5-Hydroxymethylfurfural reduces skeletal muscle superoxide production and modifies force production in rats exposed to hypobaric hypoxia.

Decreased blood-tissue oxygenation at high altitude (HA) increases mitochondrial oxidant production and reduces exercise capacity. 5-Hydroxymethylfurfural (5-HMF) is an antioxidant that increases hemoglobin's binding affinity for oxygen. For these reasons, we hypothesized that 5-HMF would improve muscle performance in rats exposed to a simulated HA of ~5500 m. A secondary objective was to measure mitochondrial activity and dynamic regulation of fission and fusion because they are linked processes impacted by HA. Fisher 344 rats received 5-HMF (40 mg/kg/day) or vehicle during exposure to sea level or HA for 72 h. Right ankle plantarflexor muscle function was measured pre- and post-exposure. Post-exposure measurements included arterial blood gas and complete blood count, flexor digitorum brevis myofiber superoxide production and mitochondrial membrane potential (ΔΨm), and mitochondrial dynamic regulation in the soleus muscle. HA reduced blood oxygenation, increased superoxide levels and lowered ΔΨm, responses that were accompanied by decreased peak isometric torque and force production at frequencies >75 Hz. 5-HMF increased isometric force production and lowered oxidant production at sea level. In HA exposed animals, 5-HMF prevented a decline in isometric force production at 75-125 Hz, prevented an increase in superoxide levels, further decreased ΔΨm, and increased mitochondrial fusion 2 protein expression. These results suggest that 5-HMF may prevent a decrease in hypoxic force production during submaximal isometric contractions by an antioxidant mechanism.

Effects of unilateral eye closure on middle ear muscle contractions.

Middle ear muscle contractions (MEMCs) are most commonly considered a response to high-level acoustic stimuli. However, MEMCs have also been observed in the absence of sound, either as a response to somatosensory stimulation or in concert with other motor activity. The relationship between MEMCs and non-acoustic sources is unclear. This study examined associations between measures of voluntary unilateral eye closure and impedance-based measures indicative of middle ear muscle activity while controlling for demographic and clinical factors in a large group of participants (N=190) with present clinical acoustic reflexes and no evidence of auditory dysfunction. Participants were instructed to voluntarily close the eye ipsilateral to the ear canal containing a detection probe at three levels of effort. Orbicularis oculi muscle activity was measured using surface electromyography. Middle ear muscle activity was inferred from changes in total energy reflected in the ear canal using a filtered (0.2 to 8 kHz) click train. Results revealed that middle ear muscle activity was positively associated with eye muscle activity. MEMC occurrence rates for eye closure observed in this study were generally higher than previously published rates for high-level brief acoustic stimuli in the same participant pool suggesting that motor activity may be a more reliable elicitor of MEMCs than acoustic stimuli. These results suggest motor activity can serve as a confounding factor for auditory exposure studies as well as complicate the interpretation of any impulsive noise damage risk criteria that assume MEMCs serve as a consistent, uniform protective factor. The mechanism linking eye and middle ear muscle activity is not understood and is an avenue for future research.

GAT inhibition preserves cerebral blood flow and reduces oxidant damage to mitochondria in rodents exposed to extreme hyperbaric oxygen.

