RESUMO
The phospholipase A2 superfamily encompasses 15 groups that are classified into: secreted PLA2 (sPLA2); cytosolic PLA2 (cPLA2); Ca2+-independent intracellular PLA2 (iPLA2); platelet-activating factor acetylhydrolase (PAF-AH); and lysosomal PLA2. Currently, approximately 700 PLA2 sequences are known, of which 200 are obtained from the venom gland of Crotalinae snakes. However, thus far, little information is available on cloning, purification and structural characterization of PLA2 from Crotalus durisssus cascavela venom gland. In the present work, we report the molecular cloning of a novel svPLA2 from C. d. cascavella (Cdc), a predominant rattlesnake subspecies in northeastern Brazil. The Cdc svPLA2 cDNA precursor is 689 nucleotides long and encodes a protein of 138 amino acid residues, with a calculated molecular mass of approximately 13,847 Da and an estimated isoelectric point of 5.14. Phylogenetic analysis of Crotalinae PLA2 reveals that Cdc PLA2 clustered with other acidic type IIA PLA2 homologues is also present in the venom of North American rattlesnakes. Hitherto, this study presents a novel PLA2 cDNA precursor from C. d. cascavella and data reported herein will be useful for further steps in svPLA2 purification and analysis.(AU)
Assuntos
Animais , Masculino , Clonagem Molecular , Venenos de Crotalídeos , Fosfolipases A2RESUMO
The phospholipase A2 superfamily encompasses 15 groups that are classified into: secreted PLA2 (sPLA2); cytosolic PLA2 (cPLA2); Ca2+-independent intracellular PLA2 (iPLA2); platelet-activating factor acetylhydrolase (PAF-AH); and lysosomal PLA2. Currently, approximately 700 PLA2 sequences are known, of which 200 are obtained from the venom gland of Crotalinae snakes. However, thus far, little information is available on cloning, purification and structural characterization of PLA2 from Crotalus durisssus cascavela venom gland. In the present work, we report the molecular cloning of a novel svPLA2 from C. d. cascavella (Cdc), a predominant rattlesnake subspecies in northeastern Brazil. The Cdc svPLA2 cDNA precursor is 689 nucleotides long and encodes a protein of 138 amino acid residues, with a calculated molecular mass of approximately 13,847 Da and an estimated isoelectric point of 5.14. Phylogenetic analysis of Crotalinae PLA2 reveals that Cdc PLA2 clustered with other acidic type IIA PLA2 homologues is also present in the venom of North American rattlesnakes. Hitherto, this study presents a novel PLA2 cDNA precursor from C. d. cascavella and data reported herein will be useful for further steps in svPLA2 purification and analysis.
Assuntos
Animais , Masculino , Clonagem Molecular , Venenos de CrotalídeosRESUMO
RESUMO Avaliou-se a eficácia de fungicidas no controle da ferrugem do feijoeiro, em condições de campo. O experimento foi conduzido em Marília, SP, com plantas de feijão do grupo Carioca. Foram realizadas 3 pulverizações a cada 14 dias, utilizando-se 300 L de calda/ha, com os ingredientes ativos: trifloxystrobin + propiconazole (75 mL/ha), prothioconazole (100 mL ha-1); azoxystrobin (50 g ha-1) + nimbus 0,25% v/v e trifloxystrobin + tebuconazole (50 + 100, 60 + 120 e 75 + 150 mL ha-1). O controle proporcionado pelos ingredientes ativos foi avaliado nas folhas, sendo o índice de infecção calculado através de escala de notas dividida em nove níveis. Constatou-se q ue todos os fungicidas e doses utilizados foram eficientes no controle da ferrugem (Uromyces appendiculatus), proporcionando mais de 90% de eficiência. Na testemunha o índice de infecção chegou a 52% na terceira avaliação.
