A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox

Despite the valuable contributions of robotics and high-throughput approaches to protein crystallization, the role of an experienced crystallographer in the evaluation and rationalization of a crystallization process is still crucial to obtaining crystals suitable for X-ray diffraction measurements. In this work, the difficult task of crystallizing the flavoenzyme l-amino-acid oxidase purified from Bothrops atrox snake venom was overcome by the development of a protocol that first required the identification of a non-amorphous precipitate as a promising crystallization condition followed by the implementation of a methodology that combined crystallization in the presence of oil and seeding techniques. Crystals were obtained and a complete data set was collected to 2.3 A resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 A, beta = 96.03 degrees. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V (M) of 2.12 A3 Da-1, corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques.

A oferta de serviços de informação por situação de vida em sítios Web de bibliotecas virtuais: em busca da qualidade no atendimento

With the advent and development of technology, mainly in the Internet, more and more electronic services are being offered to customers in all areas of business, especially in the offering of information services, as in virtual libraries. This article proposes a new opportunity to provide services to virtual libraries customers, presenting a methodology for the implementation of electronic services oriented by these customers' life situations. Through analytical observations of some national virtual libraries sites, it could be identified that the offer of services considering life situations and relationship interest situations can promote the service to their customers, providing greater satisfaction and, consequently, improving quality in the offer of information services. The visits to those sites and the critical analysis of the data collected during these visits, supported by bibliographic researches results, have enabled the description of this methodology, concluding that the provision of services on an isolated way or in accordance with the user's profile on sites of virtual libraries is not always enough to ensure the attendance to the needs and expectations of its customers, which suggests the offering of these services considering life situations and relationship interest situations as a complement that adds value to the business of virtual library. This becomes relevant when indicates new opportunities to provide virtual libraries services with quality, serving as a guide to the information providers managers, enabling the offering of new means to access information services by such customers, looking for pro - activity and services integration, in order to solve definitely real problems.

Bee communities (Hymenoptera: Anthophila) of the "Cerrado" ecosystem in Sao Paulo State, Brazil

Five surveys of the bee communities in four "Cerrado" ecosystem reserves in Sao Paulo State were compared for species richness and similarity. These areas are fragment vegetation reser-reserves located in the Cerrado Corumbata Reserve (Corumbata), Jata Ecological Park (Luiz Antonio), Cajuru (Cajuru), and Vassununga State Park - ""Gleba de Cerrado de Pe-de-Gigante"" (Santa Rita do Passa Quatro). The methodology consisted of capturing bees foraging on flowers along transects, though with small differences between surveys. These ""cerrado"" areas have a large number of species of native bees, which are important pollinators in several Brazilian ecosystems. The community of bees varied among these different fragments. Based on 500 individuals (standardized by rarefaction), Cajuru, Corumbata 1 and Corumbata 2 were the areas with highest species richness, and Jata and Pe-de-Gigante had the lowest species richness in the bee communities. The bee faunas of Corumbata 2 and Pe-de-Gigante had the highest similarity, forming a group with the bee fauna of Cajuru. The bee faunas of Corumbata 1 and Jata were isolated from this group. We found that the bee species richness and similarity found in these ""cerrado"" areas cannot be explained by general factors such as the size of the fragment, the species richness of plants and the distance between the areas. Therefore, we suppose that local factors that differ among areas, such as interactions between populations, and competition and interference from surrounding areas influence and determine bee species richness and similarity in these reserves.

Etiology of congenital hypothyroidism using thyroglobulin and ultrasound combination

Methods currently employed to establish the etiology of congenital hypothyroidism include thyroid ultrasound and scintigraphic exams. Thyroglobulin is a protein almost exclusively secreted by thyroid tissue and indirectly reflects the amount of follicular cells. Even though thyroglobulin is easy to measure, it has been not frequently used because of discordant results to distinguish mainly athyreosis and ectopy (dysgenesis). Knowing the differences in inheritance and prognosis of thyroid dysgenesis and dyshormonogenesis, it is important to define the etiology of CH, combining tools that are easy, fast and available in most medical centers. Our objective was to evaluate and compare color Doppler ultrasound and serum thyroglobulin with radionuclide scan to define the etiology of congenital hypothyroidism. We evaluated 38 children above 3 years-old off-treatment that performed serum thyroglobulin by immunofluorometric assay, color Doppler ultrasound and radionuclide study. On color Doppler ultrasound, 11 patients had athyreosis, 5 ectopic glands, being I associated to hemiagenesis. Twenty one had topic thyroid (3 goiters, 10 normal, 8 hypoplastic). Hemiagenesis and cystic lesion were not revealed by radionuclide scan. We observed substantial agreement between color Doppler ultrasound and radionuclide scan (kappa=0.745, p<0.0001). Serum thyroglobulin in athyreosis ranged from <1.0 to 18.7 mu g/L. Patients with ectopic glands showed wider thyroglobulin range (4.5 to 123 mu g/L, median 28.4 mu g/L). Only one patient showed thyroglobulin deficiency. By using color Doppler ultrasound and serum thyroglobulin levels as valuable combined tools, we established the etiology of congenital hypothyroidism limiting excessive and harmful exams in children, like radionuclide scan.

