Isolamento de uma quitinase extraída do cefalotórax do camarão marinho Litopenaeus schmitti (BURKENROAD, 1936) : avaliação de suas atividades antimicrobiana e larvicida

Chitinases are enzymes involved in degradation of chitin and are present in a range of organisms, including those that do not contain chitin, such as bacteria, viruses, plants and animals, and play important physiological and ecological roles. Chitin is hydrolyzed by a chitinolytic system classified as: endo-chitinases, exo-chitinases and N-acetyl-b-D-glucosaminidases. In this study a Litochitinase1 extracted from the cephalotorax of the shrimp Litopenaeus Schmitt was purified 987.32 times using ionexchange chromatography DEAE-Biogel and molecular exclusion Sephacryl S-200. These enzyme presented a molecular mass of about 28.5 kDa. The results, after kinetic assay with the Litochitinase1 using as substrate p-nitrophenyl-N-acetyl-b-Dglucosaminideo, showed apparent Km of 0.51 mM, optimal activity at pH ranging from 5.0 to 6.0, optimum temperature at 55°C and stability when pre-incubated at temperatures of 25, 37, 45, 50 and 55°C. The enzyme showed a range of stability at pH 4.0 to 5.5. HgCl2 inhibited Litochitinase1 while MgCl2 enhances its activity. Antimicrobial tests showed that Litochitinase1 present activity against gram-negative bacterium Escherichia coli in the 800 μg/mL concentration. The larvicidal activity against Aedes aegypti was investigated using crude extracts, F-III (50-80%) and Litochitinase1 at 24 and 48 hours. The results showed larvicidal activity in all these samples with EC50 values of 6.59 mg/mL for crude extract, 5.36 mg/mL for F-III and 0.71 mg/mL for Litochitinase1 at 24 hours and 3.22 and 0.49 mg/mL for the F-III and Litochitinase1 at 48 hours, respectively. Other experiments confirmed the presence of chitin in the midgut of Aedes aegypti larvae, which may be suffering the action of Litochitinase1 killing the larvae, but also the absence of contaminating proteins as serine proteinase inhibitors and lectins in the crude extract, F-III and Litochitinase1, indicating that the death of the larvae is by action of the Litochitinase1. We also observed that the enzymes extracted from intestinal homogenate of the larvae no have activity on Litochitinase1. These results indicate that the enzyme can be used as an alternative to control of infections caused by Escherichia coli and reducing the infestation of the mosquito vector of dengue.

Coordenação entre as respostas da catalase e da ascorbato peroxidade em folhas de feijão caupi submetidas a diferentes fontes de peróxido de hidrogênio

The plants are often exposed to variations in environmental conditions that may trigger metabolic disturbances leading to a consequent loss in productivity of crops. These stressful conditions usually induce an accumulation of reactive oxygen species (ROS) in the cell, a condition known how oxidative stress. Among these species, hydrogen peroxide (H2O2) is an important molecule involved in numerous signaling mechanisms. The present study aimed to understand the relationship between the different enzymatic mechanisms of elimination of H2O2 by catalase (CAT) and ascorbate peroxidase (APX) in leaf tissues of seedlings of the species Vigna unguiculata L. Walp, under conditions of oxidative stress induced by application of CAT inhibitor, 3-amino-1,2,4-triazole (3-AT), and H2O2 itself on the roots. Three experiments were conducted. The first experiment was performed applying the compound 3-AT (5 mM) during the time (hours). In the second experiment, seedlings were exposed to different concentrations of H2O2 (2.5, 5.0, 7.5, 10 mM) for 48 h. The third strategy included the pre-treatment with H2O2 (2.5 mM) for 24 h, followed by subsequent treatment with the inhibitor 3-AT and recovery control condition. Treatment with 3-AT causes a strong inhibition of CAT activity in leaf tissues accompanied by an increase of activity of APX. However a decrease in oxidative damage to lipids is not observed as indicated by TBARS. It was observed that activity of APX is directly linked to the content of peroxide. Inductions in the activities of CAT and APX were observed mainly in the seedlings treated with 2.5 mM H2O2. This can be associated with a decrease in oxidative damage to lipids. In contrast, one same tendency was not observed in treatments with higher concentrations of this ROS. These results suggest that the concentration of 2.5 mM H2O2 can induce responses antioxidants later in seedling cowpea. This concentration when applied as pre-treatment for 24 h promoted an induction systems removers CAT and APX, both in activity and in terms of gene expression. However this increment was not observed in the recovered plants and the plants subsequently subjected to 3-AT. Additionally, the pretreatment was not sufficient to attenuate the inhibition of CAT activity and oxidative damage to lipids caused by the subsequent application of this inhibitor. The results showed that the application of 3-AT and H2O2 in the root systems of seedlings of cowpea promote changes in the parameters analyzed in leaf tissues that indicate a direct response to the presence of these factors or systemic signaling mecanisms. H2O2 appears to activate the responses of two antioxidant systems in this study thar does not promote greater protection in case of additional treatment with 3-AT. This demonstrates the importance of the CAT system. In this work, complete results indicate that there is a difference between the signaling and the effects caused by exposure to H2O2 and by treatment with 3-AT

