COMPARAÇÃO DA UTILIZAÇÃO DAS TÉCNICAS WATSU E RELAXAMENTO AQUÁTICO EM FLUTUAÇÃO ASSISTIDA NOS SINTOMAS DE ANSIEDADE, DEPRESSÃO E PERCEPÇÃO DA DOR

Este estudo teve como objetivo comparar o efeito do método Watsu e do relaxamento aquático em flutuação assistida em piscina, no tratamento da dor crônica. Também objetiva avaliar o efeito dos sintomas de depressão e ansiedade como co-fatores nesses tratamentos. A amostra é constituída por 23 indivíduos de ambos os sexos, acima de 18 anos. No método Watsu são atendidas 13 pessoas, sendo 11 mulheres e 02 homens. Na técnica de relaxamento assistido são atendidas 10 pessoas, sendo 07 mulheres e 03 homens, entre pacientes que procuram tratamento fisioterápico no centro clínico Escola de Fisioterapia da UMESP, com dor crônica. Os dados são colhidos através de entrevista estruturada. Aplicada escala associada para avaliar percepção de dor, questionário de avaliação de sintomas de depressão Beck, de Ansiedade Idate-Estado e Ansiedade Idate-Traço. Devido ao pequeno tamanho da amostra, as comparações entre os dois tipos de tratamento (Watsu e relaxamento) e, como os resultados obtidos são equivalentes em ambos os procedimentos, optou se por apresentar os resultados apenas das análises por métodos paramétricos (teste t de média, regressão linear e análise de variância). Não são observadas diferenças significativas entre os dois grupos em relação aos escores de ansiedade e depressão antes e após a realização da pesquisa. Em relação à comparação entre os dois grupos pesquisados, quanto à percepção de dor, observou-se que tanto o método de Watsu quanto as técnicas de relaxamento mostram um efeito significativo na redução da dor. Esta pesquisa sugere que o método Watsu é tão eficaz para o controle da dor quanto o método de relaxamento, porém, o grupo de pacientes submetidos ao método Watsu é constituído por pessoas com níveis de intensidade de dor iniciais maiores do que o grupo de relaxamento. Com isso, pode-se supor que a demanda por eficácia clinica é maior para o método Watsu. Outro achado interessante é que os níveis de ansiedade ou depressão presentes nos participantes não parecem influenciar a resposta ao efeito do tratamento sobre a dor. Novos estudos do tipo duplo-cego controlados são necessários para, além de confirmar a eficácia do método, ajudar a entender quais detalhes dos procedimentos da técnica Watsu são mais eficazes para cada tipo de dor e de estado afetivo do paciente.

Cascaded Bayesian processes: an account of bias in orientation perception

Following adaptation to an oriented (1-d) signal in central vision, the orientation of subsequently viewed test signals may appear repelled away from or attracted towards the adapting orientation. Small angular differences between the adaptor and test yield 'repulsive' shifts, while large angular differences yield 'attractive' shifts. In peripheral vision, however, both small and large angular differences yield repulsive shifts. To account for these tilt after-effects (TAEs), a cascaded model of orientation estimation that is optimized using hierarchical Bayesian methods is proposed. The model accounts for orientation bias through adaptation-induced losses in information that arise because of signal uncertainties and neural constraints placed upon the propagation of visual information. Repulsive (direct) TAEs arise at early stages of visual processing from adaptation of orientation-selective units with peak sensitivity at the orientation of the adaptor (theta). Attractive (indirect) TAEs result from adaptation of second-stage units with peak sensitivity at theta and theta+90 degrees , which arise from an efficient stage of linear compression that pools across the responses of the first-stage orientation-selective units. A spatial orientation vector is estimated from the transformed oriented unit responses. The change from attractive to repulsive TAEs in peripheral vision can be explained by the differing harmonic biases resulting from constraints on signal power (in central vision) versus signal uncertainties in orientation (in peripheral vision). The proposed model is consistent with recent work by computational neuroscientists in supposing that visual bias reflects the adjustment of a rational system in the light of uncertain signals and system constraints.

