Pre-exercise hot water immersion increased circulatory heat shock proteins but did not alter muscle damage markers or endurance capacity after eccentric exercise.

Pre-exercise passive heating attenuates muscle damage caused by eccentric exercise in rats where the induction of heat shock proteins (HSPs) confers a myoprotective effect. We investigated whether pre-exercise hot water immersion (HWI) confers similar benefits in humans. Eleven recreational male athletes were immersed in 41°C water up to 60 min or until rectal temperatures reached 39.5°C. After a 6 h rest, the participants performed an eccentric downhill run for 1 h at -4% gradient to induce muscle damage. An endurance capacity test at 75% VO2max was conducted 18 h later. The control trial was similar except that participants were immersed at 34°C. Blood samples were collected to assess HSPs levels, creatine kinase, and lactate dehydrogenase activities. Plasma eHSP70 was higher post-immersion in HWI trials (1.3 ± 0.4 vs 1.1 ± 0.4; p = 0.005). Plasma eHSP27 was higher before (p = 0.049) and after (p = 0.015) endurance test in HWI. Leukocytic p-HSP27 was increased 18 h after HWI (0.97 ± 0.14 vs 0.67 ± 0.11; p = 0.04). Creatine kinase and lactate dehydrogenase activities were increased by 3-fold and 1.5-fold, respectively, after endurance test in HWI but did not differ across trials (p > 0.05). Mean heart rates were higher during eccentric run and endurance test in HWI as compared to control (p < 0.05). Endurance capacity was similar between trials (57.3 ± 11.5 min vs 55.0 ± 13.5 min; p = 0.564). Pre-exercise heating increased the expression of plasma eHSPs and leukocytic p-HSP27 but did not reduce muscle damage nor enhance endurance capacity.

Effects of Heat Exposure and Ice Slurry Ingestion on Risk-Taking Behavior in Healthcare Workers.

PURPOSE: Healthcare workers (HCWs) wearing personal protective equipment (PPE) experience physiological strain that can impair motor and psychological functions, potentially affecting patient care. We assessed the effects of heat exposure on maximal strength and risk-taking behavior amongst PPE-wearing HCWs and the efficacy of ice slurry to alleviate adverse effects. METHODS: 17 HCWs completed two experimental trials in a crossover design, consuming 5 g × kg-1 of body mass of ambient drink (AMB) or ice slurry (ICE) before donning PPE and undergoing 2-h of simulated decontamination exercise (wet-bulb globe temperature (WBGT): 25.9 ± 0.8 °C, PPE microenvironment WBGT: 29.1 ± 2.1 °C). Body core temperature (Tc), heart rate (HR), chest skin temperature (Tsk), ratings of perceived exertion (RPE), thermal sensation (RTS), maximal voluntary contraction (MVC), risk-taking behavior (Balloon Analogue Risk-Taking task; BART) and salivary cortisol were assessed. RESULTS: Pre- to post-drinking ∆Tc was greater in ICE (-0.2 ± 0.1 °C) than AMB (-0.0 ± 0.1 °C, P = 0.003). Post-drinking RTS was lower in ICE (2.7 ± 1.2) than AMB (4.1 ± 0.4, P < 0.001). ICE and AMB had similar Tc and HR (both P > 0.05), but Tsk was lower in ICE than AMB (P = 0.049). A lower MVC (30.3 ± 6.7 kg vs 27.4 ± 4.9 kg, P = 0.001) and higher BART adjusted total pump count (472 ± 170 pumps vs 615 ± 174 pumps, P = 0.017) was observed pre- to post-trial in AMB but absent in ICE (both P > 0.05). Salivary cortisol was similar between trials (P = 0.42). CONCLUSIONS: Heat-exposed PPE-wearing HCWs had impaired maximal strength and elevated risk-taking behavior. This may increase the risk of avoidable workplace accidents that can jeopardize HCWs and patient care. Ice slurry ingestion alleviated these heat-related impairments, suggesting its potential as an ergogenic aid.

External auricle temperature enhances ear-based wearable accuracy during physiological strain monitoring in the heat.