ABSTRACT The efficacy of fungicides for the controlof bean rust was evaluated, under field conditions. The experiment was conducted in Marília, SP (Brasil), using bean plants of the 'Carioca' group. Three sprayings were applied, each 14 days, with spray volume of 300 liters ha-1, with the following active ingredients: trifloxystrobin + propiconazole (75 mL ha-1), prothioconazole (100 mL ha-1), azoxystrobin (50 g ha-1) + nimbus 0,25% v/v and trifloxystrobin + tebuconazole (50 + 100, 60 + 120 and 75 + 150 mL ha-1). The disease control provided by active ingredients was evaluated on leaves, with its infection index calculated from a 9-degree disease index. All the fungicides and their doses used were efficient in controlling leaf bean rust (Uromyces appendiculatus), reaching more than 90% efficiency. In the control treatment the leaf infection index reached 52% at the third evaluation.
RESUMO
The first low-molecular-mass metalloprotease presenting prothrombin activating activity was purified from Bothrops insularis venom and named insularinase A. It is a single-chain protease with a molecular mass of 22 639 Da. cDNA sequence analysis revealed that the disintegrin domain of the precursor protein is post-translationally processed, producing the mature insularinase A. Analysis of its deduced amino acid sequence showed a high similarity with several fibrin(ogen)olytic metalloproteases and only a moderate similarity with prothrombin activators. However, SDS-PAGE of prothrombin after activation by insularinase A showed fragment patterns similar to those generated by group A prothrombin activators, which convert prothrombin into meizothrombin independently of the prothrombinase complex. In addition, insularinase A activates factor X and hydrolyses fibrinogen and fibrin. Chelating agents fully inhibit all insularinase A activities. Insularinase A induced neither detachment nor apoptosis of human endothelial cells and was also not able to trigger an endothelial proinflammatory cell response. Nitric oxide and prostacyclin levels released by endothelial cells were significantly increased after treatment with insularinase A. Our results show that, although its primary structure is related to class P-I fibrin(ogen)olytic metalloproteases, insularinase A is functionally similar to group A prothrombin activators.
Assuntos
Masculino , Humanos , Animais , Camundongos , Bothrops/classificação , Bothrops/metabolismo , Protrombina/metabolismo , Venenos de Crotalídeos/farmacologia , Venenos de Crotalídeos/isolamento & purificação , Venenos de Crotalídeos/química , Afibrinogenemia/metabolismo , Fator X/metabolismo , Sequência de AminoácidosRESUMO
Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with 131I by the chloramine T method, and separated in a Sephacryl S-200 column. The efficiency of labeling was 68%. Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (s.c.) with 0.3 ml (2.35 x 10(5) cpm/mouse) of 131I-venom (N = 42), 131I-albumin or 131I (controls, N = 28 each). Thirty minutes and 1, 3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% NaCl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption in the skin (51%) within the first 30 min compared to albumin (20.1%) and free iodine (8.2%). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach (3rd h), small intestine (3rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3rd h postinjection (1.98%). There was a low binding of labeled venom in the liver (< 0.54%), thyroid (< 0.11%) and lungs (< 0.08%), and no iodinated venom was detected in brain, heart, diaphragm, spleen or bladder. The low venom binding observed in most internal organs, comparable to that of albumin, suggests that B. erythromelas venom does not specifically target most internal organs. That is, the systemic effects of envenomation are mainly due to an indirect action.
Assuntos
Bothrops , Venenos de Crotalídeos/farmacocinética , Albuminas , Animais , Venenos de Crotalídeos/química , Venenos de Crotalídeos/toxicidade , Injeções Subcutâneas , Radioisótopos do Iodo/farmacocinética , Masculino , Camundongos , RatosRESUMO
Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with I by the chloramine T method, and separated in a Sephacrylr S-200 column. The efficiency of labeling was 68 percent. Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05 percent KI over a period of 10 days prior to the experiment, were inoculated dorsally(sc) with 03. ml (2.35 x 10(5) cpm/mouse) of I-venom (N=42), I-albumin or I (controls, N=28 each). Thirty minutes and 1, 3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85 percent NaCl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption in the skin (51 percent) within the first 30 min compared to albumin (20.1 percent) and free iodine (8.2 percent). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach (3rd h), small intestine (3rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0 percent by 12-18 h. Albumin tended to concentrate in muscle until the 3rd h postinjection (1.98 percent). There was a low binding of labeled venom is the liver (<0.54 percent), thyroid (<0.11 percent) and lungs (<0.08 percent), and no iodinated venom was detected in brain, heart, diaphragm, spleen or bladder. The low venom binding observed in most internal organs, comparable to that of albumin, suggests that B. erythromelas venom does not specifically target most internal organs. That is, the systemic effects of envenomation are mainly due to an indirect action.
