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

Determinantes genéticos da hanseníase em uma população do Rio Grande do Norte

Background: Leprosy can cause severe disability and disfigurement and is still a major health in different parts of the world. Only a subset of those individuals exposed to the pathogen will go on to develop clinical disease and there is a broad clinical spectrum amongst leprosy patients. The outcome of infection is in part due to host genes that influence control of the initial infection and the host´s immune response to that infection. Aim: Evaluate if polymorphisms type SNP in the 17q118q21 chromosomic region contribute to development of leprosy in Rio Grande do Norte population. Material and methods: A sample composed of 215 leprosy patients and 229 controls drawn from the same population were genotyped by using a Snapshot assay for eight genes (NOS2A, CCL18, CRLF3, CCL23, TNFAIP1, STAT5B, CCR7 and CSF3) located in chromosomic region 17q118q21. The genotype and allele frequency were measured and statistical analysis was performed by chi-square in SPSS version 15 and graph prism pad version 4 software. Results: Ours results indicated that the markers NOS2A8277, NOS2A8rs16949, CCR78rs11574663 and CSF38rs2227322 presented strong association with leprosy and their risk genotype were GG, TT, AA and GG respectively. The risk genotypes for all markers associated to leprosy presented recessive inheritance standard. When we compared the interaction among the markers in different combination we find that the marker NOS2A8277 associated with CCR78rs11574663 presented highest risk probability to development of leprosy. When we evaluated the haplotype of the risk markers it was found a haplotype associated with increase of the protection (CSF38rs22273228CC, CCR78 rs115746638GA, NOS2A8rs169498CT and NOS2A82778GA). The association of the clinical forms paucibacilary and multibacilary with markers showed that to the markers NOS2A8 2778GG, CCR78rs115746638AA and CSF38rs22273228GG there were a strong influence to migration to multibacilary pole and to marker NOS2A8rs169498TT the high proportion was found to the paucibacilary form. Conclusions: Changes in the genes NOS2A, CCR7 and CSF3 can influence the immune response against Mycobacterium leprae. The combination among these polymorphisms alters the risk probability to develop leprosy. The markers type SNP associated to development of the leprosy also are linked to clinical forms and its severity being the polymorphism NOS2A8rs169498TT associated with paucibacilar form and the polymorphisms NOS2A82778GG, CCR78rs115746638AA and CSF38rs22273228GG associated to multibacilar form

Avaliação de megadoses de retinol palmitato no pós-parto imediato sobre o retinol do leite de puérperas atendidas no hospital josé pedro bezerra, Natal-RN

Vitamin A deficiency (VAD) is a serious public health problem in developing countries, and as a therapeutic and prophylactic measure retinil palmitate is being supplemented. Nevertheless its efficacy has been questioned. The objective of the study was to evaluate the supplementation of two retinil palmitate megadosis on the serum retinol levels of post partum healthy mothers from Dr. José Pedro Bezerra (Hospital Santa Catarina) hospital, Natal - RN. The enrolled women (n=199) were randomly distributed into three studied groups and supplemented with retinil palmitate immediately after delivery with a single 200,000 IU dose (group S1), two 200,000 IU dose (group S2) with 24h difference between the doses, or no supplementation (group C). Among women selected, 143 remained until the end of the study. The influence of vitamin A dietary intake was evaluated during pregnancy and after 30 days of delivery. The average intake of the population was reasonable, but a high prevalence of inadequate intake was found. Retinol in colostrums and mature milk was determined by high performance liquid chromatography (HPLC). The retinol average in colostrums and mature milk in the supplemented and control groups were adequate according to the reference values. In colostrums, women from groups C, S1 and S2 presented retinol averages by milk volume of 94.8 ± 40.2 µg/dL, 92.2 ± 50.0 µg/dL and 91.8 ± 53.7 µg/dL, respectively. No difference was found between these averages (p=0.965), this was also seen when the values where expressed as µg/g of fat (p=0.905). After 30 days of delivery, retinol per milk volume differed between the control group (36.6 ± 17.5 µg/dL) and groups supplemented with 200,000 IU (51.0 ± 28.8 µg/dL) or 400,000 IU (55.2 ± 31.6 µg/dL) of retinil palmitate (p<0,05). Nevertheless, when S1 and S2 groups where compared, no significant difference was found (p=0.97). Considering retinol/g of fat, the means were 12.7 ± 6.7 µg/g, 15.6 ± 8.3 µg/g and 17.2 ± 8.9 µg/g for groups C, S1 and S2, respectively, with significant difference between groups S2 and C (p=0,01). Subclinical VAD prevalence showed a serious public health problem in the study population (32% in colostrums and 31.5% in mature milk). When analyzing the groups separately, the group which received two doses (200,000 IU + 200,000 IU) presented the lowest VAD prevalence (20.7%). Retinil palmitate supplementations of 200,000 IU and 400,000 IU (divided in two doses) in the immediate post partum showed no significant difference. Nevertheless, the 400,000 IU (divided in two doses) supplementation showed a reduction in VAD