Oxygen breathing at elevated partial pressures (PO2's) at or more than 3 atmospheres absolute (ATA) causes a reduction in brain γ-aminobutyric acid (GABA) levels that impacts the development of central nervous system oxygen toxicity (CNS-OT). Drugs that increase brain GABA content delay the onset of CNS-OT, but it is unknown if oxidant damage is lessened because brain tissue PO2 remains elevated during hyperbaric oxygen (HBO2) exposures. Experiments were performed in rats and mice to measure brain GABA levels with or without GABA transporter inhibitors (GATs) and its influence on cerebral blood flow, oxidant damage, and aspects of mitochondrial quality control signaling (mitophagy and biogenesis). In rats pretreated with tiagabine (GAT1 inhibitor), the tachycardia, secondary rise in mean arterial blood pressure, and cerebral hyperemia were prevented during HBO2 at 5 and 6 ATA. Tiagabine and the nonselective GAT inhibitor nipecotic acid similarly extended HBO2 seizure latencies. In mice pretreated with tiagabine and exposed to HBO2 at 5 ATA, nuclear and mitochondrial DNA oxidation and astrocytosis was attenuated in the cerebellum and hippocampus. Less oxidant injury in these regions was accompanied by reduced conjugated microtubule-associated protein 1A/1B-light chain 3 (LC3-II), an index of mitophagy, and phosphorylated cAMP response element binding protein (pCREB), an initiator of mitochondrial biogenesis. We conclude that GABA prevents cerebral hyperemia and delays neuroexcitation under extreme HBO2, limiting oxidant damage in the cerebellum and hippocampus, and likely lowering mitophagy flux and initiation of pCREB-initiated mitochondrial biogenesis.

The HO-1/CO System and Mitochondrial Quality Control in Skeletal Muscle.

Inducible heme oxygenase (HO)-1 catalyzes the breakdown of heme to biliverdin, iron, and carbon monoxide (CO). CO binds to cytochrome c oxidase and alters mitochondrial redox balance and coordinately regulates mitochondrial quality control (MQC) during oxidant stress and inflammation. The hypothesis presented is that the skeletal muscle HO-1/CO system helps modulate components in the MQC cycle during metabolic stress.

The Impact of Synchronized Cochlear Implant Sampling and Stimulation on Free-Field Spatial Hearing Outcomes: Comparing the ciPDA Research Processor to Clinical Processors.

OBJECTIVES: Bilateral cochlear implant (BiCI) listeners use independent processors in each ear. This independence and lack of shared hardware prevents control of the timing of sampling and stimulation across ears, which precludes the development of bilaterally-coordinated signal processing strategies. As a result, these devices potentially reduce access to binaural cues and introduce disruptive artifacts. For example, measurements from two clinical processors demonstrate that independently-running processors introduce interaural incoherence. These issues are typically avoided in the laboratory by using research processors with bilaterally-synchronized hardware. However, these research processors do not typically run in real-time and are difficult to take out into the real-world due to their benchtop nature. Hence, the question of whether just applying hardware synchronization to reduce bilateral stimulation artifacts (and thereby potentially improve functional spatial hearing performance) has been difficult to answer. The CI personal digital assistant (ciPDA) research processor, which uses one clock to drive two processors, presented an opportunity to examine whether synchronization of hardware can have an impact on spatial hearing performance. DESIGN: Free-field sound localization and spatial release from masking (SRM) were assessed in 10 BiCI listeners using both their clinical processors and the synchronized ciPDA processor. For sound localization, localization accuracy was compared within-subject for the two processor types. For SRM, speech reception thresholds were compared for spatially separated and co-located configurations, and the amount of unmasking was compared for synchronized and unsynchronized hardware. There were no deliberate changes of the sound processing strategy on the ciPDA to restore or improve binaural cues. RESULTS: There was no significant difference in localization accuracy between unsynchronized and synchronized hardware (p = 0.62). Speech reception thresholds were higher with the ciPDA. In addition, although five of eight participants demonstrated improved SRM with synchronized hardware, there was no significant difference in the amount of unmasking due to spatial separation between synchronized and unsynchronized hardware (p = 0.21). CONCLUSIONS: Using processors with synchronized hardware did not yield an improvement in sound localization or SRM for all individuals, suggesting that mere synchronization of hardware is not sufficient for improving spatial hearing outcomes. Further work is needed to improve sound coding strategies to facilitate access to spatial hearing cues. This study provides a benchmark for spatial hearing performance with real-time, bilaterally-synchronized research processors.

Iron deficiency and high-intensity running interval training do not impact femoral or tibial bone in young female rats.

