Efeitos do Exercício Físico e da Terapia Estrogênica Sobre a Reatividade Vascular de Aorta de Ratas Espontaneamente Hipertensas Ovariectomizadas: Papel do Sistema Renina-angiotensina

As doenças cardiovasculares representam as principais causas de morbidade e mortalidade no mundo, inclusive em mulheres no período pós-menopausa. Com a diminuição na produção dos estrógenos, observa-se o aparecimento e a elevação de vários fatores que podem aumentar o risco de desenvolvimento dessas doenças. Dentre esses fatores, o aumento da atividade ou up regulation do Sistema Renina-Angiotensina (SRA) tem se destacado como um importante mediador na fisiopatologia de várias doenças, tal qual, a hipertensão. Apesar de vários estudos experimentais relatarem efeitos benéficos da reposição hormonal com estrogênio, especificamente 17β-estradiol (E2), sobre a redução do risco cardiovascular, os resultados de estudos clínicos são inconclusivos. Desta forma, modificações no estilo de vida se fazem necessárias, como a incorporação da prática regular de exercícios físicos. Muitos estudos têm demonstrado que o exercício físico pode influenciar positivamente sobre os principais fatores de risco cardiovascular, inclusive em mulheres na pós-menopausa. O objetivo do presente estudo foi analisar os efeitos do treinamento físico crônico de natação e da terapia estrogênica sobre a reatividade vascular de aorta de ratas espontaneamente hipertensas (SHR) ovariectomizadas com foco na modulação efetuada pelo SRA. Os experimentos foram conduzidos com ratas SHR Sham e ovariectomizadas, as quais foram divididas aleatoriamente em cinco grupos: SHAM (S), ovariectomizadas (OVX), ovariectomizadas+TE2 (OE2), ovariectomizadas+natação (ON) e ovariectomizadas TE2+N (OE2+N). A reposição com E2 foi realizada por meio de injeções s.c. contendo 5 µg de 17β-estradiol três vezes por semana. O protocolo de treinamento de natação foi realizado por sessenta minutos diários, de forma continua, cinco vezes por semana. Tanto a terapia quanto o treinamento tiveram duração de oito semanas. Quarenta e oito horas após a última sessão de treinamento e/ou tratamento, as ratas tiveram a Pressão Arterial Sistólica (PAS) aferida e após sacrificadas, o sangue foi coletado para dosagem de angiotensina II (Ang II). Para o estudo funcional de reatividade de aorta, avaliou-se a resposta vasoconstritora à angiotensina II e vasodilatadora à Ang-(1-7), ainda efetuando bloqueios farmacológicos para elucidar o mecanismo de ação. A avaliação da expressão das proteínas do SRA em aorta foi realizada por Western Blotting. Os resultados encontrados demonstram que, o grupo ON e OE2+N apresentaram aumento nos níveis plasmáticos de Ang II, que não foi acompanhado por aumento na PAS. A ovariectomia causou um aumento da resposta vasoconstritora a Ang II e diminuição da vasodilatação de Ang-(1-7), que foi prevenida pelo exercício físico ou pela sua associação com a TE2. Além disso, o abrandamento da resposta vasoconstritora a Ang II, assim como o aumento da vasodilatação a Ang-(1-7) parece ter ocorrido por um mecanismo envolvendo o receptor AT2 e Mas, que tiveram sua expressão aumentada no grupo ON e OE2+N. No grupo ON a eficiência funcional destes receptores foi auxiliada pelo aumento na capacidade anti-oxidante, efetuada pela superóxido dismutase. Pôde-se concluir que tanto o treinamento físico quanto a reposição com E2 exercem efeitos cardioprotetores, e a prática regular do exercício físico pode ser uma excelente alternativa à terapia estrogênica em mulheres na pós-menopausa, haja vista que a associação da TE2 ao exercício não promoveu efeitos somatórios

