Evaluation of physical activity calorie equivalent (PACE) labels' impact on energy purchased in cafeterias: A stepped-wedge randomised controlled trial.

BACKGROUND: A recent meta-analysis suggested that using physical activity calorie equivalent (PACE) labels results in people selecting and consuming less energy. However, the meta-analysis included only 1 study in a naturalistic setting, conducted in 4 convenience stores. We therefore aimed to estimate the effect of PACE labels on energy purchased in worksite cafeterias in the context of a randomised study design. METHODS AND FINDINGS: A stepped-wedge randomised controlled trial (RCT) was conducted to investigate the effect of PACE labels (which include kcal content and minutes of walking required to expend the energy content of the labelled food) on energy purchased. The setting was 10 worksite cafeterias in England, which were randomised to the order in which they introduced PACE labels on selected food and drinks following a baseline period. There were approximately 19,000 workers employed at the sites, 72% male, with an average age of 40. The study ran for 12 weeks (06 April 2021 to 28 June 2021) with over 250,000 transactions recorded on electronic tills. The primary outcome was total energy (kcal) purchased from intervention items per day. The secondary outcomes were: energy purchased from non-intervention items per day, total energy purchased per day, and revenue. Regression models showed no evidence of an overall effect on energy purchased from intervention items, -1,934 kcals per site per day (95% CI -5,131 to 1,262), p = 0.236, during the intervention relative to baseline, equivalent to -5 kcals per transaction (95% CI -14 to 4). There was also no evidence for an effect on energy purchased from non-intervention items, -5 kcals per site per day (95% CI -513 to 504), p = 0.986, equivalent to 0 kcals per transaction (95% CI -1 to 1), and no clear evidence for total energy purchased -2,899 kcals per site (95% CI -5,810 to 11), p = 0.051, equivalent to -8 kcals per transaction (95% CI -16 to 0). Study limitations include using energy purchased and not energy consumed as the primary outcome and access only to transaction-level sales, rather than individual-level data. CONCLUSION: Overall, the evidence was consistent with PACE labels not changing energy purchased in worksite cafeterias. There was considerable variation in effects between cafeterias, suggesting important unmeasured moderators. TRIAL REGISTRATION: The study was prospectively registered on ISRCTN (date: 30.03.21; ISRCTN31315776).

Is altering the availability of healthier vs. less-healthy options effective across socioeconomic groups? A mega-analysis.

BACKGROUND: Availability interventions have been hypothesised to make limited demands on conscious processes and, as a result, to be less likely to generate health inequalities than cognitively-oriented interventions. Here we synthesise existing evidence to examine whether the impact of altering the availability of healthier vs. less-healthy options differs by socioeconomic position. METHODS: Individual-level data (21,360 observations from 7,375 participants) from six studies (conducted online (n = 4) and in laboratories (n = 2)) were pooled for mega-analysis. Multilevel logistic regressions analysed the impact of altering the availability of healthier options on selection of a healthier (rather than a less-healthy) option by socioeconomic position, assessed by (a) education and (b) income. RESULTS: Participants had over threefold higher odds of selecting a healthier option when the available range was predominantly healthier compared to selections when the range offered was predominantly less-healthy (odds ratio (OR): 3.8; 95%CIs: 3.5, 4.1). Less educated participants were less likely to select healthier options in each availability condition (ORs: 0.75-0.85; all p < 0.005), but there was no evidence of differences in healthier option selection by income. Compared to selections when the range offered was predominantly less-healthy, when predominantly healthier options were available there was a 31% increase in selecting healthier options for the most educated group vs 27% for the least educated. This modest degree of increased responsiveness in the most educated group appeared only to occur when healthier options were predominant. There was no evidence of any differential response to the intervention by income. CONCLUSION: Increasing the proportion of healthier options available increases the selection of healthier options across socioeconomic positions. Availability interventions may have a slightly larger beneficial effect on those with the highest levels of education in settings when healthier options predominate.

Impact of decreasing the proportion of higher energy foods and reducing portion sizes on food purchased in worksite cafeterias: A stepped-wedge randomised controlled trial.