Body core temperature (Tc) monitoring is crucial for minimizing heat injury risk. However, validated strategies are invasive and expensive. Although promising, aural canal temperature (Tac) is susceptible to environmental influences. This study investigated whether incorporation of external auricle temperature (Tea) into an ear-based Tc algorithm enhances its accuracy during multiple heat stress conditions. Twenty males (mean ± SD; age = 25 ± 3 years, BMI = 21.7 ± 1.8, body fat = 12 ± 3%, maximal aerobic capacity (VO2max) = 64 ± 7 ml/kg/min) donned an ear-based wearable and performed a passive heating (PAH), running (RUN) and brisk walking trial (WALK). PAH comprised of immersion in hot water (42.0 ± 0.3 °C). RUN (70 ± 3%VO2max) and WALK (50 ± 10%VO2max) were conducted in an environmental chamber (Tdb = 30.0 ± 0.2 °C, RH = 71 ± 2%). Several Tc models, developed using Tac, Tea and heart rate, were validated against gastrointestinal temperature. Inclusion of Tea as a model input improved the accuracy of the ear-based Tc algorithm. Our best performing model (Trf3) displayed good group prediction errors (mean bias error = - 0.02 ± 0.26 °C) but exhibited individual prediction errors (percentage target attainment ± 0.40 °C = 88%) that marginally exceeded our validity criterion. Therefore, Trf3 demonstrates potential utility for group-based Tc monitoring, with additional refinement needed to extend its applicability to personalized heat strain monitoring.

Elevated brain temperature under severe heat exposure impairs cortical motor activity and executive function.

BACKGROUND: Excessive heat exposure can lead to hyperthermia in humans, which impairs physical performance and disrupts cognitive function. While heat is a known physiological stressor, it is unclear how severe heat stress affects brain physiology and function. METHODS: Eleven healthy participants were subjected to heat stress from prolonged exercise or warm water immersion until their rectal temperatures (Tre) attained 39.5°C, inducing exertional or passive hyperthermia, respectively. In a separate trial, blended ice was ingested before and during exercise as a cooling strategy. Data were compared to a control condition with seated rest (normothermic). Brain temperature (Tbr), cerebral perfusion, and task-based brain activity were assessed using magnetic resonance imaging techniques. RESULTS: Tbr in motor cortex was found to be tightly regulated at rest (37.3°C ± 0.4°C (mean ± SD)) despite fluctuations in Tre. With the development of hyperthermia, Tbr increases and dovetails with the rising Tre. Bilateral motor cortical activity was suppressed during high-intensity plantarflexion tasks, implying a reduced central motor drive in hyperthermic participants (Tre = 38.5°C ± 0.1°C). Global gray matter perfusion and regional perfusion in sensorimotor cortex were reduced with passive hyperthermia. Executive function was poorer under a passive hyperthermic state, and this could relate to compromised visual processing as indicated by the reduced activation of left lateral-occipital cortex. Conversely, ingestion of blended ice before and during exercise alleviated the rise in both Tre and Tbr and mitigated heat-related neural perturbations. CONCLUSION: Severe heat exposure elevates Tbr, disrupts motor cortical activity and executive function, and this can lead to impairment of physical and cognitive performance.

The effects of low and normal dose ice slurry ingestion on endurance capacity and intestinal epithelial injury in the heat.

OBJECTIVES: Compare the effects of ice slurry ingestion at low and normal doses on endurance capacity and exertional heat stress-induced gastrointestinal perturbations. DESIGN: Randomised, cross-over design. METHODS: Twelve physically active males completed four treadmill running trials, ingesting ice slurry (ICE) or ambient drink (AMB) at 2 g·kg-1 (Normal; N) or 1 g·kg-1 (Low; L) doses every 15-min during exercise and 8 g·kg-1 (N) or 4 g·kg-1 (L) pre- and post-exercise. Pre-, during and post-exercise serum intestinal fatty-acid binding protein ([I-FABP]) and lipopolysaccharide ([LPS]) concentrations were determined. RESULTS: Pre-exercise gastrointestinal temperature (Tgi) was lower in L + ICE than L + AMB (p < 0.05), N + ICE than N + AMB (p < 0.001) and N + ICE than L + ICE (p < 0.001). Higher rate of Tgi rise (p < 0.05) and lower estimated sweat rate (p < 0.001) were observed in N + ICE than N + AMB. Rate of Tgi rise was similar at low dose (p = 0.113) despite a lower estimated sweat rate in L + ICE than L+AMB (p < 0.01). Time-to-exhaustion was longer in L + ICE than L + AMB (p < 0.05), but similar between N + ICE and N + AMB (p = 0.142) and L + ICE and N + ICE (p = 0.766). [I-FABP] and [LPS] were similar (p > 0.05). CONCLUSIONS: L + ICE elicited a lower heat dissipation compensatory effect with similar endurance capacity as N + ICE. Ice slurry conferred no protection against exertional heat stress-induced gastrointestinal perturbations.

Mobile application-assisted graded exercise practical: a remote teaching strategy to promote motivation and experiential learning in exercise physiology.

