Context-specific modulation of cocaine-induced locomotor sensitization and ERK and CREB phosphorylation in the rat nucleus accumbens

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Stress induces behavioral sensitization, increases nicotine-seeking behavior and leads to a decrease of CREB in the nucleus accumbens

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

A novel mutation (Q40P) in PAX8 associated with congenital hypothyroidism and thyroid hypoplasia: Evidence for phenotypic variability in mother and child

Congenital hypothyroidism associated with thyroid hypoplasia can be caused by several genetic defects, including mutations in the TSH beta -subunit, the TSH receptor, the G(A)alpha -subunit, and the transcription factor PAX8. Four girls with sporadic congenital hypothyroidism and hypoplastic thyroid glands were analyzed for mutations in PAX8 and TTF2 (FKHL15). Mutations in the coding region of the TSH beta -subunit gene, the TSH receptor gene, and exons 8 and 9 of G(mu)alpha had been excluded previously. Serum TSH concentrations were 150 mU/liter or more, TG levels were within normal limits, and thyroid autoantibodies were absent. Technetium scintigraphies did not reveal the presence of thyroid tissue, but ultrasonography documented hypoplastic, normally located glands.One patient was found to harbor a heterozygous transversion 119A -->C in exon 3 of PAX8 replacing a conserved glutamine by proline in the paired box domain (Q40P). Analysis of her family members revealed that her mother, who has a thyroid gland of normal size and mild, adult-onset autoimmune hypothyroidism, is also heterozygous for this mutation. Functional analyses of the PAX8 Q40P mutation showed impaired binding to a PAX8 response element and absent transactivation of a thyroid peroxidase promoter luciferase reporter gene.These findings confirm the important role of PAX8 in the development of the thyroid, but they indicate that PAX8 gene mutations may have a variable penetrance or expressivity. The absence of mutations in the coding sequences of the analyzed genes in the three other patients supports the concept that the pathogenesis of congenital hypothyroidism associated with thyroid hypoplasia is diverse.

Genotyping of AR and PSA polymorphisms in a patient with Klinefelter syndrome, non-Hodgkin lymphoma, and adenocarcinoma of the prostate

It has been hypothesized that the AR (androgen receptor) gene binds the two PSA (prostate-specific antigen) alleles with differing affinities and may differentially influence prostate cancer risk. In this article, we report a case of adenocarcinoma of the prostate in a 56-year-old man with Klinefelter syndrome (47,XXY) and non-Hodgkin lymphoma, as well as the AR and PSA genotype. AR and PSA gene polymorphisms were analyzed by polymerase chain reaction-based methods using DNA from peripheral white blood cells and the prostate cancer. We determined the methylation status of the AR gene on the X chromosome. The patient presents with the AG genotype for the ARE-I (androgen response element) region of the PSA gene. We detect the presence of two short AR alleles with 19 and 11CAG repeats each. Unmethylated alleles were demonstrated for both. The shorter allele was inactive in more than 60% of total DNA in both control blood and prostate cancer cells. The presence of short AR alleles and the G allele of the PSA gene may contribute to the development of prostate cancer in a 47,XXY patient. (C) 2004 Elsevier B.V. All rights reserved.

A systematic approach to identify STRE-binding proteins of the gsn glycogen synthase gene promoter in Neurospora crassa