BACKGROUND: Overconsumption of energy from food is a major contributor to the high rates of overweight and obesity in many populations. There is growing evidence that interventions that target the food environment may be effective at reducing energy intake. The current study aimed to estimate the effect of decreasing the proportion of higher energy (kcal) foods, with and without reducing portion size, on energy purchased in worksite cafeterias. METHODS AND FINDINGS: This stepped-wedge randomised controlled trial (RCT) evaluated 2 interventions: (i) availability: replacing higher energy products with lower energy products; and (ii) size: reducing the portion size of higher energy products. A total of 19 cafeterias were randomised to the order in which they introduced the 2 interventions. Availability was implemented first and maintained. Size was added to the availability intervention. Intervention categories included main meals, sides, cold drinks, snacks, and desserts. The study setting was worksite cafeterias located in distribution centres for a major United Kingdom supermarket and lasted for 25 weeks (May to November 2019). These cafeterias were used by 20,327 employees, mainly (96%) in manual occupations. The primary outcome was total energy (kcal) purchased from intervention categories per day. The secondary outcomes were energy (kcal) purchased from nonintervention categories per day, total energy purchased per day, and revenue. Regression models showed an overall reduction in energy purchased from intervention categories of -4.8% (95% CI -7.0% to -2.7%), p < 0.001 during the availability intervention period and a reduction of -11.5% (95% CI -13.7% to -9.3%), p < 0.001 during the availability plus size intervention period, relative to the baseline. There was a reduction in energy purchased of -6.6% (95% CI -7.9% to -5.4%), p < 0.001 during the availability plus size period, relative to availability alone. Study limitations include using energy purchased as the primary outcome (and not energy consumed) and the availability only of transaction-level sales data per site (and not individual-level data). CONCLUSIONS: Decreasing the proportion of higher energy foods in cafeterias reduced the energy purchased. Decreasing portion sizes reduced this further. These interventions, particularly in combination, may be effective as part of broader strategies to reduce overconsumption of energy from food in out-of-home settings. TRIAL REGISTRATION: ISRCTN registry ISRCTN87225572.

Increasing the proportion of healthier foods available with and without reducing portion sizes and energy purchased in worksite cafeterias: protocol for a stepped-wedge randomised controlled trial.

BACKGROUND: Overconsumption of energy from food contributes to high rates of overweight and obesity in many populations. A promising set of interventions tested in pilot studies in worksite cafeterias, suggests energy intake may be reduced by increasing the proportion of healthier - i.e. lower energy - food options available, and decreasing portion sizes. The current study aims to assess the impact on energy purchased of i. increasing the proportion of lower energy options available; ii. combining this with reducing portion sizes, in a full trial. METHODS: A stepped-wedge randomised controlled trial in 19 worksite cafeterias, where the proportion of lower energy options available in targeted food categories (including main meals, snacks, and cold drinks) will be increased; and combined with reduced portion sizes. The primary outcome is total energy (kcal) purchased from targeted food categories using a pooled estimate across all sites. Follow-up analyses will test whether the impact on energy purchased varies according to the extent of intervention implementation. DISCUSSION: This study will provide the most reliable estimate to date of the effect sizes of two promising interventions for reducing energy purchased in worksite cafeterias. TRIAL REGISTRATION: The study was prospectively registered on ISRCTN (date: 24.05.19; TRN: ISRCTN87225572; doi: https://doi.org/10.1186/ISRCTN87225572).

Proteomic Analysis After Status Epilepticus Identifies UCHL1 as Protective Against Hippocampal Injury.

Brief, non-harmful seizures (preconditioning) can temporarily protect the brain against prolonged, otherwise injurious seizures. Following focal-onset status epilepticus (SE) in preconditioned (tolerance) and sham-preconditioned (injury) mice, we screened for protein changes using a proteomic approach and identified several putative candidates of epileptic tolerance. Among SE-induced changes to both proteomic screens, proteins clustered in key regulatory pathways, including protein trafficking and cytoskeletal regulation. Downregulation of one such protein, ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1), was unique to injury and not evident in tolerance. UCHL1 inhibition decreased hippocampal ubiquitin, disrupted UPS function, interfered with seizure termination and exacerbated seizure-induced cell death. Though UCHL1 transcription was maintained after SE, we observed downregulation of the pro-translational antisense Uchl1 (AsUchl1) and confirmed that both AsUchl1 and rapamycin can increase UCHL1 expression in vivo. These data indicate that the post-transcriptional loss of UCHL1 following SE is deleterious to neuronal survival and may contribute to hyperexcitability, and are suggestive of a novel modality of rapamycin therapy.

