A Recombinant Protein Based on Trypanosoma cruzi P21 Enhances Phagocytosis

Background: P21 is a secreted protein expressed in all developmental stages of Trypanosoma cruzi. The aim of this study was to determine the effect of the recombinant protein based on P21 (P21-His(6)) on inflammatory macrophages during phagocytosis. Findings: Our results showed that P21-His(6) acts as a phagocytosis inducer by binding to CXCR4 chemokine receptor and activating actin polymerization in a way dependent on the PI3-kinase signaling pathway. Conclusions: Thus, our results shed light on the notion that native P21 is a component related to T. cruzi evasion from the immune response and that CXCR4 may be involved in phagocytosis. P21-His(6) represents an important experimental control tool to study phagocytosis signaling pathways of different intracellular parasites and particles.

Focal adhesion kinase governs cardiac concentric hypertrophic growth by activating the AKT and mTOR pathways

The heart responds to sustained overload by hypertrophic growth in which the myocytes distinctly thicken or elongate on increases in systolic or diastolic stress. Though potentially adaptive, hypertrophy itself may predispose to cardiac dysfunction in pathological settings. The mechanisms underlying the diverse morphology and outcomes of hypertrophy are uncertain. Here we used a focal adhesion kinase (FAK) cardiac-specific transgenic mice model (FAK-Tg) to explore the function of this non-receptor tyrosine kinase on the regulation of myocyte growth. FAK-Tg mice displayed a phenocopy of concentric cardiac hypertrophy, reflecting the relative thickening of the individual myocytes. Moreover, FAK-Tg mice showed structural, functional and molecular features of a compensated hypertrophic growth, and preserved responses to chronic pressure overload. Mechanistically, FAK overexpression resulted in enhanced myocardial FAK activity, which was proven by treatment with a selective FAK inhibitor to be required for the cardiac hypertrophy in this model. Our results indicate that upregulation of FAK does not affect the activity of Src/ERK1/2 pathway, but stimulated signaling by a cascade that encompasses PI3K, AKT, mTOR, S6K and rpS6. Moreover, inhibition of the mTOR complex by rapamycin extinguished the cardiac hypertrophy of the transgenic FAK mice. These findings uncover a unique role for FAK in regulating the signaling mechanisms that governs the selective myocyte growth in width, likely controlling the activity of PI3K/AKT/mTOR pathway, and suggest that FAK activation could be important for the adaptive response to increases in cardiac afterload. This article is part of a Special Issue entitled "Local Signaling in Myocytes". (C) 2011 Elsevier Ltd. All rights reserved.

Mild Stunting is Associated with Higher Blood Pressure in Overweight Adolescents

Background: Studies have shown that pre/postnatal undernutrition leads to higher risk of non communicable diseases such as diabetes, hypertension and obesity in adulthood. Objetive: To determine whether overweight adolescents with mild stunting [height-for-age Z scores (HAZ) in the range <-1 to >=-2] have higher blood pressure than overweight individuals with normal stature (HAZ >=-1). Methods: Participants were classified as mildly stunted or of normal stature, and further stratified according to body mass index-for-age percentiles as overweight, normal or underweight. Systolic (SBP) and diastolic (DPB) blood pressures were determined according to guidelines, and abdominal fat was analyzed by dual energy X-ray absorptiometry. Results: Mild stunted overweight individuals showed higher DBP values (p=0.001) than their underweight counterparts (69.75 +/- 12.03 and 54.46 +/- 11.24 mmHg, respectively), but similar to those of normal BMI. No differences were found in DBP values of normal, overweight and underweight individuals among the normal stature groups. An increase in SBP (p=0.01) among mild stunted individuals was found when those with overweight were compared to their underweight and normal BMI counterparts (114.70 +/- 15.46, 97.38 +/- 10.87 and 104.72 +/- 12.24 mmHg, respectively). Although no differences were observed in the means of SBP between mild stunting and normal stature groups, a significant intercept was found (p=0.01), revealing higher SBP among stunted individuals. There was a correlation between SBP and abdominal fat (r=0.42, rho=0.02) in the stunted group. Conclusions: Stunted individuals with overweight showed higher SBP than those of normal stature and overweight. These findings confirm that mild stunting increase the risk of future hypertension and alterations are evident at early age. (Arq Bras Cardiol 2012;98(1):6-12)

