent in magnetic resonance imaging and as a targeted drug delivery system with good biocompatibility. At present, one of the biggest concerns about the use of SPIONs remains around its toxicity. For this reason, it is important to establish the safe upper limit for each use. In the present study, SPION coated with cross-linked and aminated dextran (CLIO-NH2) were synthesized by the co-precipitation method and characterized by transmission electron microscopy (TEM) and light scattering dynamics. The analysis of the elementary composition of the solution at the end of the synthesis was also performed. The magnetite core size was 5.5 ? 1.4 nm, on a concentration of 10 mg Fe-NPs/ml. We have evaluated the effects of different CLIO-NH2 doses (20, 50, 100, 140 and 200 mg/kg) as a function of time after exposure (one, 16, 24 and 48 hours) on AChE activity and ache expression in zebrafish brain. At tested concentrations, only the animals exposed to 200 mg/kg, and assessed 24 h after administration of the nanoparticles, have shown decreased AChE activity. These values returned to control levels after 48 h of exposition, indicating a transitory toxical effect. The RT-qPCR results suggested that inhibition of brain AChE is not directly related with the transcriptional control, and it was probably due to post-transcriptional events. Once ACh is recognized to play an important role in the regulation of locomotor control, we further evaluated parameters of zebrafish swimming activity. CLIO-NH2 at 200mg/kg, evaluated after 24 h, also impaired all the tested parameters of zebrafish swimming activity, i.e. decreased traveled distance, mean speed, number of line crossings, and turn angle. We further investigated the iron accumulation in zebrafish brain by ICP-MS, and a significant higher level of ferric iron was found in zebrafish brains exposed to CLIO-NH2. In summary, the results presented herein provide further experimental evidence that exposure to high doses of dextran-coated iron oxide nanoparticles can be transiently neurotoxic.

Nanopart?culas superparamagn?ticas de ?xido de ferro (SPIONs) s?o de grande interesse na nanomedicina, devido ? sua capacidade para agir, simultaneamente, como um agente de contraste em imagem por resson?ncia magn?tica e como um sistema de entrega de f?rmacos espec?ficos, possuindo boa biocompatibilidade. Atualmente, uma das maiores preocupa??es com a utiliza??o de SPIONs permanece em torno da sua toxicidade e, por esta raz?o, ? importante estabelecer um limite de seguran?a para sua utiliza??o. Neste estudo, SPIONs revestidas com dextran-aminado (CLIO-NH2) foram sintetizados pelo m?todo de co-precipita??o e caracterizadas por microscopia eletr?nica de transmiss?o…

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Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM)

Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES)

O objetivo deste trabalho foi avaliar os efeitos da dieta de cafeteria desde a lacta??o nos par?metros nutricionais e na a??o da risperidona nos comportamentos de ansiedade, locomo??o, mem?ria e intera??o social. Foram utilizadas 14 ninhadas de ratos da linhagem Wistar (Rattus novergicus) alojados em gaiolas individuais, sob condi??es padr?o. Os ratos machos de cada ninhada formaram, da lacta??o at? a fase adulta, os grupos: Controle (CTRL) ? receberam ra??o padr?o e ?gua ad libitum (n = 42); Cafeteria (CAF) ? receberam dieta de cafeteria e ?gua ad libitum (n = 42). Entre o 114? e o 119? dia de vida, os animais foram subdividos (n = 21) para receberem o tratamento com salina (CTRL-S e CAF-S) ou risperidona (CTRL-R e CAF-R). Nesse per?odo foram realizados os testes comportamentais do labirinto em cruz elevado, campo aberto, teste de reconhecimento de objetos e intera??o social. No 119? os animais foram colocados em jejum, para que, no 120? dia fossem anestesiados e eutanasiados por exsanguina??o. Foram avaliados: consumo de ra??o, ingest?o cal?rica, peso corporal, ganho de peso, coeficiente de efici?ncia alimentar (CEA), comprimento naso-anal e ?ndice de massa corporal (IMC); peso dos ?rg?os e do tecido adiposo abdominal; teores de colesterol total e fra??es, triacilglicerol e glicemia do soro; teor de lip?dios, colesterol total e triacilglicerol do f?gado e das fezes; estado redox do f?gado e de diferentes por??es do enc?falo (c?rtex pr?-frontal e hipocampo). Foi utilizada an?lide de vari?ncia (ANOVA) seguida de Newman Keuls quando necess?rio (P<0,05). O grupo CAF demonstrou menor ingest?o alimentar e maior ingest?o cal?rica, devido ? densidade energ?tica da dieta. O maior consumo cal?rico se configurou em maior ganho de peso, CEA, IMC e ac?mulo de tecido adiposo abdominal. O grupo CAF tamb?m obteve eleva??o dos n?veis de triacilglicerol plasm?ticoa, al?m de lip?dios e triacilglicerol hep?tico, que foram classificados em esteatose hep?tica. Ao mesmo tempo, houve uma rela??o ruim entre as fra??es do colesterol…

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CAPES

FAPERJ

The use of supercritical fluid extraction of natural products has been shown to be a viable and promising alternative to conventional methodologies. Due to the characteristics of supercritical extraction, which allow to obtain better quality products with no chemical waste, this extraction technique has been widely used in the obtaining of bioactive molecules from microalgae, as antioxidants, fibers, phytosterols, nutrients, among others. This work aimed to study the technical feasibility of supercritical extraction of oil from the microalgae Desmodesmus sp. using carbon dioxide as solvent. Conventional method, using hexane as solvent, was used in order to compare the extraction of oils from microalgal biomass study and the results show the efficiency and selectivity of supercritical fluid extraction in respect to the fatty acids concentration. The best yields of oil occurred at the operational conditions of 300 bar and 100 ?C, 500 bar and 70 ?C and 440 bar and 90 ?C, reaching values of 3.342%, 3.103% and 2.586%, respectively. The residual biomass after supercritical fluid extraction was assessed in order to evaluate the residual potential and the results indicated that it is a promising one, due to its high concentration in carbohydrates and proteins. The analysis by gas chromatography indicated the myristic acid (C14:0), palmitic (C16:0), stearic (C18:0), oleic (C18:1) and ?-linolenic (C18:3) as the main fatty acids present in the Desmodesmus sp. oil. Simple models were used to model the extraction curves, Esqu?vel et al. (1999), Sesti & Reverchon Osseo (1994) and Zekovi? (2003) and all of them were able to represent supercritical extraction of oil from microalgae Desmodesmus sp.

A utiliza??o de fluidos supercr?ticos na extra??o de produtos naturais tem se mostrado uma alternativa vi?vel e promissora em rela??es aos m?todos convencionais. Devido ?s caracter?sticas da extra??o supercr?tica, que permitem a obten??o de produtos de melhor qualidade e sem res?duos qu?micos, por isso essa t?cnica de extra??o vem sendo muito utilizada na obten??o de mol?culas bioativas a partir de microalgas, que se destaca por apresentar antioxidantes, fibras, fitoester?is, nutrientes, entre outros. O presente trabalho teve como objetivo estudar a viabilidade t?cnica da extra??o supercr?tica do ?leo da microalga Desmodesmus sp., empregando-se di?xido de carbono como solvente. Foi utilizado m?todo convencional, soxhlet com hexano como solvente, a fim de comparar com a extra??o supercr?tica e os resultados mostraram a efici?ncia e seletividade da extra??o supercr?tica quanto ? concentra??o de ?cidos graxos.…

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES

Adenylosuccinate lyase (ASL) belongs to aspartase/fumarase superfamily of enzymes which share a general acid-base catalytic mechanism with ?-elimination of fumarate as common product. ASL is involved in both de novo and salvage pathways of purine biosynthesis. Cloning, expression, and a method to obtain homogeneous recombinant ASL from Leishmania braziliensis (LbASL) are described. Mass spectrometry analysis of recombinant LbASL, oligomeric state determination and multiple sequence alignment are presented. Steady-state kinetics of LbASL showed a Michaelis-Menten pattern. Isothermal titration calorimetry binding assays suggested that LbASL follows a Uni-Bi ordered kinetic mechanism, in which release of fumarate is followed by AMP to yield free enzyme. Initial velocity data for the reverse reaction and the Haldane relationship allowed calculation of an unfavorable equilibrium constant for LbASL-catalyzed chemical reaction. The activation energy and thermodynamic activation parameters were estimated. Solvent kinetic isotope effects V/K and V suggest a modest contribution of solvent proton transference during the rate-limiting step of the reaction. Proton inventory data show that the modest normal effect on V arises from a single protonic site, and the transition state fractionation factor value of 0.74 suggests participation of solvent proton transfer in transition-state vibrations perpendicular to the reaction coordinate. pH-rate profiles for kcat and kcat/KM suggested amino acid residues involved in, respectively, catalysis and substrate binding. A model of LbASL was built to provide a structural basis for the experimental data. A better understanding of the mode of action of LbASL is useful for the rational design of antileishmaniasis agents.