Differential DNA methylation profiles of coding and non-coding genes define hippocampal sclerosis in human temporal lobe epilepsy.

Temporal lobe epilepsy is associated with large-scale, wide-ranging changes in gene expression in the hippocampus. Epigenetic changes to DNA are attractive mechanisms to explain the sustained hyperexcitability of chronic epilepsy. Here, through methylation analysis of all annotated C-phosphate-G islands and promoter regions in the human genome, we report a pilot study of the methylation profiles of temporal lobe epilepsy with or without hippocampal sclerosis. Furthermore, by comparative analysis of expression and promoter methylation, we identify methylation sensitive non-coding RNA in human temporal lobe epilepsy. A total of 146 protein-coding genes exhibited altered DNA methylation in temporal lobe epilepsy hippocampus (n = 9) when compared to control (n = 5), with 81.5% of the promoters of these genes displaying hypermethylation. Unique methylation profiles were evident in temporal lobe epilepsy with or without hippocampal sclerosis, in addition to a common methylation profile regardless of pathology grade. Gene ontology terms associated with development, neuron remodelling and neuron maturation were over-represented in the methylation profile of Watson Grade 1 samples (mild hippocampal sclerosis). In addition to genes associated with neuronal, neurotransmitter/synaptic transmission and cell death functions, differential hypermethylation of genes associated with transcriptional regulation was evident in temporal lobe epilepsy, but overall few genes previously associated with epilepsy were among the differentially methylated. Finally, a panel of 13, methylation-sensitive microRNA were identified in temporal lobe epilepsy including MIR27A, miR-193a-5p (MIR193A) and miR-876-3p (MIR876), and the differential methylation of long non-coding RNA documented for the first time. The present study therefore reports select, genome-wide DNA methylation changes in human temporal lobe epilepsy that may contribute to the molecular architecture of the epileptic brain.

Comparison of short-term effects of midazolam and lorazepam in the intra-amygdala kainic acid model of status epilepticus in mice.

Benzodiazepines remain as the first-line treatment for status epilepticus (SE), but debate continues as to the choice and delivery route of pharmacotherapy. Lorazepam is currently the preferred anticonvulsant for clinical use, but midazolam has become a popular alternative, particularly as it can be given by nonintravenous routes. Anticonvulsants are also commonly used to terminate SE in animal models. Here, we aimed to compare the efficacy of midazolam with that of lorazepam in an experimental model of focal-onset SE. Status epilepticus was induced by intra-amygdala microinjection of kainic acid in 8week old C57Bl/6 mice. Forty minutes later, mice were treated with an intraperitoneal injection of either lorazepam or midazolam (8mg/kg). Electroencephalogram (EEG) activity, histology, and behavioral tests assessing recovery of function were evaluated and compared between groups. Intraperitoneal injection of either lorazepam or midazolam resulted in similar patterns of reduced EEG epileptiform activity during 1-hour recordings. Damage to the hippocampus and presentation of postinsult anxiety-related behavior did not significantly differ between treatment groups at 72h. However, return of normal behaviors such as grooming, levels of activity, and the evaluation of overall recovery of SE mice were all superior at 24h in animals given midazolam compared with lorazepam. Our results indicate that midazolam is as effective as lorazepam as an anticonvulsant in this model while also providing improved animal recovery after SE. These data suggest that midazolam might be considered by researchers as an anticonvulsant in animal models of SE, particularly as it appears to satisfy the requirements of refining procedures involving experimental animals at early time-points after SE.

ThermoCyte: an inexpensive open-source temperature control system for in vitro live-cell imaging.