Efeito anti-inflamatório do heparinóide isolado do camarão Litopenaeus vannamei sobre a peritonite aguda

In recent years the heparin has been the subject of several studies that aim to expand its use as a therapeutic agent, due to its ability to modulate the activity of various proteins that play important roles in the regulation of pathophysiological processes. In several experiments and preclinical trials, heparin has demonstrated an anti-inflammatory role. However, its clinical use is limited, due to its strong anticoagulant activity and hemorrhagic complications. For this reason, considerable efforts have been employed in discovery of heparin analogous (heparinoid) with reduced side effects, that retain the anti-inflammatory properties of heparin. In this context, a heparinoid obtained from the head of Litopenaeus vannamei shrimp, which presents a structural similarity to heparin, showed, in previous studies, anti-inflammatory activity in a model of acute peritonitis with reduced anticoagulant effect in vitro and low hemorrhagic activity. Thus, the present work had as objective to evaluate the effect the heparinoid of the cephalothorax of gray shrimp on the acute inflammatory response in different times (3 or 6 hours after the induction of inflammatory stimulus), using the model of acute peritonitis induced in mice. It was also analyzed the HL effect over the activity of elastase, an enzyme involved in leukocyte recruitment. Furthermore to check if the different doses of heparin and heparinoid change the hemostatic balance in vivo, was assessed the effect of these compounds on the plasma clotting time in animals submitted to inflammation. The results show that in 3 hours, all doses of heparinoid were able to prevent efficiently in the acute inflammatory process without any anticoagulant effects, unlike the extrapolation dose of heparin, which has induced a large hemorrhage due its high anticoagulant activity. However, 6 hours after induction of inflammation, only the dosages of 0.1 and 1.0 μg/Kg of heparin and 1.0 μg/Kg of heparinoid kept anti-migratory effect, without changing of the hemostatic balance. These results indicate that the anti-migratory effect of theses compounds depends on the dosage and time of inflammatory stimulus. The HL and heparin were also able to inhibit the activity of the enzyme elastase. The discovery of this bioactive compound in the cephalothorax of shrimps can arouse great interest in biotechnology, since this compound could be useful as a structural model interesting for the development of new therapeutic agents for peritonitis

O ensino de conceitos de física quântica no ensino médio utilizando experimentos de baixo custo

Dissertação (mestrado)—Universidade de Brasília, Instituto de Física, Programa de Pós-Graduação em Física, Mestrado Nacional Profissional em Ensino de Física, 2015.

O compromisso de casar e o desejo de amar : experimentos existenciais entre jovens abastados na cidade de Nova Délhi

Tese (doutorado)—Universidade de Brasília, Instituto de Ciências Sociais, Departamento de Antropologia, Programa de Pós-graduação em Antropologia Social, 2015.

Microbiologia do ciclo do nitrogênio em solos do cerrado

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Microbiana, 2016.