Antiretroviral resistance among HIV type 1-infected women first exposed to antiretrovirals during pregnancy: Plasma versus PBMCs

Resistance-associated mutations (RAMs) in plasma samples from HIV-1-infected women who received antiretroviral (ARV) prophylaxis during pregnancy was assessed and correlated with the detection of RAMs in peripheral blood mononuclear cells (PMBCs). The study population was composed of HIV-1-infected women enrolled in a prospective cohort study in Latin America and the Caribbean (NISDI Perinatal Study) as of March 1, 2005, who were diagnosed with HIV-1 infection during the current pregnancy, who received ARVs during pregnancy for prevention of mother-to-child transmission of HIV-1, and who were followed through at least the 6-12 week postpartum visit. Plasma samples collected at enrollment during pregnancy and at 6-12 weeks postpartum were assayed for RAMs. Plasma results were compared to previously described PBMC results from the same study population. Of 819 enrolled subjects, 197 met the eligibility criteria. Nucleic acid amplification was accomplished in 123 plasma samples at enrollment or 6-12 weeks postpartum, and RAMs were detected in 22 (17.9%; 95% CI: 11.7-25.9%). Previous analyses had demonstrated detection of RAMs in PBMCs in 19 (16.1%). There was high concordance between RAMs detected in plasma and PBMC samples, with only eight discordant pairs. The prevalence of RAMs among these pregnant, HIV-1-infected women is high (>15%). Rates of detection of RAMs in plasma and PBMC samples were similar.

Occurrence of Neospora caninum antibodies in capybaras (Hydrochaeris hydrochaeris) from Sao Paulo State, Brazil

Capybara (Hydrochaeris hydrochaeris) is a large rodent distributed throughout tropical America. Antibodies to Neospora caninum in 213 feral capybaras from 11 counties of the State of Sao Paulo, Brazil. were assessed using the indirect immunofluorescent antibody test (titer >= 1:25) and found in 20 (9.4%), with titers of 1:25 in 4, 1:50 in 7, and 1:100 in 9 animals. This is the first report of occurrence of N. caninum antibodies in capybaras.

Determination and Molecular Analysis of the Complete Genome Sequence of Two Wild-Type Rabies Viruses Isolated from a Haematophagous Bat and a Frugivorous Bat in Brazil

The complete genome sequences of two Brazilian wild-type rabies viruses (RABV), a BR-DR1 isolate from a haematophagous bat (Desmodus rotundus) and a BR-AL1 isolate from a frugivorous bat (Artibeus lituratus), were determined. The genomes of the BR-DR1 and RR-AL1 had 11,923 and 11,922 nt, respectively, and both encoded the five standard genes of rhabdoviruses. The complete nucleotide sequence identity between the BR-DR1 and BR-AL1 isolates was 97%. The BR-DR1 and BR-AL1 isolates had some conserved functional sites revealed by the fixed isolates, whereas both isolates had unique amino acid substitutions in the antigenic region IV of the nucleocapsid gene. Therefore, it is speculated that both isolates were nearly identical in virologic character. According to our phylogenetic analysis based on the complete genomes, both isolates belonged to genotype 1, and to the previously defined ""vampire bat-related RABV lineage"" which consisted of mainly D. rotundus- and A. lituratus- isolates; however, a branch pattern with high bootstrap values suggested that BR-DR1 was more closely related to the 9001FRA isolate, which was collected from a dog bitten by a bat in French Guiana, than to BR-AL1. This result suggests that the vampire bat-related RABV lineage includes Brazilian vampire bat and Brazilian frugivorous bat RABV and is further divided into Brazilian vampire bat and Brazilian frugivorous bat RABV sub-lineages. The phylogenetic analysis based on the complete genomes was valuable in discriminating among very closely related isolates.

