Clonagem e expressão do gene que codifica o inibidor de quimotripsina de Erythrina velutina WILLD. - caracterização e avaliação de seu potencial farmacológico

Proteinases are enzymes distributed widely founded in several organisms and perform many different functions, from maintaining homeostasis to the worsening of some diseases such as cancer, autoimmune diseases and infections. The proteins responsible of controlling the action of these enzymes are the inhibitors, that are classified based on their target proteases and are founded since simple organisms, such as bacteria, to higher organisms, such as larger plants and mammals. Plant proteinase inhibitors act by reducing or inactivating the activity of target proteases, thus, these proteins have been studied as potential tools in the treatment of diseases related to protease activities. In this context, an inhibitor of chymotrypsin from Erythrina velutina, called EvCI was previously purified and it was observed that this protein plays in vitro anticoagulant activity and anti-inflammatory activity in in vivo model. Aiming to reduce the environmental impact caused by the purification EvCI in high amounts and to facilitate the process of obtaining this protein, the recombinant chymotrypsin inhibitor from Eryhrina velutina was produced after cloning and expression in Escherichia coli. The bacteria were grown in LB medium and after induction of the expression this material was subjected to procedures for cell lysis and the product was applied on Nickel-affinity column. The proteins adsorbed were digested by thrombin and applied on Chymotrypsin-Sepharose affinity column, obtaining the purified inhibitor, named recEvCI. After electrophoresis, the recombinant inhibitor showed an approximately molecular mass of 17 kDa, and reduced the chymotrypsin and elastase activities in vitro. The recombinant inhibitor was sequenced and was found similar amino acids residues when compared to other inhibitors deposited in the database, with some modifications. recEvCI showed high stability under pH variations and reducing conditions, maintaining its activity around 80%. This protein increased the blood coagulation time in vitro by acting on the intrinsic pathway and did not show cytotoxicity against strains of mouse 3T3 fibroblasts and RAW 264.7 macrophages. recEvCI showed microbicide activity related to release of nitric oxide and consequently the activation of macrophages, futhermore having proinflammatory effects assessed by increased release of TNF-α. These results indicate that recEvCI can be biotechnologically used as a new tool in the control of coagulation-related diseases as well as can be an activating agent of the immune system in immunosuppressed individuals

Clonagem e expressão do gene que codifica o inibidor de quimotripsina de Erythrina velutina WILLD. - caracterização e avaliação de seu potencial farmacológico

Proteinases are enzymes distributed widely founded in several organisms and perform many different functions, from maintaining homeostasis to the worsening of some diseases such as cancer, autoimmune diseases and infections. The proteins responsible of controlling the action of these enzymes are the inhibitors, that are classified based on their target proteases and are founded since simple organisms, such as bacteria, to higher organisms, such as larger plants and mammals. Plant proteinase inhibitors act by reducing or inactivating the activity of target proteases, thus, these proteins have been studied as potential tools in the treatment of diseases related to protease activities. In this context, an inhibitor of chymotrypsin from Erythrina velutina, called EvCI was previously purified and it was observed that this protein plays in vitro anticoagulant activity and anti-inflammatory activity in in vivo model. Aiming to reduce the environmental impact caused by the purification EvCI in high amounts and to facilitate the process of obtaining this protein, the recombinant chymotrypsin inhibitor from Eryhrina velutina was produced after cloning and expression in Escherichia coli. The bacteria were grown in LB medium and after induction of the expression this material was subjected to procedures for cell lysis and the product was applied on Nickel-affinity column. The proteins adsorbed were digested by thrombin and applied on Chymotrypsin-Sepharose affinity column, obtaining the purified inhibitor, named recEvCI. After electrophoresis, the recombinant inhibitor showed an approximately molecular mass of 17 kDa, and reduced the chymotrypsin and elastase activities in vitro. The recombinant inhibitor was sequenced and was found similar amino acids residues when compared to other inhibitors deposited in the database, with some modifications. recEvCI showed high stability under pH variations and reducing conditions, maintaining its activity around 80%. This protein increased the blood coagulation time in vitro by acting on the intrinsic pathway and did not show cytotoxicity against strains of mouse 3T3 fibroblasts and RAW 264.7 macrophages. recEvCI showed microbicide activity related to release of nitric oxide and consequently the activation of macrophages, futhermore having proinflammatory effects assessed by increased release of TNF-α. These results indicate that recEvCI can be biotechnologically used as a new tool in the control of coagulation-related diseases as well as can be an activating agent of the immune system in immunosuppressed individuals

Rubiaceae em um remanescente de floresta atlântica no Rio Grande do Norte, Brasil

Empresa de Pesquisa Agropecuária do Rio Grande do Norte

Padrões distintos de congruência climática em duas espécies invasoras de prosopis em zonas semi-áridas da América do Sul

