Obtenção de dispersões de complexos polieletrolíticos à base de quitosana e poli(ácido metacrílico) e análise de adsorção de albumina bovina sérica

Dispersions composed of polyelectrolyte complexes based on chitosan and poly(methacrylic acid), PMAA, were obtained by the dropping method and template polymerization. The effect of molecular weight of PMAA and ionic strength on the formation of chitosan/poly(methacrylic acid), CS/PMAA, complexes was evaluated using the dropping method. The increase in molecular weight of PMAA inhibited the formation of insoluble complexes, while the increase in ionic strength first favored the formation of the complex followed by inhibiting it at higher concentrations. The polyelectrolyte complexation was strongly dependent on macromolecular dimensions, both in terms of molecular weight and of coil expansion/contraction driven by polyelectrolyte effect. The resultant particles from dropping method and template polymerization were characterized as having regions with different charge densities: chitosan predominating in the core and poly(methacrylic acid) at the surface, the particles being negatively charged, as a consequence. Albumin was adsorbed on templatepolymerized CS/PMAA complexes (after crosslinking with glutardialdehyde) and pH was controlled in order to obtain two conditions: (i) adsorption of positively charged albumin, and (ii) adsorption of albumin at its isoelectric point. Adsorption isotherms and zeta potential measurements showed that albumin adsorption was controlled by hydrogen bonding/van der Waals interactions and that brushlike structures may enhance adsorption of albumin on these particles

Análise da estrutura genética populacional da piraúna (cephalopholis fulva: serranidae) ao longo da costa e ilhas oceânicas brasileiras

Brazil has about 8,500 km of coastline and on this scale, fishing is a historically important source of animal protein for human consumption. The national fishing background shows a growth of marine fishery production until 1985 and within this period it was recorded a steady decline. From the year 2003 fishing statistics aim to some "recovery" of the total fisheries production, which probably is related to a change in industry practice. The target of commercial fishing became smaller species with low commercial value, but very abundants. The coney, Cephalopholis fulva (Serranidae), is one of these species that have been suffering a greater fishing pressure in recent years. In order to provide data about the current situation of the genetic diversity of these populations, several molecular markers have been being used for this purpose. The prior knowledge of genetic variability is crucial for management and biodiversity conservation. To this end, the control region sequences (dloop) of mtDNA from Cephalopholis fulva (Serranidae) from five geographical points of the coast of Brazil (Ceará, Rio Grande do Norte, Bahia and Espírito Santo) and the Archipelago of Fernando de Noronha (FN) were sequenced and their genetic diversity analyzed. The FST values were very low (0.0246 to 0.000), indicating high gene flow between the sampled spots. The indices h and indicate a secondary contact between previously allopatric lineages differentiated or large and stable populations with long evolutionary history. Tests of Tajima and Fu showed expansion for all populations. In contrast, the mismatch distribution and SSD indicated expansion just for coastal populations. Unlike other species of the Atlantic which have been deeply affected by events on later Pleistocene, the population-genetic patterns of C. fulva may be related to recent events occurred approximately 130,000 years ago. Moreover, the data presented by geographical samples of the specie C. fulva showed high genetic diversity, also indicating the absence of deleterious effects of over-exploitation on this specie, as well as evidence of complete panmixia between all sampled populations

Os efeitos das queimadas na Amazônia em nível celular e molecular

The Amazon holds over half of the planet's remaining tropical forests and comprises the largest biodiversity in the world, accounting for approximately 60 % of the Brazilian territory. However, deforestation fires in the region causes serious problems to exposed human. The aim of this study was to evaluate the chemical compounds as well as the cellular and molecular effects after exposure to organic material extracted from particulate matter less than 10 µm (PM10) in the Amazon region. As for the chemical composition, n-alkanes analysis showed a prevalence of anthropogenic influence during the fires in the region. In addition, there was a predominance of monosaccharides from biomass burning markers. Also, the Polycyclic Aromatic Hydrocarbons (PAH) and their derivatives have also been identified in samples collected in the Amazon. By using the PAH concentrations was possible to calculate the BaP-equivalent and it was found that the dibenz(a) anthracene contributes with 83% to potential carcinogenic risk. As for the potential mutagenic risk, the benzo (a) pyrene is the HPA that has a major contribution in this analysis. It may be noted that the retene was the most abundant PAH. This compound was genotoxic and cause death by necrosis in the human lung cells. In biological tests, the data showed that organic PM10 is capable of causing genetic damage in both plant cells and in human lung cells. This damage cause an arrest in the G1 phase of the cell cycle exposed, increasing the expression of p53 and p21. Additionally, the PM10 caused cell death by apoptosis, increasing the foci of histone - H2AX. Given these results, it is important to emphasize the reduction and better control of biomass burning in the Amazon region thus improving the quality of health of the population being exposed. As clearly stated recently by the World Health Organization, the reduction of air pollution could save millions of lives annually.

