Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

GOMES, Carlos E. M. et al. Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly). Plant Physiology and Biochemistry (Paris), v. 43, n. 12, p. 1095-1102, 2005.ISSN 0981-9428. DOI:10.1016/j.plaphy.2005.11.004.

Comparative study between the effects of hyaluronic acid and acid galactan purified from eggs of the mollusk Pomacea sp in wound healing

To compare the effect of hyaluronic acid (HA) and of AG on the healing of intestine wounds. Methods: The semi-purified extract of the eggs of the mollusc was obtained by fractionation with ammonium sulfate and purification for ion-exchange chromatography. The obtained galactans were eluted in water (neutral galactan) and in 0.1 and 0.2M NaCl (acidic galactans). The in vivo study was performed with 45 “Wistar” rats, separated in three groups (n=15). Solutions containing HA 1%, GA 1% or saline solution 0,9%, was placed topically on the sutures of wounds in the small intestine of the rats. After 05, 10 and 21 days the animals were sacrificed and biopsy of the healing tissue was done. Results: The hystologic grading was more significant for HA and AG groups when compared to the group C. AG stimulated the appearance of macrophages, giant cells and increase in the concentration of collagen in the area of the wound when compared to HA. Conclusion: The topical use of GA in intestinal wounds promoted the anticipation of events that are important in the wound healing

Purificação e caracterização parcial de uma B-D glicosidade de Artemia franciscana com ação sobre celobiose e lactose

-D-glucosidase (EC 3.2.1.21) is one of the most interesting glycosidases, especially for hydrolysis cellobiose releasing glucose, is last step degradation of cellulose. This function makes the -D-glucosidase is of great interest as a versatile industrial biocatalyst, being critical to various bio-treatment / biorefinery processes, such as bioethanol production. Hen in the report, a -D-glucosidase was extracts from protein extracted of the invertebrate marine Artemia franciscana was purified and characterized with a combination of precipitation with ammonium sulfate (0 - 30%, 30 to 50%, 50 to 80%), the fraction saturated in the range of 30 to 50% (called F-II) was applied in a molecular exclusion chromatography, in Sephacryl S-200, the fractions corresponding to the first peak of activity of -D-glucosidase were gathered and applied in a chromatography of ion exchange in Mono Q; the third peak this protein obtained chromatography, which coincides with the peak of activity of -D-glucosidase was held and applied in a gel filtration chromatography Superose 12 where the first peak protein, which has activity of -D-glucosidase was rechromatography on Superose 12. This enzyme is probably multimerica, consisting of three subunit molecular mass of 52.7 kDa (determined by SDS-PAGE) with native molecular mass of 157 kDa (determined by gel filtration chromatography on Superose 12 under the system FPLC). The enzyme was purified 44.09 times with a recovery of 1.01%. Using up p-nitrophenyl-β-D-glucopiranoside as substrate obtained a Km apparent of 0.229 mM and a Vmax of 1.109 mM.60min-1.mL-1mM. The optimum pH and optimum temperature of catalysis of the synthetic substrate were 5.0 and 45 °C, respectively. The activity of the -D-glucosidase was strongly, inhibited by silver nitrate and N- etylmaleimide, this inhibition indicates the involvement of radical sulfidrila the hydrolysis of synthetic substrate. The -D-glucosidase of Artemia franciscana presented degradativa action on celobiose, lactose and on the synthetic substrate -nitrophenyl-β-D-glucopiranoside indicating potential use of this enzyme in the industry mainly for the production of bioethanol (production of alcohol from the participating cellulose), and production hydrolysate milk (devoid of milk lactose)

Atividade β-D-N-Acetilglucosaminidásica de enzimas imobilizadas extraídas da Artemia franciscana e possíveis aplicações biotecnológicas