Caracterização parcial e atividades farmacológicas do extrato rico em polissacarídeos sulfatados da angiosperma marinha Halodule wrightii

Sulfated polysaccharides (PS) are biomolecules with a great biotechnological potential. There are few data about PS from high plants. In addition, pharmacological activities of PS from plants have not been carrying out. The aim of this work was extract PS from the angiosperm Halodule wrightii and study their anticoagulant and antioxidant activities. Histological analysis showed the presence of the PS manly in the roots. A polysaccharide-rich extract was obtained from H. wrightii by proteolysis followed by methanol and TCA precipitation. Chemical, infra-red analysis and agarose gel electrophoresis in 1.3 diaminopropane acetate buffer confirmed the presence of sulfated polysaccharides made by glucose, galactose, xylose and sulfate residues in the proportion 1: 0,9: 1: 1. In addition polyacrilamide electrophoresis have shown that extract is mainly compose by 11kDa sulfated polysaccharides. Pharmacological analysis have shown total antioxidant capacity (CAT) that resulted in 15,21 μg for equivalent of ascorbic acid, scavenging activity of the DPPH radical with 41,36 % of scavenging, activity of reducing power with the maximum of 0,290 nm (50 % of vitamin C activity) and scavenging activity superoxide radical (O2-) with a maximum of 32,23 %. Chelating activity of metal less than 4% and scavenging activity of the radical hydroxyl (OH-) less than 2%. Time of activated partial tromboplastin (aPTT) doubling the time of coagulation from 20μg of and protrombin time (PT) was not present. The data indicate that PS from Halodule wrightii could be considered for future applications in medicine, food production or cosmetic industry

Preferência alimentar e identificação das principais fontes de repasto sanguíneo de fêmeas Lutzomyia (Diptera: Psychodidae) em áreas endêmicas para leishmaniose visceral na grande Natal

Leishmaniasis are endemic diseases wild spread in the New and Old World, caused by the flagelated protozoan Leishmania. In the New World, the distribution of different forms of leishmaniasis is mostly in tropical regions. In the State of Rio Grande do Norte, Northeast Brazil, 85% of the captured sand flies fauna is Lutzomyia longipalpis. The distribution of the sand fly vector in the state overlaps with the disease distribution, where the presence of sand flies is associated with presence of animals shelters. The aim of this study was to analyse the blood meal preference of sand flies vector from the genus Lutzomyia spp. in laboratory conditions, to verify the vector life cicle at different temperatures sets and to identify the main blood meal source in endemic areas for visceral leishmaniasis (VL) at peri-urban regions of Natal. Sand flies samples were collected from the municipalities of São Gonçalo do Amarante and Nísia Floresta where female sand flies were grouped for the colony maintenance in the laboratory and for the analysis of the preferred source of sand fly blood meal in natural environment. The prevalence of blood meal preference and oviposition for the females sand flies was 97% for Cavia porcellus with oviposition of 19 eggs/female; 97% for Eqqus caballus with 19 eggs/female; 98% for human blood with 14 eggs/female; 71.3% for Didelphis albiventris with 8.4 eggs/female; 73% for Gallus gallus with 14 eggs/female; 86% for Canis familiaris with 10.3 eggs/female; 81.4% for Galea spixii with 26 eggs/female; 36% for Callithrix jachus with 15 eggs/female; 42.8% for Monodelphis domestica with 0% of oviposition. Female sand flies did not take a blood meal from Felis catus. Sand flies life cycle ranged from 32-40 days, with 21-50 oviposition rates approximately. This study also showed that at 32°C the life cycle had 31 days, at 28° C it had 50 days and at 22°C it increased to 79 days. Adjusting the temperature to 35°C the eggs did not hatch, thus blocking the life cycle. A total of 1540 sand flies were captured, among them, 1.310 were male and 230 were female. Whereas 86% of the sand flies captured were Lu. longipalpis as compared to 10.5% for Lu. evandroi and, 3.2% for L. lenti and 0.3% for Lu whitmani. The ratio between female and male sandfly was approximately 6 males to 1 female. In Nísia Floresta, 50.7% of the collected females took their blood meal from armadillo, 12.8% from human. Among the female sand flies captured in São Gonçalo do Amarante, 80 of them were tested for the Leishmania KDNA infectivity where 5% of them were infected with Leishmania chagasi. Female Lutzomyia spp. showed to have an opportunistic blood meal characteristic. The behavioral parameters seem to have a higher influence in the oviposition when compared to the level of total proteins detected in the host s bloodstream. A higher Lu. longipalpis life cycle viability was observed at 28°C. The increase of temperature dropped the life cycle time, which means that the life cycle is modified by temperature range, source of blood meal and humidity. Lu longipalpis was the most specie found in the inner and peridomiciliar environment. In Nísia Floresta, armadillos were the main source of blood meal for Lutzomyia spp. At São Gonçalo do Amarante, humans were the main source of blood meal due to CDC nets placed inside their houses

