Diabetes drugs activate neuroprotective pathways in models of neonatal hypoxic-ischemic encephalopathy.

Hypoxic-ischaemic encephalopathy (HIE) arises from diminished blood flow and oxygen to the neonatal brain during labor, leading to infant mortality or severe brain damage, with a global incidence of 1.5 per 1000 live births. Glucagon-like Peptide 1 Receptor (GLP1-R) agonists, used in type 2 diabetes treatment, exhibit neuroprotective effects in various brain injury models, including HIE. In this study, we observed enhanced neurological outcomes in post-natal day 10 mice with surgically induced hypoxic-ischaemic (HI) brain injury after immediate systemic administration of exendin-4 or semaglutide. Short- and long-term assessments revealed improved neuropathology, survival rates, and locomotor function. We explored the mechanisms by which GLP1-R agonists trigger neuroprotection and reduce inflammation following oxygen-glucose deprivation and HI in neonatal mice, highlighting the upregulation of the PI3/AKT signalling pathway and increased cAMP levels. These findings shed light on the neuroprotective and anti-inflammatory effects of GLP1-R agonists in HIE, potentially extending to other neurological conditions, supporting their potential clinical use in treating infants with HIE.

Exposure to urban particulate matter (UPM) impairs mitochondrial dynamics in BV2 cells, triggering a mitochondrial biogenesis response.

World Health Organisation data suggest that up to 99% of the global population are exposed to air pollutants above recommended levels. Impacts to health range from increased risk of stroke and cardiovascular disease to chronic respiratory conditions, and air pollution may contribute to over 7 million premature deaths a year. Additionally, mounting evidence suggests that in utero or early life exposure to particulate matter (PM) in ambient air pollution increases the risk of neurodevelopmental impairment with obvious lifelong consequences. Identifying brain-specific cellular targets of PM is vital for determining its long-term consequences. We previously established that microglial-like BV2 cells were particularly sensitive to urban (U)PM-induced damage including reactive oxygen species production, which was abrogated by a mitochondrially targeted antioxidant. Here we extend those studies to find that UPM treatment causes a rapid impairment of mitochondrial function and increased mitochondrial fragmentation. However, there is a subsequent restoration of mitochondrial and therefore cell health occurring concomitantly with upregulated measures of mitochondrial biogenesis and mitochondrial load. Our data highlight that protecting mitochondrial function may represent a valuable mechanism to offset the effects of UPM exposure in the neonatal brain. KEY POINTS: Air pollution represents a growing risk to long-term health especially in early life, and the CNS is emerging a target for airborne particulate matter (PM). We previously showed that microglial-like BV2 cells were vulnerable to urban (U)PM exposure, which impaired cell survival and promoted reactive oxygen species production. Here we find that, following UPM exposure, BV2 mitochondrial membrane potential is rapidly reduced, concomitant with decreased cellular bioenergetics and increased mitochondrial fission. However, markers of mitochondrial biogenesis and mitochondrial mass are subsequently induced, which may represent a cellular mitigation strategy. As mitochondria are more vulnerable in the developing brain, exposure to air pollution may represent a greater risk to lifelong health in this cohort; conversely, promoting mitochondrial integrity may offset these risks.

Leflunomide Treatment Does Not Protect Neural Cells following Oxygen-Glucose Deprivation (OGD) In Vitro.

Neonatal hypoxia-ischemia (HI) affects 2-3 per 1000 live births in developed countries and up to 26 per 1000 live births in developing countries. It is estimated that of the 750,000 infants experiencing a hypoxic-ischemic event during birth per year, more than 400,000 will be severely affected. As treatment options are limited, rapidly identifying new therapeutic avenues is critical, and repurposing drugs already in clinical use offers a fast-track route to clinic. One emerging avenue for therapeutic intervention in neonatal HI is to target mitochondrial dysfunction, which occurs early in the development of brain injury. Mitochondrial dynamics are particularly affected, with mitochondrial fragmentation occurring at the expense of the pro-fusion protein Optic Atrophy (OPA)1. OPA1, together with mitofusins (MFN)1/2, are required for membrane fusion, and therefore, protecting their function may also safeguard mitochondrial dynamics. Leflunomide, an FDA-approved immunosuppressant, was recently identified as an activator of MFN2 with partial effects on OPA1 expression. We, therefore, treated C17.2 cells with Leflunomide before or after oxygen-glucose deprivation, an in vitro mimic of HI, to determine its efficacy as a neuroprotection and inhibitor of mitochondrial dysfunction. Leflunomide increased baseline OPA1 but not MFN2 expression in C17.2 cells. However, Leflunomide was unable to promote cell survival following OGD. Equally, there was no obvious effect on mitochondrial morphology or bioenergetics. These data align with studies suggesting that the tissue and mitochondrial protein profile of the target cell/tissue are critical for taking advantage of the therapeutic actions of Leflunomide.