In the USA, as many as 20 % of recruits sustain stress fractures during basic training. In addition, approximately one-third of female recruits develop Fe deficiency upon completion of training. Fe is a cofactor in bone collagen formation and vitamin D activation, thus we hypothesised Fe deficiency may be contributing to altered bone microarchitecture and mechanics during 12-weeks of increased mechanical loading. Three-week old female Sprague Dawley rats were assigned to one of four groups: Fe-adequate sedentary, Fe-deficient sedentary, Fe-adequate exercise and Fe-deficient exercise. Exercise consisted of high-intensity treadmill running (54 min 3×/week). After 12-weeks, serum bone turnover markers, femoral geometry and microarchitecture, mechanical properties and fracture toughness and tibiae mineral composition and morphometry were measured. Fe deficiency increased the bone resorption markers C-terminal telopeptide type I collagen and tartate-resistant acid phosphatase 5b (TRAcP 5b). In exercised rats, Fe deficiency further increased bone TRAcP 5b, while in Fe-adequate rats, exercise increased the bone formation marker procollagen type I N-terminal propeptide. In the femur, exercise increased cortical thickness and maximum load. In the tibia, Fe deficiency increased the rate of bone formation, mineral apposition and Zn content. These data show that the femur and tibia structure and mechanical properties are not negatively impacted by Fe deficiency despite a decrease in tibiae Fe content and increase in serum bone resorption markers during 12-weeks of high-intensity running in young growing female rats.

Effects of Caffeine on Exercise Duration, Critical Velocity, and Ratings of Perceived Exertion During Repeated-Sprint Exercise in Physically Active Men.

Caffeine improves short-to-moderate distance running performance, but the effect of caffeine on repeated sprints are equivocal. This research determined if caffeine improved exercise tolerance during repeated-sprint exercise. iCV is a running velocity that distinguishes intermittent running velocities (velocities ≤ iCV) that are sustainable from those resulting in a predictable time to exhaustion (velocities > iCV). Seven physically active men (age = 21.6 ± 1.5 years, body mass = 72.8 ± 5.1 kg, VO2max = 56.9 ± 9.8 mL/kg/min) ingested caffeine (5 mg/kg) or placebo (crossover design) 60 min prior to an intermittent critical velocity (iCV) test. The treadmill grade and velocity at VO2max (vVO2max) were used for iCV testing, and consisted of 3 bouts (10 sec running and 10 sec passive rest) at 130, 110 and 120% vVO2max. Each bout continued until volitional exhaustion and was separated by 20 min of passive rest. Total distance and duration were recorded to determine exercise tolerance using the iCV model. Caffeine ingestion increased running duration at 110% vVO2max (p = 0.02), but not at 120 (p = 0.93) and 130% vVO2max (p = 0.14). Caffeine did not improve iCV model parameters. A single dose of caffeine consumed 60 min before repeated-sprints can improve performance at 110% vVO2max, but not at higher velocities.

Skeletal muscle mitochondrial fragmentation and impaired bioenergetics from nutrient overload are prevented by carbon monoxide.

Nutrient excess increases skeletal muscle oxidant production and mitochondrial fragmentation that may result in impaired mitochondrial function, a hallmark of skeletal muscle insulin resistance. This led us to explore whether an endogenous gas molecule, carbon monoxide (CO), which is thought to prevent weight gain and metabolic dysfunction in mice consuming high-fat diets, alters mitochondrial morphology and respiration in C2C12 myoblasts exposed to high glucose (15.6 mM) and high fat (250 µM BSA-palmitate) (HGHF). Also, skeletal muscle mitochondrial morphology, distribution, respiration, and energy expenditure were examined in obese resistant (OR) and obese prone (OP) rats that consumed a high-fat and high-sucrose diet for 10 wk with or without intermittent low-dose inhaled CO and/or exercise training. In cells exposed to HGHF, superoxide production, mitochondrial membrane potential (ΔΨm), mitochondrial fission regulatory protein dynamin-related protein 1 (Drp1) and mitochondrial fragmentation increased, while mitochondrial respiratory capacity was reduced. CO decreased HGHF-induced superoxide production, Drp1 protein levels and mitochondrial fragmentation, maintained ΔΨm, and increased mitochondrial respiratory capacity. In comparison with lean OR rats, OP rats had smaller skeletal muscle mitochondria that contained disorganized cristae, a normal mitochondrial distribution, but reduced citrate synthase protein expression, normal respiratory responses, and a lower energy expenditure. The combination of inhaled CO and exercise produced the greatest effect on mitochondrial morphology, increasing ADP-stimulated respiration in the presence of pyruvate, and preventing a decline in resting energy expenditure. These data support a therapeutic role for CO and exercise in preserving mitochondrial morphology and respiration during metabolic overload.