Biologia do caranguejo-uçá, Ucides cordatus (Crustacea: Decapoda: Brachyura), na Baía da Babitonga, Santa Catarina, Brasil

O objetivo deste estudo é caracterizar pela primeira vez alguns aspectos da reprodução do caranguejo-uçá em manguezais da Baía da Babitonga (Santa Catarina). Além disso, a densidade e o tamanho do estoque deste recurso pesqueiro foram também estimados. Os exemplares foram coletados mensalmente, de maio de 2002 a abril de 2003, em duas áreas distintas: Iperoba e Palmital; um total de 2265 espécimes (1623 machos e 642 fêmeas) foi analisado. Os machos com gônadas maturas foram registrados durante todo o ano, enquanto as fêmeas com gônadas maturas ocorreram em apenas cinco meses. As fêmeas ovígeras foram registradas apenas em dezembro e janeiro. O etograma do fenômeno de migração reprodutiva (andada) esteve em concordância com a maior atividade de caranguejos associada às luas cheias e novas, com maior intensidade em dezembro e janeiro, relacionados ao verão austral. A densidade total no Manguezal de Iperoba foi de 2,05 ± 0,97 ind./m², não diferindo significativamente daquela registrada para o Manguezal do Palmital (2,06 ± 1,08 ind./m²) (p < 0,05). A média global para a estimativa de densidade na Baia da Babitonga foi de 2,05 ± 1,00 ind./m², correspondendo a 1,42 ± 0,89 ind./m² com base nas galerias abertas e 0,64 ± 0,63 ind./m² para as galerias fechadas.

Biologia da traca do tomateiro sob condicoes de laboratorio.

Estudaram-se, no CPATSA-EMBRAPA em Petrolina, PE, sob condicoes de laboratorio, os aspectos biologicos de Scrobipalpula absoluta (Meyrick 1917) (Lepidoptera,Gelechiidae), importante praga do tomateiro, Iniciou-se a criacao com larvas coletadas no campo, em cultura de tomate, acondicionadas, em placas de Petri, sobre papel de filtro e folhas de tomateiro, variedade IPA-2. Os adultos foram mantidos em frascos de vidro revestidos com papel sulfite, tampados com tecido branco de nailon preso com elastico e alimentados, com uma solucao de sacarose a 10%, embebida em algodao. Obtiveram-se os seguintes dados: periodo de pre-ovoposicao 2,35 dias; numero de ovos por femea 55,16; viabilidade de ovos 44,46%, periodo de incubacao 4,30 dias; periodo de oviposicao 3,77 dias; periodo larval 10,95 dias com quatro instares; viabilidade larval 20,97%; periodo pupal 6,15 dias; viabilidade pupal 68,19%; longevidade de adultos machos e femeas 9,69 e 11,52 dias, respectivamente; ciclo biologico (ovo-morte do adulto) 38,12 dias.

Aplicação da metodologia de planejamento de experimentos para formulação de massas cerâmicas para telhas

Rio Grande do Norte State stands out as one great producer of structural clay of the brazilian northeastern. The Valley Assu ceramic tiles production stands out obtained from ilitics ball clays that abound in the region under study. Ceramics formulation and the design of experiments with mixture approach, has been applied for researchers, come as an important aid to decrease the number of experiments necessary to the optimization. In this context, the objective of this work is to evaluate the effects of the formulation, temperature and heating rate in the physical-mechanical properties of the red ceramic body used for roofing tile fabrication of the Valley Assu, using design of mixture experiments. Four clays samples used in two ceramics industry of the region were use as raw material and characterized by X-ray diffraction, chemical composition, differential thermal analysis (DTA), thermogravimetric analysis (TGA), particle size distribution analysis and plasticity techniques. Afterwards, they were defined initial molded bodies and made specimens were then prepared by uniaxial pressing at 25 MPa before firing at 850, 950 and 1050 ºC in a laboratory furnace, with heating rate in the proportions of 5, 10 e 15 ºC/min. The following tecnologicals properties were evaluated: linear firing shrinkage, water absorption and flexural strength. Results show that the temperature 1050 ºC and heating rate of 5 ºC/min was the best condition, therefore presented significance in all physical-mechanical properties. The model was accepted as valid based of the production of three new formulations with fractions mass diferents of the initial molded bodies and heated with temperature at 1050 ºC and heating rate of 5 ºC/min. Considering the formulation, temperature and heating rate as variables of the equations, another model was suggested, where from the aplication of design of experiments with mixtures was possible to get a best formulation, whose experimental error is the minor in relation to the too much formulations

