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)

Drying study of siloxane-PPG nanocomposites

This is a study of the structural transformations occurring in hybrid siloxane-polypropyleneglycol (PPG) nanocomposites, with different PPG molecular weight, along the drying process. The starting materials are wet gels obtained by the sol-gel procedure using as precursor the 3-(trietoxysilyl)propylisocyanate (IsoTrEOS) and polypropylenglycol bis(2-amino-propyl-ether) (NH2-PPG-NH2). The shrinkage and mass loss measurements were performed using a temperature-controlled chamber at 50 degreesC. The nanostructural evolution of samples during drying was studied in situ by small angle x-ray scattering (SAXS). The experimental results demonstrate that the drying process is highly dependent on the molecular weight of polymer. After the initial drying stage, the progressive emptying of pores leads to the formation of a irregular drying front in gels prepared from PPG of high molecular weight, like 4000 g/mol. As a consequence, an increase of the SAXS intensity due to the increase of electronic density contrast between siloxane clusters and polymeric matrix is observed. For hybrids containing PPG of low molecular weight, the pore emptying process is fast, leading to a regular drying front, without isolated nanopockets of solvents. SAXS intensity curves exhibit a maximum, which was associated to the existence of spatial correlation of the silica clusters embedded in the polymeric matrix. The spatial correlation is preserved during drying. These results also reveal that the structural transformation during drying is governed by capillary forces and depends on the entanglement of polymer chains.

Evaluation of varying amounts of thermal cycling on bond strength and permanent deformation of two resilient denture liners

Statement of problem. Two problems found in prostheses with resilient liners are bond failure to the acrylic resin base and increased permanent deformation due to material aging.Purpose. This in vitro study evaluated the effect of varying amounts of thermal cycling on bond strength and permanent deformation of 2 resilient denture liners bonded to an acrylic resin base.Material and methods. Plasticized acrylic resin (PermaSoft) or silicone (Softliner) resilient lining materials were processed to a heat-polymerized acrylic resin (QC-20). One hundred rectangular specimens (10 X 10-mm(2) cross-sectional area) and 100 cylindrically-shaped specimens (12.7-mm diameter X 19.0-mm height) for each liner/resin combination were used for the tensile and deformation tests, respectively. Specimen shape and liner thickness were standardized. Specimens were divided into 9 test groups (n=10) and were thermal cycled for 200, 500, 1000, 1500, 2000, 2500, 3000, 3500, and 4000 cycles. Control specimens (n=10) were stored for 24 hours in water at 37degreesC. Mean bond strength, expressed as stress at failure (MPa), was determined with a tensile test using a universal testing machine at a crosshead speed of 5 mm/min. Analysis of failure mode, expressed as a percent (%), was recorded as either cohesive, adhesive, or both, after observation. Permanent deformation, expressed as a percent (%), was determined using ADA specification no. 18. Data from both tests were examined with a 2-way analysis of variance and a Tukey test (alpha=.05).Results. For the tensile test, Softliner specimens submitted to different thermal cycling regimens demonstrated no significantly different bond strength values from the control; however, there was a significant difference between the PermaSoft control group (0.47 +/- 0.09 MPa [mean +/- SD]) and the 500 cycle group (0.46 +/- 0.07 MPa) compared to the 4000 cycle group (0.70 +/- 0.20 MPa) (P<.05). With regard to failure type, the Softliner groups presented adhesive failure (100%) regardless of specimen treatment. PermaSoft groups presented adhesive (53%), cohesive (12%), or a combined mode of failure (35%). For the deformation test, there was no significant difference among the Softliner specimens. However, a significant difference was observed between control and PermaSoft specimens after 1500 or more cycles (1.88% +/- 0.24%) (P<.05).Conclusions. This in vitro study indicated that bond strength and permanent deformation of the 2 resilient denture liners tested varied according to their chemical composition.