SMALL INTERFERING RNA TARGETING FOCAL ADHESION KINASE PREVENTS CARDIAC DYSFUNCTION IN ENDOTOXEMIA

Sepsis and septic shock are associated with cardiac depression. Cardiovascular instability is a major cause of death in patients with sepsis. Focal adhesion kinase (FAK) is a potential mediator of cardiomyocyte responses to oxidative and mechanical stress. Myocardial collagen deposition can affect cardiac compliance and contractility. The aim of the present study was to determine whether the silencing of FAK is protective against endotoxemia-induced alterations of cardiac structure and function. In male Wistar rats, endotoxemia was induced by intraperitoneal injection of lipopolysaccharide (10 mg/kg). Cardiac morphometry and function were studied in vivo by left ventricular catheterization and histology. Intravenous injection of small interfering RNA targeting FAK was used to silence myocardial expression of the kinase. The hearts of lipopolysaccharide-injected rats showed collagen deposition, increased matrix metalloproteinase 2 activity, and myocyte hypertrophy, as well as reduced 24-h +dP/dt and -dP/dt, together with hypotension, increased left ventricular end-diastolic pressure, and elevated levels of FAK (phosphorylated and unphosphorylated). Focal adhesion kinase silencing reduced the expression and activation of the kinase in cardiac tissue, as well as protecting against the increased collagen deposition, greater matrix metalloproteinase 2 activity, and reduced cardiac contractility that occur during endotoxemia. In conclusion, FAK is activated in endotoxemia, playing a role in cardiac remodeling and in the impairment of cardiac function. This kinase represents a potential therapeutic target for the protection of cardiac function in patients with sepsis.

FERM domain interaction with myosin negatively regulates FAK in cardiomyocyte hypertrophy

Focal adhesion kinase (FAK) regulates cellular processes that affect several aspects of development and disease. The FAK N-terminal FERM (4.1 protein-ezrin-radixin-moesin homology) domain, a compact clover-leaf structure, binds partner proteins and mediates intramolecular regulatory interactions. Combined chemical cross-linking coupled to MS, small-angle X-ray scattering, computational docking and mutational analyses showed that the FAK FERM domain has a molecular cleft (similar to 998 angstrom(2)) that interacts with sarcomeric myosin, resulting in FAK inhibition. Accordingly, mutations in a unique short amino acid sequence of the FERM myosin cleft, FP-1, impaired the interaction with myosin and enhanced FAK activity in cardiomyocytes. An FP-1 decoy peptide selectively inhibited myosin interaction and increased FAK activity, promoting cardiomyocyte hypertrophy through activation of the AKT-mammalian target of rapamycin pathway. Our findings uncover an inhibitory interaction between the FAK FERM domain and sarcomeric myosin that presents potential opportunities to modulate the cardiac hypertrophic response through changes in FAK activity.

Tests of one Brazilian facial reconstruction method using three soft tissue depth sets and familiar assessors

Facial reconstruction is a method that seeks to recreate a person's facial appearance from his/her skull. This technique can be the last resource used in a forensic investigation, when identification techniques such as DNA analysis, dental records, fingerprints and radiographic comparison cannot be used to identify a body or skeletal remains. To perform facial reconstruction, the data of facial soft tissue thickness are necessary. Scientific literature has described differences in the thickness of facial soft tissue between ethnic groups. There are different databases of soft tissue thickness published in the scientific literature. There are no literature records of facial reconstruction works carried out with data of soft tissues obtained from samples of Brazilian subjects. There are also no reports of digital forensic facial reconstruction performed in Brazil. There are two databases of soft tissue thickness published for the Brazilian population: one obtained from measurements performed in fresh cadavers (fresh cadavers' pattern), and another from measurements using magnetic resonance imaging (Magnetic Resonance pattern). This study aims to perform three different characterized digital forensic facial reconstructions (with hair, eyelashes and eyebrows) of a Brazilian subject (based on an international pattern and two Brazilian patterns for soft facial tissue thickness), and evaluate the digital forensic facial reconstructions comparing them to photos of the individual and other nine subjects. The DICOM data of the Computed Tomography (CT) donated by a volunteer were converted into stereolitography (STL) files and used for the creation of the digital facial reconstructions. Once the three reconstructions were performed, they were compared to photographs of the subject who had the face reconstructed and nine other subjects. Thirty examiners participated in this recognition process. The target subject was recognized by 26.67% of the examiners in the reconstruction performed with the Brazilian Magnetic Resonance Pattern, 23.33% in the reconstruction performed with the Brazilian Fresh Cadavers Pattern and 20.00% in the reconstruction performed with the International Pattern, in which the target-subject was the most recognized subject in the first two patterns. The rate of correct recognitions of the target subject indicate that the digital forensic facial reconstruction, conducted with parameters used in this study, may be a useful tool. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

