p38 MAPK regulates IL-1β induced IL-6 expression through mRNA stability in osteoblasts

Osteoblast-derived IL-6 functions in coupled bone turnover by supporting osteoclastogenesis favoring bone resorption instead of bone deposition. Gene regulation of IL-6 is complex occurring both at transcription and post-transcription levels. The focus of this paper is at the level of mRNA stability, which is important in IL-6 gene regulation. Using the MC3T3-E1 as an osteoblastic model, IL-6 secretion was dose dependently decreased by SB203580, a p38 MAPK inhibitor. Steady state IL-6 mRNA was decreased with SB203580 (2 μM) ca. 85% when stimulated by IL-1β (1-5 ng/ ml). These effects require de novo protein synthesis as they were inhibited by cycloheximide. p38 MAPK had minor effects on proximal IL-6 promoter activity in reporter gene assays. A more significant effect on IL-6 mRNA stability was observed in the presence of SB203580. Western blot analysis confirmed that SB203580 inhibited p38 MAP kinase, in response to IL-1β in a dose dependent manner in MC3T3-E1 cells. Stably transfected MC3T3-E1 reporter cell lines (MC6) containing green fluorescent protein (GFP) with the 3′untranslated region of IL-6 were constructed. Results indicated that IL-1β, TNFα, LPS but not parathyroid hormone (PTH) could increase GFP expression of these reporter cell lines. Endogenous IL-6 and reporter gene eGFP-IL-6 3′UTR mRNA was regulated by p38 in MC6 cells. In addition, transient transfection of IL-6 3′UTR reporter cells with immediate upstream MAP kinase kinase-3 and -6 increased GFP expression compared to mock transfected controls. These results indicate that p38 MAPK regulates IL-1β-stimulated IL-6 at a post transcriptional mechanism and one of the primary targets of IL-6 gene regulation is the 3′UTR of IL-6.

Fator de crescimento derivado de plaquetas humanas obtido por uma metodologia rápida e de baixo custo

Human platelet-derived growth factor (PDGF) was purified from lysates of clinically outdated human platelets by ionic exchange chromatography in CM-Sepharose. The eluated fraction was submitted to the Immunoblot/Slot Blot assay using anti-PDGF-AA and anti-PDGF-BB polyclonal antibodies and was evaluated as to its biological activity through the test of [H 3]-thymidine incorporation in NIH/3T3 cell line fibroblasts in culture. The Immunoblot/Slot Blot assay using anti-PDGF-AA and anti-PDGF-BB antibodies proved the presence of the PDGF in chromatographic cationic fraction. The comparison of biological activities between fiblobrast stimulation assay using recombinant PDGF-AB and partially purified PDGF was demonstrated in 165.796 and 157.567 cpm, respectively. This result, proved the potent mitogenic effect of partially purified PDGF and consequently their evidence about the wound healing activity.

Avaliação do nível de multicaminho no posicionamento relativo GPS e sua atenuação através da análise de multiresolução

The multipath effect affects the differential and relative positioning, even that one involving short baselines. So it is necessary to detect this effect, check the caused error level, and mainly, its removal. This paper aims at analysing and comparing some useful components in the detection of this effect. These components are the Signal to Noise Ratio (SNR), the values of MP1 and MP2 obtained from the TEQC software that indicates the multipath level in the carriers L1 and L2, the multipath repeatability in consecutive days and the elevation angle and the azimuth of the satellites. For this purpose, an experiment is carried out, comparing such components in the presence and the absence of reflector objects that cause the multipath. Not only there is clear multipath repeatability in the residuals, but it also appears in the measures SNR, MP1 and MP2, reaching up 99% of correlation. For reduction, at least, of the high frequency multipath effect, the Multi-Resolution Analysis using wavelets is applied in the double differences (DD) measures. Some statistical tests were accomplished, which indicate results improvement, and mainly, larger reliability in the solution of the ambiguities, reaching up 49% of improvement concerning the Ratio test without applying the proposed method.

