Recovery from Strongyloides venezuelensis infection in Lewis rats is associated with a strong Th2 response

P>In this study, we investigated the characteristics of the infection and subsequent immunity induced by Strongyloides venezuelensis in Lewis rats. Animals were infected with 4000 L3 of S. venezuelensis and number of eggs per gram of faeces indicated an acute phase around day 8 and a recovery phase around day 32 after infection. A strong Th2 polarization during recovery phase was ascertained by a significant increase in IgG1 and IgE compared with that in the acute period. A shift in the cytokine profile confirmed these findings. A predominant production of IFN-gamma during the acute phase was followed by IL-10 production during recovery. Together these findings show that experimental infection of Lewis rats with S. venezuelensis presents a kinetics of parasite establishment and immunity similar to that described in other models of helminthic infection.

Comparison of the susceptibility of Santa Ines and Ile de France ewes to nematode parasitism around parturition and during lactation

Breeds of sheep with resistance to nematode infection often display a reduced periparturient rise (PPR) in faecal egg counts (FEC) when compared with susceptible sheep. This trial was carried out to compare the resistance of Santa Ines, an indigenous Brazilian breed of sheep, and Ile de France ewes to natural infections by gastrointestinal nematodes, especially in the periparturient period. Twelve Santa lnes and 10 Ile de France ewes were estrus synchronized and then mated (December 2001). Lambing occurred from 30 April to 19 June 2002 and the lambs were weaned at 60 days of age. Faecal egg counts, packed cell volume (PCV), total plasma protein levels and peripheral eosinophils counts were determined from November 2001 to September 2002. To prevent deaths, individual treatment with anthelmintics was provided to ewes with FEC higher than 4000 eggs per gram (EPG) or with PCV lower than 21%. The percentage of Santa Ines ewes treated with anthelmintics was lower than that of Ile de France sheep. Eight of the 10 Ile de France ewes were treated, with one of them requiring four treatments, one requiring three treatments, and two requiring two treatments. The remaining four received a single treatment. In contrast, only 5 of the 12 Santa lnes ewes required treatment and for each this was done only once. The mean values of PCV were in the normal range in the Santa Ines ewes. In contrast, mean PCVs of Ile de France ewes reached a low of 24.5% in the second month after lambing and were significantly lower than Santa Ines 1 month before lambing (P < 0.05) and 2 months after lambing (P < 0.07). The mean plasma protein level was significantly lower in the Ile de France ewes in the second month after lambing (P < 0.01) coinciding with a reduction in PCV. The ewes of both breeds had high mean numbers of eosinophils in blood, which increased even further post-weaning when a marked reduction in FEC occurred. Haemonchus larvae were the most numerous genus in faecal cultures for both breeds followed by Trichostrongylus, Oesophagostomum and Cooperia. A periparturient rise in FEC was observed in both breeds. However, Santa Ines ewes showed a higher capacity to support the infection in an environment contaminated with large number of infective larvae of gastrointestinal nematodes. (C) 2004 Elsevier B.V. All rights reserved.

Resistance of Santa Ines, Suffolk and Ile de France sheep to naturally acquired gastrointestinal nematode infections

A study was conducted to assess the breed resistance against nematode infections in Santa Ines, Ile de France and Suffolk male lambs over a 9-month period in São Paulo state, Brazil. Lambs were born during the winter (year 2000) and were weaned at 2 months of age. The animals were then housed and treated with anthelmintics to eliminate natural infections by gastrointestinal nematodes. In late October 2000, lambs were placed in a paddock, where they stayed until August of the following year. Fecal and blood samples were taken from each animal every 2 weeks. on the same day, a pasture sample was collected to determine the number of infective larvae on the herbage. To prevent deaths, individual treatment with anthelmintics was provided to lambs with fecal egg counts (FEC) higher than 4000 eggs per gram (EPG) or with a packed cell volume (PCV) lower than 21%. In August 2001, all animals were slaughtered and the worms present in samples of the gastrointestinal contents were identified and counted. Most of the Suffolk and Ile de France sheep received three to six anthelmintic treatments over a period of 7 months, while most of the Santa Ines were not treated. Reductions in PCV and plasma protein values associated with high FEC and worm burdens were recorded, particularly, in Suffolk and Ile de France lambs. Haemonchus contortus and Oesophagostomum columbianum burdens and number of nodular lesions caused in the large intestine by O. columbianum larvae were significantly lower in Santa Ines sheep. All three breeds showed similar Trichostrongylus colubriformis worm burdens. The relative resistance of Santa Ines young male sheep was superior to that of Suffolk and Ile de France sheep. (C) 2004 Elsevier B.V. All rights reserved.