Assuntos
Animais , Ratos , Masculino , Bothrops , Venenos de Crotalídeos/farmacocinética , Albuminas , Venenos de Crotalídeos/química , Venenos de Crotalídeos/toxicidade , Injeções Subcutâneas , CamundongosRESUMO
The effects of a plant regulator were studied for rangpur lime seedlings sprayed at 14-day intervals, from the 70th day until the 210th. The seedlings were sprayed with: T1 = 0; T2 = 25; T3 = 50; T4 = 100; T5 = 150 ppm of gibberellic acid. The applications of gibberellic acid on the seedlings of rangpur lime rootstocks resulted in significant increases in plant height (T5 = 150 ppm) and stem diameter (T2 = 25 ppm and T3 = 50 ppm). The seedlings reached the buddable size in shorter time through the positive effects of the plant regulator. The stem diameter was higher with gibberellic acid at concentrations of 25 and 50 ppm. Shortening this time would benefit nurserymen by reducing various production inputs and costs.
O presente trabalho foi instalado em condições de campo com plântulas de limão `Cravo'(Citrus limonia Osbeck), em delineamento inteiramente casualizado, com três repetições no esquema de parcelas sub-divididas, onde os tratamentos principais foram 5 concentrações de ácido giberélico: T1 = 0, T2 = 25, T3 = 50, T4 = 100 e T5 = 150 ppm, parceladas em quatro pulverizações. Os tratamentos secundários foram as coletas do material, realizadas dos 70 aos 210 dias, em intervalos de 14 dias. Pelos resultados obtidos, o fitorregulador promoveu efeito favorável com relação ao incremento do comprimento do caule, sendo o tratamento de 150 ppm superior aos demais. O diâmetro do caule atingiu maiores dimensões com aplicação de ácido giberélico 50 e 25 ppm.
RESUMO
The effects of a plant regulator were studied for rangpur lime seedlings sprayed at 14-day intervals, from the 70th day until the 210th. The seedlings were sprayed with: T1 = 0; T2 = 25; T3 = 50; T4 = 100; T5 = 150 ppm of gibberellic acid. The applications of gibberellic acid on the seedlings of rangpur lime rootstocks resulted in significant increases in plant height (T5 = 150 ppm) and stem diameter (T2 = 25 ppm and T3 = 50 ppm). The seedlings reached the buddable size in shorter time through the positive effects of the plant regulator. The stem diameter was higher with gibberellic acid at concentrations of 25 and 50 ppm. Shortening this time would benefit nurserymen by reducing various production inputs and costs.
O presente trabalho foi instalado em condições de campo com plântulas de limão `Cravo'(Citrus limonia Osbeck), em delineamento inteiramente casualizado, com três repetições no esquema de parcelas sub-divididas, onde os tratamentos principais foram 5 concentrações de ácido giberélico: T1 = 0, T2 = 25, T3 = 50, T4 = 100 e T5 = 150 ppm, parceladas em quatro pulverizações. Os tratamentos secundários foram as coletas do material, realizadas dos 70 aos 210 dias, em intervalos de 14 dias. Pelos resultados obtidos, o fitorregulador promoveu efeito favorável com relação ao incremento do comprimento do caule, sendo o tratamento de 150 ppm superior aos demais. O diâmetro do caule atingiu maiores dimensões com aplicação de ácido giberélico 50 e 25 ppm.