A β-D-N-acetilglucosaminidase extracted and partially isolated from crustacean Artemia franciscana by ammonium sulfate precipitation and filtration gel chromatography Bio Gel A 1.5m. the enzyme was immobilized on ferromagnetic Dacron yielding a insoluble active derivative with 5.0 units/mg protein and 10.35% of the soluble enzyme activity. β-D-N-acetilglucosaminidase-ferromagnetic Dacron was easily removed from the reaction mixture by a magnetic field, it was reused for ten times without loss in its activity. The ferromagnetic Dacron was better activated at pH 5.0. The particles visualized at scanning electron microscope (SEM) had presented different sizes, varying between 721nm and 100µm. Infra red confirmed immobilization on support, as showed by primary amino peaks at 1640 and 1560 cm-1 . The immobilize enzyme presented Km of 2.32 ± 0.48 mM and optimum temperature of 50°C. Bought presented the same thermal stable of the soluble enzyme and larger enzymatic activity at pH 5.5. β-D-N-acetilglucosaminidase-Dacron ferromagnético showed sensible for some íons as the silver (AgNO3), with loss of activity. The β-D-N acetilglucosaminidase activity for mercury chloride (HgCl2), whom is one of the most toxic substance joined in nature, it was presented activity already diminished at 0,01mM and lost total activity at 4mM, indicating sensitivity for this type of metal. β-D-N-acetilglucosaminidase-ferromagnetic Dacron showed degradative capacity on heparan sulfate, the enzyme still demonstrated degradative capacity on heparan sulphate, suggesting a possible application to produce fractions of this glycosaminoglycan

Efeito inseticida de vicilinas isoladas de sementes de erythrina velutina em condições de semi-campo para moscas das frutas (ceratitis capitata)

Chitin-binding vicilins from legume seeds (Erythrina velutina. Canavalia ensiformes and Phaseolus vulgares) were isolated by ammonium sulfate followed by affinity chromatography on a chitin column. Effect of these vicilins on female adults of Ceratitis capitata was examined by bioassay and in a semi-field assay model. Mechanism of action of the vicilins was determined by in vivo digestibility and chitin affinity. Among the tested vicilins, E. velutina when added to diet caused strong effect on mortality at 10% dose. This insecticidal property was tested in a semi-field assay which showed the same effect observed in laboratory conditions, where doses of 10% and 15% were lethal to female adults of C. capitata. These deleterious effects were not only associated to the binding to chitin structures present in peritrophic membrane, but principally to its low digestibility in the C. capitata digestive tract. This fact was confirmed because chiting binding proteins as WGA and the other tested vicilins were not toxic to female adults of C. capitata due susceptibility of these proteins to digestive enzymes of the insects. By other side EvV was more resistant to digestive enzymes, causing deleterious effects on female adults of C. capitata. These results showed that EvV may be part of the pest management programs or an alternative in plant improvement program in the population control of this fruticulture pest

Avaliação das atividades anti-inflamatória, anticoagulante e antiproliferativa do inibidor de quimotripsina das sementes de erythrina velutina (EvCI)

Studies indicate that several components were isolated from medicinal plants, which have antibacterial, antifungal, antitumor and anti-inflammatory properties. Sepsis is characterized by a systemic inflammation which leads to the production of inflammatory mediators exacerbated by excessive activation of inflammatory cells and disseminated intravascular coagulation (DIC), in which the human neutrophil elastase plays an important role in its pathogenesis. Several epidemiological studies suggest that components of plants, especially legumes, can play a beneficial role in reducing the incidence of different cancers. A chymotrypsin inhibitor of Kunitz (Varela, 2010) was purified from seeds of Erythrina velutina (Mulungu) by fractionation with ammonium sulfate, affinity chromatography on Trypsin-Sepharose, Chymotrypsin-Sepharose and ion exchange chromatography on Resource Q 1 ml (GE Healthcare) in system FPLC / AKTA. The inhibitor, called EvCI, had a molecular mass of 17 kDa determined by SDS-PAGE. The purified protein was able to inhibit human neutrophil elastase (HNE), with an IC50 of 3.12 nM. The EvCI was able to inhibit both pathways of HNE release stimulated by PAF and fMLP (75.6% and 65% respectively). The inhibitor also inhibited leukocyte migration in septic mice about 87% and prolonged the time of coagulation and inhibition factor Xa. EvCI showed neither hemolytic activity nor cytotoxicity. EvCI showed a selective antiproliferative effect to HepG2 cell lines with IC50 of 0.5 micrograms per milliliter. These results suggest EvCI as a molecule antagonist of PAF / fMLP and a potential use in fighting inflammation related disorders, disseminated intravascular coagulation (DIC) and cancer