Análise das mutações C288Y, S65C e H63D e frequência alélica do gene HFE em pacientes com hiperferritinemia, em uma cidade do Nordeste, Brasil

Background & Aims: HFE-associated Hereditary Hemochromatosis (HH) is one of the most frequent autosomal recessive disease in the caucasian population, caused by the high absorption and deposition of iron in several organs. This accumulation results in several clinical complications such as cirrhosis, arthritis, cardiopathies, diabetes, sexual disorders and skin darkening. Although most of the cases are homozygous individuals for the C282Y mutation, another two mutations, H63D and S65C, have been reported to be associated with milder forms of the disease. The objective is to avaluate the distribution of C282Y, H63D and S65C mutations in the HFE gene in patients with suspected HH in the state of Rio Grande do Norte, Brazil. Methods: Samples of peripheral blood were taken from 335 patients originating from Natal-RN, a city in northeastern Brazil with suspected of HH and which were screened for the HFE gene C282Y, H63D and S65C mutations, using molecular genetics assays (Polymerase Chain Reaction- Restriction Fragments Length Polymorphism). The main criterion for including such patients in the study was the increasing of persistent serum ferritin in individuals aged between 18 and 70 or older, both males and females. As to the exclusion criteria, individuals holding hemolytical anemia, talassemy and previously report of blood transfusion did not take part of the study. Results: Out of the 335 patients studied, 143 patients showed absence of mutation and 195 showed some kind of mutation in the HFE gene: 07/335 (2,08%) were homozigous C282Y, 25/335 heterozygous C282Y, 25/335 (7,46%) were homozigous H63D, 115/335 (34,32%) heterozygous H63D, 5/335 (1,48%) heterozygous S65D, 11/ 335 (3,28%) and were double heterozygous (H63D/C282Y). None patients were Homozygous S65D and S65D heterozygous (S65D/H63D and S65D/C282Y). Conclusions. The distribution of the HFE gene C282Y, H63D and S65C mutations found in our group matches the tendencies observed in other European countries. Due to the high prevalence of hemochromatosis, its seriousness and easy treatment, the genetic diagnosis of HH has become a dream, especially in the high risk group.

Filogenia molecular das enzimas isocitrato liase e malato sintase e sua evolução em Viridiplanta

The plant metabolism consists of a complex network of physical and chemical events resulting in photosynthesis, respiration, synthesis and degradation of organic compounds. This is only possible due to the different kinds of responses to many environmental variations that a plant could be subject through evolution, leading also to conquering new surroundings. The glyoxylate cycle is a metabolic pathway found in glyoxysomes plant, which has unique role in the seedling establishment. Considered as a variation of the citric acid cycle, it uses an acetyl coenzyme A molecule, derived from lipids beta-oxidation to synthesize compounds which are used in carbohydrate synthesis. The Malate synthase (MLS) and Isocitrate lyase (ICL) enzyme of this cycle are unique and essential in regulating the biosynthesis of carbohydrates. Because of the absence of decarboxylation steps as rate-limiting steps, detailed studies of molecular phylogeny and evolution of these proteins enables the elucidation of the effects of this route presence in the evolutionary processes involved in their distribution across the genome from different plant species. Therefore, the aim of this study was to establish a relationship between the molecular evolution of the characteristics of enzymes from the glyoxylate cycle (isocitrate lyase and malate synthase) and their molecular phylogeny, among green plants (Viridiplantae). For this, amino acid and nucleotide sequences were used, from online repositories as UniProt and Genbank. Sequences were aligned and then subjected to an analysis of the best-fit substitution models. The phylogeny was rebuilt by distance methods (neighbor-joining) and discrete methods (maximum likelihood, maximum parsimony and Bayesian analysis). The identification of structural patterns in the evolution of the enzymes was made through homology modeling and structure prediction from protein sequences. Based on comparative analyzes of in silico models and from the results of phylogenetic inferences, both enzymes show significant structure conservation and their topologies in agreement with two processes of selection and specialization of the genes. Thus, confirming the relevance of new studies to elucidate the plant metabolism from an evolutionary perspective