Sex-specific behavioural deficits in adulthood following acute activation of the GABAA receptor in the neonatal mouse.

INTRODUCTION: Sex differences exist in the prevalence of neurodevelopmental disorders (NDDs). Part of the aetiology of NDDs has been proposed to be alterations in the balance between excitatory and inhibitory neurotransmission, leading to the question of whether males and females respond differently to altered neurotransmitter balance. We investigated whether pharmacological alteration of GABAA signalling in early development results in sex-dependent changes in adult behaviours associated with NDDs. METHODS: Male and female C57BL/6J mice received intraperitoneal injections of 0.5mg/kg muscimol or saline on postnatal days (P) 3-5 and were subjected to behavioural testing, specifically open field, light dark box, marble burying, sucralose preference, social interaction and olfactory habituation/dishabituation tests between P60-90. RESULTS: Early postnatal administration of muscimol resulted in reduced anxiety in the light dark box test in both male and female adult mice. Muscimol reduced sucralose preference in males, but not females, whereas female mice showed reduced social behaviours. Regional alterations in cortical thickness were observed in the weeks following GABAA receptor activation, pointing to an evolving structural difference in the brain underlying adult behaviour. CONCLUSIONS: We conclude that activation of the GABAA receptor in the first week of life resulted in long-lasting changes in a range of behaviours in adulthood following altered neurodevelopment. Sex of the individual affected the nature and severity of these abnormalities, explaining part of the varied pathophysiology and neurodevelopmental diagnosis that derive from excitatory/inhibitory imbalance.

Athletes and Experimental Pain: A Systematic Review and Meta-Analysis.

The evidence that athletes respond to and report indices of experimental pain differently to non-athlete populations was analysed. Databases screened were SPORTDiscus, PubMED, PsycArticles, the Cochrane Library (Cochrane Database of Systematic Reviews), Web of Science, Scopus, and CINAHL. Studies that compared experimentally induced pain responses (threshold, tolerance, intensity, unpleasantness, bothersomeness, and effect on performance) in athletes and controls were included. Meta-analyses were performed where appropriate and effects were described as standardised mean differences, pooled using random effects models. Thirty-six studies (2,492 participants) met the inclusion criteria comprising 19 pain tolerance, 17 pain threshold, 21 pain intensity, 5 pain unpleasantness, 2 performance in pain and 1 bothersomeness study. Athletes demonstrated greater pain tolerance (g = .88 [95% confidence interval [CI] .65, .13]) and reported less pain intensity (g = -.80, [95% CI -1.13, -.47]) compared to controls; they also had higher pain threshold but with smaller effects (g = .41, [95% CI .08, .75]). Differences for unpleasantness did not reach statistical significance but the effects were large (g = -1.23 [95% CI -2.29, .18]). Two studies reported that performance in pain was better in contact athletes than non-athletes, and one concluded that athletes find pain less bothersome than controls. There were considerable inconsistencies in the methods employed that were reflected in the meta-analyses' findings. Sub-group analyses of tolerance and intensity were conducted between endurance, contact, and other athlete groups, but were not significant. The data suggest that athletic participation is associated with altered pain responses, but mechanisms remain unclear and more transparent methods are recommended.This study was registered on the PROSPERO site in January 2019 (ref ID: CRD42019119611). PERSPECTIVE: This review examined differences in pain outcomes (threshold, tolerance, intensity, unpleasantness, bothersomeness) and the effect of pain on performance, in athletes versus controls. Meta-analyses revealed athletes had higher threshold and tolerance and found pain less intense than controls; there was some evidence of differences in bothersomeness and performance.