Correcão ionosférica utilizando os MAPAS: Globais do TEC DO IGS: Avaliação no posicionamento por ponto na região brasileira

One of the main drawbacks of the GPS accuracy for L1 users is the error due to ionosphere. This error depends on the total electron content presents in the ionosphere, as well as of the carrier frequency. Some models have been developed to correct GPS observables of the systematic error due to the ionosphere. The model more known and used is the Klobuchar model, which corrected 50-60% of the ionospheric error approximately. Alternatively, IGS (International GNSS Service) also has developed a model called Global Ionospheric Map (GIM). These maps, in format IONEX, are available in the site of the IGS, and one of the applications of them is to correct the GPS observables of the error due to ionosphere. This work aims at evaluating the quality of GPS point positioning using the IGS ionospheric model in the Brazilian region. Tests carried out had shown an average improvement in the horizontal and vertical determination of 72% and 26%, respectively, when GIM is used in the point positioning.

Clonagem do abacateiro variedade Duke 7 (Persea americana Mill.) por alporquia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Degradação ruminal e síntese de proteína microbiana em bovines alimentados com silagem de sorgo contendo tanino suplementado com concentrado ou uréia

This work aimed to determine ruminal parameters of dry mater (DM), neutral detergent fiber (NDF) and protein synthesis in the rumen of animals fed diets based on sorghum with different tannin levels. In situ degradation was evaluated by rumen incubation of level 1 (L1) and level 2 (L2) sorghum silages during 0, 6, 24, 48 and 96 hours. A duplicated 4 × 4 Latin Square was used. Potential degradability (PD) of DM was similar among silages, whereas the effective degradability (ED) decreased when the diets were supplemented with concentrate. Microbial nitrogen flux and microbial synthesis efficiency were not affected by roughage source. The synthesis efficiency, expressed in organic matter and crude protein digested in rumen, was higher in L1 tannin diets supplemented with concentrate. There was not relationship between the presence of tannins and the parameters of ruminal degradation.

Stress distribution in ceramic restorations over natural tooth using finite element analysis. lithium disilicate x aluminum oxide material

Background: Data on stress distribution in tooth-restoration interface with different ceramic restorative materials are limited. The aim of this chapter was to assess the stress distribution in the interface of ceramic restorations with laminate veneer or full-coverage crown with two different materials (lithium dissilicate and densely sintered aluminum oxide) under different loading areas through finite element analysis. Materials and Methods: Six two-dimensional finite element models were fabricated with different restorations on natural tooth: laminate veneer (IPS Empress, IPS Empress Esthetic and Procera AllCeram) or full-coverage crown (IPS e.max Press and Procera AllCeram). Two different loading areas (L) (50N) were also determined: palatal surface at 45° in relation to the long axis of tooth (L1) and perpendicular to the incisal edge (L2). A model with higid natural tooth was used as control. von Mises equivalent stress (σ vM) and maximum principal stress (σ max) were obtained on Ansys software. Results: The presence of ceramic restoration increased σ vM and σ max in the adhesive interface, mainly for the aluminum oxide (Procera AllCeram system) restorations. The full-coverage crowns generated higher stress in the adhesive interface under L1 while the same result was observed for the laminate veneers under L2. Conclusions: Lithium dissilicate and densely sintered aluminum oxide restorations exhibit different behavior due to different mechanical properties and loading conditions. © 2011 Nova Science Publishers, Inc.

A single multiplex SNaPshot reaction of 42 SNPs to classify admixture populations into mitochondrial DNA haplogroups