Practical classes are critical instructional activities in facilitating learning and motivation in health sciences education. With increasing pedagogical activities being conducted in virtual or remote settings, this study assessed how a remote practical assisted by physiological monitoring smartphone applications impacted student motivation and the achievement of intended learning outcomes in exercise physiology teaching. A total of 24 students (out of 30; 80%) were surveyed via a mixed-methods questionnaire containing 27 closed-ended, and 3 the traditional in-class practical in randomized order. Unpaired Student's t tests were performed for comparisons between interventions with a significance level set at P < 0.05. Students reported that both remote and in-class practicals strongly facilitated the achievement of learning outcomes. Self-reported scores for student satisfaction and perceived achievement of learning outcomes were similar between the two practical methodologies. Student motivation scores assessed using the Lab Motivation Scale revealed that students were more motivated during the remote practical, particularly in the effort domain (P < 0.05). This was in line with the identified themes from the qualitative responses that indicated that the remote practical was more engaging than the in-person practical, with greater opportunities for experiential learning and class involvement being the main factors underlying these findings. Taken together, remote practicals can be critical aspects of a blended learning curriculum that encourages student engagement and experiential learning. With further advancements in physiological monitoring wearables and smartphone technologies, remote practicals can be potential alternatives to traditional in-person practicals in exercise physiology teaching.NEW & NOTEWORTHY Remote practical classes, supported by physiological monitoring smartphone applications, were assessed for their utility in facilitating learning and raising student motivation in health sciences education in this study. A comparison of remote practicals with traditional in-class practicals revealed that a remote practical is an effective method for reinforcing physiology learning objectives with the added advantage of increased student motivation. The added value of remote practicals may be attributed to more experiential learning opportunities and increased engagement levels.

Efficacy of Isothermic Conditioning over Military-Based Heat Acclimatization and Interval Training in Tropical Native Males.

PURPOSE: We compared the effectiveness of three field-based training programs, namely military-based heat acclimatization (MHA), isothermic conditioning (IC) and interval training (IT), in inducing physiological adaptations in tropical natives. METHODS: Fifty-one untrained tropical native males (mean ± standard deviation: age, 25 ± 2 yr; body mass index, 23.6 ± 3.2 kg·m -2 ; body fat, 19% ± 5%; 2.4-km run time, 13.2 ± 0.9 min) donned the Full Battle Order attire (22 kg) and performed a treadmill route march heat stress test in an environmental chamber (dry bulb temperature, 29.9°C ± 0.5°C; relative humidity, 70% ± 3%). Heat stress tests were conducted before (PRE) and after (POST) a 2-wk training intervention consisting of either a MHA ( n = 17, 10 sessions of military-based heat acclimatization), IC ( n = 17, 10 sessions with target gastrointestinal temperature ( Tgi ) ≥ 38.5°C) or IT ( n = 17, six sessions of high-intensity interval training) program. Tgi , HR, mean weighted skin temperature ( Tsk ), physiological strain index (PSI) and thigh-predicted sweat sodium concentration ([Na + ]) were measured and analyzed by one-factor and two-factor mixed design ANOVA with a 0.05 level of significance. RESULTS: Field-based IC induced a greater thermal stimulus than MHA ( P = 0.029) and IT ( P < 0.001) during training. Reductions in mean exercise Tgi (-0.2°C [-0.3°C, 0.0°C]; P = 0.009) , PSI (-0.4 [-0.7, -0.1]; P = 0.015) and thigh-predicted sweat [Na + ] (-9 [-13, -5 mmol·L -1 ]; P < 0.001) were observed in IC but not MHA and IT (all P > 0.05). Resting HR (MHA, -4 bpm [-7, 0 bpm]; P = 0.025; IC, -7 bpm [-10, -4 bpm]; P < 0.001; IT, -4 bpm [-8, -1 bpm]; P = 0.008) and mean exercise HR (MHA, -4 [-8, 0 bpm]; P = 0.034; IC, -11 bpm [-15, -8 bpm]; P < 0.001, IT = -5 bpm [-9, -1 bpm]; P = 0.012) were lowered in all groups after training. Isothermic conditioning elicited a greater attenuation in mean exercise HR and thigh-predicted sweat [Na + ] relative to MHA (both P < 0.05). No between-group differences were observed when comparing MHA and IT (all P > 0.05). CONCLUSIONS: Isothermic conditioning induced a more complete heat-adapted phenotype relative to MHA and IT. Interval training may serve as a time efficient alternative to MHA.

Evaluation of the Possible Anticonvulsant Effect of Δ9-Tetrahydrocannabinolic Acid in Murine Seizure Models.