Curcumin Ameliorates Heat-Induced Injury through NADPH Oxidase-Dependent Redox Signaling and Mitochondrial Preservation in C2C12 Myoblasts and Mouse Skeletal Muscle.

BACKGROUND: Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and the mitochondrial electron transport chain are the primary sources of reactive oxygen species (ROS). Previous studies have shown that severe heat exposure damages mitochondria and causes excessive mitochondrial ROS production that contributes to the pathogenesis of heat-related illnesses. OBJECTIVES: We tested whether the antioxidant curcumin could protect against heat-induced mitochondrial dysfunction and skeletal muscle injury, and characterized the possible mechanism. METHODS: Mouse C2C12 myoblasts and rat flexor digitorum brevis (FDB) myofibers were treated with 5 µM curcumin; adult male C57BL/6J mice received daily curcumin (15, 50, or 100 mg/kg body weight) by gavage for 10 consecutive days. We compared ROS levels and mitochondrial morphology and function between treatment and nontreatment groups under unheated or heat conditions, and investigated the upstream mechanism and the downstream effect of curcumin-regulated ROS production. RESULTS: In C2C12 myoblasts, curcumin prevented heat-induced mitochondrial fragmentation, ROS overproduction, and apoptosis (all P < 0.05). Curcumin treatment for 2 and 4 h at 37°C induced increases in ROS levels by 42% and 59% (dihydroethidium-derived fluorescence), accompanied by increases in NADPH oxidase protein expression by 24% and 32%, respectively (all P < 0.01). In curcumin-treated cells, chemical inhibition and genetic knockdown of NADPH oxidase restored ROS to levels similar to those of controls, indicating NADPH oxidase mediates curcumin-stimulated ROS production. Moreover, curcumin induced ROS-dependent shifting of the mitochondrial fission-fusion balance toward fusion, and increases in mitochondrial mass by 143% and membrane potential by 30% (both P < 0.01). In rat FDB myofibers and mouse gastrocnemius muscles, curcumin preserved mitochondrial morphology and function during heat stress, and prevented heat-induced mitochondrial ROS overproduction and tissue injury (all P < 0.05). CONCLUSIONS: Curcumin regulates ROS hormesis favoring mitochondrial fusion/elongation, biogenesis, and improved function in rodent skeletal muscle. Curcumin may be an effective therapeutic target for heat-related illness and other mitochondrial diseases.

Carbon Monoxide and Exercise Prevents Diet-Induced Obesity and Metabolic Dysregulation Without Affecting Bone.