Lectina da esponja marinha Tedania ignis: purificação, caracterização e interação com leishmanias

Lectin obtained from the marine sponge Tedania ignis was purified and characterized by extraction of soluble proteins (crude extract) in 50mM Borax, pH 7.5. The purification procedure was carried out by crude extract precipitation with ammonium sulfate 30% (FI). The precipitated was resuspended in the same buffer and fractionated with acetone 1.0 volume (F1.0). A lectin was purified from this specific fraction by using an affinity chromatography Sepharose 6B. This lectin preferentially agglutinated human erythrocytes from B type previously treated with papain enzyme. The hemagglutinating activity lectin was dependent of divalent Mn2+ cation and was inhibited by the carbohydrates galactose, xylose and fructose. SDS-PAGE analysis indicated a molecular mass of the lectin around 45 kDa. This protein showed stability until 40°C for 1 h. Further, it showed activity between pH 2.5 and 11.5, with an enhanced activity at pH 7.5. Leishmania chagasi promastigotes stained with Coomassie brilliant blue R-250 were agglutinated by F1,0 and in the presence of galactose this interaction was abolished. These results show that this lectin could be implicated in defense procedures and it will can be used as biological tools in studies with this protozoon

Purificação e caracterização de uma ß-N-acetillhexosaminadase extraída do mamífero marinho Sotalia fluviatilis

This report shows 2232 times purification of a βNAcetylhexosaminidase from hepatic extracts from the sea mammal Sotalia fluviatilis homogenate with final recovery of 8,4%. Sequenced steps were utilized for enzyme purification: ammonium sulfate fractionation, Biogel A 1.5 m, chitin, DEAESepharose and hydroxyapatite chromatographies. The protein molecular mass was estimated in 10 kDa using SDSPAGE and confirmed by MALDITOF. It was found to have an optimal pH of 5.0 and a temperature of 60°C. Using pnitrophenylNAcetylβDglycosaminide apparent Km and Vmax values were of 2.72 mM and 0.572 nmol/mg/min, respectively. The enzyme was inhibited by mercury chloride (HgCl2) and sodium dodecil sulfate (SDS)

Purificação e caracterização parcial de duas n-acetil-B-hexosaminidases do equinoderma marinho echinometra lucunter

Two b-N-acetylhexosaminidases (F11 e F15) were purified from Echinometra lucunter gonads extracts. The purified enzymes were obtained using ammonium sulfate fractionation, followed by gel filtration chromatographies (Sephacryl S-200, Sephadex G-75 and Sephacryl S-200). The F11 fraction was purified 192.47 -fold with a 28.5% yield, and F15 fraction 85.41 -fold with a 32.3% yield. The molecular weights of the fractions were 116 kDa for F11 and 42 kDa for F15 using SDS-PAGE. In Sephacryl S-200, F15 was 84 kDa, indicating that it is a dimeric protein. When p-nitrophenyl-β-D-glycosaminide was used as substrate, we determined an apparent Km of 0.257 mM and Vmax of 0.704 for F11 and for F15 the Km was 0.235 mM and Vmax of 0.9 mM of product liberated by hour. Both enzymes have optimum pH and temperature respectively at 5.0 and 45 °C. The enzymes showed inhibition by silver nitrate, while the glucuronic acid was a potent activator. The high inhibition of F15 by N-etylmaleimide indicates that sulphydril groups are involved in the catalysis of synthetic substrate

Influência da chuva na eficácia do glyphosate em mistura com adjuvantes na dessecação de plantas daninhas