A 15-year study on the treatment of undernourished children at a nutrition rehabilitation centre (CREN), Brazil

Objective: To build a life table and determine the factors related to the time of treatment of undernourished children at a nutrition rehabilitation centre (CREN), Sao Paulo, Brazil. Design: Nutritional status was assessed from weight-for-age, height-for-age and BMI-for-age Z-scores, while neuropsychomotor development was classified according to the milestones of childhood development. Life tables, Kaplan-Meier survival curves and Cox multiple regression models were employed in data analysis. Setting: CREN (Centre of Nutritional Recovery and Education), Sao Paulo, Brazil. Subjects: Undernourished children (n 228) from the southern slums of Sao Paulo who had received treatment at CREN under a day-hospital regime between the years 1994 and 2009. Results: The Kaplan-Meier curves of survival analysis showed statistically significant differences in the periods of treatment at CREN between children presenting different degrees of neuropsychomotor development (log-rank = 6.621; P = 0.037). Estimates based on the multivariate Cox model revealed that children aged >= 24 months at the time of admission exhibited a lower probability of nutritional rehabilitation (hazard ratio (HR) = 0.49; P = 0.046) at the end of the period compared with infants aged up 12 months. Children presenting slow development were better rehabilitated in comparison with those exhibiting adequate evolution (HR = 4.48; P = 0.023). No significant effects of sex, degree of undernutrition or birth weight on the probability of nutritional rehabilitation were found. Conclusions: Age and neuropsychomotor developmental status at the time of admission to CREN are critical factors in determining the duration of treatment.

Secondary succession impairment in restored mangroves

In this work it was hypothesized that secondary succession on sites that have been managed by single planting of mangrove species is compromised by residual stressors, which could reduce the ecosystem's structural development and lower its functions. Forest structure and environmental characteristics of three planted mangrove stands are compared with reference sites. Structural attributes showed significant differences in the comparison of planted and reference stands. Avicennia schaueriana was the dominant species within both natural regeneration and old-growth stands in terms of basal area (99.2 and 99.4 %, 69.6 and 84.5 %, and 59.0 and 87.1 % for Itacorubi, Saco Grande, and Ratones, respectively). Restoration stands were dominated by Laguncularia racemosa (80.6 and 94.2 % for Saco Grande and Ratones, respectively), except at one site (Itacorubi), where A. schaueriana prevailed (99.7 %). Even though restoration and regeneration stands at Itacorubi showed similar species composition and dominance, cohort sorting revealed an inferior regeneration potential in the restoration stand. Multiple correlation analysis indicated that variables related to elevation disruptions (p (w) = 0.521) were the environmental drivers responsible for the differences observed in forest structure. At restoration sites an impaired pattern of secondary succession was observed, indicating that single species plantings may be ineffective if characteristics of the site, as well as of the area surrounding it, are not considered. The inadequate management of restoration sites can therefore have implications for both immediate and long-term large-scale ecosystem services.

New structures in the continuum of C-15 populated by two-neutron transfer

The C-13(O-18,O-16)C-15 reaction has been studied at 84 MeV incident energy. The ejectiles have been detected at forward angles and C-15 excitation energy spectra have been obtained up to about 20 MeV. Several known bound and resonant states of C-15 have been identified together with two unknown structures at 10.5 MeV (FWHM = 2.5 MeV) and 13.6 MeV (FWHM = 2.5 MeV). Calculations based Oil the removal of two uncorrelated neutrons from the projectile describe a significant part of the continuum observed in the energy spectra. In particular the structure at 10.5 MeV is dominated by a resonance of C-15 near the C-13 + n + n threshold. Similar structures are found in nearby nuclei such as C-14 and Be-11. (c) 2012 Elsevier BM. All rights reserved.

A baixa estatura leve está associada ao aumento da pressão arterial em adolescentes com sobrepeso