OBJECTIVE: Carbon monoxide (CO) may counteract obesity and metabolic dysfunction in rodents consuming high-fat diets, but the skeletal effects are not understood. This study investigated whether low-dose inhaled CO (250 ppm) with or without moderate intensity aerobic exercise (3 h/wk) would limit diet-induced obesity and metabolic dysregulation and preserve bone health. METHODS: Obesity-resistant (OR) rats served as controls, and obesity-prone (OP) rats were randomized to sedentary, sedentary plus CO, exercise, or CO plus exercise. For 10 weeks, OP rats consumed a high-fat, high-sucrose diet, whereas OR rats consumed a low-fat control diet. Measurements included indicators of obesity and metabolism, bone turnover markers, femoral geometry and microarchitecture, bone mechanical properties, and tibial morphometry. RESULTS: A high-fat, high-sucrose diet led to obesity, hyperinsulinemia, and hyperleptinemia, without impacting bone. CO alone led only to a modest reduction in weight gain. Exercise attenuated weight gain and improved the metabolic profile; however, bone fragility increased. Combined CO and exercise led to body mass reduction and a metabolic state similar to control OR rats and prevented the exercise-induced increase in bone fragility. CONCLUSIONS: CO and aerobic exercise training prevent obesity and metabolic sequelae of nutrient excess while stabilizing bone physiology.

Generalizability of clinically measured acoustic reflexes to brief sounds.

Middle ear muscle contractions (MEMC) can be elicited in response to high-level sounds, and have been used clinically as acoustic reflexes (ARs) during evaluations of auditory system integrity. The results of clinical AR evaluations do not necessarily generalize to different signal types or durations. The purpose of this study was to evaluate the likelihood of observing MEMC in response to brief sound stimuli (tones, recorded gunshots, noise) in adult participants (N = 190) exhibiting clinical ARs and excellent hearing sensitivity. Results revealed that the presence of clinical ARs was not a sufficient indication that listeners will also exhibit MEMC for brief sounds. Detection rates varied across stimulus types between approximately 20% and 80%. Probabilities of observing MEMC also differed by clinical AR magnitude and latency, and declined over the period of minutes during the course of the MEMC measurement series. These results provide no support for the inclusion of MEMC as a protective factor in damage-risk criteria for impulsive noises, and the limited predictability of whether a given individual will exhibit MEMC in response to a brief sound indicates a need to measure and control for MEMC in studies evaluating pharmaceutical interventions for hearing loss.

Increased Antiseizure Effectiveness with Tiagabine Combined with Sodium Channel Antagonists in Mice Exposed to Hyperbaric Oxygen.

Hyperbaric oxygen (HBO2) is acutely toxic to the central nervous system, culminating in EEG spikes and tonic-clonic convulsions. GABA enhancers and sodium channel antagonists improve seizure latencies in HBO2 when administered individually, while combining antiepileptic drugs from different functional classes can provide greater seizure latency. We examined the combined effectiveness of GABA enhancers (tiagabine and gabapentin) with sodium channel antagonists (carbamazepine and lamotrigine) in delaying HBO2-induced seizures. A series of experiments in C57BL/6 mice exposed to 100% oxygen at 5 atmospheres absolute (ATA) were performed. We predicted equally effective doses from individual drug-dose response curves, and the combinations of tiagabine + carbamazepine or lamotrigine were tested to determine the maximally effective combined doses to be used in subsequent experiments designed to identify the type of pharmacodynamic interaction for three fixed-ratio combinations (1:3, 1:1, and 3:1) using isobolographic analysis. For both combinations, the maximally effective combined doses increased seizure latency over controls > 5-fold and were determined to interact synergistically for fixed ratios 1:1 and 3:1, additive for 1:3. These results led us to explore whether the benefits of these drug combinations could be extended to the lungs, since a centrally mediated mechanism is believed to mediate hyperoxic-induced cardiogenic lung injury. Indeed, both combinations attenuated bronchoalveolar lavage protein content by ~ 50%. Combining tiagabine with carbamazepine or lamotrigine not only affords greater antiseizure protection in HBO2 but also allows for lower doses to be used, minimizing side effects, and attenuating acute lung injury.

Effects of vitamin D supplementation on salivary immune responses during Marine Corps basic training.