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

Purificação e caracterização parcial de uma Fucana C de Dictyota mentrualis e estudo do seu efeito antiinflamatório e nociceptivo

In recent years, sulfated polysaccharides from marine algae have emerged as an important class of natural biopolymers with potential application in human and veterinary health care, while taking advantage of the absence of potential risk of contamination by animal viruses. Among these, fucans isolated from the cell walls of marine brown alga have been study due to their anticoagulant, antithrombotic, anti-inflammatory and antiviral activities. These biological effects of fucans have been found to depend on the degree of sulfation and molecular size of the polysaccharide chains. In the present study, we examined structural features of a fucan extracted from brown alga Dictyota menstrualis and its effect on the leukocyte migration to the peritoneum. The sulfated polysaccharides were extracted from the brown seaweed by proteolytic digestion, followed by sequential acetone precipitation producing 5 fractions. Gel lectrophoresis using 0.05 M 1,3-diaminopropane-acetate buffer, pH 9.0, stained with 0.1% toluidine blue, showed the presence of sulfated polysaccharides in all fractions. The chemical analyses demonstrated that all fractions are composed mainly of fucose, xylose, galactose, uronic acid, and sulfate. Electrophoresis in agarose gel in three different buffers demonstrated that the fraction 2.0v have only one population of fucan. This compound was purify by exclusion molecular. It has shown composition of fucose, xilose, sulfate and uronic acid in molar ration of 1.0: 1.7: 1.1: 0.5 respectively. The effect of this heterofucan on the leukocyte migration was observed 6h after zymozan (mg/g) administration into the peritoneum. The heterofucan showed higher antimigratory activity, it decrease the migration of leukocyte in 83.77% to peritoneum. The results suggest that this fucan is a new antimigratory compound with potential pharmacological appications

Avaliação da suplementação materna com megadose de vitamina a sobre os níveis de retinol e alfa-tocoferol no colostro

The vitamins A and E are recognizably important in the initial stages of life and the newborn depends on nutritional adequacy of breast milk to meet their needs. These vitamins share routes of transport to the tissues and antagonistic effects have been observed in animals after supplementation with vitamin A. This study aimed to verify the effect of maternal supplementation with vitamin A megadose (200,000 UI) in the immediate post-partum on the concentration of alpha-tocopherol in colostrum. Healthy parturient women attended at a public maternity natalensis were recruited for the study and divided into two groups: control (n = 37) and supplemented (n = 36). Blood samples of colostrum and milk were collected until 12 hours after delivery. The women of the supplemented group was administered a retynil palmitate capsule and 24 hours after the first collection was obtained the 2nd sample of colostrum in two groups for analysis of retinol and alpha-tocopherol in milk. The mean retinol concentration of 50,7 ± 14,4 μg/dL (Mean ± standard deviation) and alpha-tocopherol of 1217.4 ± 959 mg/dL in the serum indicate the nutritional status biochemical appropriate. Supplementation with retynil palmitate resulted in increase not only retinol levels in the colostrum of the supplemented group (p = 0.002), but also the concentration of alpha-tocopherol (p = 0.04), changing from 1456.6 ± 1095.8 mg/dL to 1804.3 ± 1432.0 mg/dL (milk 0 and 24 respectively) compared to values in the control group, 984.6 ± 750.0 mg/dL and 1175.0 ± 730.8 mg/dL. The women had different responses to supplementation, influenced by baseline levels of retinol in colostrum. Those with previous by low levels of retinol in colostrum (<60 mg/dL) had increased the concentration of alpha-tocopherol in milk, whereas those with adequate levels (> 60 mg/dL), showed a reduction after supplementation. Supplementation with retinol palmitate is an important intervention in situations of high risk for vitamin A deficiency, when considering the need to maternal supplementation, since the excess vitamin can offer unfavorable interactions between nutrients essential for the mother-child group