Ceramide and reactive oxygen species (ROS) as signal transduction molecules in inflammation

Reactive oxygen species (ROS) and the sphingolipid ceramide are each partly responsible for the intracellular signal transduction of a variety of physiological, pharmacological or environmental agents. Furthermore, the enhanced production of many of these agents, that utilise ROS and ceramide as signalling intermediates, is associated with the aetiologies of several vascular diseases (e.g. atherosclerosis) or disorders of inflammatory origin (e.g. rheumatoid arthritis; RA). Excessive monocyte recruitment and uncontrolled T cell activation are both strongly implicated in the chronic inflammatory responses that are associated with these pathologies. Therefore the aims of this thesis are (1) to further elucidate the cellular responses to modulations in intracellular ceramide/ROS levels in monocytes and T cells, in order to help resolve the mechanisms of progression of these diseases and (2) to examine both existing agents (methotrexate) and novel targets for possible therapeutic manipulation. Utilising synthetic, short chain ceramide to mimic the cellular responses to fluctuations in natural endogenous ceramide or, stimulation of CD95 to induce ceramide formation, it is described here that ceramide targets and manipulates two discrete sites responsible for ROS generation, preceding the cellular responses of growth arrest in U937 monocytes and apoptosis in Jurkat T-cells. In both cell types, transient elevations in mitochondrial ROS generation were observed. However, the prominent redox altering effects appear to be the ceramide-mediated reduction in cytosolic peroxide, the magnitude of which dictates in part the cellular response in U937 monocytes, Jurkat T-cells and primary human peripheral blood resting or PHA-activated T-cells in vitro. The application of synthetic ceramides to U937 monocytes for short (2 hours) or long (16 hours) treatment periods reduced the membrane expression of proteins associated with cell-cell interaction. Furthermore, ceramide treated U937 monocytes demonstrated reduced adhesion to 5 or 24 hour LPS activated human umbilical vein endothelial cells (HUVEC) but not resting HUVEC. Consequently it is hypothesised that the targeted treatment of monocytes from patients with cardiovascular diseases with short chain synthetic ceramide may reduce disease progression. Herein, the anti-inflammatory and immunosuppressant drug, methotrexate, is described to require ROS production for the induction of cytostasis or cytotoxicity in U937 monocytes and Jurkat T-cells respectively. Further, ROS are critical for methotrexate to abrogate monocyte interaction with activated HUVEC in vitro. The histological feature of RA of enhanced infiltration, survivability and hyporesponsiveness of T-cells within the diseased synovium has been suggested to arise from aberrant signalling. No difference in the concentrations of endogenous T-cell ceramide, the related lipid diacylglycerol (DAG) and cytosolic peroxide ex vivo was observed. TCR activation following PHA exposure in vitro for 72 hours did not induced maintained perturbations in DAG or ceramide in T-cells from RA patients or healthy individuals. However, T-cells from RA patients failed to upregulate cytosolic peroxide in response to PHA, unlike those from normals, despite expressing identical levels of the activation marker CD25. This inability to upregulate cytosolic peroxide may contribute to the T-cell pathology associated with RA by affecting the signalling capacity of redox sensitive biomolecules. These data highlight the importance of two distinctive cellular pools of ROS in mediating complex biological events associated with inflammatory disease and suggest that modulation of cellular ceramides represents a novel therapeutic strategy to minimise monocyte recruitment.

Beta frequency neuronal network activity in the primary motor cortex

In this study I investigated the mechanisms of neuronal network oscillatory activity in rat M1 using pharmacological manipulations and electrical stimulation protocols, employing the in vitro brain slice technique in rat and magnetoencephalography (MEG) in man. Co-application of kainic acid and carbachol generated in vitro beta oscillatory activity in all layers in M1. Analyses indicated that oscillations originated from deep layers and indicated significant involvement of GABAA receptors and gap junctions. A modulatory role of GABAB, NMDA, and dopamine receptors was also evident. Intracellular recordings from fast-spiking (FS) GABAergic inhibitory cells revealed phase-locked action potentials (APs) on every beta cycle. Glutamatergic excitatory regular-spiking (RS) and intrinsically-bursting (IB) cells both received phase locked inhibitory postsynaptic potentials, but did not fire APs on every cycle, suggesting the dynamic involvement of different pools of neurones in the overall population oscillations. Stimulation evoked activity at high frequency (HFS; 125Hz) evoked gamma oscillations and reduced ongoing beta activity. 20Hz stimulation promoted theta or gamma oscillations whilst 4Hz stimulation enhanced beta power at theta frequency. I also investigated the modulation of pathological slow wave (theta and beta) oscillatory activity using magnetoencephalography. Abnormal activity was suppressed by sub-sedative doses of GABAA receptor modulator zolpidem and the observed desynchronising effect correlated well with improved sensorimotor function. These studies indicate a fundamental role for inhibitory neuronal networks in the patterning beta activity and suggest that cortical HFS in PD re-patterns abnormally enhanced M1 network activity by modulating the activity of FS cells. Furthermore, pathological oscillation may be common to many neuropathologies and may be an important future therapeutic target.

