T- and L-type Ca 2+ currents in freshly dispersed smooth muscle cells from the human proximal urethra

The purpose of the present study was to characterise Ca 2+ currents in smooth muscle cells solated from biopsy samples taken from the proximal urethra of patients undergoing surgery for bladder or prostate cancer. Cells were studied at 37°C using the amphotericin B perforated-patch onfiguration of the patch-clamp technique. Currents were recorded using Cs + -rich pipette solutions to block K + currents. Two components of current, with electrophysiological and pharmacological properties typical of T- and L-type Ca 2+ currents, were present in these cells. When steady-state inactivation curves for the L current were fitted with a Boltzmann equation, this yielded a VÎ of _45 ± 5 mV. In contrast, the T current inactivated with a VÎ of _80 ± 3 mV. The L currents were reduced in a concentration-dependent manner by nifedipine (ED50 = 159 ± 54 nM) and Ni 2+ (ED50 = 65 ± 16 mM) but were enhanced when external Ca 2+ was substituted with Ba 2+ . The T current was little affected by TTX, reduction in external Na + , application of nifedipine at concentrations below 300 nM or substitution of external Ca 2+ with Ba 2+ , but was reduced by Ni 2+ with an ED50 of 6 ± 1 mM. When cells were stepped from _100 to _30 mV in Ca 2+ -free conditions, small inward currents could be detected. These were enhanced 40-fold in divalent-cation-free solution and blocked in a concentration-dependent manner by Mg 2+ with an ED50 of 32 ± 16 mM. These data support the idea that human urethral myocytes possess currents with electrophysiological and pharmacological properties typical of T- and L-type Ca 2+ currents

Contribution of K v 2.1 channels to the delayed rectifier current in freshly dispersed smooth muscle cells from rabbit urethra

We have characterized the native voltage-dependent K + (K v ) current in rabbit urethral smooth muscle cells (RUSMC) and compared its pharmacological and biophysical properties with K v 2.1 and K v 2.2 channels cloned from the rabbit urethra and stably expressed in HEK 293 cells (HEK Kv2.1 and HEK Kv2.2 ). RUSMC were perfused with Hanks' solution at 37°C and studied using the patch clamp technique with K + -rich pipette solutions. Cells were bathed in 100 nM penitrem A (Pen A) to block large conductance Ca 2+ -activated K + (BK) currents and depolarized to +40 mV for 500 ms to evoke K v currents. These were unaffected by margatoxin, κ-dendrotoxin or α-dendrotoxin (100 nM, n=3-5), but were blocked by stromatoxin-1 (ScTx, IC 50 ~130 nM), consistent with the idea that the currents were carried through K v 2 channels. RNA was detected for K v 2.1 K v 2.2 and the silent subunit K v 9.3 in urethral smooth muscle. Immunocytochemistry showed membrane staining for both K v 2 subtypes and K v 9.3 in isolated RUSMC. HEK Kv2.1 and HEK Kv2.2 currents were blocked in a concentration dependent manner by ScTx with estimated IC 50 values of ~150 nM (K v 2.1, n=5) and 70 nM (K v 2.2, n=6). The mean V 1/2 of inactivation of the USMC K v current was – 56±3 mV (n=9). This was similar to the HEK Kv2.1 current (–55 ± 3 mV, n=13) but significantly different from the HEK Kv2.2 currents (-30 ± 3 mV, n=11). Action potentials (AP) evoked from RUSMC studied under current clamp mode were unaffected by ScTx. However when ScTx was applied in the presence of Pen A, the AP duration was significantly prolonged. Similarly, ScTx increased the amplitude of spontaneous contractions threefold, but only after Pen A application. These data suggest that K v 2.1 channels contribute significantly to the K v current in RUSMC

Barriers and facilitators to cooking from 'scratch' using basic or raw ingredients: A qualitative interview study

BACKGROUND: Previous research has highlighted an ambiguity in understanding cooking related terminology and a number of barriers and facilitators to home meal preparation. However, meals prepared in the home still include convenience products (typically high in sugars, fats and sodium) which can have negative effects on health. Therefore, this study aimed to qualitatively explore: (1) how individuals define cooking from 'scratch', and (2) their barriers and facilitators to cooking with basic ingredients. METHODS: 27 semi-structured interviews were conducted with participants (aged 18-58 years) living on the island of Ireland, eliciting definitions of 'cooking from scratch' and exploring the reasons participants cook in a particular way. The interviews were professionally transcribed verbatim and Nvivo 10 was used for an inductive thematic analysis. RESULTS: Our results highlighted that although cooking from 'scratch' lacks a single definition, participants viewed it as optimal cooking. Barriers to cooking with raw ingredients included: 1) time pressures; (2) desire to save money; (3) desire for effortless meals; (4) family food preferences; and (5) effect of kitchen disasters. Facilitators included: 1) desire to eat for health and well-being; (2) creative inspiration; (3) ability to plan and prepare meals ahead of time; and (4) greater self-efficacy in one's cooking ability. CONCLUSIONS: Our findings contribute to understanding how individuals define cooking from 'scratch', and barriers and facilitators to cooking with raw ingredients. Interventions should focus on practical sessions to increase cooking self-efficacy; highlight the importance of planning ahead and teach methods such as batch cooking and freezing to facilitate cooking from scratch