Vitamin D's role in regulating immune responses may increase during periods of elevated psychological and physiological stress. Due to the high demands placed on US Marine Corps recruits undergoing 12 weeks of basic military training, we hypothesized that vitamin D status would be related to markers of innate mucosal immunity, and daily vitamin D supplementation would augment immune responses during training. Males (n = 75) and females (n = 74) entering recruit basic training during the summer and winter volunteered to participate in a randomized, double-blind, placebo-controlled study. Subjects received either 1000 IU vitamin D3  + 2000 mg calcium/d (n = 73) or placebo (n = 76) for 12 weeks. Saliva samples were collected pre-training, during (weeks 4 and 8), and post-training (week 12) in order to determine salivary SIgA and cathelicidin (indices of mucosal immunity) and α-amylase (indicator of stress). Initial (baseline) and post-training serum 25(OH)D levels were measured. Results were as follows: serum 25(OH)D levels were 37% higher in recruits entering training in summer compared with winter. A positive relationship was observed between baseline 25(OH)D levels and SIgA secretion rates (-SR). When stress levels were high during summer training, baseline 25(OH)D levels contributed to an increase in salivary secretory immunoglobulin A secretion rates (SIgA-SR) and cathelicidin-SR, the latter only in males. Vitamin D supplementation contributed to the changes in SIgA-SR and cathelicidin-SR, specifically SIgA-SR was higher in the treatment group. These data highlight the importance of vitamin D and mucosal immune responses during arduous basic military training when stress levels are increased.

Auditory motion tracking ability of adults with normal hearing and with bilateral cochlear implants.

Adults with bilateral cochlear implants (BiCIs) receive benefits in localizing stationary sounds when listening with two implants compared with one; however, sound localization ability is significantly poorer when compared to normal hearing (NH) listeners. Little is known about localizing sound sources in motion, which occurs in typical everyday listening situations. The authors considered the possibility that sound motion may improve sound localization in BiCI users by providing multiple places of information. Alternatively, the ability to compare multiple spatial locations may be compromised in BiCI users due to degradation of binaural cues, and thus result in poorer performance relative to NH adults. In this study, the authors assessed listeners' abilities to distinguish between sounds that appear to be moving vs stationary, and track the angular range and direction of moving sounds. Stimuli were bandpass-filtered (150-6000 Hz) noise bursts of different durations, panned over an array of loudspeakers. Overall, the results showed that BiCI users were poorer than NH adults in (i) distinguishing between a moving vs stationary sound, (ii) correctly identifying the direction of movement, and (iii) tracking the range of movement. These findings suggest that conventional cochlear implant processors are not able to fully provide the cues necessary for perceiving auditory motion correctly.

Calcium and vitamin D supplementation and bone health in Marine recruits: Effect of season.

Stress fractures are common overuse injuries caused by repetitive bone loading. These fractures are of particular concern for military recruits and athletes resulting in attrition in up to 60% of recruits that sustain a fracture. Army and Navy recruits supplemented with daily calcium and vitamin D (Ca + D) demonstrated improved bone strength and reduced stress fractures. The aim of the current study was to evaluate whether Ca + D supplementation improves measures of bone health in recruits undergoing United States Marine Corps initial military training (IMT), and whether the effect of supplementation on indices of bone health varied by season. One-hundred ninety-seven Marine recruits (n = 107 males, n = 90 females, mean age = 18.9 ±â€¯1.6 y) were randomized to receive either Ca + D fortified snack bars (2000 mg Ca and 1000 IU vitamin D per day) or placebo divided into twice daily doses during 12 weeks of IMT. Anthropometrics, fasted blood samples, and peripheral quantitative computed tomography (pQCT) scans of the tibial metaphysis and diaphysis were collected upon entrance to- and post-training (12 weeks later). Half of the volunteers entered training in July and the other half started in February. Time-by-group interactions were observed for vitamin D status (25OHD) and the bone turnover markers, BAP, TRAP and OCN. 25OHD increased and BAP, TRAP and OCN all decreased in the Ca + D group (p < .05). Training increased distal tibia volumetric BMD (+1.9 ±â€¯2.8%), BMC (+2.0 ±â€¯3.1%), and bone strength index (BSI; +4.0 ±â€¯4.0%) and diaphyseal BMC (+1.0 ±â€¯2.2%) and polar stress strain index (SSIp; +0.7 ±â€¯2.1%) independent of Ca + D supplementation (p < .05 for all). When analyzed by season, change in BSI was greater in the Ca + D group as compared to placebo in the summer iteration only (T*G; p < .05). No other effects of supplementation on bone tissue were observed. When categorized by tertile of percent change in BSI, recruits demonstrating the greatest changes in BSI and 25OHD entered training with the lowest levels of 25OHD (p < .05). Over all, these results suggest that Ca + D supplementation reduced some markers of bone formation and resorption and the decline in 25OHD over training in volunteers that started training in the summer was prevented by supplementation. Baseline 25OHD and trajectory may impact bone responses to IMT, but little effect of Ca + D supplementation was observed at the investigated doses.