Hand somatosensory subcortical and cortical sources assessed by functional source separation:an EEG study

We propose a novel electroencephalographic application of a recently developed cerebral source extraction method (Functional Source Separation, FSS), which starts from extracranial signals and adds a functional constraint to the cost function of a basic independent component analysis model without requiring solutions to be independent. Five ad-hoc functional constraints were used to extract the activity reflecting the temporal sequence of sensory information processing along the somatosensory pathway in response to the separate left and right median nerve galvanic stimulation. Constraints required only the maximization of the responsiveness at specific latencies following sensory stimulation, without taking into account that any frequency or spatial information. After source extraction, the reliability of identified FS was assessed based on the position of single dipoles fitted on its retroprojected signals and on a discrepancy measure. The FS positions were consistent with previously reported data (two early subcortical sources localized in the brain stem and thalamus, the three later sources in cortical areas), leaving negligible residual activity at the corresponding latencies. The high-frequency component of the oscillatory activity (HFO) of the extracted component was analyzed. The integrity of the low amplitude HFOs was preserved for each FS. On the basis of our data, we suggest that FSS can be an effective tool to investigate the HFO behavior of the different neuronal pools, recruited at successive times after median nerve galvanic stimulation. As FSs are reconstructed along the entire experimental session, directional and dynamic HFO synchronization phenomena can be studied.

Knowledge mediation and overlapping in interfirm networks

Purpose – The literature on interfirm networks devotes scant attention to the ways collaborating firms combine and integrate the knowledge they share and to the subsequent learning outcomes. This study aims to investigate how motorsport companies use network ties to share and recombine knowledge and the learning that occurs both at the organizational and dyadic network levels. Design/methodology/approach – The paper adopts a qualitative and inductive approach with the aim of developing theory from an in-depth examination of the dyadic ties between motorsport companies and the way they share and recombine knowledge. Findings – The research shows that motorsport companies having substantial competences at managing knowledge flows do so by getting advantage of bridging ties. While bridging ties allow motorsport companies to reach distant and diverse sources of knowledge, their strengthening and the formation of relational capital facilitate the mediation and overlapping of that knowledge. Research limitations/implications – The analysis rests on a qualitative account in a single industry and does not take into account different types of inter-firm networks (e.g. alliances; constellations; consortia etc.) and governance structures. Cross-industry analyses may provide a more fine-grained picture of the practices used to recombine knowledge and the ideal composition of inter-firm ties. Practical implications – This study provides some interesting implications for scholars and managers concerned with the management of innovation activities at the interfirm level. From a managerial point of view, the recognition of the different roles played by network spanning connections is particularly salient and raises issues concerning the effective design and management of interfirm ties. Originality/value – Although much of the literature emphasizes the role of bridging ties in connecting to diverse pools of knowledge, this paper goes one step further and investigates in more depth how firms gather and combine distant and heterogeneous sources of knowledge through the use of strengthened bridging ties and a micro-context conducive to high quality relationships.

Effect of aspect ratio on thermal convection in open and closed systems

Internally heated fluids are found across the nuclear fuel cycle. In certain situations the motion of the fluid is driven by the decay heat (i.e. corium melt pools in severe accidents, the shutdown of liquid metal reactors, molten salt and the passive control of light water reactors) as well as normal operation (i.e. intermediate waste storage and generation IV reactor designs). This can in the long-term affect reactor vessel integrity or lead to localized hot spots and accumulation of solid wastes that may prompt local increases in activity. Two approaches to the modeling of internally heated convection are presented here. These are based on numerical analysis using codes developed in-house and simulations using widely available computational fluid dynamics solvers. Open and closed fluid layers at around the transition between conduction and convection of various aspect ratios are considered. We determine optimum domain aspect ratio (1:7:7 up to 1:24:24 for open systems and 5:5:1, 1:10:10 and 1:20:20 for closed systems), mesh resolutions and turbulence models required to accurately and efficiently capture the convection structures that evolve when perturbing the conductive state of the fluid layer. Note that the open and closed fluid layers we study here are bounded by a conducting surface over an insulating surface. Conclusions will be drawn on the influence of the periodic boundary conditions on the flow patterns observed. We have also examined the stability of the nonlinear solutions that we found with the aim of identifying the bifurcation sequence of these solutions en route to turbulence.

