Comparison of Transcriptomic Changes in Survivors of Exertional Heat Illness with Malignant Hyperthermia Susceptible Patients.

Exertional heat illness (EHI) is an occupational health hazard for athletes and military personnel-characterised by the inability to thermoregulate during exercise. The ability to thermoregulate can be studied using a standardised heat tolerance test (HTT) developed by The Institute of Naval Medicine. In this study, we investigated whole blood gene expression (at baseline, 2 h post-HTT and 24 h post-HTT) in male subjects with either a history of EHI or known susceptibility to malignant hyperthermia (MHS): a pharmacogenetic condition with similar clinical phenotype. Compared to healthy controls at baseline, 291 genes were differentially expressed in the EHI cohort, with functional enrichment in inflammatory response genes (up to a four-fold increase). In contrast, the MHS cohort featured 1019 differentially expressed genes with significant down-regulation of genes associated with oxidative phosphorylation (OXPHOS). A number of differentially expressed genes in the inflammation and OXPHOS pathways overlapped between the EHI and MHS subjects, indicating a common underlying pathophysiology. Transcriptome profiles between subjects who passed and failed the HTT (based on whether they achieved a plateau in core temperature or not, respectively) were not discernable at baseline, and HTT was shown to elevate inflammatory response gene expression across all clinical phenotypes.

Investigating the genetic susceptibility to exertional heat illness.

BACKGROUND: We aimed to identify rare (minor allele frequency ≤1%), potentially pathogenic non-synonymous variants in a well-characterised cohort with a clinical history of exertional heat illness (EHI) or exertional rhabdomyolysis (ER). The genetic link between malignant hyperthermia (MH) and EHI was investigated due to their phenotypic overlap. METHODS: The coding regions of 38 genes relating to skeletal muscle calcium homeostasis or exercise intolerance were sequenced in 64 patients (mostly military personnel) with a history of EHI, or ER and who were phenotyped using skeletal muscle in vitro contracture tests. We assessed the pathogenicity of variants using prevalence data, in silico analysis, phenotype and segregation evidence and by review of the literature. RESULTS: We found 51 non-polymorphic, potentially pathogenic variants in 20 genes in 38 patients. Our data indicate that RYR1 p.T3711M (previously shown to be likely pathogenic for MH susceptibility) and RYR1 p.I3253T are likely pathogenic for EHI. PYGM p.A193S was found in 3 patients with EHI, which is significantly greater than the control prevalence (p=0.000025). We report the second case of EHI in which a missense variant at CACNA1S p.R498 has been found. Combinations of rare variants in the same or different genes are implicated in EHI. CONCLUSION: We confirm a role of RYR1 in the heritability of EHI as well as ER but highlight the likely genetic heterogeneity of these complex conditions. We propose defects, or combinations of defects, in skeletal muscle calcium homeostasis, oxidative metabolism and membrane excitability are associated with EHI.

Thermoregulation and markers of muscle breakdown in malignant hyperthermia susceptible volunteers during an acute heat tolerance test.

OBJECTIVES: The study was undertaken to compare the thermal and biochemical responses to a heat tolerance test (HTT) of malignant hyperthermia (MH) susceptible individuals, volunteers who have suffered heat illness (HI) and control volunteers. METHODS: Three groups of male volunteers (n=6 in each group) were recruited to the study: MHS - civilian volunteers previously diagnosed as MH susceptible; EHI - military volunteers with a history of exertional HI; CON - military volunteers with no history of HI or MH. For the HTT, volunteers walked on a treadmill at 60% maximal oxygen uptake in a hot environment. Measurements were made of core and skin temperatures, heat flow, whole body sweat rate and serum lactate, creatine kinase and myoglobin concentrations. RESULTS: There were no differences in deep body temperature, oxygen uptake or serum lactate and creatine kinase concentrations between the three groups. One MHS volunteer and two EHI volunteers failed to achieve thermal balance with rectal temperature continuing to rise throughout the test and reaching 39.5°C, the rectal temperatures of the other volunteers plateaued at a mean (SD) of 38.7 (0.4)°C demonstrating thermal tolerance on this test. Serum myoglobin concentration and the increase in serum myoglobin was higher in MHS than EHI and CON Post HHT (P<0.05). CONCLUSION: MH susceptibility does not always predispose an individual to heat intolerance during an acute HTT, but does appear to increase muscle breakdown. The inclusion of serum myoglobin measurements to a HTT may help to distinguish patients that are potentially MHS, and who otherwise demonstrate thermal tolerance.