Epidemiologia e caracterização molecular dos vírus Dengue circulantes no Rio Grande do Norte, 2013-2014

Dengue is an acute infectious disease, usually transmitted by Aedes aegypti mosquitoes. The etiologic agents belong to the family Flaviviridae, genus Flavivirus, and occur as four antigenically related but distinct serotypes designated DENV-1, 2, 3, and 4. In Brazil, the disease represents a national public health problem. The purpose of the present study was to describe epidemiological aspects of dengue in the State of Rio Grande do Norte, from 2013 to 2014. A total of 483 blood or serum samples, collected from January 2013 to December 2014, were studied by RT-PCR for viral detection and typing. The infection was confirmed in 36.44% (176/483) of the cases studied. This study detected the circulation of three serotypes of dengue virus in Rio Grande do Norte, DENV-1, 2, and 4. The predominant serotype in 2013-2014 was the DENV-4, representing 83.51% (81/97) and 68.35% (54/79) of the positive cases, respectively. Regarding the spatial distribution, most of the cases occurred in Natal and Caicó, with 9.28% (9/97) and 18.99% (15/79), respectively. The months with the biggest viral circulation in RN were March 2013 and May 2014. The female gender was the most affected, representing 69.07% (67/97) in 2013 and 54.43% (43/79) in 2014. The most affected age groups were 21-30 years (2013) and 11-20 years (2014) with 25.77% (25/97) and 20.25% (16/79) positive cases, respectively. Phylogenetic analysis indicated that genotype V (DENV-1) and genotype II (DENV-4) circulated in the State. Our results provide information about the dynamics of DENV in the Rio Grande do Norte, important for the development of disease control strategies.

Construção de bibliotecas de cDNAs subtraídos a partir das sementes de Mamona submetidas à estresse hídrico e análise in silico de transcritos potencialmente expressos sob condições de estresse: Metalotioneina, proteína reprimida por auxina de 12.5 kDa e Glutelina tipo-A3