Proper motions of thermally emitting isolated neutron stars measured with Chandra

The remarkable astrometric capabilities of the Chandra Observatory offer the possibility to measure proper motions of X-ray sources with an unprecedented accuracy in this wavelength range. We recently completed a proper motion survey of three of the seven thermally emitting radio-quiet isolated neutron stars (INSs) discovered in the ROSAT all-sky survey. These INSs (RXJ0420.0-5022, RXJ0806.4-4123 and RXJ1308.6+2127) either lack an optical counterpart or have one so faint that ground based or space born optical observations push the current possibilities of the instrumentation to the limit. Pairs of ACIS observations were acquired 3 to 5 years apart to measure the displacement of the sources on the X-ray sky using as a reference the background of extragalactic or remote Galactic X-ray sources. We derive 2 sigma upper limits of 123 mas yr(-1) and 86 mas yr(-1) on the proper motion of RXJ0420.0-5022 and RXJ0806.4-4123, respectively. RXJ1308.6+2127 exhibits a very significant displacement (similar to 9 sigma) yielding mu = 220 +/- 25 mas yr(-1), the second fastest measured among all ROSAT-discovered INSs. The source is probably moving away rapidly from the Galactic plane at a speed which precludes any significant accretion of matter from the interstellar medium. Its transverse velocity of similar to 740 (d/700 pc) km s(-1) might be the largest of all ROSAT INSs and its corresponding spatial velocity lies among the fastest recorded for neutron stars. RXJ1308.6+2127 is thus a middle-aged (age similar to 1 My) high velocity cooling neutron star. We investigate its possible origin in nearby OB associations or from a field OB star. In most cases, the flight time from birth place appears significantly shorter than the characteristic age derived from spin down rate. Overall, the distribution in transverse velocity of the ROSAT INSs is not statistically different from that of normal radio pulsars.

Two New Species of Ocotea (Lauraceae) from the Brazilian Restinga

Two new species of Ocotea Aubl. (Lauraceae) from the restinga of Brazil are described and illustrated. Ocotea arenicola L. C. S. Assis & Mello-Silva and O. ramosissima L. C. S. Assis & Mello-Silva are discovered from Espirito Santo and Bahia states, respectively. Their diagnostic features are indicated, and their relationships to other Ocotea species are discussed.

Crystallization, data collection and data processing of maltose-binding protein (MalE) from the phytopathogen Xanthomonas axonopodis pv. citri

Maltose-binding protein is the periplasmic component of the ABC transporter responsible for the uptake of maltose/maltodextrins. The Xanthomonas axonopodis pv. citri maltose-binding protein MalE has been crystallized at 293 Kusing the hanging-drop vapour-diffusion method. The crystal belonged to the primitive hexagonal space group P6(1)22, with unit-cell parameters a = 123.59, b = 123.59, c = 304.20 angstrom, and contained two molecules in the asymetric unit. It diffracted to 2.24 angstrom resolution.

Assessing sample representativeness for inorganic chemical investigation of invertebrates by INAA

The quality of environmental studies depends on the utilization of adequate sampling protocol and analytical method for obtaining reliable results and minimizing analytical uncertainties. In order to demonstrate the applicability of INAA for determining chemical element composition of invertebrates, this work evaluated sample representativeness in terms of subsampling and sample size. Br, Co, Fe, K, Na, Sc and Zn could be determined in very small samples despite increasing of analytical uncertainties. Special attention should be directed to invertebrate species with small structures because of the high chemical variation observed among different sample sizes tested.

Treatment of Obese Adolescents: The Influence of Periodization Models and ACE Genotype

The aims of the present study were to compare the effects of two periodization models on metabolic syndrome risk factors in obese adolescents and verify whether the angiotensin-converting enzyme (ACE) genotype is important in establishing these effects. A total of 32 postpuberty obese adolescents were submitted to aerobic training (AT) and resistance training (RT) for 14 weeks. The subjects were divided into linear periodization (LP, n = 16) or daily undulating periodization (DUP, n = 16). Body composition, visceral and subcutaneous fat, glycemia, insulinemia, homeostasis model assessment of insulin resistance (HOMA-IR), lipid profiles, blood pressure, maximal oxygen consumption (VO(2max)), resting metabolic rate (RMR), muscular endurance were analyzed at baseline and after intervention. Both groups demonstrated a significant reduction in body mass, BMI, body fat, visceral and subcutaneous fat, total and low-density lipoprotein cholesterol, blood pressure and an increase in fat-free mass, VO(2max), and muscular endurance. However, only DUP promoted a reduction in insulin concentrations and HOMA-IR. It is important to emphasize that there was no statics difference between LP and DUP groups; however, it appears that there may be bigger changes in the DUP than LP group in some of the metabolic syndrome risk factors in obese adolescents with regard to the effect size (ES). Both periodization models presented a large effect on muscular endurance. Despite the limitation of sample size, our results suggested that the ACE genotype may influence the functional and metabolic characteristics of obese adolescents and may be considered in the future strategies for massive obesity control.