Boundary Terms and Junction Conditions for Generalized Scalar-Tensor Theories

We compute the boundary terms and junction conditions for Horndeski's panoptic class of scalar-tensor theories, and write the bulk and boundary equations of motion in explicitly second order form. We consider a number of special subclasses, including galileon theories, and present the corresponding formulae. Our analysis opens up of the possibility of studying tunnelling between vacua in generalized scalar-tensor theories, and braneworld dynamics. The latter follows because our results are independent of spacetime dimension.Comment: 13 pages, Equation corrected. Thanks to Tsutomu Kobayashi for informing us of the typ

The Imperfect Fluid behind Kinetic Gravity Braiding

We present a standard hydrodynamical description for non-canonical scalar field theories with kinetic gravity braiding. In particular, this picture applies to the simplest galileons and k-essence. The fluid variables not only have a clear physical meaning but also drastically simplify the analysis of the system. The fluid carries charges corresponding to shifts in field space. This shift-charge current contains a spatial part responsible for diffusion of the charges. Moreover, in the incompressible limit, the equation of motion becomes the standard diffusion equation. The fluid is indeed imperfect because the energy flows neither along the field gradient nor along the shift current. The fluid has zero vorticity and is not dissipative: there is no entropy production, the energy-momentum is exactly conserved, the temperature vanishes and there is no shear viscosity. Still, in an expansion around a perfect fluid one can identify terms which correct the pressure in the manner of bulk viscosity. We close by formulating the non-trivial conditions for the thermodynamic equilibrium of this imperfect fluid.Comment: 23 pages plus appendices. New version includes extended discussion on diffusion and dynamics in alternative frames, as well as additional references. v3 reflects version accepted for publication in JHEP: minor comments added regarding suitability to numerical approache

Increased light, moderate, and severe clear-air turbulence in response to climate change

Anthropogenic climate change is expected to strengthen the vertical wind shears at aircraft cruising altitudes within the atmospheric jet streams. Such a strengthening would increase the prevalence of shear instabilities, which generate clear-air turbulence. Climate modelling studies have indicated that the amount of moderate-or-greater clear-air turbulence on transatlantic flight routes in winter will increase significantly in future as the climate changes. However, the individual responses of light, moderate, and severe clear-air turbulence have not previously been studied, despite their importance for aircraft operations. Here we use climate model simulations to analyse the transatlantic wintertime clear-air turbulence response to climate change in five aviation-relevant turbulence strength categories. We find that the probability distributions for an ensemble of 21 clear-air turbulence diagnostics generally gain probability in their right-hand tails when the atmospheric carbon dioxide concentration is doubled. By converting the diagnostics into equivalent eddy dissipation rates, we find that the ensemble-average airspace volume containing light clear-air turbulence increases by 59% (with an intra-ensemble range of 43–68%), light-to-moderate by 75% (39–96%), moderate by 94% (37–118%), moderate-to-severe by 127% (30–170%), and severe by 149% (36–188%). These results suggest that the prevalence of transatlantic wintertime clear-air turbulence will increase significantly in all aviation-relevant strength categories as the climate changes

Defining the Molecular Character of the Developing and Adult Kidney Podocyte

BACKGROUND: The podocyte is a remarkable cell type, which encases the capillaries of the kidney glomerulus. Although mesodermal in origin it sends out axonal like projections that wrap around the capillaries. These extend yet finer projections, the foot processes, which interdigitate, leaving between them the slit diaphragms, through which the glomerular filtrate must pass. The podocytes are a subject of keen interest because of their key roles in kidney development and disease. METHODOLOGY/PRINCIPAL FINDINGS: In this report we identified and characterized a novel transgenic mouse line, MafB-GFP, which specifically marked the kidney podocytes from a very early stage of development. These mice were then used to facilitate the fluorescent activated cell sorting based purification of podocytes from embryos at E13.5 and E15.5, as well as adults. Microarrays were then used to globally define the gene expression states of podocytes at these different developmental stages. A remarkable picture emerged, identifying the multiple sets of genes that establish the neuronal, muscle, and phagocytic properties of podocytes. The complete combinatorial code of transcription factors that create the podocyte was characterized, and the global lists of growth factors and receptors they express were defined. CONCLUSIONS/SIGNIFICANCE: The complete molecular character of the in vivo podocyte is established for the first time. The active molecular functions and biological processes further define their unique combination of features. The results provide a resource atlas of gene expression patterns of developing and adult podocytes that will help to guide further research of these incredible cells