Production, purification and characterization of recombinant, full-length human claudin-1

The transmembrane domain proteins of the claudin superfamily are the major structural components of cellular tight junctions. One family member, claudin-1, also associates with tetraspanin CD81 as part of a receptor complex that is essential for hepatitis C virus (HCV) infection of the liver. To understand the molecular basis of claudin-1/CD81 association we previously produced and purified milligram quantities of functional, full-length CD81, which binds a soluble form of HCV E2 glycoprotein (sE2). Here we report the production, purification and characterization of claudin-1. Both yeast membrane-bound and detergent-extracted, purified claudin-1 were antigenic and recognized by specific antibodies. Analytical ultracentrifugation demonstrated that extraction with n-octyl-ß-d-glucopyranoside yielded monodispersed, dimeric pools of claudin-1 while extraction with profoldin-8 or n-decylphosphocholine yielded a dynamic mixture of claudin-1 oligomers. Neither form bound sE2 in line with literature expectations, while further functional analysis was hampered by the finding that incorporation of claudin-1 into proteoliposomes rendered them intractable to study. Dynamic light scattering demonstrated that claudin-1 oligomers associate with CD81 in vitro in a defined molar ratio of 1:2 and that complex formation was enhanced by the presence of cholesteryl hemisuccinate. Attempts to assay the complex biologically were limited by our finding that claudin-1 affects the properties of proteoliposomes. We conclude that recombinant, correctly-folded, full-length claudin-1 can be produced in yeast membranes, that it can be extracted in different oligomeric forms that do not bind sE2 and that a dynamic preparation can form a specific complex with CD81 in vitro in the absence of any other cellular components. These findings pave the way for the structural characterization of claudin-1 alone and in complex with CD81.

Metabolic induction and early responses of mouse blastocyst developmental programming following maternal low protein diet affecting life-long health

Previously, we have shown that a maternal low protein diet, fed exclusively during the preimplantation period of mouse development (Emb-LPD), is sufficient to induce by the blastocyst stage a compensatory growth phenotype in late gestation and postnatally, correlating with increased risk of adult onset cardiovascular disease and behavioural dysfunction. Here, we examine mechanisms of induction of maternal Emb-LPD programming and early compensatory responses by the embryo. Emb-LPD induced changes in maternal serum metabolites at the time of blastocyst formation (E3.5), notably reduced insulin and increased glucose, together with reduced levels of free amino acids (AAs) including branched chain AAs leucine, isoleucine and valine. Emb-LPD also caused reduction in the branched chain AAs within uterine fluid at the blastocyst stage. These maternal changes coincided with an altered content of blastocyst AAs and reduced mTORC1 signalling within blastocysts evident in reduced phosphorylation of effector S6 ribosomal protein and its ratio to total S6 protein but no change in effector 4E-BP1 phosphorylated and total pools. These changes were accompanied by increased proliferation of blastocyst trophectoderm and total cells and subsequent increased spreading of trophoblast cells in blastocyst outgrowths. We propose that induction of metabolic programming following Emb-LPD is achieved through mTORC1signalling which acts as a sensor for preimplantation embryos to detect maternal nutrient levels via branched chain AAs and/or insulin availability. Moreover, this induction step associates with changes in extra-embryonic trophectoderm behaviour occurring as early compensatory responses leading to later nutrient recovery. © 2012 Fleming et al.