Introduction: The cannabinoid Δ9-tetrahydrocannabinolic acid (Δ9-THCA) has long been suggested in review articles and anecdotal reports to be anticonvulsant; yet, there is scant evidence supporting this notion. The objective of this study was to interrogate the anticonvulsant potential of Δ9-THCA in various seizure models-the Scn1a+/- mouse model of Dravet syndrome, the 6-Hz model of psychomotor seizures and the maximal electroshock (MES) model of generalized tonic-clonic seizures. Materials and Methods: We examined the effect of acute Δ9-THCA treatment against hyperthermia-induced seizures, and subchronic treatment on spontaneous seizures and survival in the Scn1a+/- mice. We also studied the effect of acute Δ9-THCA treatment on the critical current thresholds in the 6-Hz and MES tests using outbred Swiss mice. Highly purified Δ9-THCA was used in the studies or a mixture of Δ9-THCA and Δ9-THC. Results: We observed mixed anticonvulsant and proconvulsant effects of Δ9-THCA across the seizure models. Highly pure Δ9-THCA did not affect hyperthermia-induced seizures in Scn1a+/- mice. A Δ9-THCA/Δ9-THC mixture was anticonvulsant in the 6-Hz threshold test, but purified Δ9-THCA and Δ9-THC had no effect. Conversely, both Δ9-THCA and Δ9-THC administered individually were proconvulsant in the MES threshold test but had no effect when administered as a Δ9-THCA/Δ9-THC mixture. The Δ9-THCA/Δ9-THC mixture, however, increased spontaneous seizure severity and increased mortality of Scn1a+/- mice. Discussion: The anticonvulsant profile of Δ9-THCA was variable depending on the seizure model used and presence of Δ9-THC. Because of the unstable nature of Δ9-THCA, further exploration of Δ9-THCA through formal anticonvulsant drug development is problematic without stabilization. Future studies may better focus on determining the mechanisms by which combined Δ9-THCA and Δ9-THC alters seizure thresholds, as this may uncover novel targets for the control of refractory partial seizures.

A Multiethnic Asian Perspective of Presumed Consent for Organ Donation: A Population-Based Perception Study.

Background: Organ shortage is still a world-wide problem, resulting in long waiting lists for kidney, liver, and heart transplant candidates across many transplant centers globally. This has resulted in the move toward presumed consent to increase deceased organ donation rates. However, there remains a paucity of literature on public attitude and barriers regarding the opt-out system, with existing studies limited to Western nations. Therefore, this study aimed to understand public sentiment and different barriers toward organ donation from the perspective of Singapore, a highly diverse and multiethnic Asian society. Methods: A cross-sectional community semi-structured interview was conducted in a public housing estate in Singapore. Pilot test was undertaken before participants were interviewed face-to-face by trained personnel. All statistical evaluations were conducted using Stata. The χ2-test compared subgroups based on patient characteristics while multivariable logistic regression identified predictors of willingness to donate/ assent. Effect estimates were quantified using odds ratio (OR). Findings: Out of 799 individuals, 85% were agreeable to organ donation after death and 81% were willing to assent to donations of family members' organs, which declined by 16% (p < 0.001) after a clinical scenario was presented. Demographic factors including ethnicity, education, marital, and employment status affected willingness to donate and assent. Knowledge correlated significantly with willingness to donate and assent. In particular, knowledge regarding brain death irreversibility had the strongest correlation (AOR 2.15; 95% CI 1.60-2.89). Conclusions: Organ donation rates remain low albeit presumed consent legislation, due to patient-level barriers, including but not limited to knowledge gaps, cultural values, religious backgrounds, and emotional impact at relatives' death. To effectively boost donor rates, it is crucial for policy makers to invest in public education and improve transplant provisions and family protocols.

Assessment of dehydration using body mass changes of elite marathoners in the tropics.

OBJECTIVES: The ACSM recommends drinking to avoid loss of body mass >2% during exercise to avert compromised performance. Our study aimed to assess the level of dehydration in elite runners following a city marathon in a tropical environment. DESIGN: Prospective cohort design. METHODS: Twelve elite runners (6 males, 6 females; age 24-41 y) had body mass measured to the nearest 0.01kg in their race attire immediately before and after the 2017 Standard Chartered Singapore Marathon 2017. Body mass change was corrected for respiratory water loss, gas exchange, and sweat retained in clothing, and expressed as % of pre-race mass (i.e. % dehydration). RESULTS: Data are expressed as means±SD (range). Dry bulb temperature and humidity were 27.9±0.1°C (27.4-28.3°C) and 79±2% (73-82%). Finish time was 155±10min (143-172min). Male runners finishing positions ranged from 2-12 out of 7627 finishers, whilst female runners placed 1-8 out of 1754 finishers. Body mass change (loss) and % dehydration for all runners were 2.5±0.5kg (1.8-3.5kg) and 4.6±0.9% (3.6-6.8%). Male runners experienced body mass loss of 2.8±0.5kg and 4.9±1.2% while females experienced body mass loss of 2.1±0.2kg and 4.3±0.6%. CONCLUSIONS: Despite experiencing dehydration (4.6% body mass loss) two-fold higher than current fluid replacement guidelines recommend (≤2%), elite male and female runners performed successfully and without medical complication in a hot weather marathon.