Estresses ambientais abióticos são fatores que causam respostas ao nível molecular, fisiológico e morfológico em plantas, dependendo também de sua intensidade e duração. É visto que algumas espécies apresentam tolerância a condições estressantes e ao mesmo tempo são fontes naturais de matéria prima para indústria. Nesse contexto encontra-se a mamona (Ricinus comunnis L.), principal fonte de óleo de rícino valorizado por suas aplicações farmacêuticas e principalmente industriais, vem sendo usada como cultura em regiões onde a disponibilidade de água é reduzida, usada como fonte de renda para agricultura da região nordeste brasileira. Visto que pouco se sabe sobre as respostas moleculares que levam essa planta a tolerar regiões secas e como as sementes, principais foco de interesse, respondem a essa escassez, nesse trabalho foram construídas duas bibliotecas de cDNAs, onde a partir de uma abordagem subtrativa, continham RNAs diferencialmente expressos em sementes de plantas mamona submetidas ao estresse hídrico durante 5 dias (biblioteca L7), e a outra RNAs diferencialmente expressos em sementes controle (biblioteca L5). A biblioteca L7 apresentou a maior variedade de transcritos com um total de 182. A maior parte das funções estabelecidas pelo sistema Gene Ontology - GO, foram direcionadas aos “Processos Metabólicos” (526), em segundo “Respostas a estímulos” (57), o terceiro termo mais abundante foram referentes a “Desenvolvimento”(26). Já na biblioteca L5, foram encontrados 91 transcritos, com maior parte de suas funções referentes a “Processos Metabólicos”(413), em segundo “Respostas a estímulos” (8) e em terceiro Regulação (6). Alguns dos transcritos da biblioteca L7 foram escolhidos para análise por repetirem-se mais de 3x e não aparecerem na biblioteca L5, o que indica uma possível regulação positiva sobre estresse. As análises sobre Metalotioneína (4x), mostraram que a sequência de proteica apresentava os domínios conservados que a caracterizava como tipo II, onde são encontrados dois domínios funcionais ricos em cisteína com posições altamente conservadas, desempenhando a função de ligar-se a metais pesados, correlacionadas assim como a atividade de eliminação EROs e defesa contra o estresse oxidativo, além de apresentar homologia com a sequência de Bruguiera gymnorhiza, uma planta de mangue adaptada a ambientes salinos. Analisamos também os transcritos da referente a proteína AUXIN-REPRESSED 12.5 KDA (3x), apontada como sendo reprimida pelo hormônio auxina e associada ao processo de dormência da semente, é descrito em uma família gênica onde vários membros pertencem as vias de resposta ao estresse. Por último, analisamos a proteína GLUTELIN TYPE-A 3 (5x), uma importante proteína de armazenamento com caráter hidrofílico, possivelmente direcionada para o vacúolo. Em nosso trabalho foi possível observar um aumento de transcritos em relação a subtração controle, possivelmente reflexo do aumento do metabolismo da semente, tanto para resposta defensiva ao estresse hídrico quanto para o amadurecimento rápido da semente onde foram observados transcritos referentes a resposta oxidativa, controle hormonal, proteínas de reserva e produção de óleo.

Filogenia molecular do gênero Morganella Zeller (Fungi, Basidiomycota) utilizando marcadores ITS

With the development and improvement of techniques for molecular studies and their subsequent application to the systematic, significant changes occurred in the classification of gasteroid fungi. The genus Morganella belongs to the family Lycoperdaceae, and is characterized mainly by lignicolous habit and presence of paracapilicium. Recent data demonstrate the discovery of new species for the group and the existence of a wide variety of species occurring in tropical ecosystems. However, the phylogenetic relationships of the genus, as well as the taxonomic classification, still require revisions to be better understood, the literature studies that address this issue are still very scarce. Thus, the objective of this study was to conduct studies of molecular phylogeny with species of the genus Morganella, to enhance understanding of the phylogeny of the group by including tropical species data. For this, the specimens used both for DNA extractions as for morphological review were obtained from Brazilian and foreign herbaria. For morphological analysis were observed characters relevant to the group's taxonomy. For phylogenetic analysis the Maximum Parsimony and Bayesian Analyzes were used, using the internal transcribed spacer (ITS) of nuclear ribosomal DNA. In phylogenetic analyzes, representatives from Morganella form a monophyletic clade with good support value and based on these results the genus should not be included as subgenus of Lycoperdon. The analysis indicated that M. pyriformis was not grouped with other representatives of Morganella, and therefore should not be included in the group as representative of Apioperdon subgenus because it is a Lycoperdon representative. Moreover, M. fuliginea, M. nuda, M. albostipitata, M. velutina, M. subincarnata are grouped with high support values within the genus Morganella. Morganella arenicola based on morphological and molecular studies does not aggregate in Morganella. Morganella nuda was grouped with M. fuliginea giving indications that can be treated as an intraspecific variation. The results of the analyzes favor to a better understanding of the species of Morganella. However, additional studies using a greater number of species, as well as other molecular markers are needed for a better understanding of the phylogenetic of Morganella.