The gene encoding glycogen synthase in Neurospora crassa (gsn) is transcriptionally down-regulated when mycelium is exposed to a heat shock from 30 to 45 degrees C. The gsn promoter has one stress response element (STRE) motif that is specifically bound by heat shock activated nuclear proteins. In this work, we used biochemical approaches together with mass spectrometric analysis to identify the proteins that bind to the STRE motif and could participate in the gsn transcription regulation during heat shock. Crude nuclear extract of heat-shocked mycelium was prepared and fractionated by affinity chromatography. The fractions exhibiting DNA-binding activity were identified by electrophoretic mobility shift assay (EMSA) using as probe a DNA fragment containing the STRE motif DNA-protein binding activity was confirmed by Southwestern analysis. The molecular mass (MM) of proteins was estimated by fractionating the crude nuclear extract by SDS-PAGE followed by EMSA analysis of the proteins corresponding to different MM intervals. Binding activity was detected at the 30-50 MM kDa interval. Fractionation of the crude nuclear proteins by IEF followed by EMSA analysis led to the identification of two active fractions belonging to the pIs intervals 3.54-4.08 and 6.77-7.31. The proteins comprising the MM and pI intervals previously identified were excised from a 2-DE gel, and subjected to mass spectrometric analysis (MALDI-TOF/TOF) after tryptic digestion. The proteins were identified by search against the MIPS and MIT N. crassa databases and five promising candidates were identified. Their structural characteristics and putative roles in the gsn transcription regulation are discussed.

Dynamic stationary response of reinforced plates by the boundary element method

A direct version of the boundary element method (BEM) is developed to model the stationary dynamic response of reinforced plate structures, such as reinforced panels in buildings, automobiles, and airplanes. The dynamic stationary fundamental solutions of thin plates and plane stress state are used to transform the governing partial differential equations into boundary integral equations (BIEs). Two sets of uncoupled BIEs are formulated, respectively, for the in-plane state ( membrane) and for the out-of-plane state ( bending). These uncoupled systems are joined to formamacro-element, in which membrane and bending effects are present. The association of these macro-elements is able to simulate thin-walled structures, including reinforced plate structures. In the present formulation, the BIE is discretized by continuous and/or discontinuous linear elements. Four displacement integral equations are written for every boundary node. Modal data, that is, natural frequencies and the corresponding mode shapes of reinforced plates, are obtained from information contained in the frequency response functions (FRFs). A specific example is presented to illustrate the versatility of the proposed methodology. Different configurations of the reinforcements are used to simulate simply supported and clamped boundary conditions for the plate structures. The procedure is validated by comparison with results determined by the finite element method (FEM).

Using of model updating considering damped frequency response function

The search for better performance in the structural systems has been taken to more refined models, involving the analysis of a growing number of details, which should be correctly formulated aiming at defining a representative model of the real system. Representative models demand a great detailing of the project and search for new techniques of evaluation and analysis. Model updating is one of this technologies, it can be used to improve the predictive capabilities of computer-based models. This paper presents a FRF-based finite element model updating procedure whose the updating variables are physical parameters of the model. It includes the damping effects in the updating procedure assuming proportional and none proportional damping mechanism. The updating parameters are defined at an element level or macro regions of the model. So, the parameters are adjusted locally, facilitating the physical interpretation of the adjusting of the model. Different tests for simulated and experimental data are discussed aiming at defining the characteristics and potentialities of the methodology.

Three-Dimensional Finite Element Analysis of Vertical and Angular Misfit in Implant-Supported Fixed Prostheses

Purpose: Three-dimensional finite element analysis was used to evaluate the effect of vertical and angular misfit in three-piece implant-supported screw-retained fixed prostheses on the biomechanical response in the peri-implant bone, implants, and prosthetic components. Materials and Methods: Four three-dimensional models were fabricated to represent a right posterior mandibular section with one implant in the region of the second premolar (2PM) and another in the region of the second molar (2M). The implants were splinted by a three-piece implant-supported metal-ceramic prosthesis and differed according to the type of misfit, as represented by four different models: Control = prosthesis with complete fit to the implants; UAM (unilateral angular misfit) = prosthesis presenting unilateral angular misfit of 100 pm in the mesial region of the 2M; UVM (unilateral vertical misfit) = prosthesis presenting unilateral vertical misfit of 100 pm in the mesial region of the 2M; and TVM (total vertical misfit) = prosthesis presenting total vertical misfit of 100 pm in the platform of the framework in the 2M. A vertical load of 400 N was distributed and applied on 12 centric points by the software Ansys, ie, a vertical load of 150 N was applied to each molar in the prosthesis and a vertical load of 100 N was applied at the 2PM. Results: The stress values and distribution in peri-implant bone tissue were similar for all groups. The models with misfit exhibited different distribution patterns and increased stress magnitude in comparison to the control. The highest stress values in group UAM were observed in the implant body and retention screw. The groups UVM and TVM exhibited high stress values in the platform of the framework and the implant hexagon, respectively. Conclusions: The three types of misfit influenced the magnitude and distribution of stresses. The influence of misfit on peri-implant bone tissue was modest. Each type of misfit increased the stress values in different regions of the system. INT J ORAL MAXILLOFAC IMPLANTS 2011;26:788-796