Live-cell imaging is a common technique in microscopy to investigate dynamic cellular behaviour and permits the accurate and relevant analysis of a wide range of cellular and tissue parameters, such as motility, cell division, wound healing responses and calcium (Ca2+) signalling in cell lines, primary cell cultures and ex vivo preparations. Furthermore, this can occur under many experimental conditions, making live-cell imaging indispensable for biological research. Systems which maintain cells at physiological conditions outside of a CO2 incubator are often bulky, expensive and use proprietary components. Here we present an inexpensive, open-source temperature control system for in vitro live-cell imaging. Our system 'ThermoCyte', which is constructed from standard electronic components, enables precise tuning, control and logging of a temperature 'set point' for imaging cells at physiological temperature. We achieved stable thermal dynamics, with reliable temperature cycling and a standard deviation of 0.42°C over 1 h. Furthermore, the device is modular in nature and is adaptable to the researcher's specific needs. This represents simple, inexpensive and reliable tool for laboratories to carry out custom live-cell imaging protocols, on a standard laboratory bench, at physiological temperature.

A finite element model of contusion spinal cord injury in rodents.

Traumatic spinal cord injuries result from high impact forces acting on the spine and are proceeded by an extensive secondary inflammatory response resulting in motor, sensory, and autonomic dysfunction. Experimental in vivo traumatic spinal cord injuries in rodents using a contusion model have been extremely useful in elucidating the underlying pathophysiology of these injuries. However, the relationship between the pathophysiology and the biomechanical factors is still not well understood. Therefore, the aim of this research is to provide a comprehensive analysis of the biomechanics of traumatic spinal cord injury in a rat contusion model. This is achieved through the development and validation of a finite element model of the thoracic rat spinal cord and subsequently simulating controlled cortical impact-induced traumatic spinal cord injury. The effects of impactor velocity, depth, and geometry on the resulting stresses and strains within the spinal cord are investigated. Our results show that increasing impactor depth results in larger stresses and strains within the spinal cord tissue as expected. Further, for the first time ever our results show that impactor geometry (spherical versus cylindrical) plays an important role in the distribution and magnitude of stresses and strains within the cord. Therefore, finite element modelling can be a powerful tool used to predict stresses and strains that occur in spinal cord tissue during trauma.

Communicating evidence about the environment's role in obesity and support for government policies to tackle obesity: a systematic review with meta-analysis.

Public support for many policies that tackle obesity by changing environments is low. This may reflect commonly held causal beliefs about obesity, namely that it is due to failures of self-control rather than environmental influences. Several studies have sought to increase public support by changing these and similar causal beliefs, with mixed results. The current review is the first systematic synthesis of these studies. Searches of PsycInfo, Medline, Web of Science, Scopus, and Open Grey yielded 20 eligible studies (N = 8977) from 11,776 abstracts. Eligible studies were controlled experiments with an intervention group that communicated information about the environment's role in obesity, and a measure of support for environment-based obesity policies. The protocol was prospectively registered on PROSPERO. Meta-analyses showed no evidence that communicating information about the environment's influence on obesity changed policy support or the belief that the environment influences obesity. A likely explanation for this null effect is the ineffectiveness of interventions that were designed to change the belief that the environment influences obesity. The possibility remains, however, that the association observed between beliefs about the causes of obesity and attitudes towards obesity policies is correlational and not causal.

Acceptability of policies to reduce consumption of red and processed meat: A population-based survey experiment.

Policies to reduce meat consumption are needed to help achieve climate change targets, and could also improve population health. Public acceptability can affect the likelihood of policy implementation. This study estimated the acceptability of policies to reduce red and processed meat consumption, and whether acceptability differed when policies were framed as benefitting health or the environment. In an online experiment, 2215 UK adults rated the acceptability of six policies, presented in a randomised order. Prior to rating policies, participants were randomised to one of two framing conditions, with policy outcomes described either as benefitting health or the environment. Regression models examined differences in the primary outcome - policy acceptability (rated on a 7-point scale) - by framing. Labels were the most accepted policy (48% support), followed by a media campaign (45%), reduced availability (40%) and providing incentives (38%). Increasing price (27%) and banning advertising (26%) were the least accepted. A substantial proportion of participants neither supported nor opposed most policies (26-33%), although this fell to 16% for increasing price. There was no evidence that framing policy benefits from a health or environment perspective influenced acceptability (-0.06, 95%CIs: 0.18,0.07). Fewer than half of the UK sample expressed support for any of six policies to reduce meat consumption, regardless of framing measures as benefitting health or the environment. Conversely, fewer than half expressed opposition, with the exception of price, suggesting considerable scope to influence public opinion in support of meat reduction measures to meet environmental and health goals.