Com o objetivo de estudar a influência do momento da chuva após a aplicação do herbicida glyphosate isolado e em mistura com adjuvantes na dessecação de plantas daninhas, foram instalados dois experimentos na Fazenda Experimental de Ensino e Pesquisa da Faculdade de Ciências Agrárias e Veterinárias, UNESP - campus de Jaboticabal-SP, em duas épocas: inverno de 2000 (junho - agosto) e verão de 2001 (janeiro-março). Os experimentos foram instalados no delineamento experimental de blocos ao acaso, em arranjo fatorial 4 (tratamentos herbicidas) x 5 (períodos livres de chuva simulada) + 1, com quatro repetições. Os tratamentos consistiram da combinação de glyphosate SAqC (360 g e.a. ha-1) isolado ou adicionado de uréia (50 g L-1 de calda) ou óleo vegetal (100 ml L-1 de calda) ou sulfato de amônio (100 g L-1 de calda), com cinco períodos de chuva simulada (1, 2, 4, 6 e > 48 horas após a aplicação), além de uma testemunha. A chuva foi simulada com um sistema de irrigação por aspersão, e a quantidade aplicada variou entre 18 e 19 mm. em ambas as épocas a chuva foi prejudicial à ação do glyphosate, principalmente nos menores períodos livres de chuva após a aplicação. Os sintomas de fitointoxicação apareceram mais rapidamente no verão. A utilização de adjuvantes na calda de pulverização não beneficiou o desempenho do herbicida glyphosate no controle das plantas daninhas, no inverno. A adição de uréia (50 g L¹) é uma boa alternativa para o controle de plantas daninhas, no verão, em situações sujeitas à chuva até duas horas após a aplicação.

ATIVIDADE MICROBIANA E PROPRIEDADES BIOQUÍMICAS do SOLO RESULTANTES da APLICAÇÃO DE GESSO AGRÍCOLA NA CULTURA do REPOLHO

Conselho Nacional do Desenvolvimento Científico e Tecnológico (CNPq)

Manejo da adubação nitrogenada paraa batata-doce: fontes e parcelamento de aplicação

Com o objetivo de avaliar o rendimento da batata-doce [Ipomoea batatas (L.) Lam.], cultivar Rainha Branca, submetida a fontes e parcelamentos da aplicação de 80 kg ha-1 de nitrogênio, instalou-se um experimento, no período de abril a agosto de 2007, na Universidade Federal da Paraíba, em Areia. O delineamento experimental utilizado foi o de blocos casualizados, em esquema fatorial 2 x 7, com duas fontes de nitrogênio (uréia e sulfato de amônio) e oito de épocas parcelamentos da aplicação (100% no plantio; 100% aos 30 dias após o plantio (DAP); 100% aos 60 DAP; 50% no plantio e 50% aos 30 DAP; 50% no plantio e 50% aos 60 DAP; 50% aos 30 e 50% aos 60 DAP; 33% no plantio, 33% aos 30 e 33% aos 60 DAP), com quatro repetições. Foram avaliados o peso médio de raízes comerciais, a produção e o número de raízes comerciais por planta e as produtividades total e comercial de raízes de batatadoce. O maior peso médio de raízes comerciais (294 g) foi obtido com o parcelamento do N, como uréia, 50% no plantio e 50% aos 30 DAP. O parcelamento de N 33% no plantio, 33% aos 30 DAP e 33% aos 60 DAP na forma sulfato de amônio foi responsável pela maior produção de raízes comerciais por planta (337 g). Para o número de raízes comercias e as produtividades total e comercial, o N na forma de sulfato de amônio parcelado 33% no plantio, 33% aos 30 DAP e 33% aos 60 DAP proporcionou os maiores valores, 1,50 raízes, 30,5 e 28,4 t ha-1, respectivamente.