Translocation-coupled DNA cleavage by the Type ISP restriction-modification enzymes

Endonucleolytic double-strand DNA break production requires separate strand cleavage events. Although catalytic mechanisms for simple dimeric endonucleases are available, there are many complex nuclease machines which are poorly understood in comparison. Here we studied the single polypeptide Type ISP restriction-modification (RM) enzymes, which cleave random DNA between distant target sites when two enzymes collide following convergent ATP-driven translocation. We report the 2.7 Angstroms resolution X-ray crystal structure of a Type ISP enzyme-DNA complex, revealing that both the helicase-like ATPase and nuclease are unexpectedly located upstream of the direction of translocation, inconsistent with simple nuclease domain-dimerization. Using single-molecule and biochemical techniques, we demonstrate that each ATPase remodels its DNA-protein complex and translocates along DNA without looping it, leading to a collision complex where the nuclease domains are distal. Sequencing of single cleavage events suggests a previously undescribed endonuclease model, where multiple, stochastic strand nicking events combine to produce DNA scission

Genome-wide association study of kidney function decline in individuals of European descent.

Genome-wide association studies (GWASs) have identified multiple loci associated with cross-sectional eGFR, but a systematic genetic analysis of kidney function decline over time is missing. Here we conducted a GWAS meta-analysis among 63,558 participants of European descent, initially from 16 cohorts with serial kidney function measurements within the CKDGen Consortium, followed by independent replication among additional participants from 13 cohorts. In stage 1 GWAS meta-analysis, single-nucleotide polymorphisms (SNPs) at MEOX2, GALNT11, IL1RAP, NPPA, HPCAL1, and CDH23 showed the strongest associations for at least one trait, in addition to the known UMOD locus, which showed genome-wide significance with an annual change in eGFR. In stage 2 meta-analysis, the significant association at UMOD was replicated. Associations at GALNT11 with Rapid Decline (annual eGFR decline of 3 ml/min per 1.73 m(2) or more), and CDH23 with eGFR change among those with CKD showed significant suggestive evidence of replication. Combined stage 1 and 2 meta-analyses showed significance for UMOD, GALNT11, and CDH23. Morpholino knockdowns of galnt11 and cdh23 in zebrafish embryos each had signs of severe edema 72 h after gentamicin treatment compared with controls, but no gross morphological renal abnormalities before gentamicin administration. Thus, our results suggest a role in the deterioration of kidney function for the loci GALNT11 and CDH23, and show that the UMOD locus is significantly associated with kidney function decline.Kidney International advance online publication, 10 December 2014; doi:10.1038/ki.2014.361

Identification of a Putative Network of Actin-Associated Cytoskeletal Proteins in Glomerular Podocytes Defined by Co-Purified mRNAs

The glomerular podocyte is a highly specialized and polarized kidney cell type that contains major processes and foot processes that extend from the cell body. Foot processes from adjacent podocytes form interdigitations with those of adjacent cells, thereby creating an essential intercellular junctional domain of the renal filtration barrier known as the slit diaphragm. Interesting parallels have been drawn between the slit diaphragm and other sites of cell-cell contact by polarized cells. Notably mutations in several genes encoding proteins localized to the foot processes can lead to proteinuria and kidney failure. Mutations in the Wilm's tumor gene (WT1) can also lead to kidney disease and one isoform of WT1, WT1(+KTS), has been proposed to regulate gene expression post-transcriptionally. We originally sought to identify mRNAs associated with WT1(+KTS) through an RNA immunoprecipitation and microarray approach, hypothesizing that the proteins encoded by these mRNAs might be important for podocyte morphology and function. We identified a subset of mRNAs that were remarkably enriched for transcripts encoding actin-binding proteins and other cytoskeletal proteins including several that are localized at or near the slit diaphragm. Interestingly, these mRNAs included those of α-actinin-4 and non-muscle myosin IIA that are mutated in genetic forms of kidney disease. However, isolation of the mRNAs occurred independently of the expression of WT1, suggesting that the identified mRNAs were serendipitously co-purified on the basis of co-association in a common subcellular fraction. Mass spectroscopy revealed that other components of the actin cytoskeleton co-purified with these mRNAs, namely actin, tubulin, and elongation factor 1α. We propose that these mRNAs encode a number of proteins that comprise a highly specialized protein interactome underlying the slit diaphragm. Collectively, these gene products and their interactions may prove to be important for the structural integrity of the actin cytoskeleton in podocytes as well as other polarized cell types