A enzima Adenilosuccinato liase (ASL) pertence a superfam?lia de enzimas aspartase/fumarase, as quais compartilham o mecanismo catal?tico…

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Tuberculosis re-emerged in the mid 80s and currently about two million people die each year due to this disease. The resurgence of tuberculosis has become a threat to public health. The high susceptibility of HIV-infected patients and the proliferation of multi-drug resistant strains have created the need to develop new therapies. However, no new class of drugs against tuberculosis was developed in the last 45 years. Thus, it becomes imminent need to develop new antituberculosis agents. As purine metabolism could be implicated in mycobacterial latency the purine nucleoside phosphorylase (EC 2.4.2.1) has been proposed as target for development of antibacterial drugs. PNP catalyzes the reversible cleavage in the presence of inorganic phosphate of N-ribosidic bonds of the purine nucleosides and deoxynucleosides, except adenosine. This reaction generates the purine base and ribose (deoxyribose)-1-phosphate. PNP is specific for purine nucleosides in the β-configuration and cleaves the glycosidic bond with inversion of configuration to produce α-ribose-1-phosphate. In this work PNP was crystallized in association with inosine, hypoxanthine and acyclovir, substrate, product and inhibitor, respectively, and data were collected using synchrotron radiation. In addition, molecular dynamics simulations and isothermal titration calorimetry experiments were used to provide detailed information on the dynamic properties and to investigate the profile and thermodynamic affinities of MtPNP associated with these molecules. With obtained results we hope to contribute to the search of new selective inhibitors for MtPNP, since differences between the mycobacterial and human enzyme binding sites have been also identified, making structure-based drug design feasible.

A tuberculose ressurgiu na metade dos anos 80 e, atualmente aproximadamente dois milh?es de pessoas morrem por ano devido a esta doen?a. O ressurgimento da tuberculose tornou-se uma amea?a ? sa?de p?blica. A alta susceptibilidade dos pacientes infectados com HIV e a prolifera??o de cepas resistentes a m?ltiplas drogas t?m criado a necessidade de se desenvolver novas terapias. No entanto, nenhuma nova classe de drogas contra a tuberculose foi desenvolvida nos ?ltimos 45 anos. Deste modo, torna-se iminente a necessidade de se desenvolver novos agentes antituberculose. Como o metabolismo de purinas pode estar implicado na lat?ncia da micobact?ria, a purina nucleos?deo fosforilase de Mycobacterium tuberculosis (MtPNP) (EC 2.4.2.1) foi proposta como alvo para o desenvolvimento de drogas antibacterianas. A PNP catalisa a clivagem revers?vel, na presen?a de fosfato inorg?nico, de liga??es N-ribos?dicas de nucleos?deos e desoxinucleos?deos de purina, com exce??o da adenosina. Esta rea??o produz base p?rica livre e ribose (desoxiribose)-1-fosfato. A rea??o se processa com invers?o de…

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This study was designed to characterize, by means of functional and molecular approaches, the relevance of kinin B1 and B2 receptors in bladder cancer. Our data clearly shows that both B1 des-Arg9-BK and B2 BK receptor agonists were able to stimulate the proliferation of the grade 3-derived bladder cancer T24 cells. Furthermore, the incubation of B1 and B2 receptor antagonists, SSR240612 and HOE140, respectively, markedly inhibited the proliferation rate of T24 cells. In contrast, only higher concentrations of BK elicited the proliferation of the grade 1 bladder cancer cell line RT4, while des-Arg9-BK incubation completely failed to induce its proliferation. Interestingly, real time PCR experiments revealed that mRNA expression of B2, and mainly B1 receptors was found superior in T24 cells, in comparison to the low malignity grade RT4 cells. Furthermore, data obtained using bladder cancer human biopsies revealed that B1 receptor expression was visibly increased in all tumoral samples or under chronic inflammation of bladder. Concerning the signaling pathways related to the mitogenic effects of kinins, we bring novel evidence showing that pharmacological inhibition of PI3Kg with AS252424 concentration-dependently reduced T24 cell proliferation induced by BK or des-Arg9-BK. Finally, the incubation of T24 cells with kinin agonists led to a marked activation of PI3K/AKT and ERK 1/2 signaling pathways, whereas p38 MAP kinase remained unaffected. Our results indicate that kinin B2, and especially B1 receptors appear to be implicated in bladder cancer progression. It is tempting to suggest that selective kinin antagonists might represent potential therapeutic alternatives for bladder cancer control.

O presente estudo teve por objetivo caracterizar, atrav?s de abordagens funcionais e moleculares, a relev?ncia dos receptores B1 e B2 para as cininas no c?ncer de bexiga. Os dados obtidos mostraram que tanto o agonista dos receptores B1, des- Arg9-BK, quanto do receptor B2, BK, foram capazes de estimular a prolifera??o das c?lulas de c?ncer de bexiga grau 3, denominadas T24. Al?m disso, a incuba??o dos antagonistas dos receptores B1 e B2, SSR240612 e HOE140, respectivamente, inibiram acentuadamente a prolifera??o das c?lulas T24. Por outro lado, apenas maiores concentra??es de BK levaram ? prolifera??o das c?lulas de c?ncer de bexiga grau 1 RT4; enquanto a incuba??o com des-Arg9-BK n?o induziu sua prolifera??o. De maneira similar, os resultados revelaram que a express?o de mRNA dos receptores B2 e, principalmente, dos receptores B1 foi superior em c?lulas T24, em compara??o com as c?lulas RT4, que apresentam baixo grau de malignidade. Al?m disso, os dados obtidos com bi?psias de c?ncer de bexiga humano revelaram que a express?o do receptor B1 foi claramente maior em todas as amostras tumorais ou, em uma bi?psia obtida de um quadro de…

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The purinergic system can induce proliferation, differentiation and death on tumor cells. Different P2 purinergic receptors are involved in preventing the growth of these cells. Although the mechanisms are not completely known, they may represent interesting therapeutic targets to control tumors. The RT4 and T24 human tumor bladder cell lines have been used as low and high cancer malignancy phenotype models, respectively. Our research group has demonstrated that quercetin increases the ADP hydrolysis and inhibits ecto-5'-nucleotidase/ CD73, exerting an anti-proliferative effect on T24 cell lines. We also demonstrate that T24 cell lines expresses the purinergic enzymes CD39L4 and ecto-5'-nucleotidase / CD73, and have low hydrolytic activity of ATPase and ADPase in contrast to a high activity AMPase. These data suggest the involvement of the enzymes and purinergic receptors on progression the tumor of the bladder, as well as other tumor types. The objective of the present study was characterize the purinergic receptors P2X (1-7) expression profile on human bladder tumor cell lines and biopsies; and investigate the inflammatory genes that are up-regulated on high-grade human tumor bladder cells compared to low grade tumor bladder cells. Human bladder cells RT4 and T24 were obtained from the ATCC. Cells were cultured in DMEN and RPMI, respectively, supplemented with 10% of fetal bovine serum (FBS), under ideal conditions of cultivation Data were analyzed by one-way analysis of variance (ANOVA), followed by Tukey Kramer test. To confirm the relevance of P2X (1-7) receptors in bladder cancer, we have also assessed their expression in human biopsies. For this purpose, upon approval by the local ethics committee, specimens of bladder tumors and normal tissues were obtained from the same patients, who have been undergone surgical resection at the Hospital S?o Lucas/PUCRS. All samples were collected and rapidly frozen in -80?C with RNA holder. Tissue specimens were ground and then sonicated in a TRIzol kit. The mRNA level was analyzed using RT-PCR analysis. Real-time PCR revealed that the mRNA expression of P2X purinergic receptors, particulary P2X4R was significantly up-regulated on both T24 and RT4 cell lines. Data from human bladder tumor biopsies comparing to bladder normal tissue, showed that P2X6 and P2X7 receptors appeared to be most often up-regulated on tumor tissues. Twelve inflammation related genes, CCL2, CCR2, TNF-α, MAPK3, IL-1β, IL-6, ADORA1, CD200, CD4, CHRNA4, EDNRA e ITGB2 were significantly up-regulated on the grade III bladder tumor cell line T24 when compared to grade I bladder tumor cell line RT4. Our results indicate that the inflammation genes overexpressed may play an important role in regulating urinary bladder cancer. We can conclude that P2X purinoreceptors and inflammation genes are differentially…