Utilização da Berberina na Identificação dos Mastócitos do Molusco Anomalocardia brasiliana

Berberine is an alkaloid used as a fluorochrome in the identification of heparin and DNA. Enerback, 1974, described the technique used until today to study granules rich in heparin of vertebrate mast cells. Santos et al., 2003, studied mast cells of the mollusk Anomalocardia brasiliana using biochemical and histological analysis. This work used the fluorescent dye berberine technique to improve characterization of these cells. Mollusk organs (ctenidium and mantle) were processed with routine histological techniques. Tissue sections were treated with berberine 0,02% in redistilled water acidified to pH 4, by the addition of citric acid for 20 minutes. The visualization was made through fluorescence microscopy with ultraviolet region emission. The mast cell fluorescence had a strong yellow color, where cell nuclei appeared more greenish. This result was very similar to the ones reported before. Mast cells are location at the epithelium surface is the same in both organs, mantle and ctenidium. The fluorescence was easily observed in the granules. Therefore, this technique showed to be good and sensitive to study mast cell of invertebrates

Análises Estruturais e atividades biológicas de exopolissacarídeo extraído do fungo comestível pleurotus Sajor-Caju e de seu derivado sulfatado quimicamente.

The exopolysaccharides are extracellular compounds produced by some species of fungi and bacteria. It is suggested that these molecules, even when in the form of complex polysaccharide-peptide, are the main bioactive molecules of many fungus. Some of the biological activities displayed by these compounds can be accentuated and others may arise when you add chemically polar or nonpolar groups to polysaccharides. The fruiting body of Pleurotus sajor-caju produces a heteropolysaccharide with antineoplastic and antimicrobial activity, but other biological activities of this polymer have not been evaluated. In this work the exopolysaccharide of Pleurotus sajor-caju was sulfated chemically and structurally characterized. We also evaluated the antiproliferative, antioxidant and anticoagulant activities from native exopolysaccharide (PN) and its sulfated derivated (PS). Polyacrylamide gel electrophoresis, infrared spectroscopy and nuclear magnetic resonance (¹³C) proved successful in sulfation of PN to obtain PS. Analysis by gas chromatography-mass spectroscopy showed that PN and PS are composed of mannose, galactose, 3-O-methyl-galactose and glucose in proportion percentage of 44,9:16,3:19,8:19 and 49, 7:14,4:17,7:18,2, respectively. The percentage of sulfate found in PS was 22.5%. Antioxidants assays revealed that the sulfation procedure affects differently the activities of exopolysaccharides, while the total antioxidant capacity, the scavenging activity of superoxide radical and ferric chelating were not affected by sulfation, on the other hand the chemical modification of PN enhanced the scavenging activity of hydroxyl radical and reducing power. PS also showed anticoagulant activity in a dose-dependent manner and clotting time was 3.0 times higher than the baseline value in APTT at 2 mg/mL. The exopolysaccharide not presented antiproliferative activity against HeLa tumor cells, but PS affects the cellular proliferation in a time-dependent manner. After 72 h, the inhibition rate of PS (2.0 mg/mL) on HeLa cells was about 60%. The results showed that PN sulfation increase some of their activities.

Efeito Da Suplementação Materna Com Palmitato De Retinila Sobre A Concentração De Retinol No Colostro Em Condições De Jejum E Pós-Prandial.

Vitamin A is important in many essential body processes and its deficiency results in serious consequences for human health. Breast milk is the only source of this vitamin for children that are exclusively breastfed. Analysis of vitamin A in mother s milk is important because its concentration is related to maternal vitamin A status and to its ingestion by the mother during pregnancy. The aim of the present study was to assess the effect of maternal supplementation with retynil palmitate on the concentration of colostrum retinol under fasting and postprandial conditions. A total of 149 nursing mothers were recruited at the Januário Cicco Maternity School (Natal, Brazil) and allocated to two groups: Comparison (n = 69) and Test (n = 80). Blood and colostrum (in fasting and postprandial conditions) samples were collected up to 24hs after delivery. Serum retinol and colostrum levels were analyzed by high-performance liquid chromatography. The serum retinol level of 41.6 ± 12.7μg/dL (mean ± standard deviation) indicates adequate biochemical nutritional status. Colostrum retinol level was not influenced by serum retinol levels under any of the conditions established. In the colostrum, the retinol concentration in the unsupplemented test group was 67.3 ± 37.7 μg/dL under fasting and 80.3 ± 35.1 μg/dL under postprandial conditions (p<0.05), showing an increase of 19.3%. In the supplemented test group the values were 102.6 ± 57.3 μg/dL and 133.4 ± 78.3 μg/dL under fasting and postprandial, respectively (p<0.05), representing an increase of 30%. Considering that under fasting conditions most of the vitamin A transported to the milk originates in the retinol binding protein (RBP), the postprandial increase in colostrum retinol suggests a different transport mechanism of retinol to maternal milk from that performed by RBP. This situation becomes more evident under supplementation conditions.