Rebreather Unit to Prolong Underwater Survival Time.

INTRODUCTION: This study investigated whether the timing of activation affects the utility of an emergency underwater rebreather unit (RBU) when submerged in cold water. METHOD: On two successive occasions, 16 male UK Royal Marines were submerged in stirred water at 12.2°C for up to 78 s. The subjects were lowered (taking 18 s) into the water in a seated position and were instructed to take a large breath in, activate the unit, breath-hold for as long as possible, exhale into the unit, and breathe normally to and from the unit for the remainder of submersion. On one occasion the subjects were instructed to activate the RBU when the water reached chest height (Condition-1) and, on the other, prior to the feet entering the water (Condition-2). Measurements were made of the duration of breath-hold, rebreathing and submersion, exhaled oxygen and carbon dioxide concentrations, skin temperature, and heart rate. RESULTS: In 16 of the 32 submersions, the breath-hold was released before the subject became fully submerged and in 8 submersions the subject requested early withdrawal from the water. Mean (SD) breath-hold duration was 14.0 (13.8) s and the duration of rebreathing was 45.9 (21.9) s. The duration of breath-hold once completely submerged was longer in Condition-1 (9.1 s) than Condition-2 (4.1 s). CONCLUSIONS: The study indicates the RBU should be activated just before the mouth becomes submerged rather than before entering the water, and that the RBU will prolong underwater stay time, thereby increasing survival prospects. House CM, Shaw AM, Roiz de Sa DG. Rebreather unit to prolong underwater stay time, thereby increasing survival prospects.

Next-generation Sequencing of RYR1 and CACNA1S in Malignant Hyperthermia and Exertional Heat Illness.

BACKGROUND: Variants in RYR1 are associated with the majority of cases of malignant hyperthermia (MH), a form of heat illness pharmacogenetically triggered by general anesthetics, and they have also been associated with exertional heat illness (EHI). CACNA1S has also been implicated in MH. The authors applied a targeted next-generation sequencing approach to identify variants in RYR1 and CACNA1S in a cohort of unrelated patients diagnosed with MH susceptibility. They also provide the first comprehensive report of sequencing of these two genes in a cohort of survivors of EHI. METHODS: DNA extracted from blood was genotyped using a "long" polymerase chain reaction technique, with sequencing on the Illumina GAII or MiSeq platforms (Illumina Inc., USA). Variants were assessed for pathogenicity using bioinformatic approaches. For further follow-up, DNA from additional family members and up to 211 MH normal and 556 MH-susceptible unrelated individuals was tested. RESULTS: In 29 MH patients, the authors identified three pathogenic and four novel RYR1 variants, with a further five RYR1 variants previously reported in association with MH. Three novel RYR1 variants were found in the EHI cohort (n = 28) along with two more previously reported in association with MH. Two other variants were reported previously associated with centronuclear myopathy. The authors found one and three rare variants of unknown significance in CACNA1S in the MH and EHI cohorts, respectively. CONCLUSIONS: Targeted next-generation sequencing proved efficient at identifying diagnostically useful and potentially implicated variants in RYR1 and CACNA1S in MH and EHI.

Validation of one-mile walk equations for the estimation of aerobic fitness in British military personnel under the age of 40 years.

OBJECTIVE: To provide an accurate estimate of peak oxygen uptake (VO2 peak) for British Royal Navy Personnel aged between 18 and 39, comparing a gold standard treadmill based maximal exercise test with a submaximal one-mile walk test. METHODS: Two hundred military personnel consented to perform a treadmill-based VO2 peak test and two one-mile walk tests round an athletics track. The estimated VO2 peak values from three different one-mile walk equations were compared to directly measured VO2 peak values from the treadmill-based test. One hundred participants formed a validation group from which a new equation was derived and the other 100 participants formed the cross-validation group. RESULTS: Existing equations underestimated the VO2 peak values of the fittest personnel and overestimated the VO2 peak of the least aerobically fit by between 2% and 18%. The new equation derived from the validation group has less bias, the highest correlation with the measured values (r = 0.83), and classified the most people correctly according to the Royal Navy's Fitness Test standards, producing the fewest false positives and false negatives combined (9%). CONCLUSION: The new equation will provide a more accurate estimate of VO2 peak for a British military population aged 18 to 39.