Exercise on Prescription: trial protocol and evaluation of outcomes

BACKGROUND: In many countries exercise prescriptions are used in an attempt to initiate a physically active lifestyle in sedentary populations. Previous studies have primarily evaluated low intensive exercise prescription interventions and found moderately positive effects on physical activity and aerobic fitness. In a highly intensive Danish exercise prescription scheme called 'Exercise on Prescription' (EoP) the general practitioners can prescribe EoP to sedentary patients with lifestyle diseases. The aim of this randomized trial is to assess the short- and long-term effects of the EoP scheme. Thus, the aim of this paper is to describe the randomized controlled trial designed for evaluating effectiveness of EoP, and to present results from validations of outcome measures. METHODS/DESIGN: EoP involves a 16-week supervised training intervention and five counselling sessions (health profiles). All patients referred to EoP were eligible for the trial and were offered participation during the baseline health profile. Comparisons between the EoP group and the control group were made at baseline, and after four and ten months. Physiological measures used were maximal oxygen uptake (VO(2)max), glycosylated haemoglobin (HbA1c), bodyweight, and BMI. Patient-reported measures used were physical activity, health-related quality of life, amount and intensity of exercise, compliance with national guidelines for physical activity, and physical fitness. The validation of the cycle ergometer test found a strong correlation between maximal work capacity and VO(2)max, and acceptable test-retest reliability at group level. Calibration of the HbA1c apparatus was stable over ten weeks with minimal use, and test-retest reliability was good. High agreement percents were found for test-retest reliability for the self-administered questionnaire. DISCUSSION: The trial is designed to provide information about the effectiveness of the EoP scheme. The trial is part of a health technology assessment of EoP, which besides the effectiveness covers the patient perspective, the organization, and the health economy. All three methods validated were found useful for the EoP trial

Efficient qubit detection using alkali earth metal ions and a double STIRAP process

We present a scheme for robust and efficient projection measurement of a qubit consisting of the two magnetic sublevels in the electronic ground state of alkali earth metal ions. The scheme is based on two stimulated Raman adiabatic passages (STIRAP) involving four partially coherent laser fields. We show how the efficiency depends on experimentally relevant parameters: Rabi frequencies, pulse widths, laser linewidths, one- and two-photon detunings, residual laser power, laser polarization and ion motion.Comment: 14 pages, 15 figure

Examining Synaptotagmin 1 Function in Dense Core Vesicle Exocytosis under Direct Control of Ca2+

We tested the long-standing hypothesis that synaptotagmin 1 is the Ca2+ sensor for fast neurosecretion by analyzing the intracellular Ca2+ dependence of large dense-core vesicle exocytosis in a mouse strain carrying a mutated synaptotagmin C2A domain. The mutation (R233Q) causes a twofold increase in the KD of Ca2+-dependent phospholipid binding to the double C2A-C2B domain of synaptotagmin. Using photolysis of caged calcium and capacitance measurements we found that secretion from mutant cells had lower secretory rates, longer secretory delays, and a higher intracellular Ca2+-threshold for secretion due to a twofold increase in the apparent KD of the Ca2+ sensor for fast exocytosis. Single amperometric fusion events were unchanged. We conclude that Ca2+-dependent phospholipid binding to synaptotagmin 1 mirrors the intracellular Ca2+ dependence of exocytosis

Synaptobrevin N-terminally bound to syntaxin–SNAP-25 defines the primed vesicle state in regulated exocytosis

Time-resolved measurements of exocytosis identify a domain of the SNARE complex required to keep vesicles readily releasable

Synaptotagmin-7 Is an Asynchronous Calcium Sensor for Synaptic Transmission in Neurons Expressing SNAP-23

