Calcium sensitivity and the Frank-Starling mechanism of the heart are increased in titin N2B region deficient mice

Previous work suggests that titin-based passive tension is a factor in the Frank-Starling mechanism of the heart, by increasing length-dependent activation (LDA) through an increase in calcium sensitivity at long sarcomere length. We tested this hypothesis in a mouse model (N2B KO model) in which titin-based passive tension is elevated as a result of the excision of the N2B element, one of cardiac titin's spring elements. LDA was assessed by measuring the active tension-pCa (-log[Ca2+]) relationship at sarcomere length (SLs) of 1.95, 2.10 and 2.30mum in WT and N2B KO skinned myocardium. LDA was positively correlated with titin-based passive tension, due to an increase in calcium sensitivity at the longer SLs in the KO. For example, at pCa 6.0 the KO:WT tension ratio was 1.28+/-0.07 and 1.42+/-0.04 at SLs of 2.1 and 2.3mum, respectively. There was no difference in protein expression or phosphorylation of sarcomeric proteins. We also measured the calcium sensitivity after PKA treating the skinned muscle and found that titin-based passive tension was also now correlated with LDA, with a slope that was significantly increased compared to no PKA treatment. Finally, we performed isolated heart experiments and measured the Frank-Starling relation (slope of developed wall stress-LV volume relation) as well as diastolic stiffness (slope of diastolic wall stress - volume relation). The FSM was more pronounced in the N2B KO hearts and the slope of the FSM correlated with diastolic stiffness. These findings support that titin-based passive tension triggers an increase in calcium sensitivity at long sarcomere length, thereby playing an important role in the Frank-Starling mechanism of the heart

Spatial accessibility and social inclusion: The impact of Portugal's last health reform

Health policies seek to promote access to health care and should provide appropriate geographical accessibility to each demographical functional group. The dispersal demand of health‐careservices and the provision for such services atfixed locations contribute to the growth of inequality intheir access. Therefore, the optimal distribution of health facilities over the space/area can lead toaccessibility improvements and to the mitigation of the social exclusion of the groups considered mostvulnerable. Requiring for such, the use of planning practices joined with accessibility measures. However,the capacities of Geographic Information Systems in determining and evaluating spatial accessibility inhealth system planning have not yet been fully exploited. This paper focuses on health‐care services planningbased on accessibility measures grounded on the network analysis. The case study hinges on mainlandPortugal. Different scenarios were developed to measure and compare impact on the population'saccessibility. It distinguishes itself from other studies of accessibility measures by integrating network data ina spatial accessibility measure: the enhanced two‐stepfloating catchment area. The convenient location forhealth‐care facilities can increase the accessibility standards of the population and consequently reducethe economic and social costs incurred. Recently, the Portuguese government implemented a reform thataimed to improve, namely, the access and equity in meeting with the most urgent patients. It envisaged,in terms of equity, the allocation of 89 emergency network points that ensured more than 90% of thepopulation be within 30 min from any one point in the network. Consequently, several emergency serviceswere closed, namely, in rural areas. This reform highlighted the need to improve the quality of the emergencycare, accessibility to each care facility, and equity in their access. Hence, accessibility measures becomean efficient decision‐making tool, despite its absence in effective practice planning. According to anapplication of this type of measure, it was possible to verify which levels of accessibility were decreased,including the most disadvantaged people, with a larger time of dislocation of 12 min between 2001 and 2011

Hyperboloidal slices for the wave equation of Kerr-Schild metrics and numerical applications

We present new results from two open source codes, using finite differencing and pseudo-spectral methods for the wave equations in (3+1) dimensions. We use a hyperboloidal transformation which allows direct access to null infinity and simplifies the control over characteristic speeds on Kerr-Schild backgrounds. We show that this method is ideal for attaching hyperboloidal slices or for adapting the numerical resolution in certain spacetime regions. As an example application, we study late-time Kerr tails of sub-dominant modes and obtain new insight into the splitting of decay rates. The involved conformal wave equation is freed of formally singular terms whose numerical evaluation might be problematically close to future null infinity.Comment: 15 pages, 12 figure

An improved formulation of the relativistic hydrodynamics equations in 2D Cartesian coordinates

A number of astrophysical scenarios possess and preserve an overall cylindrical symmetry also when undergoing a catastrophic and nonlinear evolution. Exploiting such a symmetry, these processes can be studied through numerical-relativity simulations at smaller computational costs and at considerably larger spatial resolutions. We here present a new flux-conservative formulation of the relativistic hydrodynamics equations in cylindrical coordinates. By rearranging those terms in the equations which are the sources of the largest numerical errors, the new formulation yields a global truncation error which is one or more orders of magnitude smaller than those of alternative and commonly used formulations. We illustrate this through a series of numerical tests involving the evolution of oscillating spherical and rotating stars, as well as shock-tube tests.Comment: 19 pages, 9 figure