Investigation into the transition to online learning due to the COVID-19 pandemic, between new and continuing undergraduate students.

Higher Education Institutions (HEIs) were required to rapidly respond to the COVID-19 pandemic, integrating online and blended learning approaches to sustain teaching and learning provision. However, limited evidence exists to understand the student experience and perception of the various methods of online learning, in particular across different levels of study (new and continuing students). Therefore, the aim of this study was to compare the experiences of online learning transition, between new first year undergraduate students and continuing second and third year students, enrolled on various undergraduate sport programmes. A total of 182 students responded to an online survey, which investigated the students' perceptions of online learning approaches. Participants were split according to level of study; [Level 3 (Foundation Year) and 4 (First Year Undergraduate) combined N = 62, Level 5 (Second Year Undergraduate), N = 51 and Level 6 (Third Year Undergraduate), N = 69]. Key findings highlight that both new and continuing students had an overall negative perception of online learning but did acknowledge that online learning provided a more flexible approach to their overall learning experience compared to face-to-face. Face-to-face teaching was deemed more engaging and sociable, in particular for the practical aspects of the programmes. Overall, there were no significant differences between the different levels of study for any of the questions asked. Although continuing students raised the difficulties of conducting practical sessions online, whereas this was not mentioned by new students. To conclude, this study provides novel insights into the experience of new and continuing students, and we advise that future blended learning strategies should consider the programme as a whole, rather than tailoring pedagogic strategies based on the level of study.

A national survey of clubs medical provision and facilities in BUCS American Football 2019-2020.

BACKGROUND: British American Football (BAF) is a developing sport in the UK, with keen growth in the British Universities and Colleges Sport (BUCS) league. Participation in BAF carries risks and so to facilitate safe participation medical care services must be evaluated. AIMS: To evaluate medical provision in BUCS American Football in the 2019-2020 season. METHODS: An online survey tool was used to collect data from BUCS BAF teams in the 2019-2020 season. Thirty-one teams (from across England, Wales and Scotland) responded to questions on facilities, provision and procedures. RESULTS: Almost 42% of teams had a regular team first aider who attended home games each week. Only 61.5% attended away games and 7.7% attended team training. Access to a first aider was not dependent upon division. Home games were more likely to be risk assessed and have an emergency action plan compared to away games. The majority of teams had access to automated external defibrillator (AED) within 100 m of the pitch, yet only 29% of staff were trained to use them. Almost 84% of teams reported carrying a designated fully charged phone (with signal). Prominent qualitative themes indicated were cost/funding as barriers to hiring qualified medical staff, lack of institutional support, unreliability of medical provision and inadequate facilities/preparation for games. CONCLUSIONS: These findings provide key information on the status of medical provision, facilities and protocols in BUCS BAF. Data reveals a lack of consistent medical personnel, particularly at training and away games, and training in emergency care.

The current state of concussion knowledge and attitudes in British American Football.

OBJECTIVES: To examine concussion knowledge and concussion attitudes of players, coaches, and support staff in British American Football (BAF). METHODS: Data from players, coaches and support staff (n = 236) were collected from across all leagues in BAF. An online survey tool was used which included the Rosenbaum Concussion Knowledge and Attitudes Survey (RoCKAS), and questions examining concussion education and perceived risk of participating in football. RESULTS: The mean score on the RoCKAS concussion knowledge was 21.0 ± 2.1 of a possible score of 25 reflecting good knowledge. Of a possible score of 65, the mean concussion attitude score was 55.6 ± 6.1 showing safe attitude. Whilst an overall safe attitude was seen, almost half of participants (45.3%) noted they would continue to play with a concussion. No relationship was found between CAI and prior concussion history. Fifty seven percent of participants agreed the benefits of playing football outweighed the risks. Forty eight percent reported that they had received no concussion-related education in the past 12 months. CONCLUSION: BAF participants have good concussion knowledge and safe attitudes. However, risky behavior is demonstrated through unsafe likelihood to report and attitude to long-term health risks. Access to the British American Football Association (BAFA) concussion policy and education was poor raising questions over what sources of information stakeholders are drawing their knowledge from. These findings can help form the foundation of educational interventions (e.g. coaching workshops) to challenge current misconceptions and improve likelihood to report concussion in BAF.