Análise da interação entre moléculas desenhadas a partir do ácido anacárdico e a proteína PPAR usando ferramentas in silico

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Molecular, 2016.

Estudo da participação de microRNAs na regulação da resposta imune inata de macrófagos murinos à infecção por Paracoccidioides brasiliensis

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Pós-Graduação em Biologia Molecular, 2016.

Toxicidade in vitro e in vivo de quantum dots de carbono recobertos com boronato

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2016.

Ecologia parasitária de isópodos e biologia reprodutiva em tibiro, Oligoplites spp (Osteichthyes: carangidae) das águas costeiras de Natal, Rio Grande do Norte

Oligoplites palometa (Cuvier) and Oligoplites saurus (Bloch & Schneider) (Osteichthyes: Carangidae) are coastal pelagic fish species. The objective of this study was to investigate the effect of parasitism by isopods and the reproductive biology of the leather jack, Oligoplites spp. A total of 113 individuals of Oligoplites spp (35 of O. saurus and 78 of O. palometa) were captured during the period between January, 2005 and July, 2007 from the coastal waters of Natal, RN. The morphometric and meristic characters were registered; macroscopic analyses were done to collect the isopod parasites and the gonads were observed to verify the reproductive aspects. The crustacean isopod parasite, Cymothoa spinipalpa (Isopoda: Cymothidae) was encountered on the tongue in the oral cavity in both fish hosts. The parasitic indices of C. spinipalpa showed high values of prevalance of 51,4% for O. saurus and 64,1% for O. palometa. The host presented significant correlations between body mass and body length of O. palometa and the number of parasites. Body lengths of female C. spinipalpa were small due to the limited space available in the oral cavity of the host. There was a significant difference in the sex ratio of O. palometa, males outnumbered the females. The isopod parasites showed specificity to the oral cavity of hosts. Macroscopic analyses of gonads showed four stages of gonadal development for both sex of O. palometa: immature, maturing, mature and spent

Aspectos da biologia reprodutiva de Eretmochelys imbricata (testudines, cheloniidae) no litoral sul do Rio Grande do Norte, Brasil

Female hawksbill turtles (Eretmochelys imbricata) nesting along the southeastcoastline of Rio Grande do Norte State, Brazil (6º13'40"S, 35º03'05"W) were captured and weighed during the four months from January to April 2007, in the course of the annual egglaying season, which extended from 06 rd November 2006 to 30 rd May 2007. In all, 99 weight measurements were performed. On first contact the females exhibited an average post-oviposition weight of 79.1 kg (range 56.2-98.9 kg, SD = 10.9 kg, n = 44 females). Those individuals which were subsequently recaptured showed a mean weight loss of 1.7 kg (range 0.7-4.5 kg, SD = 1.0 kg, n = 39 sets of measurements on 20 females) in the interval between two consecutive post-ovipositions, separated by a maximum time interval of 17 days. In the cases where the female aborted the nesting process, the pre-oviposition weight was measured. The clutch weight, that is to say, the weight loss between consecutive pre-oviposition and post-oviposition measurements (separated by a maximum time interval of 3 days), was found to be 5.2 kg (range 4.3-6.0 kg, SD = 0.9 kg, n = 6 sets of measurements). This value is significantly higher (t-test, p<0.001) than the loss between two consecutive post-oviposition measurements with the same female. The mean recovery in body weight, that is to say, the average gain in weight between successive post-oviposition and pre-oviposition captures of the same individual (separated by a time interval of 12 to 17 days), was found to be 3.0 kg (range 1.9-4.3 kg, SD = 1.0 kg, n = 4 sets of measurements) Although the small sample size makes it unwise to generalise, the recovery in body weight was found to be always significantly lower (t-test, p<0.005) than the clutch weight. This fact is in agreement with the observed weight loss tendency throughout the breeding season for this species. Considering the clutch weight and the internidal recovery in body weight we found that the total weight loss of the adult hawksbill females after three to five nesting events varied from 10.4% (range 8.7-11.9%, SD = 1.6%, n = 3) to 14.1% (range 11.8-15.4%, SD = 1.3%, n = 6) in relation to their initial pre-oviposition weight. If there were no body weight recovery during the internesting interval we estimate that a female that nests three to five times in the course of the season would lose from 19% to 31% of its initial weight. We emphasise that our clutch weight estimate was performed by weighing the females and not by multiplying the number of eggs in the nest by their average unit weight. In this way, our measurements take into account the loss of liquid during the oviposition. Despite the unequivocal evidence of body weight recovery during the internidal interval, it is not clear if the cause of this process is rehydration or feeding