Multiple resistance to anthelmintics by Haemonchus contortus and Trichostrongylus colubriformis in sheep in Brazil

The objective of this study was to determine the level of resistance of Haemonchus contortus and Trichostrongylus colubriformis in sheep to levamisole albendazole ivermectin moxidectin closantel and trichlorfon The parasites were isolated from sheep naturally infected by gastrointestinal nematodes and were then kept in monospecifically infected lambs for production of infective larvae (L3) of both species Forty-two lambs at three months of age were simultaneously artificially infected with 4000 L3 of H contortus and 4000 L3 of T colubriformis The animals were allocated Into seven groups with six animals each that received one of the following treatments Group 1-control no treatment Group 2-moxidectin (0 2 mg/kg body weight (BW)) Group 3-closantel (10 mg/kg BW) Group 4-trichlorfon (100 mg/kg BW) Group 5-levamisole phosphate (4 7 mg/kg BW) Group 6-albendazole (5 0 mg/kg BW) and Group 7-ivermectin (0 2 mg/kg BW) Nematode fecal egg counts (FEC) were carried out on the day of treatment and again at 3 7 10 and 14 days post-treatment on the same occasions composite fecal cultures were prepared for each group for production of L3 which were identified into genus The animals were sacrificed for worm counts at 14 days after treatment The efficacy of each treatment was calculated from the arithmetic mean of the FEC or worm burden of the treated group compared with the values of the control group Only trichlorfon and moxidectin treatments resulted in a significant reduction of H contortus recorded at necropsy (73% and 45% respectively) Moxidectin reduced T colubriformis worm burdens by 82% and albendazole by 19% All other anthelmintics resulted in no significant reduction in the numbers of worms found at necropsy In conclusion the Isolates of H contortus and T colubriformis showed multiple resistance to all groups of anthelmintics tested This is the first report based on the controlled efficacy test to show resistance of T colubriformis to macrocyclic lactones in Brazil (C) 2010 Elsevier B.V. All rights reserved

Initiating structural studies of Lys49-PLA2 homologues complexed with an anionic detergent, a fatty acid and a natural lipid

Lys49-Phospholipase A(2) (Lys49-PLA(2) - EC 3.1.1.4) homologues damage membranes by a Ca2+-independent mechanism which does not involve catalytic activity. Both MjTX-II from Bothrops moojeni and BthTX-I from Bothrops jararacussu are dimeric in solution and in the crystalline states, and a model for the Ca2+-independent membrane damaging mechanism has been suggested in which flexibility at the dimer interface region pert-nits quaternary structural transitions between open and closed membrane bound dimer conformations which results in the perturbation of membrane phospholipids and disruption of the bilayer structure [1]. With the aim of gaining insights into the structural determinants involved in protein/lipid association, we report here the crystallization and preliminary X-ray analysis of the (i) MjTX-II/SDS complex at a resolution of 2.78Angstrom, (ii) MjTX-II/STE complex at a resolution of 1.8 Angstrom and (W) BthTX-I/DMPC complex at 2.72Angstrom. These complexes were crystallized by the hanging drop vapour-diffusion technique in (i) HEPES buffer (pH 7.5) 1.8M ammonium sulfate with 2% (w/v) polyethyleneglycol 400, in (ii) 0.6-0.8 M sodium citrate as the precipitant (pH 6.0-6.5) and in (iii) sodium citrate buffer (pH 5.8) and PEG 4000 and 20% isopropanol, respectively. Single crystals of these complexes have been obtained and X-ray diffraction data have been collected at room temperature using a R-AXIS IV imaging plate system and graphite monochromated Cu Kalpha X-ray radiation generated by a Rigaku RU300 rotating anode generator for (i) and (W) and using using a Synchrotron Radiation Source (Laboratorio Nacional de Luz Sincrotron, LNLS, Campinas, Brazil) for (ii).

Crystallization and preliminary X-ray analysis of bucain, a novel toxin from the Malayan krait Bungarus candidus

Bucain is a three-finger toxin, structurally homologous to snake-venom muscarinic toxins, from the venom of the Malayan krait Bungarus candidus. These proteins have molecular masses of approximately 6000-8000 da and encompass the potent curaremimetic neurotoxins which confer lethality to Elapidae and Hydrophidae venoms. Bucain was crystallized in two crystal forms by the hanging-drop vapour-diffusion technique in 0.1 M sodium citrate pH 5.6, 15% PEG 4000 and 0.15 M ammonium acetate. Form I crystals belong to the monoclinic system space group C2, with unit-cell parameters a = 93.73, b = 49.02, c = 74.09 Angstrom, beta = 111.32degrees, and diffract to a nominal resolution of 1.61 Angstrom. Form II crystals also belong to the space group C2, with unit-cell parameters a = 165.04, b = 49.44, c = 127.60 Angstrom, beta = 125.55degrees, and diffract to a nominal resolution of 2.78 Angstrom. The self-rotation function indicates the presence of four and eight molecules in the crystallographic asymmetric unit of the form I and form II crystals, respectively. Attempts to solve these structures by molecular-replacement methods have not been successful and a heavy-atom derivative search has been initiated.