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Programa de Demanda Social (CAPES)

A ovinocultura ?? uma atividade pecu??ria em crescente expans??o no Brasil, sendo a regi??o Nordeste a de maior potencial para o aumento de produ????o. Contudo, a sanidade dos rebanhos ainda ?? deficiente, havendo uma eminente necessidade de aumentar o monitoramento do rebanho, seja atrav??s de medidas profil??ticas ou pelo diagn??stico precoce de enfermidades. As mensura????es s??ricas de metab??litos s??o m??todos tradicionais e seguros, que auxiliam os cl??nicos na monitora????o dos rebanhos, utilizando-se como base os intervalos de refer??ncia. O presente estudo tem como objetivo padronizar t??cnicas de mensura????o dos metab??litos s??ricos glicose, ureia, cloretos, magn??sio, f??sforo e c??lcio em microplacas, para desenvolvimento de ensaios mais baratos e pr??ticos, auxiliando na determina????o de intervalos de refer??ncia regionais. Foram utilizados soros de 40 ovinos das ra??as Dorper e Santa In??s clinicamente sadios. As mensura????es foram feitas utilizando kits comerciais, atrav??s de ensaios convencionais e usando t??cnicas propostas como refer??ncias. Os ensaios em microplaca foram escolhidos de acordo com suas respectivas correla????es de Pearson, sendo posteriormente testadas para defini????o de repetitividade, reprodutibilidade e linearidade. Todos os ensaios desenvolvidos e padronizados apresentaram performance confi??vel, representando uma possibilidade de emprego em experimentos para determina????o de valores de refer??ncia, bem como para animais com dif??cil obten????o de amostras sangu??neas.

Sheep breeding is an expanding livestock industry in Brazil, and the Northeast region has the highest potential for this kind of economic activity. However, herd???s health maintenance is still deficient, and there is a perceived need to increase health monitoring, through prophylactic or disease early diagnosis measures. Serum measurements of metabolites are traditional and safe methods which assist clinicians in monitoring of flocks through the use of reference ranges. The present study aimed to standardize the measurement of the serum metabolites glucose, urea, chloride, magnesium, phosphorus and calcium in microplates, to develop cheaper and practical testing techniques. Sera from 40 Dorper and Santa In??s clinically healthy sheep were used, and measurements were made using commercial kits. Microplates assays were chosen according to their respective Pearson correlations and subsequently tested for repeatability,…

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior

About half of planet population depend on rice. In Brazil, this cereal crop is widely spread, and influenced by harsh climate conditions such as Humid Tropics one. In this region, certainly nitrogen is the most limiting nutrient. However, low insolation, acid and highly Al3+ satured soils still endangers the culture. In these soils, ammonium can be the single plant s Nitrogen source for plants. Its assimilatory pathway is economic and fast, although it presents strong integration with C metabolism. This work intended to recognize ammonium assimilation integrated with carbon metabolism. Two rice varieties were used: one adapted to Humid Tropics environment - Piau? - and an improved variety represented by IAC-47, selected under high N levels. Assays were carried on with 26 days after emergency plants, cultivated in Hoagland and Arnon (1950) modified solution with N-NH4 + 0.1 mM and 1.0 mM N-NH4 +. Plasmalem (P)-H+ATPases; vacuole (V)-H+ATPases; vacuole (V)-H+PPases activity as well as glutamine sinthetase (GS), glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH) were analysed. Moreover, it was determined soluble sugars, amino-N, N-NH4 +, total-N, fresh and dry weight, and nutrition solution pH and N-NH4 + variation. Also, seminal root length was measured as well as glycolysis was analyzed through phosphofructokinase (PFK) activity at plants roots tissue. P-H+ATPases activity was higher in IAC-47 roots. On the other hand, at Piaui roots, enhances at microsomal protein were observed, and V-H+ATPases presented higher activity as well as root development was high. This allowed the adapted variety to absorb N like the improved one. However, both varieties accumulated fresh weight in shoots at treatment 1.0 mM N-NH4 + and showed roots/shoots decrease at the same treatment. Moreover, amino N and NH4 + concentration was higher at Piau? shoots under 1.0 mM N-NH4 +. Both Piau? shoots and roots cultivated under 1.0 mM NNH4 + accumulated less soluble sugars. Also, Piaui shoots had higher a+b chorophyl concentration. PFK activity was enhanced at every plant, what accompanied ammonium levels. Piau? plants always presented low sugar oxidation levels. It also seemed to concentrate its sugars. Probably it happened to later availability for grain fullfil. To ensure this, it has high ammonium and amino-N pools. Nitrogen absorbed by 1.0 mM N-NH4 + Piau? seemed to be substrate for GDH-amination, wich activity was superior. Despite this, GDH deamination was high. In this way, Piau? could have GDH activity as an anaplerotic pathway to regulate C skeletons need. Meanwhile, GS increases were observed at IAC-47, toghether with GOGAT increases at 1.0 mM N-NH4 + shoots. IAC-47 assimilated ammonium at GS/GOGAT pathway, …

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CAPES

Nowadays, there is an interest in natural alternatives for insect control pests of stored grain, including use of essential oils. Thus, the objective of this work was to study the Cymbopogon nardus and C. citratus essential oils effect on reproductive cycle of Callosobruchus maculatus and some aspects regarding to xenobiotic and lipids metabolism. Thus, Cymbopogon citratus and C. nardus essential oils were extracted by hydrodistillation and analyzed by gas chromatography-mass spectrometry. Then, essential oils fumigants effects were tested at different concentrations (0.1; 0.2; 0.4; 0.6; 0.8 and 1?L cm-3) on C. maculatus and then evaluated some aspects of reproductive cycle, such as, lethal concentration to kill half of population members (LC50), oviposition and emergency of new adults. Essential oils fumigant effects were also tested to evaluated crude protein, total activity of some enzymes (acetylcholinesterase, glutathione S-transferase, carboxiesterase, alpha and beta-esterases) and lipid fractions (with virgin and copulated females separately), at concentration, 0.4 and 0.1 ?L, and periods, 12 and 6 hours, for C. citratus and C. nardus, respectively. Results obtained after essential oils chemical analysis showed main compounds, neral (36.6%), geranial (50.5%) to C. citratus, citronellal (50.3%) and nerol (10.8%) for C. nardus. Essential oils fumigant effects promoted increased mortality in C. nardus assay (LC50 = 0.12 ?L cm-3); both essential oils decreased oviposition and emergence of new adults. Crude extracts protein analysis showed that C. citratus and C. nardus essential oils promoted increase in females protein content and decrease in males. C. maculatus females resistance enzymes under C. citratus essential oil influence, showed increase activity for alfa-esterase, carboxylesterase and decrease for beta-esterase, acetylcholinesterase and glutathione S-transferase; in males were observed decrease in activity for acetylcholinesterase, carboxylesterase and increase for alfa-esterase and glutathione S-transferase; C. maculatus females under C. nardus essential oil influence promoted decrease in activity enzyme for acetylcholinesterase, carboxylesterase, alpha and beta-esterase; in males were observed decrease in activity for alpha-esterase and increase for carboxylesterase. In lipid fractions there was decrease in triacylglycerols content for females copulated under fumigant effect of C. citratus essential oil.