Severe Lactic Acidosis Treated With Prolonged Hemodiafiltration in a Disseminated Histoplasmosis

Infections with Histoplasma are rarely seen in immunocompromized patients. We report the case of a renal transplant recipient who presented with disseminated histoplasmosis 3.5 years after transplant He presented severe lactic acidosis (LA), sepsis complicated by circulatory failure, renal failure, and liver dysfunction. We describe the successful use of continuous venovenous hemodiafiltration (CVVHDF) with regional citrate anticoagulation, treatment that stabilized our patient until infectious focus was identified and treated. The lactate was decreasing, concomitant with hemodynamic improvement, with reduction and suspension of the norepinephrine. The serum lactate level normalized 52 hours after CVVHDF initiated (from 28.9 to 2.2 mmol/L). Continuous renal replacement therapy was safely applied and can be recommended as an efficient method on adjuvant treatment of hyperlactatemia. ASAIO Journal 2009; 55:123-125.

MicroRNAs 29 are involved in the improvement of ventricular compliance promoted by aerobic exercise training in rats

Soci UPR, Fernandes T, Hashimoto NY, Mota GF, Amadeu MA, Rosa KT, Irigoyen MC, Phillips MI, Oliveira EM. MicroRNAs 29 are involved in the improvement of ventricular compliance promoted by aerobic exercise training in rats. Physiol Genomics 43: 665-673, 2011. First published March 29, 2011; doi:10.1152/physiolgenomics.00145.2010.-MiRNAs regulate cardiac development, hypertrophy, and angiogenesis, but their role in cardiac hypertrophy (CH) induced by aerobic training has not previously been studied. Aerobic training promotes physiological CH preserving cardiac function. This study assessed involvement of miRNAs-29 in CH of trained rats. Female Wistar rats (n = 7/group) were randomized into three groups: sedentary (S), training 1 (T1), training 2 (T2). T1: swimming sessions of 60 min/5 days/wk/10 wk. T2: similar to T1 until 8th wk. On the 9th wk rats swam 2x/day, and on the 10th wk 3x/day. MiRNAs analysis was performed by miRNA microarray and confirmed by real-time PCR. We assessed: markers of training, CH by ratio of left ventricle (LV) weight/body wt and cardiomyocytes diameter, pathological markers of CH (ANF, skeletal alpha-actin, alpha/beta-MHC), collagen I and III (COLIAI and COLIIIAI) by real-time PCR, protein collagen by hydroxyproline (OH-proline) concentration, CF and CH by echocardiography. Training improved aerobic capacity and induced CH. MiRNAs-1, 133a, and 133b were downregulated as observed in pathological CH, however, without pathological markers. MiRNA-29c expression increased in T1 (52%) and T2 (123%), correlated with a decrease in COLIAI and COLIIIAI expression in T1 (27%, 38%) and T2 (33%, 48%), respectively. MiRNA-29c was inversely correlated to OH-proline concentration (r = 0.61, P = 0.05). The E/A ratio increased in T2, indicating improved LV compliance. Thus, these results show that aerobic training increase miR-29 expression and decreased collagen gene expression and concentration in the heart, which is relevant to the improved LV compliance and beneficial cardiac effects, associated with aerobic high performance training.

Morphological and mechanical properties of hybrid matrices of polysiloxane-polyvinyl alcohol prepared by sol-gel technique and their potential for immobilizing enzyme

Hybrid matrices of polysiloxane-polyvinyl alcohol (POS-PVA) were prepared by sol-gel technique using different concentrations of the organic component (polyvinyl alcohol, PVA) in the synthesis medium. The goal was to prepare carriers for immobilizing enzyme by taking into consideration properties as hardness, mean pore diameter, specific surface area and pore size distribution. The matrices were activated with sodium metaperiodate to render functional groups for binding the lipase from Candida rugosa, used here as a study model. Results showed that low proportion of PVA gave POS-PVA with low surface area and pore volume, although with higher hardness. The chemical activation decreased the pore volume and increased the pore size with a decrease on the surface area of about 60-75%. The matrices for enzyme immobilization were chosen considering the best combination of high surface area and hardness. Thus, the POS-PVA prepared with 5.56 x 10(-5) M of PVA with a surface area of 123 m(2)/g and hardness of 71 HV (50 gf 30 s) was shown to be suitable to immobilize the lipase, with an immobilization yield of about 40%. (c) 2008 Elsevier B.V. All rights reserved.