The effect of hydrothermal alteration on the Sr and Nd isotopic signatures of the Barra do Itapirapua Carbonatite, Southern Brazil

The Cretaceous Barra do Itapirapua carbonatite in southern Brazil experienced extensive postmagmatic hydrothermal alteration. In this article, Sr and Nd isotope ratios of coexisting samples of hydrothermally overprinted and of preserved, nonoverprinted carbonatite are presented. Hydrothermal alteration caused strong REE enrichment, leading to the formation of minerals of the bastnaesite group. In the overprinted carbonatite, Nd contents reach 4000 ppm, two orders of magnitude higher than in the fresh carbonatite, but epsilon(Nd) varies only within a range of 3.4 units. In contrast, Sr was leached from the carbonatite during the postmagmatic alteration; hence values of around 10,000 ppm in the fresh carbonatite drop to about 1000 ppm in the overprinted samples. Leaching is accompanied by a variation of Sr isotopic composition toward more radiogenic values, resulting in an increase of 15 units in epsilon(Sr). Variation of Sr isotopic composition is related to postmagmatic alteration and is decoupled from the variation of Nd isotopic composition, ruling out heterogeneities in the mantle source as the main cause of isotopic variability in the data set. Furthermore, this cannot be explained by bulk crustal contamination. A two-step model is proposed in which (1) a REE-rich, carbonatite-derived hydrothermal fluid overprinted the pristine carbonatite, causing REE-enrichment with a relative small change of isotopic composition; and (2) crust-derived hydrothermal fluids percolated the cooling carbonatite, leaching the original Sr from the carbonatite and introducing a more radiogenic Sr isotopic signature. The amounts of carbonatite-derived Nd with primitive, carbonatite-like Nd isotope ratios introduced during the first stage of hydrothermal alteration are high enough to buffer the effect of crust-derived Nd on the Nd isotopic composition of the overprinted carbonatite.

Colorimetric enzymatic assay of L-malic acid using dehydrogenase from baker's yeast

A colorimetric method has been developed and optimized to measure L-malic acid in samples of fruit juices and wine. This method is based on oxidation of the analyte, catalyzed by malate dehydrogenase (MDH) from dry baker's yeast, and in combination with the reduction of a tetrazolium salt (MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). In the present study, the method exhibited sensitivity in the range of 500-4000 mu M of L-malic acid in the reaction cuvette, with the lower detection limit of 6.7-10(-2) g/L, the upper limit of 53.6.10(-2) g/L and a maximum standard deviation of only 2.5 % for the analyzed samples. The MDH activity from baker's yeast was also optimized, the enzyme showed a high stability at pH=8.0-9.0 and the activity was maintained completely at temperatures up to 40 degrees C for 1 hour. The results show that the colorimetric method using enzymatic preparations from dry baker's yeast is a simple and low-cost method with possibility of wide application.

Poly(glutamic acid) nanofibre modified glassy carbon electrode: Characterization by atomic force microscopy, voltammetry and electrochemical impedance

Glassy carbon electrodes (GCE) were modified with poly(glutamic acid) acid films prepared using three different procedures: glutamic acid monomer electropolymerization (MONO), evaporation of poly(glutamic acid) (PAG) and evaporation of a mixture of poly(glutamic acid)/glutaraldehyde (PAG/GLU). All three films showed good adherence to the electrode surface. The performance of the modified GCE was investigated by cyclic voltammetry and differential pulse voltammetry, and the films were characterized by atomic force microscopy (AFM) and electrochemical impedance spectroscopy (EIS). The three poly(glutamic acid) modified GCEs were tested using the electrochemical oxidation of ascorbic acid and a decrease of the overpotential and the improvement of the oxidation peak current was observed. The PAG modified electrode surfaces gave the best results. AFM morphological images showed a polymeric network film formed by well-defined nanofibres that may undergo extensive swelling in solution, allowing an easier electron transfer and higher oxidation peaks. (C) 2007 Elsevier Ltd. All rights reserved.

Peak Separation by Derivative Spectroscopy Applied to FTIR Analysis of Hydrolized Silica

Reliable spectral analysis is only achieved if the spectrum is thoroughly investigated in regard to all hidden and overlapped peaks. This paper describes the steps undertaken to find and separate such peaks in the range of 3000 to 4000 cm(-1) in the case of three different infrared absorption spectra of the glass surface of hydrolyzed silica optical fibers. Peak finding was done by the analysis of the second and fourth derivatives of the digital data, coupled with the available knowledge of infrared spectroscopy of silica-water interaction in the investigated range. Peak separation was accomplished by curve fitting with four different models. The model with the best fit was described by a sum of pure Gaussian peaks. Shoulder limit and detection limit maps were used to validate the revealed spectral features.