A functional eEF2K-eEF2 pathway in the NAc is critical for the expression of cocaine-induced psychomotor sensitisation and conditioned place preference.

Recent evidence links synaptic plasticity and mRNA translation, via the eukaryotic elongation factor 2 kinase (eEF2K) and its only known substrate, eEF2. However, the involvement of the eEF2 pathway in cocaine-induced neuroadaptations and cocaine-induced behaviours is not known. Knock-in (KI) mice and shRNA were used to globally and specifically reduce eEF2K expression. Cocaine psychomotor sensitization and conditioned place preference were used to evaluate behavioural outcome. Changes in eEF2 phosphorylation were determined by western blot analyses. No effect was observed on the AMPA/NMDA receptor current ratio in the ventral tegmental area, 24 h after cocaine injection in eEF2K-KI mice compared with WT. However, development and expression of cocaine psychomotor sensitization were decreased in KI mice. Phosphorylated eEF2 was decreased one day after psychomotor sensitization and returned to baseline at seven days in the nucleus accumbens (NAc) of WT mice, but not in eEF2K-KI mice. However, one day following cocaine challenge, phosphorylated eEF2 decreased in WT but not KI mice. Importantly, specific targeting of eEF2K expression by shRNA in the NAc decreased cocaine condition place preference. These results suggest that the eEF2 pathway play a role in cocaine-induced locomotor sensitization and conditioned place preference.

Montelukast reduces grey matter abnormalities and functional deficits in a mouse model of inflammation-induced encephalopathy of prematurity.

Encephalopathy of prematurity (EoP) affects approximately 30% of infants born < 32 weeks gestation and is highly associated with inflammation in the foetus. Here we evaluated the efficacy of montelukast, a cysteinyl leukotriene receptor antagonist widely used to treat asthma in children, to ameliorate peripheral and central inflammation, and subsequent grey matter neuropathology and behaviour deficits in a mouse model of EoP. Male CD-1 mice were treated with intraperitoneal (i.p.) saline or interleukin-1beta (IL-1ß, 40 µg/kg, 5 µL/g body weight) from postnatal day (P)1-5 ± concomitant montelukast (1-30 mg/kg). Saline or montelukast treatment was continued for a further 5 days post-injury. Assessment of systemic and central inflammation and short-term neuropathology was performed from 4 h following treatment through to P10. Behavioural testing, MRI and neuropathological assessments were made on a second cohort of animals from P36 to 54. Montelukast was found to attenuate both peripheral and central inflammation, reducing the expression of pro-inflammatory molecules (IL-1ß, IL-6, TNF) in the brain. Inflammation induced a reduction in parvalbumin-positive interneuron density in the cortex, which was normalised with high-dose montelukast. The lowest effective dose, 3 mg/kg, was able to improve anxiety and spatial learning deficits in this model of inflammatory injury, and alterations in cortical mean diffusivity were not present in animals that received this dose of montelukast. Repurposed montelukast administered early after preterm birth may, therefore, improve grey matter development and outcome in EoP.

Differential Effects of Urban Particulate Matter on BV2 Microglial-Like and C17.2 Neural Stem/Precursor Cells.

Air pollution affects the majority of the world's population and has been linked to over 7 million premature deaths per year. Exposure to particulate matter (PM) contained within air pollution is associated with cardiovascular, respiratory, and neurological ill health. There is increasing evidence that exposure to air pollution in utero and in early childhood is associated with altered brain development. However, the underlying mechanisms for impaired brain development are not clear. While oxidative stress and neuroinflammation are documented consequences of PM exposure, cell-specific mechanisms that may be triggered in response to air pollution exposure are less well defined. Here, we assess the effect of urban PM exposure on two different cell types, microglial-like BV2 cells and neural stem/precursor-like C17.2 cells. We found that, contrary to expectations, immature C17.2 cells were more resistant to PM-mediated oxidative stress and cell death than BV2 cells. PM exposure resulted in decreased mitochondrial health and increased mitochondrial ROS in BV2 cells which could be prevented by MitoTEMPO antioxidant treatment. Our data suggest that not only is mitochondrial dysfunction a key trigger in PM-mediated cytotoxicity but that such deleterious effects may also depend on cell type and maturity.