Atualmente, existe o interesse em alternativas naturais para o controle de insetos pragas de gr?os armazenados, entre elas pode-se citar a utiliza??o dos ?leos essenciais. Deste modo, o…

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Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do RJ, FAPERJ, Brasil.

The amount of waste produced daily by dairy cattle is one of the major problems in intensive management systems. Disposal of waste from animal facilities has become a challenge for breeders and specialists because it involves technical, health and economic aspects. In this sense, the main objective of this work was to evaluate the performance of a subsurface horizontal flow system (SAC), cultivated with vetiver grass (Chrysopogon zizanioides), in the treatment of milk wastewater. The work was developed in the area of Integrated Agroecological Production System (SIPA), also known as "Agroecological Farm of Km 47", Serop?dica-RJ. The evaluations were carried out by means of physical-chemical analyzes carried out in the wastewater in and out of this treatment unit, measurement of the mass and height production of the vetiver grass, as well as nutritional analyzes of its aerial part. For chemical-physical analyzes, the removal efficiency of oxygen, chemical oxygen demand, ammoniacal nitrogen, nitrite, nitrate, Kjeldahl total nitrogen, phosphorus, total suspended solids, turbidity and color were calculated, calculated at 27.12% , 18.27%, 32.41%, 26.92%, 9.12%, 11.43%, 2%, 27.56%, 30.50% and 20.08%, respectively. The nitrogen, phosphorus, potassium, calcium and magnesium exports were quantified for the nutrient analysis of the vetiver grass, all of them in unit kg.t-1 of dry mass, being calculated in 23.73, 5.2, 25 , 43, 2.7 and 1.58. Based on the monitoring carried out over the period in which the work was carried out, it can be concluded that the system of flooding built of subsurface horizontal flow, cultivated with vetiver grass, presented good performance in the treatment of milkwaste residues, highlighting It is also due to the low cost of installation, low maintenance and to be exempt in the consumption of electric energy, being indicated for small and medium units of agricultural systems.

A quantidade de dejetos produzidos diariamente por bovinos de leite ? um dos maiores problemas em sistemas de manejo intensivo. A disposi??o dos res?duos das instala??es para animais tem se constitu?do num desafio para criadores e especialistas, pois envolve aspectos t?cnicos, sanit?rios e econ?micos. Neste sentido, este trabalho teve como principal objetivo avaliar o desempenho de um sistema de alagado constru?do (SAC) de fluxo horizontal subsuperficial, cultivado com capim vetiver (Chrysopogon zizanioides), no tratamento da ?gua residu?ria de bovinocultura de leite. O trabalho foi desenvolvido na ?rea do Sistema Integrado de…

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The present work aimed the quantitative and structural characterization of microorganism activity by the quantification of phospholipids using the PLFA (Phospholipid - Derived Fatty Acids) technique in a soil from Cerrado biome under crop-farming system determining the relation between the changes in the tillage and the activity and diversity of microorganisms in the soil. These factors were related with physical and chemical soil variables. In this study, a typical acric Red Latosol with clayey texture under crop-farming integration was targeted, being samples collected from one year formation pasture, three years formation pasture and forest as reference. The sampling was performed on March and August 2013. Multivariate analysis using permutation test (based on 1000 permutations) and the Principal Component analysis regarding Instrumental Variables (PCAV) were used. The tillage and the management applied on the soil under crop-farming system affected the distribution, the structure and the physiological conditions of the microbial population in the soil. The distribution of these organisms was higher in pasture areas comparing to forest area, where organisms presenting opportunistic habit were predominant in contrast to forest area where the dominant presence was of strategist organisms. The difference in the time of pasture introduction has not influenced the structure of the microbial community, once that the distribution was the same in both studied areas. The stress index were higher in forest site than in pasture sites in both periods, probably due to the naturally low fertility level of this soil as much as its high acidity. The soil fertility data could explain the microbial variation under rainy and dry seasons. The organic matter in pasture sites was similar independent of the formation time. In the forest area, there is a higher contrast regarding the organic matter availability, which can explain the high heterogeneity on this area comparing to pasture areas.

Este trabalho teve como objetivo a caracteriza??o quantitativa e estrutural da atividade dos microrganismos a partir da quantifica??o de fosfol?pideos pela t?cnica de extra??o PLFA (Phospholipid - Derived Fatty Acids) em solo do bioma cerrado sob sistema integra??o lavoura-pecu?ria, determinando v?nculos entre mudan?as da cobertura com atividade e diversidade de microrganismos no solo. Estes valores foram relacionados com vari?veis f?sicas e qu?micas do solo. No presente estudo foi avaliada ?rea de Latossolo Vermelho ?crico t?pico de textura argilosa sob uso em sistema de integra??o lavoura-pecu?ria, sendo coletadas amostras em pasto com um ano de…

Atualmente, os combust�veis f�sseis representam 80% da produ��o energ�tica global, sendo que os restantes 20% correspondem �s energias renov�veis. Desta forma, � imperiosa a procura de fontes alternativas de energia sustent�vel de modo a reduzir as emiss�es de gases com efeito de estufa, nomeadamente o di�xido de carbono, que atinge j� os 400 ppmv na atmosfera. A biomassa algal tem sido apresentada como uma alternativa para a redu��o da concentra��o de CO2 na atmosfera, j� que tem a capacidade de sequestrar 1,83 kg CO2 por 1 kg de biomassa produzida. Para al�m disso, a biomassa algal pode constituir uma fonte de biocombust�veis e outros produtos de maior valor acrescentado. No entanto, a implementa��o industrial da produ��o e explora��o da biomassa algal depara-se ainda com v�rios desafios, nomeadamente a redu��o de custos e recursos nos processos upstream e downstream, onde os v�rios estudos t�m sido centrados. Com vista a contribuir para o esclarecimento de algumas das quest�es acima apresentadas, foram abordados aspectos da produ��o de biomassa e principalmente as quest�es da desconstru��o da biomassa da esp�cie Chlorella vulgaris. A produ��o de biomassa algal foi avaliada a partir de duas s�ries de ensaios de crescimento algal onde v�rias vari�veis f�sicas e bioqu�micas foram testadas. A concentra��o m�xima de biomassa obtida foi de 0,7 g/L, utilizando um meio de refer�ncia (meio Bristol normal modificado) e alimentando ar a um caudal de 1,5 L.L-1.min-1. Noutra s�rie de ensaios em que o caudal de ar foi aumentado para 3 L.L-1.min-1 a produ��o de biomassa caiu para cerca de 0,1 g/L, indiciando problemas com a transfer�ncia de massa e/ou stress mec�nico. Relativamente � disrup��o celular, foram testados v�rios m�todos f�sicos e f�sico-qu�micos, nomeadamente ultrassons, moagem, tratamento t�rmico e �cido, aplicados a biomassa seca e h�mida. Nos hidrolisados analisaram-se os teores em a��cares, prote�na e l�pidos. Dos v�rios m�todos testados em biomassa h�mida, o que apresentou melhores resultados foi a hidr�lise �cida (pH=1, Temperatura: 121�C, tempo: 1h) com uma recupera��o de 29,7% de a��cares e 21,3% de prote�nas. A secagem pr�via das amostras (altamente intensiva em energia) n�o contribuiu para um aumento significativo da extens�o da disrup��o celular. O incremento foi de 51,9% para 53,3% (a��cares e prote�na) para a biomassa seca, o que est� dentro do erro experimental e claramente n�o justifica o consumo energ�tico envolvido na secagem. Por outro lado, a composi��o do meio influenciou a composi��o das c�lulas. O refor�o dos nutrientes azoto e f�sforo conduziu � redu��o da s�ntese de a��cares e potenciou a s�ntese de prote�nas. O teor de l�pidos recuperados da biomassa rondou os 22%. Os ensaios realizados com maior quantidade de biomassa, necess�ria para a determina��o dos l�pidos, permitiu evidenciar a presen�a de um res�duo s�lido, de cerca de 20%, que dever� ser objeto de estudos futuros.