Synchronization of neurotransmitter release with the presynaptic action potential is essential for maintaining fidelity of information transfer in the central nervous system. However, synchronous release is frequently accompanied by an asynchronous release component that builds up during repetitive stimulation, and can even play a dominant role in some synapses. Here, we show that substitution of SNAP-23 for SNAP-25 in mouse autaptic glutamatergic hippocampal neurons results in asynchronous release and a higher frequency of spontaneous release events (mEPSCs). Use of neurons from double-knock-out (SNAP-25, synaptotagmin-7) mice in combination with viral transduction showed that SNAP-23-driven release is triggered by endogenous synaptotagmin-7. In the absence of synaptotagmin-7 release became even more asynchronous, and the spontaneous release rate increased even more, indicating that synaptotagmin-7 acts to synchronize release and suppress spontaneous release. However, compared to synaptotagmin-1, synaptotagmin-7 is a both leaky and asynchronous calcium sensor. In the presence of SNAP-25, consequences of the elimination of synaptotagmin-7 were small or absent, indicating that the protein pairs SNAP-25/synaptotagmin-1 and SNAP-23/synaptotagmin-7 might act as mutually exclusive calcium sensors. Expression of fusion proteins between pHluorin (pH-sensitive GFP) and synaptotagmin-1 or -7 showed that vesicles that fuse using the SNAP-23/synaptotagmin-7 combination contained synaptotagmin-1, while synaptotagmin-7 barely displayed activity-dependent trafficking between vesicle and plasma membrane, implying that it acts as a plasma membrane calcium sensor. Overall, these findings support the idea of alternative syt∶SNARE combinations driving release with different kinetics and fidelity

Regulation of Ca2+ channels by SNAP-25 via recruitment of syntaxin-1 from plasma membrane clusters

SNAP-25 regulates Ca(2+) channels, with potentially important consequences for diseases involving an aberrant SNAP-25 expression level. How this regulation is executed mechanistically remains unknown. We investigated this question in mouse adrenal chromaffin cells and found that SNAP-25 inhibits Ca(2+) currents, with the B-isoform being more potent than the A-isoform, but not when syntaxin-1 is cleaved by botulinum neurotoxin C. In contrast, syntaxin-1 inhibits Ca(2+) currents independently of SNAP-25. Further experiments using immunostaining showed that endogenous or exogenous SNAP-25 expression recruits syntaxin-1 from clusters on the plasma membrane, thereby increasing the immunoavailability of syntaxin-1 and leading indirectly to Ca(2+) current inhibition. Expression of Munc18-1, which recruits syntaxin-1 within the exocytotic pathway, does not modulate Ca(2+) channels, whereas overexpression of the syntaxin-binding protein Doc2B or ubMunc13-2 increases syntaxin-1 immunoavailability and concomitantly down-regulates Ca(2+) currents. Similar findings were obtained upon chemical cholesterol depletion, leading directly to syntaxin-1 cluster dispersal and Ca(2+) current inhibition. We conclude that clustering of syntaxin-1 allows the cell to maintain a high syntaxin-1 expression level without compromising Ca(2+) influx, and recruitment of syntaxin-1 from clusters by SNAP-25 expression makes it available for regulating Ca(2+) channels. This mechanism potentially allows the cell to regulate Ca(2+) influx by expanding or contracting syntaxin-1 clusters

Freshening increases the susceptibility to heat stress in intertidal mussels (Mytilus edulis) from the Arctic.

Funder: William Demant FondenFunder: DANCEAFunder: Familien Hede Nielsens Fond; Id: http://dx.doi.org/10.13039/501100007438Funder: Selskabet for Arktisk Forskning og TeknologiFunder: Aage V. Jensens Fond; Id: http://dx.doi.org/10.13039/501100002721Temperatures in the Arctic are increasing at a faster pace than at lower latitudes resulting in range expansion of boreal species. In Greenland, the warming also drives accelerating melt of the Greenland Ice Sheet resulting in more meltwater entering Greenland fjords in summer. Our aim was to determine if increasing summer temperatures combined with lower salinity can induce the expression of stress-related proteins, for example, heat shock protein, in boreal intertidal mussels in Greenland, and whether low salinity reduces the upper thermal limit at which mortality occurs. We conducted a mortality experiment, using 12 different combinations of salinity and air temperature treatments during a simulated tidal regime, and quantified the change in mRNA levels of five stress-related genes (hsp24, hsp70, hsp90, sod and p38) in surviving mussels to discern the level of sublethal stress. Heat-induced mortality occurred in mussels exposed to an air temperature of 30°C and mortality was higher in treatments with lowered salinity (5 and 15‰), which confirms that low habitat salinity decreases the upper thermal limit of Mytilus edulis. The gene expression analysis supported the mortality results, with the highest gene expression found at combinations of high temperature and low salinity. Combined with seasonal measurements of intertidal temperatures in Greenland, we suggest heat stress occurs in low salinity intertidal area, and that further lowered salinity in coastal water due to increased run-off can make intertidal bivalves more susceptible to summer heat stress. This study thus provides an example of how different impacts of climate warming can work synergistically to stress marine organisms