Can sieve-element plastids in Panicum maximum (Poaceae) leaves act in the blockage of injured sieve-tube elements?

The ultrastructural features and the plastid changes caused by sample preparation were studied in sieve elements of Panicum maximum leaves. Samples of expanded leaves, taken near the ligule region, were fixed and processed by common light and transmission electron microscopy methods. In mature sieve-tube elements, the protoplast is electron-translucent and plastids are the most frequent organelles. Mitochondria and smooth endoplasmic reticulum segments are also visible and occupy a parietal position within the cell. The plastids are globular and show electron-dense proteinaceous inclusions in the stroma. The protein crystals are predominantly cuneate, but thin crystalloids and amorphous and/or filamentous proteins also occur. The presence of intact plastids plus others in different phases of plastid envelope rupture were interpreted as evidence that this rupture is a normal event in response to injury. This plastid envelope rupture is possibly activated by the release of pressure in the sieve-tube element. After plastid membrane vesiculation, the stroma and the protein crystals are dispersed within the sieve-element ground cytoplasm. The vesicles originating from the plastid envelope move to one cell pole, while protein crystalloids move to the opposite pole and agglomerate in the sieve-plate region. Our findings indicate that these protein crystalloids, which deposit in the sieve plate, may act in sieve-plate pores occlusion, preventing the release of phloem sap, similar to the role of P-protein in dicotyledons. (c) 2008 Elsevier GmbH. All rights reserved.

SIRT1 Deacetylase Activity and the Maintenance of Protein Homeostasis in Response to Stress: An Overview

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

EFFECTS of SELENIUM SUPPLEMENTATION on CATTLE ANTI-RABIES HUMORAL IMMUNE RESPONSE and LEVELS of SERUM SELENIUM and CORTISOL

This study evaluated the effect of different concentrations of selenium (Se) supplementation on cattle anti-rabies humoral immune response, serum Se concentrations and cortisol levels. Sixty uncastrated male Nelore calves from 10 to 12 months grazing on Brachiaria decumbens forage were studied. The animals were assigned to one of four groups (n = 15 each), which received non-supplemented diets (Gc) or supplemented with daily and individual Selenium ( Se) concentrations of 3.6 mg (G(3.6)), 5.4 mg (G(5.4)) or 6.4 mg (G(6.4)). The calves were immunized on day 0 with one dose of commercial liquid inactivated rabies vaccination. on days 15, 30, 60, 90 and 120, the cattle underwent the same stressing procedures used for vaccination in the corral. Cattle blood samples were collected after vaccination and stressing procedures to determine serum Se levels, rabies antibody titers and serum cortisol. Se levels were also determined in forage samples collected from the paddocks in which the cattle were held. Se concentration in B. decumbens was 0.04 mg of Se/kg dry matter. Baseline Se levels obtained on day 0 were higher in Gc than in G(5.4) and G(6.4) (P = 0.005). Serum Se levels decreased in Gc throughout the experiment (P < 0.004), increased in G(3.6) (P < 0.000) and G(5.4) (P < 0.000) and were kept high from day 60 on in group G(6.4) (P < 0.002). Rabies antibody titers did not differ among control and supplemented groups. However, 120 days after vaccination rabies antibody titers were kept above protective levels (>= 0.5 UI/mL) only in group G(3.6) (P < 0.00002), whereas they dropped in the other groups (P < 0.05). Serum cortisol levels did not differ among the experimental groups (P = 0.79), reached peak levels on day 90 and returned close to baseline levels on day 120. Se and cortisol levels were not markedly correlated. Serum cortisol and rabies antibody titers were correlated only in group G(6.4), on day 60 (R = 0.513; P = 0.05) and 120 (R = 0.644; P = 0.009). Serum Se and rabies antibody titers were correlated only in group G(6.4), on day 60 (R = -0.580; P = 0.023). In conclusion: a) the profile of Se variation is different among groups receiving different concentrations of this element; b) the supplementation dosage of 3.6 mg Se/animal/day is efficient to treat/prevent marginal Se deficiency; c) individual supplementation with daily concentrations of 3.6 mg Se enhances the maintenance of rabies antibody titers in cattle; d) individual supplementation with daily concentrations of 3.6; 5.4 and 6.4 mg Se are ineffective in reducing serum cortisol; e) repeated cattle handling in corrals stress animals that adapt to these procedures, although serum cortisol does not return to baseline levels by 120 days; and f) the stress generated by repeated management in cattle in the corral does not diminish antibody titers after vaccination against rabies.