Induction of Mitochondrial Fragmentation and Mitophagy after Neonatal Hypoxia-Ischemia.

Hypoxia-ischemia (HI) leads to immature brain injury mediated by mitochondrial stress. If damaged mitochondria cannot be repaired, mitochondrial permeabilization ensues, leading to cell death. Non-optimal turnover of mitochondria is critical as it affects short and long term structural and functional recovery and brain development. Therefore, disposal of deficient mitochondria via mitophagy and their replacement through biogenesis is needed. We utilized mt-Keima reporter mice to quantify mitochondrial morphology (fission, fusion) and mitophagy and their mechanisms in primary neurons after Oxygen Glucose Deprivation (OGD) and in brain sections after neonatal HI. Molecular mechanisms of PARK2-dependent and -independent pathways of mitophagy were investigated in vivo by PCR and Western blotting. Mitochondrial morphology and mitophagy were investigated using live cell microscopy. In primary neurons, we found a primary fission wave immediately after OGD with a significant increase in mitophagy followed by a secondary phase of fission at 24 h following recovery. Following HI, mitophagy was upregulated immediately after HI followed by a second wave at 7 days. Western blotting suggests that both PINK1/Parkin-dependent and -independent mechanisms, including NIX and FUNDC1, were upregulated immediately after HI, whereas a PINK1/Parkin mechanism predominated 7 days after HI. We hypothesize that excessive mitophagy in the early phase is a pathologic response which may contribute to secondary energy depletion, whereas secondary mitophagy may be involved in post-HI regeneration and repair.

Mitochondrial dynamics in the neonatal brain - a potential target following injury?

The impact of birth asphyxia and its sequelae, hypoxic-ischaemic (HI) brain injury, is long-lasting and significant, both for the infant and for their family. Treatment options are limited to therapeutic hypothermia, which is not universally successful and is unavailable in low resource settings. The energy deficits that accompany neuronal death following interruption of blood flow to the brain implicate mitochondrial dysfunction. Such HI insults trigger mitochondrial outer membrane permeabilisation leading to release of pro-apoptotic proteins into the cytosol and cell death. More recently, key players in mitochondrial fission and fusion have been identified as targets following HI brain injury. This review aims to provide an introduction to the molecular players and pathways driving mitochondrial dynamics, the regulation of these pathways and how they are altered following HI insult. Finally, we review progress on repurposing or repositioning drugs already approved for other indications, which may target mitochondrial dynamics and provide promising avenues for intervention following brain injury. Such repurposing may provide a mechanism to fast-track, low-cost treatment options to the clinic.

Concussion Reporting and Safeguarding Policy Development in British American Football: An Essential Agenda.

The objective of this study was to examine concussion reporting and safeguarding policy in British American Football (BAF). Data were collected via an online survey tool. The data presented are part of a broader study that examined injury profiles, concussion reporting behaviors, and medical provision in BAF. Concussion-like symptoms were found in over half (58.8%) of the participants. Of those, 36.4% reported they had previously been formally diagnosed with a concussion while playing BAF. Just under half of the participants (44.7%) had suspected they had had a concussion, although it was not formally diagnosed, and 23.5% of the participants had previously hidden concussion symptoms. Fifty-eight percent of the teams reported they did not have a regular game-day medic, with a range of hired medical personnel who attended the games. Prominent barriers to hiring a medic included budget, institutional support shortfall, and lack of medic reliability and game knowledge. BAF is a developing sport with a clear vision for growth of participation. Yet, the current concussion and medical provision policies do not address the sport's welfare needs. Through discussion of these policies in the context of this study's findings, we highlight vital areas of concern in policy and practice that the British American Football Association needs to address in their medical and concussion policies.

Early life exposure to air pollution impacts neuronal and glial cell function leading to impaired neurodevelopment.