RESUMO
The effects of growth regulators (gibberellins and cytokmins) and potassium nitrate were evaluated for Citrus amblycarpa seed germination and rootstock growth. First an experiment was performed in a germinator, using as a medium for seed germination filter paper wetted with distiled water. Seeds were treated with growth regulators and potassium nitrate during 24 hours. The results obtained indicate that the treatment of 100 mg/1 of GA4 + GA7 + phenylmethylaminepurine was the best, with a percentage of germination of 88% at 21 days. Based on the results obtained in the germinator, seeds of Citrus amblycarpa were seeded in soil and when the seedlings were 6 cm high they were sprayed with 50 mg/1 of GA4 + GA7 + phenylmethylaminepurine; 100 mg/1 of GA4 + GA7 + phenylmethylaminepurine and 150 mg/1 of GA4 + GA7 + phenylmethylaminepurine. Dry weights of leaves and stems, leaf area and height of plants were evaluated. The analysis of these parameters showed that the GA4 + GA7 + phenylmethylaminepurine at 50 mg/1 was the best treatment.
Foram avaliados os efeitos de fitorreguladores (giberelinas e citocininas) e nitrato de potássio (KNO3), na germinação de sementes e na promoção do crescimento de "seedlings" do porta-enxerto 'Nasnaran' (Citrus amblycarpa Ochese). Realizou-se primeiramente o experimento em germinador, contendo papel de filtro umedecido com água destilada e gerbox, como meio para a germinação das sementes. Os tratamentos foram realizados mediante imersão das sementes, em soluções preparadas com os fítorreguladores e KNO3, durante 24 horas. Através dos resultados obtidos, concluiu-se que o tratamento com 100 mg/1 de GA4 + GA7 + fenilmetilaminopurina, apresentou a maior porcentagem de germinação (88%), num tempo médio de 21 dias. Tomando como base o resultado do germinador, sementes de Citrus amblycarpa foram semeadas em canteiros e quando as plantas atingiram a altura de 6,0 cm receberam 2 pulverizações com 0; 50; 100 e 150 mg/1 de GA4 + GA7 + fenilmetilami-nopurina, com intervalo de 21 dias. Avaliou-se a matéria seca total, caule e folhas, área foliar e altura das plantas, sendo que a análise desses parâmetros mostrou que o GA4 + GA7 + fenilmetUaminopurina na concentração de 50 mg/1 foi o tratamento que apresentou os melhores resultados.
RESUMO
In order to evaluate qualitative and nutritional aspects of fig-trees with respect to six irrigation and six nitrogen levels, at Ilha Solteira, SP, Brazil, an experiment was carried out in randomized blocks, with subdivided plots, and four replications. Results showed that in four dates during harvest, only the first analysis (January 2, 1991) showed influence of nitrogen fertilization on fruit soluble solids (brix). There was no significant effect of treatments on pulp/peel relation for the four harvestings. In relation to leaf macronutrient concentration at flowering, water supply influenced N, P and Ca concentrations, and nitrogen influenced only Ca concentration. For an average of 10 t.ha-1 of mature fruit and 1.3 t.ha-1 of immature fruit production, there was a nutrient export of about 65 kg.ha-1 of N; 10 kg.ha-1 of P2O5; 44 kg.ha-1 of K2O; 35 kg.ha-1 of Ca and 9 kg.ha-1 of Mg.
Para avaliar aspectos qualitativos e nutricionais da figueira em relação a seis níveis de irrigação e de nitrogênio, desenvolveu-se um experimento em blocos ao acaso, com parcelas subdivididas e com quatro repetições, em Ilha Solteira,SP. Os resultados mostraram que em quatro datas de colheita, apenas a primeira análise (2 de janeiro de 91) mostrou influência da fertilização nitrogenada sobre os sólidos solúveis (brix) dos frutos. Não houve efeito significativo dos tratamentos sobre a relação polpa/casca, nas quatro colheitas. Com relação à concentração de macronutrientes nas folhas na época de florescimento, o suprimento de água influenciou as concentrações de N, P e Ca e a aplicação de nitrogênio influenciou apenas a concentração de Ca. Para uma produção média de 10 t.ha-1 de frutos maduros e de 1,3 t.ha-1 de frutos verdes, observou-se uma exportação de aproximadamente 65 kg.ha-1 de N; 10 kg.ha-1 de P2O5 ; 44 kg.ha-1 de K2O; 35 kg.ha-1 de Ca e 9 kg.ha-1 de Mg.