Currently, fossil fuels account for 80% of global energy production, and the remaining 20% correspond to renewable…

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CAPES

Medidas relativas aos espectros de Absor????o da Albumina S??rica Bovina com varia????o do comprimento de onda de varredura entre 300 a 500nm demonstraram um aumento nas intensidades da banda de absor????o da ASB, ap??s adi????o de aliquotas dos ligantes (??- lapachonas, cumarinas e Rutinas), sobretudo em 280nm, que ?? o comprimento de onda de absor????o caracter??stico dos dois res??duos de triptofano presentes na ASB. Medidas de Dicro??smo Circular confirmam a presen??a das bandas de absor????o caracter??sticas dos res??duos de triptofano, cujos m??ximos de absor????o est??o em 208 e 222nm nas diferentes temperaturas estudadas. As modifica????es obtidas nos espectros de Dicro??smo Circular da ASB a partir da adi????o de al??quotas das esp??cies ligantes deram origem ao efeito ocorrido na disposi????o planar original da ??-h??lice presente na estrutura secund??ria da ASB. A emiss??o fluorescente de ASB nas temperaturas de 298K, 303K, 310K e para os sistemas ASB/??-lapachonas e ASB/rutinas e 288K, 293K, 298K para o sistema ASB/Cumarinas, mostrou um efeito supressivo sobre o espectro de emiss??o da ASB, sobretudo em 345 nm, emiss??o do res??duo de triptofano da ASB, ap??s a adi????o dos ligantes. Os valores para os par??metros termodin??micos ??Go, ??Ho, ??So para os sistema ASB/??-lapachonas, ASB/4-metil-7-hidroxicumarinas e ASB/Rutinas n??o sofreram grandes varia????es como uma fun????o do ligante. A partir destes valores pode-se concluir que o tipo de intera????o predominante entre ASB/ligante foi hidrof??bica. A sobreposi????o entre os espectros de absor????o do supressor e de emiss??o da ASB possibilitou determinar para cada par ASB/supressor a dist??ncia cr??tica de intera????o e o raio de Foster, os quais mostraram a probabilidade da exist??ncia do fen??meno de Transfer??ncia da Energia Ressonante de Fluoresc??ncia (FRET) em todos os casos. No sistema ASB/??-lapachona e ASB/3-??cido sulf??nico ??-lapachona obtivemos respectivamente para um valor m??dio relativo as tr??s temperaturas de trabalho, temos: R0= 2,69 nm em rela????o a r = 3,02 e R0 = 2,32nm em rela????o a r = 3,32. J?? para o par 7-hidroxicumarina e 4-metil-7- hidroxicumarina, temos: R0= 3,53 e r = 3,83nm; R0= 3,70 nm e r = 4,33 nm. Por fim, no par ASB/Rutina e ASB/Rutina-metilada: R0= 3,43nm e r =…

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A tuberculose prevalece como uma das principais causas de morte no mundo ocasionadas por um ?nico agente infeccioso, o Mycobacterium tuberculosis. A enzima Chiquimato Quinase (MtSK) codificada pelo gene aroK de M. tuberculosis, quinta enzima da via do chiquimato. catalisa a transfer?ncia de um grupo fosfato do ATP para o carbono 3 do grupo hidroxila do chiquimato, formando chiquimato-3-fosfato e ADP. A interrup??o do gene aroK demonstrou que a enzima MfSK ? essencial para a viabilidade do M. tuberculosis. Neste trabalho, apresentamos a purifica??o da enzima MtSK at? a homogeneidade, an?lise por espectrometria de massa, sequenciamento de amino?cidos da por??o N-Terminal, determina??o do estado oligom?rico da enzima em solu??o, cin?tica em estado estacion?rio, espectroscopia de fluoresc?ncia e calorimetria de titula??o isot?rmica. Os resultados encontrados sugerem que a rea??o catalisada pela enzima monom?rica MtSK segue um mecanismo em equil?brio-r?pido aleat?rio para a adi??o dos substratos e uma libera??o ordenada dos produtos, onde a libera??o do chiquimato-3-fosfato ? seguida pela libera??o do ADP, resultando na enzima na sua forma livre. Os resultados de calorimetria demonstraram uma grande diferen?a de calor gerada na associa??o da enzima livre aos seus ligantes, revelando assinaturas termodin?micas de intera??es n?o-covalentes para cada processo de liga??o. Estes resultados ajudaram a compreender melhor o modo de a??o da enzima MtSK e podem subsequentemente serem ?teis para guiar o desenho racional de inibidores relacionados a esta enzima e que possam ser avaliados posteriormente como anti-TB.

Submitted by Jorge Silva ([email protected]) on 2017-05-02T21:37:15Z No. of bitstreams: 1 2016 - Tatiane Cristina dos Santos Bonfim.pdf: 2465996 bytes, checksum: 94f74695ec8fd235bfbcd7f4a82aec87 (MD5)

Made available in DSpace on 2017-05-02T21:37:15Z (GMT). No. of bitstreams: 1 2016 - Tatiane Cristina dos Santos Bonfim.pdf: 2465996 bytes, checksum: 94f74695ec8fd235bfbcd7f4a82aec87 (MD5) Previous issue date: 2016-02-25

CNPq

The interaction between intermediate hosts and helminths can cause metabolic and reproductive changes in the host snail, they start using their reserves to maintain its vital functions, immune system activation, repair of tissue damage and also to supply the necessary energy for the parasites development. Investigations of co-infection by different helminths can provide essential information about the biology of their co-existence. Our aims are investigate the reproductive changes, evaluate the activity of lactate dehydrogenase and the concentrations of glucose in the hemolymph and glycogen in the digestive gland complex and cephalopedal mass, and also verify histological and histochemical changes of Biomphalaria glabrata experimentally co-infected with Echinostoma paraensei and Angiostrongylus cantonensis. Five groups of snails i.e. uninfected, with single and double infections, either E. paraensei first (E+A) or A. cantonensis first (A+E) were followed; three times a week during four weeks the numbers of egg masses, eggs and hatched snails were counted. Histological and histochemical samples of the ovotestis, albumen gland, digestive gland complex and cephalopedal mass were collected after four weeks as well samples for the biochemical analysis. The number of egg masses/snail, eggs/snail and hatched snails showed significant differences comparing the control group to all infected groups, especially in group E + A, with the majority of values of parameters analyzed lower than 50% of those observed for control snails. A significant decrease in glucose levels and glycogen content, in contrast an increase of lactate dehydrogenase activity was observed in the infected snails. These results indicate intense energy demand that takes the snail increasing the anaerobic degradation of carbohydrates in the infected snails to obtain energy in an attempt to maintain homeostasis. The histological analysis showed that presence of both parasites in the all organs analyzed and the parasites were associated with cell disorganization.