Public support for policies to improve population and planetary health: A population-based online experiment assessing impact of communicating evidence of multiple versus single benefits.

BACKGROUND: Effective interventions for reducing the consumption of products that harm population and planetary health often lack public support, impeding implementation. Communicating evidence of policies' effectiveness can increase public support but there is uncertainty about the most effective ways of communicating this evidence. Some policies have multiple benefits such as both improving health and the environment. This study assesses whether communicating evidence of multiple versus single benefits of a policy increases its support. METHOD: Participants (n = 4616) nationally representative of the British population were randomised to one of 24 groups in an online experiment with a 4 × 3 × 2 between-subjects factorial design. The messages that participants viewed differed according to the evidence they communicated (no message, effectiveness for changing behaviour, effectiveness for changing behaviour + one policy benefit, effectiveness for changing behaviour + three policy benefits), type of policy (taxation, availability) and the target behaviour (consumption of energy-dense food, alcohol, or meat). The primary outcome was policy support. RESULTS: In a full factorial ANOVA, there was a significant main effect of communicating evidence of effectiveness on policy support, which was similar across policies and behaviours. Communicating three benefits increased support relative to communicating one benefit (d = 0.15; p = 0.01). Communicating one benefit increased support compared to providing evidence for changing behaviour alone (d = 0.13; p = 0.004) or no message (d = 0.11 p = 0.022). CONCLUSION: Communicating evidence of a policy's benefits increases support for policy action across different behaviours and policies. Presenting multiple benefits of policies enhances public support.

Release probability increases towards distal dendrites boosting high-frequency signal transfer in the rodent hippocampus.

Dendritic integration of synaptic inputs involves their increased electrotonic attenuation at distal dendrites, which can be counterbalanced by the increased synaptic receptor density. However, during network activity, the influence of individual synapses depends on their release fidelity, the dendritic distribution of which remains poorly understood. Here, we employed classical optical quantal analyses and a genetically encoded optical glutamate sensor in acute hippocampal slices of rats and mice to monitor glutamate release at CA3-CA1 synapses. We find that their release probability increases with greater distances from the soma. Similar-fidelity synapses tend to group together, whereas release probability shows no trends regarding the branch ends. Simulations with a realistic CA1 pyramidal cell hosting stochastic synapses suggest that the observed trends boost signal transfer fidelity, particularly at higher input frequencies. Because high-frequency bursting has been associated with learning, the release probability pattern we have found may play a key role in memory trace formation.

Communicating Evidence about the Causes of Obesity and Support for Obesity Policies: Two Population-Based Survey Experiments.

Public support for numerous obesity policies is low, which is one barrier to their implementation. One reason for this low support is the tendency to ascribe obesity to failings of willpower as opposed to the environment. Correlational evidence supports this position. However, the experimental evidence is mixed. In two experimental studies, participants were randomised to receive no message, messages about the environment's influence on obesity (Study 1 & 2), or messages about the environment's influence on human behaviour (Study 1). We investigated whether communicating these messages changed support for obesity policies and beliefs about the causes of obesity. Participants were recruited from nationally representative samples in Great Britain (Study 1 & 2) and the USA (Study 2) (total n = 4391). Study 2 was designed to replicate existing research. Neither study found evidence that communicating the messages increased support for obesity policies or strengthened beliefs about the environment's role in obesity. Study 2, therefore, did not replicate two earlier experimental studies. Instead, the studies reported here suggest that people's beliefs about the causes of obesity are resistant to change in response to evidence and are, therefore, not a promising avenue to increase support for obesity policies.

Publisher Correction: Astrocytes mediate neurovascular signaling to capillary pericytes but not to arterioles.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

LTP Induction Boosts Glutamate Spillover by Driving Withdrawal of Perisynaptic Astroglia.