A two-stage model of orientation integration for Battenberg-modulated micropatterns

The visual system pools information from local samples to calculate textural properties. We used a novel stimulus to investigate how signals are combined to improve estimates of global orientation. Stimuli were 29 × 29 element arrays of 4 c/deg log Gabors, spaced 1° apart. A proportion of these elements had a coherent orientation (horizontal/vertical) with the remainder assigned random orientations. The observer's task was to identify the global orientation. The spatial configuration of the signal was modulated by a checkerboard pattern of square checks containing potential signal elements. The other locations contained either randomly oriented elements (''noise check'') or were blank (''blank check''). The distribution of signal elements was manipulated by varying the size and location of the checks within a fixed-diameter stimulus. An ideal detector would only pool responses from potential signal elements. Humans did this for medium check sizes and for large check sizes when a signal was presented in the fovea. For small check sizes, however, the pooling occurred indiscriminately over relevant and irrelevant locations. For these check sizes, thresholds for the noise check and blank check conditions were similar, suggesting that the limiting noise is not induced by the response to the noise elements. The results are described by a model that filters the stimulus at the potential target orientations and then combines the signals over space in two stages. The first is a mandatory integration of local signals over a fixed area, limited by internal noise at each location. The second is a taskdependent combination of the outputs from the first stage. © 2014 ARVO.

A template model predicts detection of sparse stimul

Contemporary models of contrast integration across space assume that pooling operates uniformly over the target region. For sparse stimuli, where high contrast regions are separated by areas containing no signal, this strategy may be sub-optimal because it pools more noise than signal as area increases. Little is known about the behaviour of human observers for detecting such stimuli. We performed an experiment in which three observers detected regular textures of various areas, and six levels of sparseness. Stimuli were regular grids of horizontal grating micropatches, each 1 cycle wide. We varied the ratio of signals (marks) to gaps (spaces), with mark:space ratios ranging from 1 : 0 (a dense texture with no spaces) to 1 : 24. To compensate for the decline in sensitivity with increasing distance from fixation, we adjusted the stimulus contrast as a function of eccentricity based on previous measurements [Baldwin, Meese & Baker, 2012, J Vis, 12(11):23]. We used the resulting area summation functions and psychometric slopes to test several filter-based models of signal combination. A MAX model failed to predict the thresholds, but did a good job on the slopes. Blanket summation of stimulus energy improved the threshold fit, but did not predict an observed slope increase with mark:space ratio. Our best model used a template matched to the sparseness of the stimulus, and pooled the squared contrast signal over space. Templates for regular patterns have also recently been proposed to explain the regular appearance of slightly irregular textures (Morgan et al, 2012, Proc R Soc B, 279, 2754–2760)

Sub-cortical sources of the somatosensory pathway are hypoactive in migraine interictally:a Functional Source Separation analysis

Background: Recent morpho-functional evidence pointed out that abnormalities in the thalamus could play a major role in the expression of migraine neurophysiological and clinical correlates. Whether this phenomenon is primary or secondary to its functional disconnection from the brainstem remains to be determined. We used a Functional Source Separation algorithm of EEG signal to extract the activity of the different neuronal pools recruited at different latencies along the somatosensory pathway in interictal migraine without aura (MO) patients. Methods: Twenty MO patients and 20 healthy volunteers (HV) underwent EEG recording. Four ad-hoc functional constraints, two sub-cortical (FS14 at brainstem and FS16 at thalamic level) and two cortical (FS20 radial and FS22 tangential parietal sources), were used to extract the activity of successive stages of somatosensory information processing in response to the separate left and right median nerve electric stimulation. A band-pass digital filter (450-750 Hz) was applied offline in order to extract high-frequency oscillatory (HFO) activity from the broadband EEG signal. Results: In both stimulated sides, significant reduced sub-cortical brainstem (FS14) and thalamic (FS16) HFO activations characterized MO patients when compared with HV. No difference emerged in the two cortical HFO activations between the two groups. Conclusions: Present results are the first neurophysiological evidence supporting the hypothesis that a functional disconnection of the thalamus from the subcortical monoaminergic system may underline the interictal cortical abnormal information processing in migraine. Further studies are needed to investigate the precise directional connectivity across the entire primary subcortical and cortical somatosensory pathway in interictal MO. Written informed consent to publication was obtained from the patient(s).