A intera??o entre hospedeiros intermedi?rios e helmintos pode causar altera??es metab?licas e reprodutivas no molusco hospedeiro, que come?am a usar suas reservas para manter as fun??es vitais, na elabora??o de resposta imunol?gica e repara??o de danos teciduais e tamb?m para compensar a drenagem de nutrientes/energia pelos parasitos em desenvolvimento. Investiga??es sobre as co-infec??es por helmintos de esp?cies diferentes podem fornecer informa??es essenciais sobre a biologia da sua co-exist?ncia, refletindo aspectos mais pr?ximos ?queles que encontramos em…

Submitted by Sandra Pereira ([email protected]) on 2017-06-20T15:46:28Z No. of bitstreams: 1 2010 - Carla Rodrigues Bahiense.pdf: 694207 bytes, checksum: 012f3bf71990c09f742e165f567b3e77 (MD5)

Made available in DSpace on 2017-06-20T15:46:28Z (GMT). No. of bitstreams: 1 2010 - Carla Rodrigues Bahiense.pdf: 694207 bytes, checksum: 012f3bf71990c09f742e165f567b3e77 (MD5) Previous issue date: 2010-07-13

Belonging to the Order Psittaciformes, Family Psittacidae, the Ara ararauna, like other parrots, is a Brazilian bird much exploited in the pet market, which makes it a frequent veterinary patient. Despite of the extreme clinical relevance of laboratory examinations, data on hematology and serum biochemistry of this species are scarce. Several factors can interfere with the birds results of hematological and biochemical values, such as age, sex, reproductive period, among others. The study aimed to recognize patterns and to determine haematological and serum biochemical parameters for the species in question, explaining possible variations related to sex, management and method of restraint. In the experiment was used 68 specimens, 33 from a commercial breeding facility and 35 from the live collection of the RIOZOO Foundation. The blood samples were collected by jugular vein, transferred into tubes containing EDTA and other with no anticoagulant. Only the birds of RIOZOO were subjected to a new collection, which occurred 10 minutes after of anesthesia with isoflurane, thus forming a group of 33 animals anesthetized and another 35 unanesthetized. Of each animal were determined trombocyte conts, erythrocyte counts, total and specific leucocyte counts, concentrations of total plasmatic protein, urea, creatinine, uric acid, cholesterol, tryglicer?des, total serum protein, albumin and globulin, and activityes of alkaline phosphatase, aspartate aminotransferase, amylase, lipase, and creatine-kinase enzymes. The results were analyzed by ANOVA and paired T tests. Females had higher trombocitometria. Non-anesthetized animals had a greater total WBC count. Weren?t significant differences between anesthetized males and females. The birds originated from RIOZOO had higher values of PCV, red blood cell count, MCV, eosinophils, basophils, urea, AST and CK, however, lower values of albumin and creatinine. The study revealed that was a significant level of discrepancies between the different groups, allowing the creation of a standard more specific hematologic, according to the individual characteristics of each patient.

Pertencente ? Ordem Psittaciforme e Fam?lia Psittacidae, a Ara ararauna, assim como os demais psitac?deos, ? uma ave brasileira muito explorada no mercado pet, o que a torna um freq?ente paciente veterin?rio. Apesar da extrema relev?ncia cl?nica dos exames laboratoriais, dados acerca da hematologia e bioqu?mica s?rica dessa esp?cie ainda s?o escassos. Importante ? considerar tamb?m que diversos fatores n?o patol?gicos podem interferir em resultados dos exames hematol?gicos e bioqu?micos das aves, como a idade,…

Submitted by Celso Magalhaes ([email protected]) on 2020-02-14T14:46:22Z No. of bitstreams: 1 2014 - Tatiana Faria Maia.pdf: 8764609 bytes, checksum: 6abee87823a8f954640b0100eade28ec (MD5)

Made available in DSpace on 2020-02-14T14:46:22Z (GMT). No. of bitstreams: 1 2014 - Tatiana Faria Maia.pdf: 8764609 bytes, checksum: 6abee87823a8f954640b0100eade28ec (MD5) Previous issue date: 2014-03-27

Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES

Brazil is the country with the greatest biological diversity , presenting the widest continental biota of the Earth. The Atlantic Forest corresponds an area of 13 % of the brazilian territory, sheltering a large portion of this diversity. The fungi are extremely important organisms for the functioning of these ecosystems. Fungi commonly isolated from soil stands out species of Aspergillus. The members of the section Nigri are among the most important in the genus Aspergillus because are common worldwide and feature economical and biotechnological potential. Their identification is considered difficult and complex due to high genetic diversity. Therefore, the taxonomy of Aspergillus section Nigri has been investigated using morphological, biochemical and molecular data. Some species are responsible for causing food deterioration and mycotoxins production, however, other are important producers of organic acids, antibiotics and extracellular enzymes. Among produced the enzymes by these fungi, we can cite cellulases, amylases, peptidases, pectinase and phytase which have applications in several industrial sectors such as textile, food and drinks, agroindustry of animal feed, sillage, pharmaceutical, etc. The objective was to identify Aspergillus section Nigri species isolated from litter and soil through morphological, biochemical and molecular characterization, as well as to evaluate the enzymatic potential of these isolates. For these study were collected 198 samples from litter and soil (0-5 and 5-10 cm depth) during the two periods, rainy and dry the Atlantic Forest Biome belonging to the Minas Gerais State. After isolation, 42 Black Aspergillus were identified using macroscopic and microscopic criteria on CYA 25 and 37 ?C, MEA 15, 25, 33, 36 and 40 ?C for a period of 10 days. For biochemical analysis, was used CREA, MEA-Boscalid for 7 days at 25 ?C, and Ehrlich test. The molecular analysis was performed by sequencing the ITS1 and ITS4 regions. The enzymatic potential was evaluated through enzymatic index, a semi -quantitative parameter used to evaluate the production of hydrolytic enzymes by fungi in solid medium. The study revealed the existence of 6 different groups, showing a good dicersity of species of Aspergillus section Nigri of. In the rainy season there was a greater richness of these species, where it was found 92.8 %, while in the dry season it was found only 7.2%. The groups I and II were identified through morphological and molecular characterization belonging to uniseriate species of Aspergillus …

Submitted by Jorge Silva ([email protected]) on 2019-11-05T17:57:43Z No. of bitstreams: 1 2015 - Marcus Vinicius Loss Sperandio.pdf: 2382015 bytes, checksum: 474c94a478b6c54deb3404f98766611b (MD5)

Made available in DSpace on 2019-11-05T17:57:43Z (GMT). No. of bitstreams: 1 2015 - Marcus Vinicius Loss Sperandio.pdf: 2382015 bytes, checksum: 474c94a478b6c54deb3404f98766611b (MD5) Previous issue date: 2015-02-26

CNPq

The soil nitrogen (N) availability is generally low, leading to frequently input of fertilizer and the search of varieties with higher nitrogen use efficiency. Among the favorable parameters include higher expression of plasma membrane pumps protons and high-affinity nitrate transporters under low N cultivation. This work aimed to identify the expression pattern of OsNRT1.1A, OsNRT1.1B and OsNRT1.1C, possible orthologs of the Arabidopsis double affinity nitrate tranporter NRT1.1 (CHL1; NPF8.9) in response to NO3- and NH4+, as well as evaluate the importance of OsA2 and OsA7 proton pumps genes to N uptake. To evaluate OsNRT1.1 (A, B and C) gene expression, two experiments were performed: the first using Nipponbare rice variety and the second using IAC-47 (improved) and Piau? (Landrace) rice varieties. The OsA2 and OsA7 gene silencing was made using the artificial micro RNA (amiRNA) followed by rice genetic transformation. The experiment to evaluate the contribution of plasma membrane H+-ATPase (PM H+-ATPase) to low NO3- levels was made using IAC-47 and Piau? rice varieties. The OsNRT1.1 gene expression analysis in the roots showed that OsNRT1.1A and OsNRT1.1B are highly expressed, while OsNRT1.1C showed low expression rate. OsNRT1.1B expression was downregulated with ressuply of 2.0 mM of NO3 – N, in the plants cultivated using NH4+ and N starving plants, however, OsNRT1.1B expression was upregulated with constant supply of 2.0 mM of NO3 – N. After rice genetic transformation, Real Time PCR analysis showed that only one amiRNA successfully downregulated the target, furthermore the lineages silencing the target were multiplied in greenhouse. The rice lineages downregulating OsA2 (osa2.2, osa2.3, osa2.4), OsA7 (osa7.1, osa7.2, osa7.3) and control plants (plants transformed with empty IRS) were cultivated using four treatments: three days N starvation, constant with 0.2 mM of NO3 – N, ressuply with 0.2 mM of NO3 – N and constant with 2.0 mM of NO3 – N. The osa7 mutants shown lower growth, meanwhile osa2 growth normally compared to control plants (IRS). Both mutants shown lower NO3- uptake when grown with constant 0.2 mM of NO3 – N compared to control IRS plants, meanwhile the other treatments did not show differences of NO3- uptake. The PM H+-ATPase activity was lower in the osa7 and osa2 mutants compared to IRS plants in all treatments used. The V-H+-ATPase activity did not show difference in the mutants osa7 and osa2, however, V-H+-PPase activity was slower only in the osa7 mutants compared to IRS control plants. The mutant plants osa2 showed lower N uptake Efficiency index, accumulating lower…