Thermoregulatory responses to ice slurry ingestion during low and moderate intensity exercises with restrictive heat loss.

OBJECTIVES: We investigated the thermoregulatory responses to ice slurry ingestion during low- and moderate-intensity exercises with restrictive heat loss. DESIGN: Randomised, counterbalanced, cross-over design. METHODS: Following a familiarisation trial, ten physically active males exercised on a motorised treadmill at low-intensity (L; 40% VO2max) or moderate-intensity (M; 70% VO2max) for 75-min, in four randomised, counterbalanced trials. Throughout the exercise bout, participants donned a raincoat to restrict heat loss. Participants ingested 2gkg-1 body mass of ambient water (L+AMB and M+AMB trials) or ice slurry (L+ICE and M+ICE trials) at 15-min intervals during exercise in environmental conditions of Tdb, 25.1±0.6°C and RH, 63±5%. Heart rate (HR), gastrointestinal temperature (Tgi), mean weighted skin temperature (Tsk), estimated sweat loss, ratings of perceived exertion (RPE) and thermal sensation (RTS) were recorded. RESULTS: Compared to L+AMB, participants completed L+ICE trials with lower ΔTgi (0.8±0.3°C vs 0.6±0.2°C; p=0.03), mean RPE (10±1 vs 9±1; p=0.03) and estimated sweat loss (0.91±0.2L vs 0.78±0.27L; p=0.04). Contrastingly, Tgi (p=0.22), Tsk (p=0.37), HR (p=0.31), RPE (p=0.38) and sweat loss (p=0.17) were similar between M+AMB and M+ICE trials. RTS was similar during both low-intensity (4.9±0.5 vs 4.7±0.3; p=0.10) and moderate-intensity exercise (5.3±0.47 vs 5.0±0.4; p=0.09). CONCLUSIONS: Per-cooling using ice slurry ingestion marginally reduced thermal strain during low-intensity but not during moderate-intensity exercise. Ice slurry may be an effective and practical heat mitigation strategy during low-intensity exercise such as in occupational and military settings, but a greater volume should be considered to ensure its efficacy.

Serine-70 phosphorylated Bcl-2 prevents oxidative stress-induced DNA damage by modulating the mitochondrial redox metabolism.

Bcl-2 phosphorylation at serine-70 (S70pBcl2) confers resistance against drug-induced apoptosis. Nevertheless, its specific mechanism in driving drug-resistance remains unclear. We present evidence that S70pBcl2 promotes cancer cell survival by acting as a redox sensor and modulator to prevent oxidative stress-induced DNA damage and execution. Increased S70pBcl2 levels are inversely correlated with DNA damage in chronic lymphocytic leukemia (CLL) and lymphoma patient-derived primary cells as well as in reactive oxygen species (ROS)- or chemotherapeutic drug-treated cell lines. Bioinformatic analyses suggest that S70pBcl2 is associated with lower median overall survival in lymphoma patients. Empirically, sustained expression of the redox-sensitive S70pBcl2 prevents oxidative stress-induced DNA damage and cell death by suppressing mitochondrial ROS production. Using cell lines and lymphoma primary cells, we further demonstrate that S70pBcl2 reduces the interaction of Bcl-2 with the mitochondrial complex-IV subunit-5A, thereby reducing mitochondrial complex-IV activity, respiration and ROS production. Notably, targeting S70pBcl2 with the phosphatase activator, FTY720, is accompanied by an enhanced drug-induced DNA damage and cell death in CLL primary cells. Collectively, we provide a novel facet of the anti-apoptotic Bcl-2 by demonstrating that its phosphorylation at serine-70 functions as a redox sensor to prevent drug-induced oxidative stress-mediated DNA damage and execution with potential therapeutic implications.

Interactions between cannabidiol and Δ9 -tetrahydrocannabinol in modulating seizure susceptibility and survival in a mouse model of Dravet syndrome.