FUNDAMENTO: Estudos têm demonstrado que a desnutrição pré/pós-natal leva a um maior risco de doenças não transmissíveis, como diabetes, hipertensão e obesidade na idade adulta. OBJETIVO: Determinar se os adolescentes com sobrepeso e desnutrição leve [escores-Z altura/idade (HAZ) na faixa de <-1 a > -2] têm pressão arterial mais elevada do que os indivíduos com sobrepeso e com estatura normal (HAZ > -1). MÉTODOS: Os participantes foram classificados como de baixa estatura leve ou de estatura normal, e estratificados de acordo com os percentis de massa corporal para a idade, como sobrepeso, peso normal ou abaixo do peso. As pressões arteriais sistólica (PAS) e diastólica (PAD) foram determinadas de acordo com as diretrizes e a gordura abdominal foi analisada por absorciometria de dupla emissão de raios-X. RESULTADOS: Indivíduos com baixa estatura leve e sobrepeso apresentaram valores mais elevados da PAD (p = 0,001) do que suas contrapartes de baixo peso (69,75 ± 12,03 e 54,46 ± 11,24 mmHg, respectivamente), mas semelhantes àqueles com IMC normal. Não foram encontradas diferenças nos valores de PAD em indivíduos normais, indivíduos com sobrepeso e com baixo peso entre os grupos de estatura normal. Foi encontrado um aumento na PAS (p = 0,01) entre os indivíduos com baixa estatura leve quando comparados os indivíduos com sobrepreso com suas contrapartes de baixo peso e IMC normal (114,70 ± 15,46, 97,38 ± 10,87 e 104,72 ± 12,24 mmHg, respectivamente). Embora não tenham sido observadas diferenças nas médias de PAS entre os grupos de baixa estatura leve e estatura normal, foi encontrado um intercepto significativo (p = 0,01), revelando maior PAS entre os indivíduos com baixa estatura leve. Houve correlação entre PAS e gordura abdominal (r = 0,42, ρ = 0,02) no grupo com baixa estatura leve. CONCLUSÃO: Indivíduos de baixa estatura leve com sobrepeso apresentaram maior PAS do que os de estatura normal e sobrepeso. Esses achados confirmam que a baixa estatura leve aumenta o risco futuro de hipertensão e essas alterações são evidentes em indivíduos jovens.

Carbono em agregados do solo sob vegetação nativa, pastagem e sistemas agrícolas no bioma Cerrado

A matéria orgânica do solo (MOS) é responsável pela fertilidade, ciclagem de nutrientes e estabilidade da estrutura, possuindo estreita relação com a agregação do solo. No entanto, ainda não é clara a relação entre as classes de agregados na dinâmica de acúmulo ou perda de MOS em função do uso e manejo do solo. Este trabalho teve como objetivo avaliar os teores de carbono (C) em agregados do solo sob quatro usos e manejos: Cerrado nativo (CE), sistema plantio direto (SPD), sistema plantio convencional (SPC) e pastagem (PA). As áreas de estudo estão localizadas no município de Rio Verde (GO), em solo classificado como Latossolo Vermelho distrófico de textura argilosa. Em amostras de solo coletadas em fevereiro de 2007 nas camadas de 0-5, 5-10 e 10-20 cm de profundidade, foram determinados: a quantidade de macroagregados (maiores que 2,0 mm), mesoagregados (maiores que 0,25 e menores que 2,00 mm) e microagregados (maiores que 0,05 e menores que 0,25 mm) estáveis em água, os índices de agregação diâmetro médio ponderado (DMP) e geométrico (DMG) e os teores de C nas classes de agregados estabelecidas. No solo avaliado observou-se predominância de macroagregados, com maior quantidade sob CE e PA, em comparação ao solo sob SPD e SPC, indicando efeito negativo do cultivo do solo na conservação de macroagregados. Contudo, os teores de C nos agregados do solo na camada de 0-20 cm foram maiores no CE e SPD em relação a PA e SPC, sugerindo que o não revolvimento e a manutenção dos resíduos culturais na superfície do solo favorecem o acúmulo de C. Com isso, foi verificado que os macro e mesoagregados podem ser mais sensíveis ao tipo de uso e manejo do solo, quando comparados aos microagregados. A agregação do solo sob PA é semelhante à do CE, embora com menores teores de C, sugerindo dinâmica de agregação diferenciada, o que merece a atenção de novas pesquisas.