Made available in DSpace on 2015-04-14T13:35:19Z (GMT). No. of bitstreams: 1 434470.pdf: 6587349 bytes, checksum: 405b1e4ec3fc9ec1beb9e4856acacdf7 (MD5) Previous issue date: 2011-08-24

Aproximadamente 1/3 da popula??o mundial est? infectada com o Mycobacterium tuberculosis, agente etiol?gico da tuberculose humana (TB). A incid?ncia global e o constante aumento da resist?ncia ?s drogas tornam evidente a urgente necessidade do desenvolvimento de novas drogas, para o tratamento, e vacinas, para a preven??o desta doen?a. As enzimas das vias metab?licas fundamentais s?o vistas como atrativos alvos moleculares definidos. A enzima guanosina monofosfato sintase (GMPS), uma amidotransferase do tipo G, catalisa a amina??o da xantosina monofosfato (XMP) ? guanosina monofosfato (GMP), em uma rea??o que ocorre em dois dom?nios, o glutamina amidotrasferase e o adenosina trifosfato (ATP) pirofosfatase, onde ocorre o ataque do intermedi?rio altamente reativo adenil-XMP, levando a forma??o de GMP. O gene guaA (Rv3396c), o qual codifica a enzima GMPS, foi identificado por homologia de sequ?ncia no genoma de M. tuberculosis H37Rv. N?s evidenciamos que a forma??o do adenil-XMP causa uma inibi??o da enzima pelo aumento dos n?veis de ATP e XMP. Al?m disso, a GMPS de M. tuberculosis (MtGMPS), apresentou um comportamento alost?rico na varia??o dos n?veis de XMP, demonstrando cooperatividade positiva e diferentes aspectos cin?ticos dos j? reportados para as enzimas de humanos e Escherichia coli. O mecanismo ping-pong foi proposto com base nos resultados evidenciados pela libera??o de pirofosfato (PPi) depois da liga??o da am?nia, corroborando com os dados previamente publicados para outras GMPSs. Nossos resultados podem ser ?teis por revelar o papel desta enzima no metabolismo de purinas do M. tuberculosis, fornecendo conhecimento para o desenvolvimento de novas drogas e vacinas para o tratamento e preven??o, respectivamente

Submitted by SBI Biblioteca Digital ([email protected]) on 2018-02-15T12:50:05Z No. of bitstreams: 1 VANDERLEI JOSE JUNIOR.pdf: 1351825 bytes, checksum: 599d050ec951a6a8e7497a0cea5d2e17 (MD5)

Made available in DSpace on 2018-02-15T12:50:05Z (GMT). No. of bitstreams: 1 VANDERLEI JOSE JUNIOR.pdf: 1351825 bytes, checksum: 599d050ec951a6a8e7497a0cea5d2e17 (MD5) Previous issue date: 2017-12-15

Introduction: Prostate cancer is one of the most prevalent diseases in the male population, occupying the second position among malignant neoplasms. There are several therapeutic options for the treatment of localized prostate cancer, ranging from conservative behaviors to interventional treatments such as radical prostatectomy or external radiotherapy, associated or not with brachytherapy. Objective: To identify the factors that can predict biochemical recurrence and to evaluate treatment toxicity. METHOD: This is a retrospective and longitudinal study of 162 patients diagnosed with prostate cancer treated with conformational external radiotherapy associated with high dose rate brachytherapy (BTATD) between 2005 and 2014. The database was used of the Radium ? Campinas Oncology Institute, collected prospectively. Results: The mean follow-up time was 57 months. No grade 3 late toxicity was observed in the gastrointestinal tract, with only 1 patient (0.6%) genitourinary tract. The only categorical variable that presented statistical significance for biochemical relapse was the Nadir PSA <1 ng / ml (p = 0.018). The biochemical recurrence rate found was 96.3%, based on the Phoenix criteria (PSA nadir + 2 ng / ml). Conclusions: This study demonstrated that in the treatment of localized prostate cancer, the association of external radiotherapy with BATD is a safe therapeutic option, with a low degree 3 late toxicity and a biochemical recurrence of only 3.7% (with HF = 95 %).

Introdu??o: O c?ncer de pr?stata ? uma das doen?as mais prevalentes na popula??o masculina, ocupando a segunda posi??o entre as neoplasias malignas. H? v?rias op??es terap?uticas para o tratamento do c?ncer de pr?stata localizado, podendo variar de condutas conservadoras ? tratamentos intervencionistas como a prostatectomia radical ou a radioterapia externa, associada ou n?o ? braquiterapia. Objetivo: Identificar os fatores que possam predizer recidiva bioqu?mica e avaliar a toxicidade do tratamento. M?todo: Tratase de um estudo retrospectivo e longitudinal, com 162 pacientes diagnosticados com c?ncer de pr?stata, tratados com radioterapia externa conformacional associada ? braquiterapia de alta taxa de dose (BATD), entre 2005 e 2014. Utilizou-se o banco de dados do Radium - Instituto de Oncologia de Campinas, coletados prospectivamente. Resultados: O tempo m?dio de seguimento foi de 57 meses. N?o foi observada toxicidade tardia grau 3 no trato gastrointestinal, sendo apenas 1 paciente (0,6%) trato genitourin?rio. A ?nica vari?vel categ?rica que apresentou signific?ncia estat?stica para recidiva bioqu?mica foi o PSA Nadir <1 ng/…

Made available in DSpace on 2015-04-14T14:51:08Z (GMT). No. of bitstreams: 1 431437.pdf: 291286 bytes, checksum: aad289e4c817a0471c68e921a07ac277 (MD5) Previous issue date: 2011-03-17

Caracter?sticas: A depress?o ? uma das doen?as mentais que mais afeta a popula??o mundialmente. Tamb?m conhecida como um transtorno do humor ? caracterizada por sentimentos de derrota, baixa autoestima e culpa, sendo duas vezes mais comum em mulheres do que em homens. Uma das suas principais conseq??ncias no organismo s?o as disfun??es cognitivas. Objetivos: N?s investigamos os efeitos do suporte cognitivo sobre a mem?ria contextual na depress?o e analisamos as caracter?sticas neuropsicol?gicas e hormonais que podem afetar a mem?ria. M?todos: Dezessete pacientes medicados com depress?o maior unipolar (idade 20- 40 anos, 14 mulheres) e 22 controles pareados por idade, g?nero e educa??o realizaram o subteste Vocabul?rio do WAIS-III (VWAIS), o Wisconsin Card Sorting Test (WCST) e um teste de mem?ria para reconhecimento de itens (objetos) e contextos (local), o qual foi aplicado com ou sem pista incidental. Amostras de salivas para an?lise de cortisol e DHEA foram coletadas as 07AM, 4PM e 10PM. Resultados: Pacientes e controles apresentaram desempenho equiparado nos escores de reconhecimento de itens, mas os pacientes apresentaram um d?ficit de mem?ria contextual na aus?ncia da pista. O mau desempenho nos testes neuropsicol?gicos (VWAIS e WCST) e baixos n?veis de cortisol e da raz?o cortisol/DHEA em pacientes n?o foram a causa relacionada a esse d?ficit. Conclus?o: D?ficits na mem?ria contextual em pacientes com depress?o maior podem ser reduzidos fornecendo suporte cognitivo adicional na codifica??o

Made available in DSpace on 2015-04-14T14:51:14Z (GMT). No. of bitstreams: 1 435930.pdf: 928390 bytes, checksum: 7d9c63e8c3ebdc0a8df50f72dcdee9ab (MD5) Previous issue date: 2011-08-26

Nanocomposites derived from carbon atoms have been the focus of interest in various industrial fields such as electronic engineering, pharmaceuticals, medical devices, cosmetics, food packaging and other, since its discovery in 1985. Fullerene C60 is a nanocomposite with 60 carbon atoms, which has been the subject of numerous studies for its ability to store atoms in the interior and accept various structural modifications. The importance of understanding all the possible effects of exposure to fullerenes is very great, since its production and its use is growing every day increasing environmental exposure to this compound. The cholinergic system has as its main neurotransmitter acetylcholine (ACh). The acetylcholinesterase (AChE, EC3.1.1.7) is an important regulatory enzyme that controls the transmission of nerve impulses across cholinergic synapses by hydrolysis of the ACh. AChE levels are controlled by the interaction of ACh with its receptors, and when the interaction is enhanced, the levels of AChE are increased. Therefore, AChE can be used as a marker of cholinergic function. In this study our goal was to determine whether intraperitoneal injections of fullerene C60 at the doses of 7.5, 15 and 30 mg/kg and the time of 6h, 12h and 24h of exposure would cause a change in the modulation of the cholinergic system. We observed that the dose of 30 mg/kg in the exposure time of 24 hours showed a 84% increase in enzyme activity compared with the vehicle control group. These results suggest a possible neurotoxic effect. Further studies should be conducted to extend these findings.