Based on climate data and occurrence records, ecological niche models (ENM) are an important opportunity to identify areas at risk or vulnerable to biological invasion. These models are based on the assumption that there is a match between the climatic characteristic of native and invaded regions predicting the potential distribution of exotic species. Using new methods to measure niche overlap, we chose two exotic species fairly common in semi-arid regions of South America, Prosopis juliflora (Sw.) D.C. and Prosopis pallida (H. ; B. ex. Willd) HBK, to test the climate matching hypothesis. Our results indicate that both species occur with little niche overlap in the native region while the inverse pattern is observed in the invaded region on South America, where both species occur with high climatic overlap. Maybe some non-climate factor act limiting the spread of P. pallida on the native range. We believe that a founder effect can explain these similarities between species niche in the invaded region once the seeds planted in Brazil came from a small region on the Native range (Piura in Peru), where both species occur sympatric. Our hypothesis of a founder effect may be evident when we look at the differences between the predictions of the models built in the native and invaded ranges. Furthermore, our results indicate that P. juliflora shows high levels of climate matching between native and invaded ranges. However, conclusions about climate matching of P. pallida should be taken with caution. Our models based on climatic variables provide multiple locations suitable for occurrence of both species in regions where they still don t have occurrence records, including places of high interest for conservation.

O Gênero Cyathus haller: pers. (Agaricales, Basidiomycota) em áreas de caatinga do Nordeste brasileiro

Fungi of the family Nidulariaceae or bird s nest fungi present a vase-shaped basidiomata with the wall surface smooth or striated lengthwise and internal structures resembling small bird eggs in a nest, the peridioles. Among the five genera that comprise the group, the more representative is Cyathus Haller: Pers. The Brazilian Northeast region shows major importance for the world s diversity by containing a Brazilian unique and exclusively phytoecological domain, the Caatinga, which lacks particular attention regarding macrofungi. Although the gradual growth of knowledge in recent years, studies are still insipient in the Caatinga domain. Between their various vegetal formations are the Brejos de Altitude , described as islands with moist perennial vegetation, low soil fertility and annual precipitation of 900 1.300 mm. This study aimed to evaluate the taxonomic richness of Nidulariaceae fungi in Caatinga s areas of Northeastern Brazil, describing and identifying species, as well as expanding the collection of Herbarium UFRN Fungos. For description of Nidulariaceae fungi was followed the proposed by specialized literature in the group. We have found and described 10 species of the genus Cyathus, namely: C. earlei Lloyd, C. gayanus Tul. & C. Tul., C. gracilis H.J. Brodie, C. intermedius (Mont.) Tul. & C. Tul., C. limbatus Tul. & C. Tul., C. montagnei Tul. & C. Tul., C. pallidus Berk. & M.A. Curtis, C. poeppigii Tul. & C. Tul., C. striatus (Huds.) Willd. and C. tríplex Lloyd. Beyond these have been described four new species to science: Cyathus calvescens R. Cruz & Baseia sp. nov., C. hortum R. Cruz & Baseia sp. nov., C. magnomuralis R. Cruz & Baseia sp. nov. and C. parvocinereus R. Cruz & Baseia sp. nov. Two samples were identified only to genus level. No member of Mycocalia J.T. Palmer, Nidula V.S. White, Nidularia Fr. and Crucibulum Tul. & C. Tul. has been found, however the results were extremely valuable for being an unprecedented work in Caatinga, and importante tool for implementation of conservation projects and sustainable utilization of this domain

Potencial de uso dos recursos vegetais em uma comunidade rural do semiárido do Rio Grande do Norte

The aim of the present study was to identify the main plant use categories of native varieties from the Caatinga biome, in the rural community of Caicó, Rio Grande do Norte state (Northeastern Brazil). Semi-structured and structured interviews were used to gather information from local specialists about the use of the plants. The uses of 69 species are described by 23 observers (woodsmen, herb doctors, healers, farmers and housewives) aged 35 years or more. These species were allocated to seven categories: medicinal, wood plants, nutritional, mystic, fuel, forage plants and domestic use. The most represented families were Fabaceae (14 spp.), Euphorbiaceae (6 spp.), Cucurbitaceae (3 spp.) and Cactaceae (3 spp.). The calculation of use- value showed that the cumaru (Amburana cearensis (Allemão) A. C. Sm) and the jurema-preta (Mimosa tenuiflora (Willd.) Poir) were the most commonly mentioned, in addition to having the largest number of uses. The data obtained confirm the potential of the plants from the Seridó region and reinforce the importance of biodiversity for rural communities, underscoring the need for local plant management

Rubiaceae em um remanescente de floresta atlântica no Rio Grande do Norte, Brasil

Empresa de Pesquisa Agropecuária do Rio Grande do Norte