Concentração do óleo de girassol em compostos insaturados utilizando destilação molecular

Vegetable oils are characterized as important raw materials in the supplying of natural substances of interest pharmaceutical, food and cosmetic industry. Sunflower oil stands out for its important composition present in unsaturated fatty acids such as oleic acid (C18:1) and linoleic (C18:2), responsible for many health benefits. The main objective of this study is obtain enriched fractions in unsaturated compounds from refined sunflower oil. The oil used in this study was characterized by the determination of some properties, like iodine number, acid number and viscosity. A transesterification was done to transform the triglycerides into their corresponding methyl esters of fatty acids. These was submitted the molecular distillation process, for present as an efficient alternative to separation and purification of these substances, using high vacuum and low temperatures. Of the esters fractions that was obtained, were analyzed by gas chromatography. The experimental design technique was used to evaluate the influence of the temperature variation of evaporation and condensation system on the percentage obtained residue. The evaporator temperature proved to be the most influential variable on the studied response. The optimized conditions for the answer was studied at 100 °C for evaporator temperature and 10 °C for the condenser temperature. The graph of "split ratio" showed that for the lowest flow feed (1 mL/min) and higher evaporator temperature (110 °C) was obtained in the largest fraction of distillate. It also used the study of the influence of evaporator temperature on the concentration of unsaturated compounds. The best operating conditions for temperature was 90 °C reached 82.21 % of unsaturated compounds. Elimination curves of the unsaturated compounds present in the distillate stream were obtained. The simulation results of the molecular distillation process of sunflower oil showed the concentration profiles for three different feed flow rates. The speed, temperature and thickness profiles of the liquid film were obtained. The speed of the film increases as the fluid flows through the walls of the evaporator, reaching a maximum on length of 0.075 m. The film thickness decreases on the route, since many compounds are volatilized. The result of the temperature profile had to be consistent with the literature reproduced, being constant after reaching the maximum operating temperature in the length of 0.15 m. This study allowed characterizing and focusing, through experimental analysis, unsaturated compounds and observing the sunflower oil´s behavior through process simulation.

Concentração do óleo de girassol em compostos insaturados utilizando destilação molecular

Vegetable oils are characterized as important raw materials in the supplying of natural substances of interest pharmaceutical, food and cosmetic industry. Sunflower oil stands out for its important composition present in unsaturated fatty acids such as oleic acid (C18:1) and linoleic (C18:2), responsible for many health benefits. The main objective of this study is obtain enriched fractions in unsaturated compounds from refined sunflower oil. The oil used in this study was characterized by the determination of some properties, like iodine number, acid number and viscosity. A transesterification was done to transform the triglycerides into their corresponding methyl esters of fatty acids. These was submitted the molecular distillation process, for present as an efficient alternative to separation and purification of these substances, using high vacuum and low temperatures. Of the esters fractions that was obtained, were analyzed by gas chromatography. The experimental design technique was used to evaluate the influence of the temperature variation of evaporation and condensation system on the percentage obtained residue. The evaporator temperature proved to be the most influential variable on the studied response. The optimized conditions for the answer was studied at 100 °C for evaporator temperature and 10 °C for the condenser temperature. The graph of "split ratio" showed that for the lowest flow feed (1 mL/min) and higher evaporator temperature (110 °C) was obtained in the largest fraction of distillate. It also used the study of the influence of evaporator temperature on the concentration of unsaturated compounds. The best operating conditions for temperature was 90 °C reached 82.21 % of unsaturated compounds. Elimination curves of the unsaturated compounds present in the distillate stream were obtained. The simulation results of the molecular distillation process of sunflower oil showed the concentration profiles for three different feed flow rates. The speed, temperature and thickness profiles of the liquid film were obtained. The speed of the film increases as the fluid flows through the walls of the evaporator, reaching a maximum on length of 0.075 m. The film thickness decreases on the route, since many compounds are volatilized. The result of the temperature profile had to be consistent with the literature reproduced, being constant after reaching the maximum operating temperature in the length of 0.15 m. This study allowed characterizing and focusing, through experimental analysis, unsaturated compounds and observing the sunflower oil´s behavior through process simulation.