Expressão, purificação e estudos estruturais da chaperona molecular Hsp65 de Mycobacterium leprae & caracterização da proteína NUDEL - nuclear distribuition element-like

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

Lumped element modeling of operational structures by inverting the mobility models

This paper is concerned with what a source precisely sees when it drives a receiver such as a continuous structural object. An equivalent lumped element system consisting of masses, springs and dampers is developed to visually represent the operational structural dynamics of a single-input structure at the driving point. The development is solely based on the mobility model of the driving point response. The mobility model is mathematically inverted to give the impedance model that is suitable for lumped element modeling. The two types of structures studied are unconstrained inertial objects and constrained resilient objects. The lumped element systems presented suggest a new view to dynamics that a single-input flexible structure in operation can be decomposed into the two subsystems: a base system of single degree of freedom (or of a mass for an inertial object) whose mass is in contact with the source and an appendage system consisting of a series of oscillators each of which is attached to the base mass. The driving point response is a result of the coupling between the two subsystems. (C) 2014 Elsevier Ltd. All rights reserved.

Lumped Element Modeling of a Flexible Manipulator System

This paper is concerned with a modeling method that can be used for an experimental identification of a dynamic system. More specifically, an equivalent lumped element system is presented to represent in a unique and exact manner a complete proportional-derivative (PD) controlled servo positioning system having a flexible manipulator. The impedance and mobility approach is used to transform the P and D control gains to an electrical spring and an electrical damper, respectively. The impedance model method is used to transform the flexible manipulator to a coupled system between a single contact mass at the driving position and a series of noncontact masses each of which is connected to the contact mass via a resilient member. This method is applicable whenever the driving point response of such a manipulator is available. Experimental work is presented to support the theory developed.

Complex modal analysis of a vertical rotor through finite element method

Natural frequencies were analyzed (axial, torsional and flexural) and frequency response of a vertical rotor with a hard disk at the edge through the classical and complex modal analysis. The mathematical modeling was based on the theory of Euler-Bernoulli beam. The equation that rules the movement was obtained through the Lagrangian formulation. The model considered the effects of bending, torsion and axial deformation of the shaft, besides the gravitational and gyroscopic effects. The finite element method was used to discretize the structure into hollow cylindrical elements with 12 degrees of freedom. Mass, stiffness and gyroscopic matrices were explained consistently. This type of tool, based on the use of complex coordinates to describe the dynamic behavior of rotating shaft, allows the decomposition of the system in two submodes, backward and forward. Thus, it is possible to clearly visualize that the orbit and direction of the precessional motion around the line of the rotating shaft is not deformed. A finite element program was developed using Matlab ®, and numerical simulations were performed to validate this model.