Increasing intention to cook from basic ingredients: A randomised controlled study

The promotion of home cooking is a strategy used to improve diet quality and health. However, modern home cooking typically includes the use of processed food which can lead to negative outcomes including weight gain. In addition, interventions to improve cooking skills do not always explain how theory informed their design and implementation. The Behaviour Change Technique (BCT) taxonomy successfully employed in other areas has identified essential elements for interventions. This study investigated the effectiveness of different instructional modes for learning to cook a meal, designed using an accumulating number of BCTs, on participant's perceived difficulty, enjoyment, confidence and intention to cook from basic ingredients. 141 mothers aged between 20 and 39 years from the island of Ireland were randomised to one of four conditions based on BCTs (1) recipe card only [control condition]; (2) recipe card plus video modelling; (3) recipe card plus video prompting; (4) recipe card plus video elements. Participants rated their enjoyment, perceived difficulty, confidence and intention to cook again pre, mid and post experiment. Repeated one-way factorial ANOVAs, correlations and a hierarchical regression model were conducted. Despite no significant differences between the different conditions, there was a significant increase in enjoyment (P < 0.001), confidence (P < 0.001) and intention to cook from basics again (P < 0.001) and a decrease in perceived difficulty (P = 0.001) after the experiment in all conditions. Intention to cook from basics pre-experiment, and confidence and enjoyment (both pre and post experiment) significantly contributed to the final regression model explaining 42% of the variance in intention to cook from basics again. Cooking interventions should focus on practical cooking and increasing participants' enjoyment and confidence during cooking to increase intention to cook from basic ingredients at home

ARIADNE - A novel optical LArTPC: technical design report and initial characterisation using a secondary beam from the CERN PS and cosmic muons

ARIADNE is a 1-ton (330 kg fiducial mass) dual-phase liquid argon (LAr) time projection chamber (TPC) featuring a novel optical readout. Four electron-multiplying charge-coupled device (EMCCD) cameras are mounted externally, and these capture the secondary scintillation light produced in the holes of a thick electron gas multiplier (THGEM). Track reconstruction using this novel readout approach is demonstrated. Optical readout has the potential to be a cost effective alternative to charge readout in future LArTPCs. In this paper, the technical design of the detector is detailed. Results of mixed particle detection using a secondary beam from the CERN PS (representing the first ever optical images of argon interactions in a dual-phase LArTPC at a beamline) and cosmic muon detection at the University of Liverpool are also presented.Comment: 58 pages, 40 figures. Changes from previous version based on pre-publication review: improved quality of various figures, improved clarity of some definitions and reduced longer sentences for better readability, fixed typos and formatting error

The role of attachment in body weight gain and weight loss in bariatric patients

Purpose: To explore the role of attachment styles in obesity. Material and methods: The present study explored differences in insecure attachment styles between an obese sample waiting for bariatric surgery (n=195) and an age, sex and height matched normal weight control group (n=195). It then explored the role of attachment styles in predicting change in BMI one year post bariatric surgery (n=143). Results: The bariatric group reported significantly higher levels of anxious attachment and lower levels of avoidant attachment than the control non obese group. Baseline attachment styles did not, however, predict change in BMI post-surgery. Conclusion: Attachment style is different in those that are already obese from those who are not. Attachment was not related to weight loss post-surgery

Vasodilation of rat skeletal muscle arteries by the novel BK channel opener GoSlo is mediated by the simultaneous activation of BK and Kv7 channels

BACKGROUND AND PURPOSE: BK channels play important roles in various physiological and pathophysiological processes and thus have been the target of several drug development programs focused on creating new efficacious BK channel openers, such as the GoSlo-SR compounds. However, the effect of GoSlo-SR compounds on vascular smooth muscle has not been studied. Therefore, we tested the hypothesis that GoSlo-SR compounds dilate arteries exclusively by activating BK channels. EXPERIMENTAL APPROACH: Experiments were performed on rat Gracilis muscle, saphenous, mesenteric and tail arteries using isobaric and isometric myography, sharp microelectrodes, digital droplet PCR and the patch-clamp technique. KEY RESULTS: GoSlo-SR compounds dilated isobaric and relaxed and hyperpolarized isometric vessel preparations and their effects were abolished after (i) functionally eliminating K channels by pre-constriction with 50 mM KCl or (ii) blocking all K channels known to be expressed in vascular smooth muscle. However, these effects were not blocked when BK channels were inhibited. Surprisingly, the K(V)7 channel inhibitor XE991 reduced their effects considerably, but neither K(V)1 nor K(V)2 channel blockers altered the inhibitory effects of GoSlo-SR. However, the combined blockade of BK and K(V)7 channels abolished the GoSlo-SR-induced relaxation. GoSlo-SR compounds also activated K(V)7.4 and K(V)7.5 channels expressed in HEK 293 cells. CONCLUSIONS AND IMPLICATIONS: This study shows that GoSlo-SR-compounds are effective relaxants in vascular smooth muscle and mediate their effects by a combined activation of BK and K(V)7.4/K(V)7.5 channels. Activation of K(V)1, K(V)2 or K(V)7.1 channels or other vasodilator pathways seem not to be involved