Genotoxicidade e composição do material particulado emitido pela queima de Biomassa: um estudo de caso em Tangará da Serra, Região da Amazônia Brasileira

Tangara da Serra is located on southwestern Mato Grosso and is found to be on the route of pollutants dispersion originated in the Legal Amazon s deforestation area. This region has also a wide area of sugarcane culture, setting this site quite exposed to atmospheric pollutants. The objective of this work was to evaluate the genotoxicity of three different concentrations of organic particulate matter which was collected from August through December / 2008 in Tangara da Serra, using micronucleus test in Tradescantia pallida (Trad-MCN). The levels of particulate matter less than 10μm (MP10) and black carbon (BC) collected on the Teflon and polycarbonate filters were determined as well. Also, the alkanes and polycyclic aromatic hydrocarbons (PAHs) were identified and quantified on the samples from the burning period by gas chromatography detector with flame ionization detection (GC-FID). The results from the analyzing of alkanes indicate an antropic influence. Among the PAHs, the retene was the one found on the higher quantity and it is an indicator of biomass burning. The compounds indene(1,2,3-cd)pyrene and benzo(k)fluoranthene were identified on the samples and are considered to be potentially mutagenic and carcinogenic. By using Trad-MCN, it was observed a significant increase on the micronucleus frequency during the burning period, and this fact can be related to the mutagenic PAHs which were found on such extracts. When the period of less burnings is analyzed and compared to the negative control group, it was noted that there was no significant difference on the micronuclei rate. On the other hand, when the higher burning period is analyzed, statistically significant differences were evident. This study showed that the Trad-MCN was sensible and efficient on evaluating the genotoxicity potencial of organic matter from biomass burning, and also, emphasizes the importance of performing a chemical composition analysis in order to achieve a complete diagnosis on environmental risk control

Novas propriedades do SKTI (Inibidor de tripsina de soja): inibição para elastase neutrofílica humana e efeitos no processo de injúria pulmonar aguda

Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 µg (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases

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

Resposta imune à saliva de flebotomíneos (diptera: psychodidae) em indivíduos residentes em áreas urbana e periurbana no nordeste do brasil

Visceral leishmaniasis (VL) has a wide geographical distribution in tropical and subtropical areas of the planet, which is a protozoan parasite of the genus Leishmania. This pathogen is transmitted to the host through the sandflies bite, with its saliva, the immune response that leads to both. In the state of Rio Grande do Norte, 85% of the sand flies captured is Lutzomyia longipalpis, but the second most abundant, Lutzomyia evandroi, it deserves emphasis because its wide distribution and eclectic behavior. The exposure of people living in endemic areas for the insect vector VL greatly increases the chances of infection. This study aimed to evaluate aspects of the epidemiological profile of VL in endemic areas of human and nonendemic in the metropolitan area of Natal, as well as verify the abundance and seasonal fluctuations of sandflies species in two counties endemic for VL. Were collected in the municipalities of Nísia Floresta, Parnamirim, São Gonçalo do Amarante and Macaíba, of which groups of females were separated for further dissection of the salivary glands and identification of species. The blood samples used were from individuals of two Natal s districts where it has never been reported cases of VL and neighborhoods of Parnamirim applicants who present cases of VL. In the municipality of Nísia Floresta, the most abundant species was L. evandroi with 38.39%, followed by L. longipalpis with 36.22%, L. walkeri 19.67% L. lenti 3.81%, L. wellcomei 1.39% and L. whitmani 0.52%. Already in Parnamirim the proportions were L. walkeri with 73.15%, L. evandroi with 10.55%, L. wellcomei 7.63%, L. longipalpis 6.37%, L. whitmani 1.46%, L. sordellii 0.52%, L. intermedia 0.21 and L. shanonni 0.1%. In both municipalities was observed higher abundance of species distributed in the initial months of the year, as February and March. The study showed that no difference in exposure to the vector of VL among individuals from endemic and non endemic area for this disease. But there are differences in exposure between individuals of L. longipalpis and L. evandroi, confirming the great powers of the first vector. It was also characterized as predominant phenotype in the population of endemic areas who had negative serologic responses to antigens of Leishmania and result in negative Montenegro skin test (DTH), indicating that much of the population hasn t been bitten by infected insects