BACKGROUND AND PURPOSE: Extracts from the cannabis plant can dramatically improve the health of children suffering from refractory epilepsies such as Dravet syndrome. These extracts typically contain cannabidiol (CBD), a phytocannabinoid with well-documented anticonvulsant effects, but may also contain Δ9 -tetrahydrocannabinol (Δ9 -THC). It is unclear whether the presence of Δ9 -THC modulates the anticonvulsant efficacy of CBD. Here, we utilized the Scn1a+/- mouse model of Dravet syndrome to examine this question. EXPERIMENTAL APPROACH: Scn1a+/- mice recapitulate core features of Dravet syndrome, including hyperthermia-induced seizures, early onset spontaneous seizures and sudden death. We assessed the effects on CBD and Δ9 -THC alone, and in combination on hyperthermia-induced seizures, spontaneous seizures and premature mortality. KEY RESULTS: Administered alone, CBD (100 mg·kg-1 i.p.) was anticonvulsant against hyperthermia-induced seizures as were low (0.1 and 0.3 mg·kg-1 i.p.) but not higher doses of Δ9 -THC. A subthreshold dose of CBD (12 mg·kg-1 ) enhanced the anticonvulsant effects of Δ9 -THC (0.1 mg·kg-1 ). Sub-chronic oral administration of Δ9 -THC or CBD alone did not affect spontaneous seizure frequency or mortality while, surprisingly, their co-administration increased the severity of spontaneous seizures and overall mortality. CONCLUSION AND IMPLICATIONS: Low doses of Δ9 -THC are anticonvulsant against hyperthermia-induced seizures in Scn1a+/- mice, effects that are enhanced by a sub-anticonvulsant dose of CBD. However, proconvulsant effects and increased premature mortality are observed when CBD and Δ9 -THC are sub-chronically dosed in combination. The possible explanations and implications of this are discussed.

The intestinal virome in children with cystic fibrosis differs from healthy controls.

Intestinal bacterial dysbiosis is evident in children with cystic fibrosis (CF) and intestinal viruses may be contributory, given their influence on bacterial species diversity and biochemical cycles. We performed a prospective, case-control study on children with CF and age and gender matched healthy controls (HC), to investigate the composition and function of intestinal viral communities. Stool samples were enriched for viral DNA and RNA by viral extraction, random amplification and purification before sequencing (Illumina MiSeq). Taxonomic assignment of viruses was performed using Vipie. Functional annotation was performed using Virsorter. Inflammation was measured by calprotectin and M2-pyruvate kinase (M2-PK). Eight CF and eight HC subjects were included (50% male, mean age 6.9 ± 3.0 and 6.4 ± 5.3 years, respectively, p = 0.8). All CF subjects were pancreatic insufficient. Regarding the intestinal virome, no difference in Shannon index between CF and HC was identified. Taxonomy-based beta-diversity (presence-absence Bray-Curtis dissimilarity) was significantly different between CF and HC (R2 = 0.12, p = 0.001). Myoviridae, Faecalibacterium phage FP Taranis and unclassified Gokushovirinae were significantly decreased in CF compared with HC (q<0.05). In children with CF (compared to HC), the relative abundance of genes annotated to (i) a peptidoglycan-binding domain of the peptidoglycan hydrolases (COG3409) was significantly increased (q<0.05) and (ii) capsid protein (F protein) (PF02305.16) was significantly decreased (q<0.05). Picornavirales, Picornaviridae, and Enterovirus were found to positively correlate with weight and BMI (r = 0.84, q = 0.01). Single-stranded DNA viruses negatively correlated with M2-PK (r = -0.86, q = 0.048). Children with CF have an altered intestinal virome compared to well-matched HC, with both taxonomic and predicted functional changes. Further exploration of Faecalibacterium phages, Gokushovirinae and phage lysins are warranted. Intestinal viruses and their functions may have important clinical implications for intestinal inflammation and growth in children with CF, potentially providing novel therapeutic targets.

Thermoregulation in the Aging Population and Practical Strategies to Overcome a Warmer Tomorrow.

As global temperatures continue to rise, improving thermal tolerance in the aged population is crucial to counteract age-associated impairments in thermoregulatory function. Impairments in reflex cutaneous vasodilation and sweating response can augment the vulnerability of older adults to heat-related injuries following exposure to heat stress. Mechanisms underlying a compromised cutaneous vasodilation are suggested to include reduced sympathetic neural drive, diminished cholinergic co-transmitter contribution, and altered second messenger signaling events. On the other hand, impairments in sweating response are ascribed to reduced sweat gland cholinergic sensitivity and altered cyclooxygenase and nitric oxide signaling. Several practical mitigation strategies such as exercise, passive heating, and behavioral adaptations are proposed as means to overcome heat stress and improve thermal tolerance in the aged. Aerobic exercise training is shown to be amongst the most effective ways to enhance thermoregulatory function. However, in elderly with limited exercise capability due to chronic diseases and mobility issues, passive heating can serve as a functional alternative as it has been shown to confer similar benefits to that of exercise training. Supplementary to exercise training and passive heating, behavioral adaptations can be applied to further enhance the heat-preparedness of the aged.