Absorção e redistribuição de nitrogênio (15N) em Citrus mitis Bl

Com a avaliação da eficiência de uso do nitrogênio, tem-se melhor entendimento dos aspectos nutricionais e respostas à adubação. O presente ensaio teve por objetivo estudar a absorção e redistribuição de nitrogênio (15N) em Citrus mitis Bl.. As fontes de fertilizante utilizadas foram: sulfato de amônio, uréia, nitrato de cálcio e nitrato de potássio. O delineamento experimental utilizado foi inteiramente casualizado, com 4 tratamentos e 3 repetições. Foram realizadas duas amostragens, aos 10 e 20 dias após a aplicação do adubo marcado, a fim de determinar os teores de N nas diferentes partes da planta. Através dos resultados, verificou-se que não houve efeito dos tratamentos sobre o peso de matéria seca e conteúdo de N nas plantas. A eficiência de absorção de N variou com a natureza do fertilizante nitrogenado e com a época de amostragem, ao passo que a redistribuição do N não foi afetada. A eficiência máxima de absorção do N variou de 14% (uréia) e 31% (sulfato de amônio), respectivamente, aos 10 e 20 dias após a aplicação do 15N.

Fontes alternativas e doses de nitrogênio no milho safrinha em sucessão à soja

A produtividade do milho safrinha pode ser aumentada com a adubação nitrogenada em cobertura mesmo quando cultivado em sucessão à soja. No entanto, existem inconsistências dos resultados especialmente quanto às fontes e doses a serem empregadas nessa modalidade de cultivo. Com o objetivo de avaliar o efeito de fontes e doses de nitrogênio em cobertura no milho safrinha, cultivado após soja no sistema plantio direto, conduziu-se o experimento em um Latossolo Vermelho distrófico, em Chapadão do Céu (GO). O delineamento experimental foi o de blocos ao acaso, em esquema fatorial 4 x 4, com quatro repetições. Os tratamentos foram constituídos pela combinação de quatro fontes (uréia, sulfato de amônio, uréia extrusada com produtos amiláceos (Amiréia® 180S) e sulfonitrato de amônio com inibidor de nitrificação (Entec® 26)) e quatro doses de nitrogênio (0; 30; 60 e 120 kg ha-1). A interação fonte x dose não foi significativa para nenhuma das variáveis avaliadas. A aplicação de nitrogênio na forma de entec proporcionou maiores teores de nitrogênio na folha do milho safrinha que o sulfato de amônio e a amiréia. A produtividade de grãos do milho safrinha foi maior quando o nitrogênio em cobertura foi fornecido na forma de sulfato de amônio, em comparação com a amiréia. A aplicação de nitrogênio em cobertura aumentou os teores de nitrogênio e enxofre na folha, altura da planta, diâmetro do colmo, número de espigas por planta e grãos por espiga, massa de 1.000 grãos e produtividade de grãos do milho safrinha, em sucessão à soja, independentemente da fonte utilizada.

Manejo químico de espécies de trapoeraba com aplicação isolada e em mistura de diferentes herbicidas

This paper evaluated the effect of different herbicides applied isolated and mixed on two species of dayflower (Commelina benghalensis and Commelina villosa). Both species were grown under pot conditions with a capacity of 14 L. Three stems with 10 cm were planted per pot. The herbicides application was made using a stationary spray system, to provide a spray volume of 200 L ha(-1), with spray tip XR 110.02VS, when the plants were with dimensions of 25-35 cm for C. benghalensis and 30-35 cm for C. villosa. The mixture saflufenacil more effective control of C. benghalensis were saflufenacil + glyphosate (48 + 720 g ha(-1)) and, saflufenacil + (glyphosate + imazethapyr) with and without the addition of ammonium sulfate (24 + 623 g ha(-1)). The combination of carfentrazone + glyphosate (20 + 720 g ha(-1)) were also effective in the C. benghalensis control. The saflufenacil mixture allowed plant control of C. villosa, where saflufenacil + glyphosate (24 + 720 g ha(-1)) with the addition of ammonium sulfate, showed a good visually control of this species. The application of 2,4-D (720 g ha(-1)) and 2,4-D + glyphosate (720 + 720 g ha(-1)) provided the best visual controls of C. benghalensis e C. villosa. All chemical treatment regardless of Commelina species reduced the dry mass of dayflower.