evidence that ca 2 cycling by the plasma membrane ca 2 atpase increases the excitability of the extracellular ca 2 sensing receptor

The extracellular Ca 2+ -sensing receptor (CaR) is a widely expressed G-protein-coupled receptor that translates information about [Ca 2+ ] in the extracellular milieu to the interior of the cell, usually via intracellular Ca 2+ signaling pathways. Using fura-2 imaging of cytoplasmic [Ca 2+ ], we observed that HEK293 cells expressing CaR oscillated readily under conditions permissive for CaR activation. Spiking was also triggered in the absence of external Ca 2+ by the CaR agonist spermine (1 mM). Oscillating cells were typically located in clusters of closely apposed cells, but Ca 2+ spiking was insensitive to the gap junction inhibitor 18α-glycyrrhetinic acid. We hypothesized that Ca 2+ signals might be amplified, in part, through a positive feedback loop in which Ca 2+ extrusion via the plasma membrane Ca 2+ -ATPase (PMCA) activates CaRs on the same cell or adjacent cells through local increases in [Ca 2+ ] out . In support of this idea, addition of exogenous Ca 2+ buffers (keeping free [Ca 2+ ] out constant) attenuated or eliminated Ca 2+ signals (manifested as oscillations), as did PMCA inhibitors (HgCl 2 , orthovanadate and Caloxin 2A1). Measurement of extracellular [Ca 2+ ] using the near membrane probe fura-C 18 revealed that external [Ca 2+ ] rose following receptor activation, sometimes displaying an oscillatory pattern. Our data suggest that PMCA-mediated cycling of Ca 2+ across the plasma membrane leads to localized increases in [Ca 2+ ] out that increase the excitability of CaR

Bone marrow fibroblasts parallel multiple myeloma progression in patients and mice: in vitro and in vivo studies

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Stability of growing vesicles

We investigate the stability of growing vesicles using the formalism of nonequilibrium thermodynamics. The vesicles are growing due to the accretion of lipids to the bilayer which forms the vesicle membrane. The thermodynamic description is based on the hydrodynamics of a water{/}lipid mixture together with a model of the vesicle as a discontinuous system in the sense of linear nonequilibrium thermodynamics. This formulation allows the forces and fluxes relevant to the dynamic stability of the vesicle to be identified. The method is used to analyze the stability of a spherical vesicle against arbitrary axisymmetric perturbations. It is found that there are generically two critical radii at which changes of stability occur. In the case where the perturbation takes the form of a single zonal harmonic, only one of these radii is physical and is given by the ratio $2 L_p / L_\gamma$, where $L_p$ is the hydraulic conductivity and $L_\gamma$ is the Onsager coefficient related to changes in membrane area due to lipid accretion. The stability of such perturbations is related to the value of $l$ corresponding to the particular zonal harmonic: those with lower $l$ are more unstable than those with higher $l$. Possible extensions of the current work and the need for experimental input are discussed.Comment: 17 pages, 3 figure

Study of the Decay phi --> eta pi0 gamma with the KLOE detector

In a sample of 5.3x10^7 phi-decays observed with the KLOE detector at the Frascati phi-factory Dafne we find 605 eta pi0 gamma events with eta --> gamma\gamma and 197 eta pi0 gamma events with eta --> pi+ pi- pi0. The decay phi --> eta pi0 gamma is dominated by the process phi --> a0 gamma. From a fit to the eta pi0 mass spectrum we find BR(phi --> ao(980) gamma)= (7.4 +- 0.7)x10^-5.Comment: 12 pages, 6 figures, submitted to Phys.Lett.

Formyl Peptide Receptor as a Novel Therapeutic Target for Anxiety-Related Disorders

Formyl peptide receptors (FPR) belong to a family of sensors of the immune system that detect microbe-associated molecules and inform various cellular and sensorial mechanisms to the presence of pathogens in the host. Here we demonstrate that Fpr2/3-deficient mice show a distinct profile of behaviour characterised by reduced anxiety in the marble burying and light-dark box paradigms, increased exploratory behaviour in an open-field, together with superior performance on a novel object recognition test. Pharmacological blockade with a formyl peptide receptor antagonist, Boc2, in wild type mice reproduced most of the behavioural changes observed in the Fpr2/3(-/-) mice, including a significant improvement in novel object discrimination and reduced anxiety in a light/dark shuttle test. These effects were associated with reduced FPR signalling in the gut as shown by the significant reduction in the levels of p-p38. Collectively, these findings suggest that homeostatic FPR signalling exerts a modulatory effect on anxiety-like behaviours. These findings thus suggest that therapies targeting FPRs may be a novel approach to ameliorate behavioural abnormalities present in neuropsychiatric disorders at the cognitive-emotional interface

Computing, Design, Art: Reflections on an Innovative Moment in History

The paper is concerned with the role of art and design in the history and philosophy of computing. It offers insights arising from research into a period in the 1960s and 70s, particularly in the UK, when computing became more available to artists and designers, focusing on John Lansdown (1929-1999) and Bruce Archer (1922-2005) in London. Models of computing interacted with conceptualisations of art, design and related creative activities in important ways

Homemade oral supplement: a proposal for the nutritional recovery of children and adolescents with cancer