Extrasynaptic actions of glutamate are limited by high-affinity transporters expressed by perisynaptic astroglial processes (PAPs): this helps maintain point-to-point transmission in excitatory circuits. Memory formation in the brain is associated with synaptic remodeling, but how this affects PAPs and therefore extrasynaptic glutamate actions is poorly understood. Here, we used advanced imaging methods, in situ and in vivo, to find that a classical synaptic memory mechanism, long-term potentiation (LTP), triggers withdrawal of PAPs from potentiated synapses. Optical glutamate sensors combined with patch-clamp and 3D molecular localization reveal that LTP induction thus prompts spatial retreat of astroglial glutamate transporters, boosting glutamate spillover and NMDA-receptor-mediated inter-synaptic cross-talk. The LTP-triggered PAP withdrawal involves NKCC1 transporters and the actin-controlling protein cofilin but does not depend on major Ca2+-dependent cascades in astrocytes. We have therefore uncovered a mechanism by which a memory trace at one synapse could alter signal handling by multiple neighboring connections.

Multiplexed calcium imaging of single-synapse activity and astroglial responses in the intact brain.

All-optical registration of neuronal and astrocytic activities within the intact mammalian brain has improved significantly with recent advances in optical sensors and biophotonics. However, relating single-synapse release events and local astroglial responses to sensory stimuli in an intact animal has not hitherto been feasible. Here, we present a multiplexed multiphoton excitation imaging approach for assessing the relationship between presynaptic Ca2+ entry at thalamocortical axonal boutons and perisynaptic astrocytic Ca2+ elevations, induced by whisker stimulation in the barrel cortex of C57BL/6 mice. We find that, unexpectedly, Ca2+ elevations in the perisynaptic astrocytic regions consistently precede local presynaptic Ca2+ signals during spontaneous brain activity associated with anaesthesia. The methods described here can be adapted to a variety of optical sensors and are compatible with experimental designs that might necessitate repeated sampling of single synapses over a longitudinal behavioural paradigm.

Self-regulatory consequences of observing others making goal progress: A longitudinal field study in weight loss groups.

OBJECTIVE: What happens when people see others making progress toward a goal that they also hold? Is it motivating or could it undermine goal pursuit because people feel that they have made progress themselves (i.e., they experience vicarious goal satiation)? METHODS: We investigated these questions in a longitudinal field context - a group weight loss programme. N = 132 participants who were overweight or obese and attended weekly weight loss classes completed questionnaires over 11 weeks to investigate the consequences of observing other people making progress toward their goal of losing weight RESULTS: Observing others making good progress was associated with participants holding stronger intentions to lose weight themselves (B = 0.04, p = .012), positive goal-related affect (B = 0.27, p = .017), and feeling that they were also making progress themselves (B = 0.22, p < .001). However, observing others making good progress was also associated with losing a smaller amount of weight over the following week (B = .13, p = .005). Mediation analyses showed a significant indirect effect of observing others making good progress, via feelings about their own progress, on changes in weight, B = .02, 95% CI [.00, .04]. CONCLUSIONS: People who view others making progress tend to be less successful at losing weight themselves over the following week. The findings suggest that this is, in part, explained by the person feeling as if they have made progress themselves; thereby providing the first demonstration of vicarious goal satiation in a field context. Statement of contribution What is already known on this subject? Many people strive toward health goals (e.g., try to lose weight) with other people. However, research into vicarious goal satiation (VGS) suggests that seeing someone else achieve their goal may lead people to believe that they have made progress themselves and undermine goal pursuit. What does this study add? A longitudinal test of VGS in a field context, namely weight loss groups. Observing other people do well was motivating for the observer. They also believed that they had made progress themselves and were less successful at losing weight.

Computed tomography confirmation of component rotation in nanosensor-balanced total knee arthroplasty.

Balanced gaps and proper rotation are felt to be essential for optimum range of motion, stability, and patellar tracking in total knee arthroplasty. The purpose of this study is to assess, using computed tomography, the rotation of femoral and tibial components in fresh-frozen human cadaver knees that have been balanced using nanosensor trials while also observing how this rotation affects measured compartment loads and requirement for ligament balancing adjustment. We found that minor degrees of rotational malalignment of the femur and tibia were common using standard instrumentation and measured resection technique. Quantitative balance and rotational congruence are aided by nanosensor guidance, and femoral malrotation of up to 8° does not appear to affect compartment loads significantly as long as rotational congruity is present.

Author Correction: Disentangling astroglial physiology with a realistic cell model in silico.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.