Tmc1 point mutation affects Ca2+ sensitivity and block by dihydrostreptomycin of the mechanoelectrical transducer current of mouse outer hair cells

The transduction of sound into electrical signals depends on mechanically sensitive ion channels in the stereociliary bundle. The molecular composition of this mechanoelectrical transducer (MET) channel is not yet known. Transmembrane channel-like protein isoforms 1 (TMC1) and 2 (TMC2) have been proposed to form part of the MET channel, although their exact roles are still unclear. Using Beethoven (Tmc1Bth/Bth) mice, which have an M412K point mutation in TMC1 that adds a positive charge, we found that Ca2+ permeability and conductance of the MET channel of outer hair cells (OHCs) were reduced. Tmc1Bth/Bth OHCs were also less sensitive to block by the permeant MET channel blocker dihydrostreptomycin, whether applied extracellularly or intracellularly. These findings suggest that the amino acid that is mutated in Bth is situated at or near the negatively charged binding site for dihydrostreptomycin within the permeation pore of the channel. We also found that the Ca2+ dependence of the operating range of the MET channel was altered by the M412K mutation. Depolarization did not increase the resting open probability of the MET current of Tmc1Bth/Bth OHCs, whereas raising the intracellular concentration of the Ca2+ chelator BAPTA caused smaller increases in resting open probability in Bth mutant OHCs than in wild-type control cells. We propose that these observations can be explained by the reduced Ca2+ permeability of the mutated MET channel indirectly causing the Ca2+ sensor for adaptation, at or near the intracellular face of the MET channel, to become more sensitive to Ca2+ influx as a compensatory mechanism

Hair cell maturation is differentially regulated along the tonotopic axis of the mammalian cochlea

Sound amplification within the mammalian cochlea depends upon specialized hair cells, the outer hair cells (OHCs), which possess both sensory and motile capabilities. In various altricial rodents, OHCs become functionally competent from around postnatal day 7 (P7), before the primary sensory inner hair cells (IHCs), which become competent at about the onset of hearing (P12). The mechanisms responsible for the maturation of OHCs and their synaptic specialization remain poorly understood. We report that spontaneous Ca2+ activity in the immature cochlea, which is generated by CaV1.3 Ca2+ channels, differentially regulates the maturation of hair cells along the cochlea. Under near‐physiological recording conditions we found that, similar to IHCs, immature OHCs elicited spontaneous Ca2+ action potentials (APs), but only during the first few postnatal days. Genetic ablation of these APs in vivo, using CaV1.3−/− mice, prevented the normal developmental acquisition of mature‐like basolateral membrane currents in low‐frequency (apical) hair cells, such as IK,n (carried by KCNQ4 channels), ISK2 and IACh (α9α10nAChRs) in OHCs and IK,n and IK,f (BK channels) in IHCs. Electromotility and prestin expression in OHCs were normal in CaV1.3−/− mice. The maturation of high‐frequency (basal) hair cells was also affected in CaV1.3−/− mice, but to a much lesser extent than apical cells. However, a characteristic feature in CaV1.3−/− mice was the reduced hair cell size irrespective of their cochlear location. We conclude that the development of low‐ and high‐frequency hair cells is differentially regulated during development, with apical cells being more strongly dependent on experience‐independent Ca2+ APs

Dominance Based Crossover Operator for Evolutionary Multi-objective Algorithms

In spite of the recent quick growth of the Evolutionary Multi-objective Optimization (EMO) research field, there has been few trials to adapt the general variation operators to the particular context of the quest for the Pareto-optimal set. The only exceptions are some mating restrictions that take in account the distance between the potential mates - but contradictory conclusions have been reported. This paper introduces a particular mating restriction for Evolutionary Multi-objective Algorithms, based on the Pareto dominance relation: the partner of a non-dominated individual will be preferably chosen among the individuals of the population that it dominates. Coupled with the BLX crossover operator, two different ways of generating offspring are proposed. This recombination scheme is validated within the well-known NSGA-II framework on three bi-objective benchmark problems and one real-world bi-objective constrained optimization problem. An acceleration of the progress of the population toward the Pareto set is observed on all problems