Elucidação molecular e micelar dos superagregados de anfotericina B

A Anfotericina B (AmB) é amplamente utilizada no tratamento de infecções fúngicas sistêmicas. No entanto, a sua utilização é limitada devido a sua elevada toxicidade aguda e crônica. Os superagregados de AmB (H-AmB) obtido pelo processo de aquecimento controlado apresentam um potencial reduzido de toxicidade quando comparados à Anfotericina B micelar não aquecida (M-AmB). Diante do exposto, o objetivo deste estudo foi avaliar as características físico-químicas do processo de formação dos superagregados após a liofilização do H-AmB, por meio de técnicas tais como a calorimetria de exploratória diferencial (DSC), termogravimetria (TG) e análise térmica diferencial (DTG), ressonância magnética nuclear (RMN), espalhamento dinâmico de luz (DLS) e difração de raios X (DRX). Os dados de DLS indicaram um tamanho de aproximadamente 260 nm deste novo sistema, sendo este tamanho compatível e viável para administração intravenosa. A análise de DRX demonstrou a formação de um novo estado cristalino do sistema micelar, podendo este estar relacionado a presença de grande proporção de desoxicolato de sódio (NaDC) na formulação. O aquecimento do NaDC resulta na formação de uma estrutura helicoidal intermolecular, promovendo desta forma o aumento da agregação micelar e consequentemente aumento de seu tamanho. Os estudos das análises térmicas demonstraram que o processo de aquecimento não influência no comportamento das amostras. Os dados de RMN da H-AmB demonstraram a presença de ácido desoxicólico além do NaDC. O ácido desoxicólico é formado após o processo de aquecimento e sugere-se que o equilíbrio entre ambas as moléculas seja responsável pela redução da toxicidade da AmB. Os resultados aqui apresentados sugerem que o processo de aquecimento controlado altera a organização estrutural das micelas, resultando na diminuição da toxicidade, na melhoria da estabilidade térmica e na manutenção da atividade.

Elucidação molecular e micelar dos superagregados de anfotericina B

A Anfotericina B (AmB) é amplamente utilizada no tratamento de infecções fúngicas sistêmicas. No entanto, a sua utilização é limitada devido a sua elevada toxicidade aguda e crônica. Os superagregados de AmB (H-AmB) obtido pelo processo de aquecimento controlado apresentam um potencial reduzido de toxicidade quando comparados à Anfotericina B micelar não aquecida (M-AmB). Diante do exposto, o objetivo deste estudo foi avaliar as características físico-químicas do processo de formação dos superagregados após a liofilização do H-AmB, por meio de técnicas tais como a calorimetria de exploratória diferencial (DSC), termogravimetria (TG) e análise térmica diferencial (DTG), ressonância magnética nuclear (RMN), espalhamento dinâmico de luz (DLS) e difração de raios X (DRX). Os dados de DLS indicaram um tamanho de aproximadamente 260 nm deste novo sistema, sendo este tamanho compatível e viável para administração intravenosa. A análise de DRX demonstrou a formação de um novo estado cristalino do sistema micelar, podendo este estar relacionado a presença de grande proporção de desoxicolato de sódio (NaDC) na formulação. O aquecimento do NaDC resulta na formação de uma estrutura helicoidal intermolecular, promovendo desta forma o aumento da agregação micelar e consequentemente aumento de seu tamanho. Os estudos das análises térmicas demonstraram que o processo de aquecimento não influência no comportamento das amostras. Os dados de RMN da H-AmB demonstraram a presença de ácido desoxicólico além do NaDC. O ácido desoxicólico é formado após o processo de aquecimento e sugere-se que o equilíbrio entre ambas as moléculas seja responsável pela redução da toxicidade da AmB. Os resultados aqui apresentados sugerem que o processo de aquecimento controlado altera a organização estrutural das micelas, resultando na diminuição da toxicidade, na melhoria da estabilidade térmica e na manutenção da atividade.

Purificação, caracterização e análise da atividade bioinseticida, de um inibidor de tripsina em sementes de catanduva (Piptadenia moniliformis)