Nanocompostos derivados de ?tomos de carbono t?m sido foco de interesse para aplica??es em v?rios campos industriais, tais como engenharia eletr?nica, produtos farmac?uticos, dispositivos m?dicos, cosm?ticos, embalagens de alimentos, entre outros, desde seu descobrimento em 1985. O fulereno C60 ? um nanocomposto com 60 ?tomos de carbono que tem sido alvo de in?meros estudos por suas propriedades como a capacidade de armazenar ?tomos em seu interior e aceitar diversas modifica??es estruturais. A import?ncia de entendermos todos os poss?veis efeitos de uma exposi??o aos fulerenos ? muito grande, j? que a sua produ??o e sua utiliza??o est?o crescendo a cada dia aumentando a exposi??o do meio ambiente a este composto. O sistema colin?rgico tem como principal neurotransmissor a acetilcolina (ACh). A acetilcolinesterase (AChE, E.C.3.1.1.7) ? uma importante enzima regulat?ria que controla a transmiss?o de impulsos nervosos atrav?s da sinapse colin?rgica pela hidr?lise da ACh. Os n?veis de AChE s?o controlados pela intera??o da ACh com seus receptores, sendo que quando a intera??o ? acentuada, aumentam os n?veis de AChE Portanto, a AChE pode ser usada como um marcador da fun??o colin?rgica. Neste estudo nosso objetivo foi verificar se inje??es intraperitoneais de…

Submitted by Sandra Pereira ([email protected]) on 2018-01-26T12:38:56Z No. of bitstreams: 1 2010 - Juberlan Silva Garcia.pdf: 6866273 bytes, checksum: 77b115c7b87deb998b7440f5ecb87940 (MD5)

Made available in DSpace on 2018-01-26T12:38:56Z (GMT). No. of bitstreams: 1 2010 - Juberlan Silva Garcia.pdf: 6866273 bytes, checksum: 77b115c7b87deb998b7440f5ecb87940 (MD5) Previous issue date: 2010-12-03

The trematode Echinostoma paraensei parasitizes the small intestine of rodents, being its natural definitive host Nectomys squamipes. In spite of the great importance of echinostomatides, there are few studies on morphology, biology and physiology of E. paraensei and its interaction with their intermediate and definitive hosts. In the present study 50 Rattus norvegicus (Wistar),adult females were used, 35 were individually infected with 150 E. paraensei metacercariae (Sumidouro strain) and 15 were maintained uninfected, as control group. Weekly, seven infected and three uninfected rodents were euthanized using CO2.The blood was collected to hematological analysis, and serum obtained by centrifugation and used to aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), bilirubin, glucose, total proteins and fractions determinations. Fragments of liver were collected to glycogen determination and histophatological analysis. Adult worms were loaded until third week of infection, being the higher number of worms loaded at first week of inection. The hepatocytes had rounded edges, with signs of impaired nuclear, mild infiltration of polymorphonuclear and sinusoids slightly expanded. The hepatocytes were vacuolated with discrete perivascular and periportal lymphocytic infiltration. The serum biochemical parameters analyzed were increased at the first week of infection, varying onward. Hematological analysis revealed the development of normocytic and normocromic anaemia with anisocytic alteration. In the white blood cells, only a leukopenia was observed at the third week of infection. The present results are discussed.

O tremat?deo Echinostoma paraensei ? um parasito de intestino delgado de roedores, sendo seu hospedeiro definitivo natural Nectomys squamipes. No presente estudo, foram utilizadas 50 f?meas adultas de Rattus norvegicus (Wistar), 35 animais infectados individualmente com 150 metacerc?rias de E. paraensei (linhagem Sumidouro) e 15 animais n?o infectados (controle). Semanalmente, sete animais infectados e tr?s animais controle foram submetidos ? eutan?sia com CO2. O sangue foi coletado para an?lise hematol?gica, e o soro obtido por centrifuga??o, para as determina??es de aspartato aminotransferase (AST), alanina aminotransferase (ALT), gamaglutamiltransferase (GGT), fosfatase alcalina (ALKP), bilirrubina, glicose, prote?nas totais e fra??es. Fragmentos de f?gado foram coletados para a determina??o de glicog?nio e para a an?lise histopatol?gica. Vermes adultos foram recuperados at? a terceira semana de infec??o, sendo o maior n?mro de…

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Made available in DSpace on 2020-07-28T18:34:10Z (GMT). No. of bitstreams: 1 2011 - Usha Vashist.pdf: 15299905 bytes, checksum: 57cd96658d7751c2b9843044f4be2010 (MD5) Previous issue date: 2011-02-23

Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES, Brasil.

Plasmodium juxtanucleare and Borrelia anserina are common pathogens in domestic and wild birds, they can cause serious damages in the birds, especially those created organically, drugs free and not confined. These pathogens are related to anemia, diarrhea, inflammation, damage to vital organs, limb paralysis, anorexia, weight loss and reduction in egg production. This mixed infection can occur naturally among birds, but this subject is poorly studied, for this reason the present survey aimed to evaluate the biological, biochemical and histopathological features of infections caused by P. juxtanucleare and B. anserina in Gallus gallus. For this experiment, initially a search for birds infected by P. juxtanucleare was made in the Seropedica City, RJ. Two hundred thirty four birds, among them ducks, widgeons, geese, turkeys and chickens were sampled. Plasmodia were isolated from blood of chickens of Silky Japan?s breed for experimental inoculations. The plasmodia were characterized based on morphology and morphometry, and the species confirmed by molecular technique. Experimental infections were performed in quails and ducks for the infectivity of the P. juxtanucleare isolate studies. Thereafter, for the biological, biochemical and histopathological studies, 48 Cobb chickens with one day old were acquired. At 60 days old the chicks were divided into four groups of 12 birds each: G1-uninfected control, G2-infected by P. juxtanucleare isolated from chicken of Silky Japan's breed of the Serop?dica City, G3- infected by B. anserina strain cryopreserved and maintained in the Laboratory of Parasitic Diseases, Epidemiology and Public Health Department, Veterinary Institute, Federal Rural University of Rio de Janeiro and G4- infected by the same strain of P. juxtanucleare and B. anserina previously cited. Over 45 days, daily, the birds were observed for the presence of clinical signs of infection. Throughout the experiment blood smears were made from birds to parasitemia and spirochaetemia evaluation. Weekly, the serum enzymes alanine transaminase (ALT), aspartate transaminase (AST), uric acid, creatinine, total proteins were analyzed and in the experiment end histological sections of liver and kidney of birds in each experimental group were evaluated. The prevalence of P. juxtanucleare in chickens, ducks, widgeons, geese and turkeys was 56%, 28%, 32%, 40% and 50%, respectively, and the parasitemias averaged 0.12%, 0.08%, 0.02% 0.03% and 0.1%, respectively. Biomolecular and morphometric analysis of parasites isolated from chickens of…