Pharmacokinetics of Phytocannabinoid Acids and Anticonvulsant Effect of Cannabidiolic Acid in a Mouse Model of Dravet Syndrome.

Cannabis sativa produces a complex mixture of many bioactive molecules including terpenophenolic compounds known as phytocannabinoids. Phytocannabinoids come in neutral forms (e.g., Δ9-tetrahydrocannabinol, THC; cannabidiol, CBD; etc.) or as acid precursors, which are dominant in the plant (e.g., Δ9-tetrahydrocannabinolic acid, THCA; cannabidiolic acid, CBDA; etc.). There is increasing interest in unlocking the therapeutic applications of the phytocannabinoid acids; however, the present understanding of the basic pharmacology of phytocannabinoid acids is limited. Herein the brain and plasma pharmacokinetic profiles of CBDA, THCA, cannabichromenic acid (CBCA), cannabidivarinic acid (CBDVA), cannabigerolic acid (CBGA), and cannabigerovarinic acid (CBGVA) were examined following intraperitoneal administration in mice. Next it was examined whether CBDA was anticonvulsant in a mouse model of Dravet syndrome (Scn1aRX/+ mice). All the phytocannabinoid acids investigated were rapidly absorbed with plasma tmax values of between 15 and 45 min and had relatively short half-lives (<4 h). The brain-plasma ratios for the acids were very low at ≤0.04. However, when CBDA was administered in an alternate Tween 80-based vehicle, it exhibited a brain-plasma ratio of 1.9. The anticonvulsant potential of CBDA was examined using this vehicle, and it was found that CBDA significantly increased the temperature threshold at which the Scn1aRX/+ mice had a generalized tonic-clonic seizure.

Coadministered cannabidiol and clobazam: Preclinical evidence for both pharmacodynamic and pharmacokinetic interactions.

OBJECTIVE: Cannabidiol (CBD) has been approved by the US Food and Drug Administration (FDA) to treat intractable childhood epilepsies, such as Dravet syndrome and Lennox-Gastaut syndrome. However, the intrinsic anticonvulsant activity of CBD has been questioned due to a pharmacokinetic interaction between CBD and a first-line medication, clobazam. This recognized interaction has led to speculation that the anticonvulsant efficacy of CBD may simply reflect CBD augmenting clobazam exposure. The present study aimed to address the nature of the interaction between CBD and clobazam. METHODS: We examined whether CBD inhibits human CYP3A4 and CYP2C19 mediated metabolism of clobazam and N-desmethylclobazam (N-CLB), respectively, and performed studies assessing the effects of CBD on brain and plasma pharmacokinetics of clobazam in mice. We then used the Scn1a+/- mouse model of Dravet syndrome to examine how CBD and clobazam interact. We compared anticonvulsant effects of CBD-clobazam combination therapy to monotherapy against thermally-induced seizures, spontaneous seizures and mortality in Scn1a+/- mice. In addition, we used Xenopus oocytes expressing γ-aminobutyric acid (GABA)A receptors to investigate the activity of GABAA receptors when treated with CBD and clobazam together. RESULTS: CBD potently inhibited CYP3A4 mediated metabolism of clobazam and CYP2C19 mediated metabolism of N-CLB. Combination CBD-clobazam treatment resulted in greater anticonvulsant efficacy in Scn1a+/- mice, but only when an anticonvulsant dose of CBD was used. It is important to note that a sub-anticonvulsant dose of CBD did not promote greater anticonvulsant effects despite increasing plasma clobazam concentrations. In addition, we delineated a novel pharmacodynamic mechanism where CBD and clobazam together enhanced inhibitory GABAA receptor activation. SIGNIFICANCE: Our study highlights the involvement of both pharmacodynamic and pharmacokinetic interactions between CBD and clobazam that may contribute to its efficacy in Dravet syndrome.

Heart Rate Measures From Wrist-Worn Activity Trackers in a Laboratory and Free-Living Setting: Validation Study.