One Kunitz-type trypsin inhibitors (PmTI) was purified from Piptadenia moniliformis seeds, a tree of the sub-family Mimosoideae, by TCA precipitation, affinity chromatography on immobilized trypsin-Sepharose, DEAE cellulose (ion exchange) and Superose 12 (molecular exclusion) column FPLC/AKTA. The inhibitor has Mr of 25 kDa by SDS-PAGE and chromatography molecular exclusion. The N-terminal sequence of this inhibitor showed high homology with other family Kunitz inhibitors. This also stable variations in temperature and pH and showed a small decrease in its activity when incubated with DDT in the concentration of 100mM for 120 minutes. The inhibition of trypsin by PmTI was competitive, with Ki of 1.57 x10-11 M. The activity of trypsin was effectively inhibited by percentage of inhibition of 100%, among enzymes tested, was not detected inhibition for the bromelain, was weak inhibitor of pancreatic elastase (3.17% of inhibition) and inhibited by 76.42% elastase of neutrophils, and inhibited in a moderate, chymotrypsin and papain with percentage of inhibition of 42.96% and 23.10% respectively. In vitro assays against digestive proteinases from Lepidoptera, Diptera and Coleoptera pests were carried out. Several degrees of inhibition were found. For Anthonomus grandis and Ceratitis capitata the inhibition was 89.93% and 70.52%, respectively, and the enzymes of Zabrotes subfasciatus and Callosobruchus maculatus were inhibited by 5.96% and 9.41%, respectively, and the enzymes of Plodia. interpunctella and Castnia licus were inhibited by 59.94% and 23.67, respectively. In vivo assays, was observed reduction in the development of larvae in 4rd instar of C. capitata, when PmTI was added to the artificial diet, getting WD50 and LD50 of 0.30% and 0.33%, respectively. These results suggest that this inhibitor could be a strong candidate to plant management programs cross transgenic

Análise toxicológica in vitro e in vivo de uma fucana antitrombótica da alga marrom Spatoglossum schdöederi

Fucan is a term used to denominate a family of sulfated polysaccharides rich in L-fucose. They are extracted mainly from the extracellular matrix of brown algae and echinoderms. The brown alga Spatoglossum schröederi (Dictyotaceae) has three heterofucans named A, B and C. Our research group have been extracted non anticoagulant heterofucan from S. schröederi which possess antithrombotic activity in vivo. However, their toxicity in vitro and in vivo has not yet been determined. For the results in toxicity in vitro, we observed that the fucan A at 20, 500 and 1000 μg/plate showed no mutagenic activity in Kado test (Microsuspension), when the bacterial strains TA97a, TA98, TA100 and TA102, with and without S9 were used. The comet assay showed that fucan A (from 20 to 1000 μg/mL) did not cause any genotoxic effect on CHO cells. There was no damage to the DNA of these cells, as evidenced by the tail length and tail moment, which were similar to that found for the negative control. The fucan A from S. schröederi was administered at 20 μg/g of rat (dose which it showed high antithrombotic activity) during two months. After that, the animals were killed and examined. The data showed that fucan A did not cause any change in biochemistry and hematological parameters, as well as, in the morphology and size of the rat s organs analyzed. In conclusion, this study indicates that fucan is a compound with potential pharmacological that has no toxicity

Análise do transcriptoma das glândulas venenosas do escorpião Tityus stigmurus

In Brazil, accidents with scorpions are considered of medical importance, not only by the high incidence, but also for the potentiality of the venom from some species in determining severe clinical conditions. Tityus stigmurus is a widely distributed scorpion species in Northeastern Brazil and known to cause severe human envenomations, inducing pain, hyposthesia, edema, erythema, paresthesia, headaches and vomiting. The present study uses a transcriptomic approach to characterize the molecular repertoire from the non-stimulated venom gland of Tityus stigmurus scorpion. A cDNA library was constructed and 540 clones were sequenced and grouped into 37 clusters, with more than one EST (expressed sequence tag) and 116 singlets. Forty-one percent of ESTs belong to recognized toxin-coding sequences, with antimicrobial toxins (AMP-like) the most abundant transcripts, followed by alfa KTx- like, beta KTx-like, beta NaTx-like and alfa NaTx-like. Our analysis indicated that 34% include other possible venom molecules , whose transcripts correspond to anionic peptides, hypothetical secreted peptides, metalloproteinases, cystein-rich peptides and lectins. Fifteen percent of ESTs are similar to cellular transcripts. Sequences without good matches corresponded to 11%. This investigation provides the first global view of cDNAs from Tityus stigmurus. This approach enables characterization of a large number of venom gland component molecules, which belong either to known or atypical types of venom peptides and proteins from the Buthidae family