Estoques de carbono e nitrogênio no solo cultivado com mamona na Caatinga

Grande parte da produção brasileira de mamona encontra-se no Nordeste, como opção de cultura para a região semiárida no bioma Caatinga. O objetivo deste estudo foi avaliar as alterações nos estoques de C e N devido à mudança de uso do solo de vegetação natural para o cultivo de mamona no bioma Caatinga. Este trabalho foi realizado na Fazenda Floresta, no município de Irecê, no centro-norte baiano. O clima da região é do tipo BSwh (Köppen) - clima semiárido de altitude. O solo foi classificado em Latossolo Vermelho de textura argilosa. As situações avaliadas foram: três áreas cultivadas com mamona com diferentes tempos de implantação: (i) com 10 anos, (ii) com 20 anos e (iii) com 50 anos; e uma área de referência (vegetação nativa de Caatinga) contígua às situações avaliadas. Os estoques de C e N foram determinados em amostras de solo coletadas em cinco minitrincheiras, nas camadas de 0-5, 5-10, 10-20 e 20-30 cm. Os resultados mostraram que o constante aporte de resíduos vegetais na Caatinga promoveu a manutenção dos estoques de C (90 Mg ha-1) e N (10 Mg ha-1) para a camada de 0-30 cm. A mudança de uso da terra para o cultivo da mamona ocasiona redução em aproximadamente 50 % nos estoques de C e N do solo em relação à vegetação nativa nos primeiros 10 anos de implantação da cultura. A meia-vida da matéria orgânica do solo (MOS) calculada para essa situação na região do semiárido foi de 4,7 anos. O fator de emissão de C do solo, devido à mudança de uso da terra após 20 anos, conforme proposto pelo método do IPCC (2006), foi de 2,47 Mg C ano-1. Por meio do conjunto dos resultados, observa-se a fragilidade do solo do bioma Caatinga no que se refere à perda de MOS devido à mudança de uso da terra.

Alpha cluster structure in 16O.

The alpha cluster phenomenon in the light nuclei structure has been the subject of a longtime investigation since the proposal of the Ikeda diagrams, however the mechanism of the cluster formation is still not completely understood. In fact, if the clusters have a fairly rigid crystal-like or a gas-like structure remains an open question. The interpretation of the Hoyle state as an α condensate brought a renewed interest to this subject, in particular to resonances analogous to the Hoyle state. In this context the study of the experimental evolution of the α-cluster phenomenon through (6Li,d) transfer reactions has been performed in São Paulo. Particularly important are the regions around the nα thresholds where the α-cluster structure states are predicted. The resonant states around the 4α threshold in the nucleus 16O are the focus of the present contribution. The 12C(6Li,d)16O reaction was measured at a bombarding energy of 25.5 MeV employing the São Paulo Pelletron-Enge-Spectrograph facility and the nuclear emulsion detection technique. Resonant states above the α threshold were measured and an energy resolution of 15-30 keV allows to define states previously unresolved. The angular distributions of the absolute cross sections were determined in a range of 4-40 degree in the center of mass system and up to 17 MeV excitation energy. The upper limit for the resonance widths in the crucial region of the 4α threshold was obtained. These values revealed to be at least a factor three smaller than the ones previously reported in the literature, indicating that the α cluster structure information on this region should be revised.

(18O,16O) Two-neutron transfer reactions for spectroscopic studies.

A systematic study of the response of different nuclei to the (18O, 16O) two-neutron transfer reaction at 84 MeV incident energy was pursued at the INFN-LNS in Catania (Italy). The experiments were performed using several solid targets from light (9Bc, 11 B, 12,13C, 16O, 28Si) to heavier ones (58,64Ni, 120Sn, 208Pb). The 16O ejectiles were detected at forward angles by the MAGNEX magnetic spectrometer and identified without the need of time of flight measurements. Exploiting the large momentum (≈ 25%) and angular (50 msr) acceptance of the spectrometer, energy spectra were obtained with a relevant yield up to about 20 MeV excitation energy. A common feature of the light nuclei spectra is the strong population of states with well known configuration of two-particle over a core and the appearance of unknown resonant structures in the continuum. These latter can reveal the excitation of a collective mode connected with the transfer of a pair. For the heavier nuclei as 66Ni a completely different behaviour is observed indicating the presence of more dissipative processes in the reaction mechanisms that hide the spectroscopic information.

In vivo assessment of specific cytotoxic T lymphocyte killing

The direct killing of target cells by cytotoxic T lymphocytes (CTLs) plays a fundamental role in protective immunity to viral, bacterial, protozoan and fungi infections, as well as to tumor cells. In vivo cytotoxic assays take into account the interaction of target and effector cells in the context of the proper microenvironment making the analysis biologically more relevant than in vitro cytotoxic assays. Thus, the development, improvement and validation of in vivo methods are necessary in view of the importance of the results they may provide. We describe and discuss in this manuscript a method to evaluate in vivo specific cytotoxic T lymphocyte killing. We used as model system mice immunized with human recombinant replication-deficient adenovirus 5 (HAd5) containing different transgenes as the trigger of a CTL-mediated immune response. To these mice, we adoptively transferred syngeneic cells labeled with different vital fluorescent dyes. Donor cells were pulsed (target) or not (control non-target) with distinct CD8 T-cell epitopes, mixed in a 1:1 ratio and injected i.v. into immunized or non-immunized recipient mice. After 18-24h, spleen cells are collected and analysed by flow cytometry. A deviation from the 1:1 ratio of control and target cell populations indicates antigen specific lysis of target cells