Determination of ambroxol in syrups using diffuse reflectance spectroscopy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

INDUÇÃO do AMADURECIMENTO DE FRUTOS CÍTRICOS em PÓS-COLHEITA COM A APLICAÇÃO DE ETHEPHON

O presente experimento foi conduzido no Departamento de Botânica -- IB/Unesp, Botucatu-SP, com o objetivo de estudar o efeito da aplicação pós-colheita de Ethephon no desverdecimento de laranjas-doces (Citrus sinensis (L.) Osbeck) cultivar 'Hamlin' e 'Baianinha', consideradas cultivares precoces. Os frutos foram imersos por diferentes tempos em solução aquosa de Ethephon mais 0,05% do adjuvante Extravon (25% de Alquilfenol - poliglicoléter). Os frutos foram colocados em caixas de madeira, sobre bancada, em temperatura ambiente, onde permaneceram por 72 horas. Os tratamentos utilizados foram os seguintes: T1- testemunha (água); T2 -- Ethephon -- 1000 mg.L-1 / 5 minutos de imersão; T3 - Ethephon -- 2000 mg.L-1 / 5 minutos de imersão; T4 -- Ethephon -- 3000 mg.L-1 / 5 minutos de imersão; T5 -- Ethephon -- 4000 mg.L-1 / 5 minutos de imersão; T6 -- Ethephon -- 1000 mg.L-1 / 10 minutos de imersão; T7 - Ethephon -- 2000 mg.L-1 / 10 minutos de imersão; T8 -- Ethephon -- 3000 mg.L-1 / 10 minutos de imersão; T9 -- Ethephon -- 4000 mg.L-1 / 10 minutos de imersão. O delineamento experimental adotado foi o de blocos ao acaso, com 9 tratamentos e 4 repetições. Após 72 horas, o experimento foi avaliado através das seguintes variáveis: teor de clorofila da casca, teor de sólidos solúveis (0Brix) e textura dos frutos, além da análise visual com a utilização de fotografias. Com isso, verificou-se que houve influência do Ethephon no processo de mudança da cor da casca de frutos cítricos em todos os tratamentos, diferenciado-se estatisticamente da testemunha, porém, na concentração de 4000 mg.L-1, houve início de necrose em alguns pontos da casca dos frutos. Assim, pode-se concluir que a melhor concentração foi a de 1000 mg.L-1 de Ethephon, para acelerar o processo de desverdecimento ou mudança da coloração da casca de frutos de laranjas precoces cultivares 'Hamlin' e 'Baianinha'.

IBA e carboidratos no enraizamento de brotações procedentes de estacas radiciais de Rubus Spp.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Genética molecular do hipotireoidismo congênito

O hipotireoidismo congênito (HC) ocorre, mundialmente, em 1/3000-4000 neonatos e pode ser classificado em permanente ou transitório. O HC primário é responsável pela maioria dos afetados, enquanto o secundário e terciário são raros. Nos países iodo-suficientes, a disgenesia tireóidea (DT) é a causa mais freqüente de HC. Os defeitos hereditários da síntese hormonal ocorrem em minoria de crianças portadoras de HC. Fatores ambientais, genéticos e auto-imunes concorrem na etiologia do HC, mas na maioria dos casos de DT a causa é obscura. Atribui-se aos genes envolvidos na ontogenia da glândula tireóidea, como os fatores de transcrição TITF1, TITF2, PAX-8 e receptor de TSH (TSHR), função patogenética na DT. Até o momento não foi descrita anormalidade no gene TITF1 como causa de HC, enquanto foram identificadas mutações no PAX-8 em cinco recém-nascidos com DT. Embora não envolvidas na DT, mutações inativadoras do TSHR podem produzir espectro de defeitos congênitos oscilando entre hipertirotropinemia com eutireoidismo e hipotireoidismo com hipoplasia glandular. A clonagem dos genes envolvidos na biossíntese dos hormônios tireóideos, como o da tireoperoxidase (TPO) e tireoglobulina (Tg), permitiu a identificação de mutações responsáveis por alguns casos de bócio e hipotireoidismo decorrente de defeito de incorporação de iodeto ou anormalidades na síntese de Tg. Recentemente, foi demonstrada a base molecular do defeito de transporte ativo de iodeto e da síndrome de Pendred, respectivamente, devidas a mutações no gene NIS (simportador de sódio e iodeto) e no gene PDS (pendrina). em conclusão, grande parte dos pacientes com HC e DT não tem esclarecida, ainda, a causa molecular desta síndrome.

Aspectos genéticos do hipotireoidismo congênito

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)