Orchestration of renewable generation in low energy buildings and districts using energy storage and load shaping

There is increasing penetration of renewable generation in buildings and districts. There are challenges in making the effective use of this generation. The objective of the ORIGIN project (Orchestration of Renewable Integrated Generation In Neighborhoods) is to shape loads so that the fraction of energy consumed that is from local renewable generation is maximized, and energy imported from outside sources is minimized. This paper presents the overall approach taken in the ORIGIN project and explores building physics aspects of solar thermal storage system orchestration. The case study districts are briefly introduced and characteristics of their generation, buildings, districts and shiftable loads described. The orchestration approach taken in ORIGIN is then presented. At the core of the ORIGIN system is the orchestration algorithm which generates informational and control outputs to shape future loads to best meet the objectives. The model based approach used to quantify thermal and electrical load shifting opportunities for pre-charging, coasting or avoiding loads, while meeting thermal comfort and other demands, is described using a solar thermal storage system as an example. The future steps for the ORIGIN project; retrofit of the ORIGIN system into existing districts and potential for other future applications is briefly discussed

Coping styles relate to health and work environment of Norwegian and Dutch hospital nurses:A comparative study

Nurses exposed to high nursing stress report no health complaints as long as they have high coping abilities. The purpose of this study was to investigate coping styles in relation to the health status and work environment of Norwegian and Dutch hospital nurses. This comparative study included a random sample of 5400 Norwegian nurses and a convenience sample of 588 Dutch nurses. Coping, health, and work environment were assessed by questionnaire in both samples and associations were investigated bivariately and multi-variately. We found that active problem-solving coping was associated with the health and work environment of Norwegian nurses but not with the health and work environment of Dutch. Passive coping (avoiding problems or waiting to see what happens) was found to relate to poor general health, poor mental health, low job control, and low job support in both Norwegian and Dutch nurses. Improvements in the nursing work environment may not only result in better mental health, but may also reduce passive coping

Modifications of the endosomal compartment in peripheral blood mononuclear cells and fibroblasts from Alzheimer’s disease patients

International audienceIdentification of blood-based biomarkers of Alzheimer’s disease (AD) remains a challenge. Neuropathological studies have identified enlarged endosomes in post-mortem brains as the earliest cellular change associated to AD. Here the presence of enlarged endosomes was investigated in peripheral blood mononuclear cells from 48 biologically defined AD patients (25 with mild cognitive impairment and 23 with dementia (AD-D)), and 23 age-matched healthy controls using immunocytochemistry and confocal microscopy. The volume and number of endosomes were not significantly different between AD and controls. However, the percentage of cells containing enlarged endosomes was significantly higher in the AD-D group as compared with controls. Furthermore, endosomal volumes significantly correlated to [C11]PiB cortical index measured by positron emission tomography in the AD group, independently of the APOE genotype, but not to the levels of amyloid-beta, tau and phosphorylated tau measured in the cerebrospinal fluid. Importantly, we confirmed the presence of enlarged endosomes in fibroblasts from six unrelated AD-D patients as compared with five cognitively normal controls. This study is the first, to our knowledge, to report morphological alterations of the endosomal compartment in peripheral cells from AD patients correlated to amyloid load that will now be evaluated as a possible biomarker

Enhancing Perception of Complex Sculptural Forms using Interactive Real-time Ray tracing

This paper looks at experiments into using real-time ray tracing to significantly enhance shape perception of complex three-dimensional digitally created structures. The author is a computational artist whose artistic practice explores the creation of intricate organic three-dimensional forms using simulation of morphogenesis. The generated forms are often extremely detailed, comprising tens of millions of cellular primitives. This often makes depth perception of the resulting structures difficult. His practice has explored various techniques to create presentable artefacts from the data, including high resolution prints, animated videos, stereoscopic installations, 3D printing and virtual reality. The author uses ray tracing techniques to turn the 3D data created from his morphogenetic simulations into visible artefacts. This is typically a time-consuming process, taking from seconds to minutes to create a single frame. The latest generation of graphics processing units offer dedicated hardware to accelerate ray tracing calculations. This potentially allows the generation of ray traced images, including self-shadowed complex structures and multiple levels of transparency, from new viewpoints at frame rates capable of real-time interaction. The author presents the results of his experiments using this technology with the aim of providing significantly enhanced perception of his generated three-dimensional structures by allowing user-initiated interaction to generate novel views, and utilizing depth cues such as stereopsis, depth from motion and defocus blurring. The intention is for these techniques to be usable to present new exhibitable works in a gallery context