Laboratory Focus on Improving the Culture of Biosafety: Statewide Risk Assessment of Clinical Laboratories That Process Specimens for Microbiologic Analysis

The Wisconsin State Laboratory of Hygiene challenged Wisconsin laboratories to examine their biosafety practices and improve their culture of biosafety. One hundred three clinical and public health laboratories completed a questionnaire-based, microbiology-focused biosafety risk assessment. Greater than 96% of the respondents performed activities related to specimen processing, direct microscopic examination, and rapid nonmolecular testing, while approximately 60% performed culture interpretation. Although they are important to the assessment of risk, data specific to patient occupation, symptoms, and travel history were often unavailable to the laboratory and, therefore, less contributory to a microbiology-focused biosafety risk assessment than information on the specimen source and test requisition. Over 88% of the respondents complied with more than three-quarters of the mitigation control measures listed in the survey. Facility assessment revealed that subsets of laboratories that claim biosafety level 1, 2, or 3 status did not possess all of the biosafety elements considered minimally standard for their respective classifications. Many laboratories reported being able to quickly correct the minor deficiencies identified. Task assessment identified deficiencies that trended higher within the general (not microbiology-specific) laboratory for core activities, such as packaging and shipping, direct microscopic examination, and culture modalities solely involving screens for organism growth. For traditional microbiology departments, opportunities for improvement in the cultivation and management of highly infectious agents, such as acid-fast bacilli and systemic fungi, were revealed. These results derived from a survey of a large cohort of small- and large-scale laboratories suggest the necessity for continued microbiology-based understanding of biosafety practices, vigilance toward biosafety, and enforcement of biosafety practices throughout the laboratory setting

Deconstructing sarcomeric structure-function relations in titin-BioID knock-in mice

Proximity proteomics has greatly advanced the analysis of native protein complexes and subcellular structures in culture, but has not been amenable to study development and disease in vivo. Here, we have generated a knock-in mouse with the biotin ligase (BioID) inserted at titin's Z-disc region to identify protein networks that connect the sarcomere to signal transduction and metabolism. Our census of the sarcomeric proteome from neonatal to adult heart and quadriceps reveals how perinatal signaling, protein homeostasis and the shift to adult energy metabolism shape the properties of striated muscle cells. Mapping biotinylation sites to sarcomere structures refines our understanding of myofilament dynamics and supports the hypothesis that myosin filaments penetrate Z-discs to dampen contraction. Extending this proof of concept study to BioID fusion proteins generated with Crispr/CAS9 in animal models recapitulating human pathology will facilitate the future analysis of molecular machines and signaling hubs in physiological, pharmacological, and disease context

Intrinsic Josephson Effect in the Layered Two-dimensional t-J Model

The intrinsic Josephson effect in the high-Tc superconductors is studied using the layered two-dimensional t-J model. The d.c.Josephson current which flows perpendicular to the t-J planes is obtained within the mean-field approximation and the Gutzwiller approximation. We find that the Josephson current has its maximum near the optimum doping region as a function of the doping rate.Comment: 4 pages, 3 figure

Foundations of character: methodological aspects of a study of character development in three- to six-year-old children with a focus on sharing behaviours

This article focuses on methodological issues arising in a study of character development, using illustrations of ‘sharing behaviours.’ Based primarily in six early years settings in southeast England the research records naturalistic observations of peer interactions for 55 children aged three to six years. Applying grounded theory to the processes of observing, analysing and interpreting evidence required a cautious and collectively reflective approach. The methodology sought to moderate the influence of the researchers' prior knowledge of ‘grand theories’ of moral development and assumptions about relevance to the observation records. The study's originality lay in the exploration of moral development without reference to any particular grand theory as an explanatory framework; and in the reluctance to be drawn to potentially simplistic rationalisations of the children's intentions on the basis of their observed behaviours. Exploring young children's subjective experiences, this research provides insights into the intricacy of this process, steering away from ‘neat’ findings and attempting to reflect the sophistication of the children's skilful and sometimes surprising negotiations of moral dilemmas. Implications for practice relate to the complexities involved in attempts to unravel the developing moral characters of young children and the practice through which this may be nurtured

Spacelike distance from discrete causal order

Any discrete approach to quantum gravity must provide some prescription as to how to deduce continuum properties from the discrete substructure. In the causal set approach it is straightforward to deduce timelike distances, but surprisingly difficult to extract spacelike distances, because of the unique combination of discreteness with local Lorentz invariance in that approach. We propose a number of methods to overcome this difficulty, one of which reproduces the spatial distance between two points in a finite region of Minkowski space. We provide numerical evidence that this definition can be used to define a `spatial nearest neighbor' relation on a causal set, and conjecture that this can be exploited to define the length of `continuous curves' in causal sets which are approximated by curved spacetime. This provides evidence in support of the ``Hauptvermutung'' of causal sets.Comment: 32 pages, 16 figures, revtex4; journal versio