Atividade antimalárica de plantas medicinais da biorregião do Araripe-CE em modelo murino - Plasmodium berghei

Malaria, also popularly known as maleita , intermittent fever, paludism, impaludism, third fever or fourth fever, is an acute infectious febrile disease, which, in human beings, is caused by four species: Plasmodium falciparum, P. vivax, P. malariae and P. ovale. Malaria, one of the main infectious diseases in the world, is the most important parasitoses, with 250 million annual cases and more than 1 million deaths per year, mainly in children younger than live years of age. The prophylactic and therapeutic arsenal against malaria is quite restricted, since all the antimalarials currently in use have some limitation. Many plant species belonging to several families have been tested in vivo, using the murine experimental model Plasmodium berghei or in vitro against P. falciparum, and this search has been directed toward plants with antithermal, antimalarial or antiinflammatory properties used in popular Brazilian bolk medicine. Studies assessing the biological activity of medicinal plant essential oils have revealed activities of interest, such as insecticidal, spasmolytic and antiplasmodic action. It has also been scientifically established that around 60% of essential oils have antifungal properties and that 35% exhibit antibacterial properties. In our investigation, essential oils were obtained from the species Vanillosmopsis arborea, Lippia sidoides and Croton zethneri which are found in the bioregion of Araripe-Ceará. The chemical composition of these essential oils was partially characterized and the presence of monoterpenes and sesquiterpenes. The acute toxicity of these oils was assessed in healthy mice at different doses applied on a single day and on four consecutive days, and in vitro cytotoxicity in HeLa and Raw cell lines was determined at different concentrations. The in vivo tests obtained lethal dose values of 7,1 mg/Kg (doses administered on a single day) and 1,8 mg/Kg (doses administered over four days) for 50% of the animals. In the in vitro tests, the inhibitory concentration for 50% of cell growth in Hela cell lines was 588 μg/mL (essential oil from C. zethneri after 48 h), from 340-555 μg/mL (essential oil from L. sidoides, after 24 and 48 h). The essential oil from V. arborea showed no cytotoxicity and none of the essential oils were cytotoxic in Raw cell lines. These data suggest a moderate toxicity in the essential XVIII oils under study, a finding that does not impede their testing in in vivo antimalarial assays. Was shown the antimalarial activity of the essential oils in mice infected with P. berghei was assessed. The three species showed antimalarial activity from 36%-57% for the essential oil from the stem of V. arborea; from 32%-82% for the essential oil from the leaves of L. sidoides and from 40%-70% of reduction for the essential oil from the leaves of C. zethneri. This is the first study showing evidence of antimalarial activity with these species from northeast Brazil. Further studies to isolate the active ingredients of these oils are needed to determine if a single active ingredient accounts for the antimalarial activity or if a complex integration of all the compounds present occurs, a situation reflected in their biological activity