BACKGROUND: Wrist-worn activity trackers are popular, and an increasing number of these devices are equipped with heart rate (HR) measurement capabilities. However, the validity of HR data obtained from such trackers has not been thoroughly assessed outside the laboratory setting. OBJECTIVE: This study aimed to investigate the validity of HR measures of a high-cost consumer-based tracker (Polar A370) and a low-cost tracker (Tempo HR) in the laboratory and free-living settings. METHODS: Participants underwent a laboratory-based cycling protocol while wearing the two trackers and the chest-strapped Polar H10, which acted as criterion. Participants also wore the devices throughout the waking hours of the following day during which they were required to conduct at least one 10-min bout of moderate-to-vigorous physical activity (MVPA) to ensure variability in the HR signal. We extracted 10-second values from all devices and time-matched HR data from the trackers with those from the Polar H10. We calculated intraclass correlation coefficients (ICCs), mean absolute errors, and mean absolute percentage errors (MAPEs) between the criterion and the trackers. We constructed decile plots that compared HR data from Tempo HR and Polar A370 with criterion measures across intensity deciles. We investigated how many HR data points within the MVPA zone (≥64% of maximum HR) were detected by the trackers. RESULTS: Of the 57 people screened, 55 joined the study (mean age 30.5 [SD 9.8] years). Tempo HR showed moderate agreement and large errors (laboratory: ICC 0.51 and MAPE 13.00%; free-living: ICC 0.71 and MAPE 10.20%). Polar A370 showed moderate-to-strong agreement and small errors (laboratory: ICC 0.73 and MAPE 6.40%; free-living: ICC 0.83 and MAPE 7.10%). Decile plots indicated increasing differences between Tempo HR and the criterion as HRs increased. Such trend was less pronounced when considering the Polar A370 HR data. Tempo HR identified 62.13% (1872/3013) and 54.27% (5717/10,535) of all MVPA time points in the laboratory phase and free-living phase, respectively. Polar A370 detected 81.09% (2273/2803) and 83.55% (9323/11,158) of all MVPA time points in the laboratory phase and free-living phase, respectively. CONCLUSIONS: HR data from the examined wrist-worn trackers were reasonably accurate in both the settings, with the Polar A370 showing stronger agreement with the Polar H10 and smaller errors. Inaccuracies increased with increasing HRs; this was pronounced for Tempo HR.

Ser70 phosphorylation of Bcl-2 by selective tyrosine nitration of PP2A-B56δ stabilizes its antiapoptotic activity.

Bcl-2 is frequently overexpressed in hematopoietic malignancies, and selective phosphorylation at ser70 enhances its antiapoptotic activity. Phospho-ser70 is dephosphorylated by specific heterotrimers of protein phosphatase 2A (PP2A). We report here that a mild pro-oxidant intracellular milieu induced by either pharmacological inhibition or genetic knockdown of superoxide dismutase 1 (SOD1) inhibits the functional holoenzyme assembly of PP2A and prevents Bcl-2 ser70 dephosphorylation. This redox-dependent regulation of Bcl-2 phosphorylation is due to nitrosative modification of B56δ, which we identify as the regulatory subunit mediating PP2A-dependent Bcl-2 dephosphorylation. Redox inhibition of PP2A results from peroxynitrite-mediated nitration of a conserved tyrosine residue within B56δ (B56δ(Y289)). Although nitrated B56δ(Y289) binds efficiently to ser70-phosphorylated Bcl-2, this specific modification inhibits the recruitment of the PP2A catalytic core (A and C subunits). Furthermore, inhibition of B56δ(Y289) nitration restores PP2A holoenzyme assembly, thereby permitting S70 dephosphorylation of Bcl-2 and inhibiting its antiapoptotic activity. More important, in primary cells derived from clinical lymphomas, Bcl-2 phosphorylation at S70 directly correlates with B56δ nitration and repression of SOD1, but inversely correlates with B56δ interaction with the PP2A-C catalytic subunit. These data underscore the role of a pro-oxidant milieu in chemoresistance of hematopoietic and other cancers via selective targeting of tumor suppressors such as PP2A.

Mitochondrial ROS and involvement of Bcl-2 as a mitochondrial ROS regulator.

Mitochondria are the major intracellular source of reactive oxygen species (ROS). While excessive mitochondrial ROS (mitoROS) production induces cell injury and death, there is accumulating evidence that non-toxic low levels of mitoROS could serve as important signaling molecules. Therefore, maintenance of mitoROS at physiological levels is crucial for cell homeostasis as well as for survival and proliferation. This review describes the various mechanisms that keep mitoROS in check, with particular focus on the role of the onco-protein Bcl-2 in redox regulation. In addition to its canonical anti-apoptotic activity, Bcl-2 has been implicated in mitoROS regulation by its effect on mitochondrial complex IV activity, facilitating the mitochondrial incorporation of GSH and interaction with the small GTPase-Rac1 at the mitochondria. We also discuss some of the plausible mechanism(s) which allows Bcl-2 to sense and respond to the fluctuations in mitoROS.