Intra- and interannual variability in seagrass carbon and nitrogen stable isotopes from south Florida, a preliminary study

δ13C and δ15N values were determined for the seagrassThalassia testudinum at four permanent seagrass monitoring stations in southFlorida, USA, through a quarterly sampling program over 3-years (1996–1998). All sites are seagrass beds with water depths of less than 6 m. Two sites are located on the Florida Bay side of the Florida Keys, and the other two sites are on the Atlantic side. The data analyzed over the 3 year study period display unique patterns associated with seasonal changes in primary productivity and potentially changes in the N and C pools. The mean carbon and nitrogenisotope values of T. testudinum from all four stations vary from −7.2 to −10.4‰ and 1.1 to 2.2‰, respectively. However, certain stations displayed anomalously depleted nitrogenisotope values (as low as −1.2‰). These values may indicate that biogeochemical processes like N fixation, ammonification and denitrification cause temporal changes in the isotopic composition of the source DIN. Both δ13C and δ15N values displayed seasonal enrichment-depletion patterns, with maximum enrichment occurring during the summer to early fall. The intra-annual variations of δ13C values from the different stations ranged from about 1 to 3.5‰; whereas variations in δ15N ranged from about 1 to 4.9‰. Certain sites showed a positive relationship between isotope values and productivity. These data indicate δ13C values display a high degree of seasonal variability as related to changes in productivity. δ15N values show clear intra-annual variations, but the observed changes do not necessarily follow a distinct seasonal cycle, indicating that changes in DIN will need further investigation.

A molecular and stable isotopic approach to investigate the importance of algal and detrital energy pathways in a freshwater marsh

The relative importance of algal and detrital energy pathways remains a central question in wetlands ecology. We used bulk stable isotope analysis and fatty acid composition to investigate the relative contributions of periphyton (algae) and floc (detritus) in a freshwater wetland with the goal of determining the inputs of these resource pools to lower trophic-level consumers. All animal samples revealed fatty acid markers indicative of both microbial (detrital) and algal origins, though the relative contributions varied among species. Vascular plant markers were in low abundance in most consumers. Detritivory is important for chironomids and amphipods, as demonstrated by the enhanced bacterial fatty acids present in both consumers, while algal resources, in the form of periphyton, likely support ephemeropteran larvae. Invertebrates such as amphipods and grass shrimp appear to be important resources for small omnivorous fish, while Poecilia latipinna appear to strongly use periphyton and Ephemeroptera larvae as food sources. Both P. latipinna and Lepomis spp. assimilated small amounts of vascular plant debris, possibly due to unintentional ingestion of floc while foraging for invertebrates and insect larvae. Physid snails, Haitia spp., were characterized by considerably different fatty acid compositions than other taxa examined, and likely play a unique role in Everglades’ food webs.

Sediment and Nutrient Deposition Associated with Hurricane Wilma in Mangroves of the Florida Coastal Everglades

The distribution of mangrove biomass and forest structure along Shark River estuary in the Florida Coastal Everglades (FCE) has been correlated with elevated total phosphorus concentration in soils thought to be associated with storm events. The passage of Hurricane Wilma across Shark River estuary in 2005 allowed us to quantify sediment deposition and nutrient inputs in FCE mangrove forests associated with this storm event and to evaluate whether these pulsing events are sufficient to regulate nutrient biogeochemistry in mangrove forests of south Florida. We sampled the spatial pattern of sediment deposits and their chemical properties in mangrove forests along FCE sites in December 2005 and October 2006. The thickness (0.5 to 4.5 cm) of hurricane sediment deposits decreased with distance inland at each site. Bulk density, organic matter content, total nitrogen (N) and phosphorus (P) concentrations, and inorganic and organic P pools of hurricane sediment deposits differed from surface (0–10 cm) mangrove soils at each site. Vertical accretion resulting from this hurricane event was eight to 17 times greater than the annual accretion rate (0.30± 0.03 cm year−1) averaged over the last 50 years. Total P inputs from storm-derived sediments were equivalent to twice the average surface soil nutrient P density (0.19 mg cm−3). In contrast, total N inputs contributed 0.8 times the average soil nutrient N density (2.8 mg cm−3). Allochthonous mineral inputs from Hurricane Wilma represent a significant source of sediment to soil vertical accretion rates and nutrient resources in mangroves of southwestern Everglades. The gradient in total P deposition to mangrove soils from west to east direction across the FCE associated with this storm event is particularly significant to forest development due to the P-limited condition of this carbonate ecosystem. This source of P may be an important adaptation of mangrove forests in the Caribbean region to projected impacts of sea-level rise.