Effects of intranasal TNFα on granulocyte recruitment and activity in healthy subjects and patients with allergic rhinitis

<p>Abstract</p> <p>Background</p> <p>TNFα may contribute to the pathophysiology of airway inflammation. For example, we have recently shown that nasal administration of TNFα produces late phase co-appearance of granulocyte and plasma exudation markers on the mucosal surface. The objective of the present study was to examine indices of granulocyte presence and activity in response to intranasal TNFα challenge.</p> <p>Methods</p> <p>Healthy subjects and patients with allergic rhinitis (examined out of season) were subjected to nasal challenge with TNFα (10 μg) in a sham-controlled and crossover design. Nasal lavages were carried out prior to and 24 hours post challenge. Nasal biopsies were obtained post challenge. Nasal lavage fluid levels of myeloperoxidase (MPO) and eosinophil cationic protein (ECP) were analyzed as indices of neutrophil and eosinophil activity. Moreover, IL-8 and α₂-macroglobulin were analyzed as markers of pro-inflammatory cytokine production and plasma exudation. Nasal biopsy numbers of neutrophils and eosinophils were monitored.</p> <p>Results</p> <p>Nasal lavage fluid levels of MPO recorded 24 hours post TNFα challenge were increased in healthy subjects (p = 0.0081) and in patients with allergic rhinitis (p = 0.0081) (<it>c.f</it>. sham challenge). Similarly, α₂-macroglobulin was increased in healthy subjects (p = 0.014) and in patients with allergic rhinitis (p = 0.0034). Lavage fluid levels of ECP and IL-8 were not affected by TNFα challenge. TNFα increased the numbers of subepithelial neutrophils (p = 0.0021), but not the numbers of eosinophils.</p> <p>Conclusion</p> <p>TNFα produces a nasal inflammatory response in humans that is characterised by late phase (i.e., 24 hours post challenge) neutrophil activity and plasma exudation.</p

Mutation of von Hippel–Lindau Tumour Suppressor and Human Cardiopulmonary Physiology

BACKGROUND: The von Hippel–Lindau tumour suppressor protein–hypoxia-inducible factor (VHL–HIF) pathway has attracted widespread medical interest as a transcriptional system controlling cellular responses to hypoxia, yet insights into its role in systemic human physiology remain limited. Chuvash polycythaemia has recently been defined as a new form of VHL-associated disease, distinct from the classical VHL-associated inherited cancer syndrome, in which germline homozygosity for a hypomorphic VHL allele causes a generalised abnormality in VHL–HIF signalling. Affected individuals thus provide a unique opportunity to explore the integrative physiology of this signalling pathway. This study investigated patients with Chuvash polycythaemia in order to analyse the role of the VHL–HIF pathway in systemic human cardiopulmonary physiology. METHODS AND FINDINGS: Twelve participants, three with Chuvash polycythaemia and nine controls, were studied at baseline and during hypoxia. Participants breathed through a mouthpiece, and pulmonary ventilation was measured while pulmonary vascular tone was assessed echocardiographically. Individuals with Chuvash polycythaemia were found to have striking abnormalities in respiratory and pulmonary vascular regulation. Basal ventilation and pulmonary vascular tone were elevated, and ventilatory, pulmonary vasoconstrictive, and heart rate responses to acute hypoxia were greatly increased. CONCLUSIONS: The features observed in this small group of patients with Chuvash polycythaemia are highly characteristic of those associated with acclimatisation to the hypoxia of high altitude. More generally, the phenotype associated with Chuvash polycythaemia demonstrates that VHL plays a major role in the underlying calibration and homeostasis of the respiratory and cardiovascular systems, most likely through its central role in the regulation of HIF