SNaPshot minisequencing reaction is in increasing use because of its fast detection of many polymorphisms in a single assay. In this work we described a highly sensitive single nucleotide polymorphisms (SNPs) typing method with detection of 42 mitochondrial DNA (mtDNA) SNPs in a single PCR and SNaPshot multiplex reaction in order to allow haplogroup classification in Latin American admixture population. We validated the panel typing 160 Brazilian individuals. DNA was extracted from blood spotted on filter paper using Chelex protocol. Forty SNPs were selected targeting haplogroup-specific mutations in Europeans, Africans and Asians (only precursors of Native Americans haplogroups A2, B2, C1, and D1) and two non-coding SNPs were chosen to increase the power of discrimination between individuals (SNPs positions 16,519 and 16,362). It was done using a modified version of a previously published multiplex SNaPshot minisequencing reaction established to resolve European haplogroups, adding SNPs targeting Africans (L0, L1, L2, L3, and L*) and Asians (A, B, C, and D) haplogroups based on SNPs described at PhyloTree.org build 2. PCR primers were designed using PerlPrimer software and checked with the Autodimer program. Thirty-three primer-pairs were used to amplify 42 SNPs. Using this panel, we were able to successfully classify 160 individuals into their correct haplogroups. Complete SNP profiles were obtained from 10. pg of total DNA. We conclude that it is possible to build and genotype more than 40 mtDNA SNPs in a single multiplex PCR and SNaPshot reaction, with sensitivity and reliability, resolving haplogroup classification in admixture populations. © 2011.

CIGALA: Challenging the solar maximum in Brazil with PolaRxS

The upcoming solar maximum, which is expected to reach its peak around May 2013, occurs at a time when our reliance on high-precision GNSS has reached unprecedented proportions. The perturbations of the ionosphere caused by increased solar activity pose a major threat to these applications. This is particularly true in equatorial regions where high exposure to solar-induced disturbances is coupled with explosive growth of precise GNSS applications. Along with the various types of solar-induced ionospheric disturbances, strong scintillations are amongst the most challenging, causing phase measurement errors up to full losses of lock for several satellites. Brazil, which heavily relies on high-precision GNSS, is one of the most affected regions due notably to the proximity to the southern crest of the ionospheric equatorial anomaly and to the South Atlantic Magnetic Anomaly. In the framework of the CIGALA project, we developed the PolaRxS™, a GNSS receiver dedicated to the monitoring of ionospheric scintillation indices not only in the GPS L1 band but for all operational and upcoming constellations and frequency bands. A network of these receivers was deployed across the whole Brazilian territory in order to first investigate and secondly to mitigate the impact of scintillation on the different signals, ensuring high precision GNSS availability and integrity in the area. This paper reports on the validation of the PolaRxS™ receiver as an ionospheric scintillation monitor and the first results of the analysis of the data collected with the CIGALA network.

ENCORE: Enhanced Galileo code receiver for surveying applications

In this paper we describe the development of a low-cost high-accuracy Galileo Code receiver, user application software and positioning algorithms for land management applications, which have been implemented using a dedicated FPGA board and dual frequency Galileo E5/L1 Radio Frequency Front-End. The current situation of rural property surveying in Brazil is described and the use of code measurements from the new Galileo signals E5 AltBOC combined with E1 MBOC for use in land management applications is explored. We explain how such approach is expected to allow delivering an absolute positioning solution which could bridge the gap between receivers of high cost/complexity/accuracy based on carrier phase and receivers of lower cost/accuracy based on pseudorange observables. The system is presented together with a detailed description of main components: the Code Receiver and the Application Software. The work presented is part of an ongoing European-Brazilian consortium effort to explore the use of new Galileo for land management applications in Brazil and sponsored by the GNSS Supervisory Authority (GSA).

Fracionamento da fertirrigação e seu efeito na produção da alface sob ambiente protegido

Buscando quantificar o efeito do parcelamento da fertirrigação no desenvolvimento da alface, esta pesquisa procurou verificar o número de parcelas de fertirrigação que apresentasse o melhor desempenho no crescimento da cultura. O experimento se deu em blocos casualizados, com três repetições, quatro tratamentos de fertirrigação e três lâminas de irrigação (L1, L2 e L3). Em um dos tratamentos de fertirrigação (F1), toda a adubação necessária se deu no início do plantio; no tratamento F2, a metade da adubação se deu no início do plantio e a outra metade no meio do ciclo da cultura, uma vez planejada a colheita em 40 dias após o plantio; o tratamento F3 foi fracionado: 1/3 da adubação no início do plantio, 1/3 depois de 13 dias e o ultimo terço após 26 dias do plantio; o tratamento F4 seguiu o mesmo raciocínio, fracionando a aplicação em 4 partes, de 10 em 10 dias. Logo, os quatro tratamentos de fertirrigação receberam a mesma dosagem de fertilizantes. Os resultados permitiram concluir que a aplicação F1 resultou no pior desempenho, com massa fresca de 135,06 gramas por planta, enquanto o melhor resultado se deu no tratamento F2, com massa fresca de 161,40 g. Em todas as lâminas, o parcelamento F3 resultou em sistema radicular mais desenvolvido. O tratamento L1 apresentou em média maior desenvolvimento das raízes, seguido do L2 e L3. O tratamento L1 apresentou menor condutividade elétrica no extrato da solução dos vasos, sendo o único tratamento que diferiu estatisticamente dos demais.