The World Health Organisation recently listed air pollution as the most significant threat to human health. Air pollution comprises particulate matter (PM), metals, black carbon and gases such as ozone (O3 ), nitrogen dioxide (NO2 ) and carbon monoxide (CO). In addition to respiratory and cardiovascular disease, PM exposure is linked with increased risk of neurodegeneration as well as neurodevelopmental impairments. Critically, studies suggest that PM crosses the placenta, making direct in utero exposure a reality. Rodent models reveal that neuroinflammation, neurotransmitter imbalance and oxidative stress are triggered following gestational/early life exposure to PM, and may be exacerbated by concomitant mitochondrial dysfunction. Gestational PM exposure (potentiated by mitochondrial impairment in the metabolically active neonatal brain) not only impacts neurodevelopment but may sensitise the brain to subsequent cognitive impairment. Having reviewed this field, we conclude that strategies are urgently required to reduce exposure to PM during this sensitive developmental period.

The role of mitochondria in cocaine addiction.

The incidence of cocaine abuse is increasing especially in the U.K. where the rates are among the highest in Europe. In addition to its role as a psychostimulant, cocaine has profound effect on brain metabolism, impacting glycolysis and impairing oxidative phosphorylation. Cocaine exposure alters metabolic gene expression and protein networks in brain regions including the prefrontal cortex, the ventral tegmental area and the nucleus accumbens, the principal nuclei of the brain reward system. Here, we focus on how cocaine impacts mitochondrial function, in particular through alterations in electron transport chain function, reactive oxygen species (ROS) production and oxidative stress (OS), mitochondrial dynamics and mitophagy. Finally, we describe the impact of cocaine on brain energy metabolism in the developing brain following prenatal exposure. The plethora of mitochondrial functions altered following cocaine exposure suggest that therapies maintaining mitochondrial functional integrity may hold promise in mitigating cocaine pathology and addiction.

Exposure to Contact Sports Results in Maintained Performance During Experimental Pain.

During pain, motor performance tends to decline. However, athletes who engage in contact sports are able to maintain performance despite the inherent pain that accompanies participation. This may be the result of being challenged rather than threatened by pain; adaptive coping strategies; habituation to pain; or finding pain less bothersome. This study aimed to measure performance of a novel motor task both in pain and not in pain within experienced contact athletes (n = 40), novice contact athletes (n = 40), and noncontact athletes (n = 40). Challenge and threat perceptions were manipulated during the pain condition and measures of pain tolerance, perception, coping styles, and bothersomeness were taken. Results indicated that contact athletes, regardless of experience, were able to maintain their performance during painful stimulation. Noncontact athletes, conversely, performed significantly worse during pain stimulation. In addition, contact athletes tended to be more challenged and the noncontact athletes more threatened within the pain condition. Experienced contact athletes demonstrated higher levels of pain tolerance and direct coping, and reported lower levels of pain bothersomeness and intensity than the other groups. The results suggest that even relatively brief exposure to contact sports may be enough to help maintain performance in pain. Being in a challenged state appears to be an important factor during performance in pain. Moreover, pain tolerance, intensity, and bothersomeness may differentiate novice and experienced athletes. PERSPECTIVE: Exposure to voluntary pain and challenge states are associated with adaptive responses to pain. Motor task performance may be maintained in individuals with more experience of sports-related pain.

Zeta Inhibitory Peptide attenuates learning and memory by inducing NO-mediated downregulation of AMPA receptors.

Zeta inhibitory peptide (ZIP), a PKMζ inhibitor, is widely used to interfere with the maintenance of acquired memories. ZIP is able to erase memory even in the absence of PKMζ, via an unknown mechanism. We found that ZIP induces redistribution of the AMPARGluA1 in HEK293 cells and primary cortical neurons, and decreases AMPAR-mediated currents in the nucleus accumbens (NAc). These effects were mimicked by free arginine or by a modified ZIP in which all but the arginine residues were replaced by alanine. Redistribution was blocked by a peptidase-resistant version of ZIP and by treatment with the nitric oxide (NO)-synthase inhibitor L-NAME. ZIP increased GluA1-S831 phosphorylation and ZIP-induced redistribution was blocked by nitrosyl-mutant GluA1-C875S or serine-mutant GluA1-S831A. Introducing the cleavable arginine-alanine peptide into the NAc attenuated expression of cocaine-conditioned reward. Together, these results suggest that ZIP may act as an arginine donor, facilitating NO-dependent downregulation of AMPARs, thereby attenuating learning and memory.