B-type natriuretic peptide-induced delayed modulation of TRPV1 and P2X3 receptors of mouse trigeminal sensory neurons

Important pain transducers of noxious stimuli are small- and medium-diameter sensory neurons that express transient receptor vanilloid-1 (TRPV1) channels and/or adenosine triphosphate (ATP)-gated P2X3 receptors whose activity is upregulated by endogenous neuropeptides in acute and chronic pain models. Little is known about the role of endogenous modulators in restraining the expression and function of TRPV1 and P2X3 receptors. In dorsal root ganglia, evidence supports the involvement of the natriuretic peptide system in the modulation of nociceptive transmission especially via the B-type natriuretic peptide (BNP) that activates the natriuretic peptide receptor-A (NPR-A) to downregulate sensory neuron excitability. Since the role of BNP in trigeminal ganglia (TG) is unclear, we investigated the expression of BNP in mouse TG in situ or in primary cultures and its effect on P2X3 and TRPV1 receptors of patch-clamped cultured neurons. Against scant expression of BNP, almost all neurons expressed NPRA at membrane level. While BNP rapidly increased cGMP production and Akt kinase phosphorylation, there was no early change in passive neuronal properties or responses to capsaicin, \u3b1,\u3b2-meATP or GABA. Nonetheless, 24 h application of BNP depressed TRPV1 mediated currents (an effect blocked by the NPR-A antagonist anantin) without changing responses to \u3b1,\u3b2-meATP or GABA. Anantin alone decreased basal cGMP production and enhanced control \u3b1,\u3b2-meATP-evoked responses, implying constitutive regulation of P2X3 receptors by ambient BNP. These data suggest a slow modulatory action by BNP on TRPV1 and P2X3 receptors outlining the role of this peptide as a negative regulator of trigeminal sensory neuron excitability to nociceptive stimuli. \ua9 2013 Vilotti et al

1000 Genomes-based meta-analysis identifies 10 novel loci for kidney function.

HapMap imputed genome-wide association studies (GWAS) have revealed >50 loci at which common variants with minor allele frequency >5% are associated with kidney function. GWAS using more complete reference sets for imputation, such as those from The 1000 Genomes project, promise to identify novel loci that have been missed by previous efforts. To investigate the value of such a more complete variant catalog, we conducted a GWAS meta-analysis of kidney function based on the estimated glomerular filtration rate (eGFR) in 110,517 European ancestry participants using 1000 Genomes imputed data. We identified 10 novel loci with p-value < 5 × 10(-8) previously missed by HapMap-based GWAS. Six of these loci (HOXD8, ARL15, PIK3R1, EYA4, ASTN2, and EPB41L3) are tagged by common SNPs unique to the 1000 Genomes reference panel. Using pathway analysis, we identified 39 significant (FDR < 0.05) genes and 127 significantly (FDR < 0.05) enriched gene sets, which were missed by our previous analyses. Among those, the 10 identified novel genes are part of pathways of kidney development, carbohydrate metabolism, cardiac septum development and glucose metabolism. These results highlight the utility of re-imputing from denser reference panels, until whole-genome sequencing becomes feasible in large samples