Prevalência de Oestrus ovis (Diptera: Oestridae) em ovinos da região Central de São Paulo, Brasil

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

The impact of adding SBAS data on GPS data processing in southeast of Brazil: Preliminary result

Nowadays, L1 SBAS signals can be used in a combined GPS+SBAS data processing. However, such situation restricts the studies over short baselines. Besides of increasing the satellite availability, SBAS satellites orbit configuration is different from that of GPS. In order to analyze how these characteristics can impact GPS positioning in the southeast area of Brazil, experiments involving GPS-only and combined GPS+SBAS data were performed. Solutions using single point and relative positioning were computed to show the impact over satellite geometry, positioning accuracy and short baseline ambiguity resolution. Results showed that the inclusion of SBAS satellites can improve the accuracy of positioning. Nevertheless, the bad quality of the data broadcasted by these satellites limits their usage. © Springer-Verlag Berlin Heidelberg 2012.

Alterações da densidade mineral óssea em vértebras de ratos wistar submetidos à ausência de carga nos membros pélvicos

The suspension of rats by the tail model is used to investigate the behavior of bone in animals unable to move around. Bone is an adaptative tissue that develops in structure and function, among other factors, in response to mechanical forces applied to it and metabolic demands that it will suffer. The absence of mechanical forces and deformation of bone that occurs causes a decrease in calcium deposition in the absence of stimuli on osteoblasts and osteocytes, favoring the action of osteoclasts, making bones weak and brittle. Therefore, the mechanical action is necessary to stimulate local bone response and thus provide growth and remodeling. The aim of this study was to evaluable by radiographic densitometry, the tail suspension for 15 and 36 days alter the bone mineral density of cervical vertebrae (C3), thoracic (T6) and lumbar (L1 and L3) of Wistar rats. Thirty Rattus norvegicus albinus, adult, male, Wistar strain, average body mass ± 350g, were divided into 3 groups: control (n = 10) - not suspended; S15 (n = 10) - suspended for 15 days and S36 (n = 10) - suspended for 36 days. For densitometric analysis vertebrae were radiographed, scanned, digitized and analyzed by the computer program ImageJ®. There was a statistically significant increase in bone mineral density in group S15, probably by the restlessness of the animals to the suspension, with a decrease in group S36, and this hypothetically is linked to the accommodation of the rats, concluding that the tail suspension altered bone mineral density in first time with a decrease over time.

Preparation and characterization of poly(ε-caprolactone) nanospheres containing the local anesthetic lidocaine

The objective of this work was to develop a modified release system for the local anesthetic lidocaine (LDC), using poly(ε-caprolactone) (PCL) nanospheres (NSs), to improve the pharmacological properties of the drug when administered by the infiltration route. In vitro experiments were used to characterize the system and investigate the release mechanism. The NSs presented a polydispersion index of 0.072, an average diameter of 449.6nm, a zeta potential of -20.1mV, and an association efficiency of 93.3%. The release profiles showed that the release of associated LDC was slower than that of the free drug. Atomic force microscopy analyses showed that the spherical structure of the particles was preserved as a function of time, as well as after the release experiments. Cytotoxicity and pharmacological tests confirmed that association with the NSs reduced the toxicity of LDC, and prolonged its anesthetic action. This new formulation could potentially be used in applications requiring gradual anesthetic release, especially dental procedures. © 2012 Wiley Periodicals, Inc.