Objective The aim of this study was to evaluate the impact of homemade oral supplements on the nutritional recovery of patients with mild or severe malnutrition or at nutritional risk. Methods Eight recipes of homemade oral supplements containing 30% to 35% of the total energy expenditure were proposed. The patients with severe malnutrition (group B) received the oral supplement for 2 weeks and the others for 4 weeks (group A). Oral supplementation with homemade supplements was compared with oral supplementation with store-bought supplements, investigated earlier with a protocol with the same design. Results Homemade oral supplements contain much lower amounts of certain micronutrients but are five times cheaper than store-bought supplements. In group A, 88% of the patients taking homemade oral supplements and 84% of the patients taking store-bought supplements responded positively to supplementation. In group B, 22% of the patients taking homemade oral supplements and 25% of the patients taking store-bought supplements recovered. The difference was not significant. The impact of store-bought supplementation on the triceps skinfold thicknesses and arm circumferences of the patients in group A was greater than that obtained with homemade supplements. In group B, the effect on triceps skinfold thickness was not significant (p=0.16). Patients taking homemade or store-bought oral supplements presented similar protein and energy intakes and improvements in nutritional status. Only the body composition of patients in group A taking store-bought oral supplements was better. Conclusion The results obtained by this study suggest that the therapeutic use of homemade oral supplements is an alternative capable of promoting the nutritional recovery of cancer patients, especially those who cannot afford store-bought supplements.Objetivo Avaliar o impacto do suplemento oral artesanal na recuperação do estado nutricional de pacientes com desnutrição leve, grave e com risco nutricional. Métodos Propuseram-se oito receitas de suplementos visando ofertar entre 30,0% e 35,0% do gasto energético total. Os pacientes com desnutrição grave (grupo B) receberam o suplemento oral por duas semanas, e os demais pacientes (grupo A), por quatro semanas. Para a comparação dos resultados obtidos com o emprego do suplemento oral artesanal, foram utilizados dados referentes a um protocolo anterior, com o mesmo desenho, entretanto, com a utilização de suplemento oral industrializado. Resultados O suplemento oral artesanal fica muito aquém no que diz respeito a alguns micronutrientes, entretanto é cinco vezes mais barato do que a preparação com o suplemento oral industrializado. Os pacientes do grupo A com suplemento oral artesanal apresentaram 88,0% de resposta positiva na semana de avaliação, enquanto os com suplemento oral industrializado tiveram 84,0%. No grupo B, foram recuperados 22,0% dos pacientes com suplemento oral artesanal e 25,0% do grupo com suplemento oral industrializado, não apresentando, portanto, diferença significante. Comparando o impacto do industrializado com o do artesanal na prega cutânea tricipital e circunferência do braço, verificou-se que o suplemento oral industrializado no grupo A apresentou melhores resultados que o suplemento oral artesanal, e no grupo B, esse efeito observado na prega cutânea não foi significante (p=0,16). Os consumos de energia e de proteína, assim como a evolução nutricional, foram semelhantes entre suplemento oral industrializado e suplemento oral artesanal. Apenas a composição corpórea no grupo A com suplemento oral industrializado apresentou melhores resultados. Conclusão Os resultados apresentados neste estudo sugerem que o emprego da terapia com suplemento artesanal seja uma opção capaz de auxiliar na recuperação nutricional de pacientes oncológicos e uma opção para populações financeiramente desfavorecidas.Hospital Samaritano de São PauloInstituto Adriana GarófoloUniversidade Federal de São Paulo (UNIFESP) Departamento de Pediatria Instituto de Oncologia PediátricaUniversidade Federal de São Paulo (UNIFESP) Departamento de PediatriaUNIFESP, Depto. de Pediatria Instituto de Oncologia PediátricaUNIFESP, Depto. de PediatriaSciEL

Allosteric mutants show that PrfA activation is dispensable for vacuole escape but required for efficient spread and <em>Listeria</em> survival <em>in vivo</em>

The transcriptional regulator PrfA controls key virulence determinants of the facultative intracellular pathogen Listeria monocytogenes. PrfA-dependent gene expression is strongly induced within host cells. While the basis of this activation is unknown, the structural homology of PrfA with the cAMP receptor protein (Crp) and the finding of constitutively activated PrfA* mutants suggests it may involve ligand-induced allostery. Here, we report the identification of a solvent-accessible cavity within the PrfA N-terminal domain that may accommodate an activating ligand. The pocket occupies a similar position to the cAMP binding site in Crp but lacks the cyclic nucleotide-anchoring motif and has its entrance on the opposite side of the β-barrel. Site-directed mutations in this pocket impaired intracellular PrfA-dependent gene activation without causing extensive structural/functional alterations to PrfA. Two substitutions, L48F and Y63W, almost completely abolished intracellular virulence gene induction and thus displayed the expected phenotype for allosteric activation-deficient PrfA mutations. Neither PrfA(allo) substitution affected vacuole escape and initial intracellular growth of L. monocytogenes in epithelial cells and macrophages but caused defective cell-to-cell spread and strong attenuation in mice. Our data support the hypothesis that PrfA is allosterically activated during intracellular infection and identify the probable binding site for the effector ligand. They also indicate that PrfA allosteric activation is not required for early intracellular survival but is essential for full Listeria virulence and colonization of host tissues