Astaxanthin but not quercetin preserves mitochondrial integrity and function, ameliorates oxidative stress, and reduces heat-induced skeletal muscle injury.

Heat stress causes mitochondrial dysfunction and increases mitochondrial production of reactive oxygen species (ROS), both of which contribute to heat-induced skeletal muscle injury. In this study, we tested whether either astaxanthin or quercetin, two dietary antioxidants, could ameliorate heat-induced skeletal muscle oxidative injury. In mouse C2C12 myoblasts exposed to 43°C heat stress, astaxanthin inhibited heat-induced ROS production in a concentration-dependent manner (1-20 µM), whereas the ROS levels remained high in cells treated with quercetin over a range of concentrations (2-100 µM). Because mitochondria are both the main source and a primary target of heat-induced ROS, we then tested the effects of astaxanthin and quercetin on mitochondrial integrity and function, under both normal temperature (37°C) and heat stress conditions. Quercetin treatment at 37°C induced mitochondrial fragmentation and decreased membrane potential (ΔΨ m ), accompanied by reduced protein expression of the master regulator of mitochondrial biogenesis peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α). It also induced cleavage of mitochondrial inner-membrane fusion protein OPA1. In contrast, astaxanthin at 37°C increased protein expression of PGC-1α and mitochondrial transcription factor A (TFAM), and maintained tubular structure and normal ΔΨm . Under 43°C heat stress conditions, whereas quercetin failed to rescue C2C12 cells from injury, astaxanthin treatment prevented heat-induced mitochondrial fragmentation and depolarization, and apoptotic cell death. We also isolated rat flexor digitorum brevis myofibers and confirmed the data from C2C12 myoblasts that astaxanthin but not quercetin preserves mitochondrial integrity and function and ameliorates heat-induced skeletal muscle injury. These results confirm that mitochondria may be a potential therapeutic target for heat-related illness and suggest that astaxanthin may potentially be an effective preventive strategy.

Human middle-ear muscles rarely contract in anticipation of acoustic impulses: Implications for hearing risk assessments.

The current study addressed the existence of an anticipatory middle-ear muscle contraction (MEMC) as a protective mechanism found in recent damage-risk criteria for impulse noise exposure. Specifically, the experiments reported here tested instances when an exposed individual was aware of and could anticipate the arrival of an acoustic impulse. In order to detect MEMCs in human subjects, a laser-Doppler vibrometer (LDV) was used to measure tympanic membrane (TM) motion in response to a probe tone. Here we directly measured the time course and relative magnitude changes of TM velocity in response to an acoustic reflex-eliciting (i.e. MEMC eliciting) impulse in 59 subjects with clinically assessable MEMCs. After verifying the presence of the MEMC, we used a classical conditioning paradigm pairing reflex-eliciting acoustic impulses (unconditioned stimulus, UCS) with various preceding stimuli (conditioned stimulus, CS). Changes in the time-course of the MEMC following conditioning were considered evidence of MEMC conditioning, and any indication of an MEMC prior to the onset of the acoustic elicitor was considered an anticipatory response. Nine subjects did not produce a MEMC measurable via LDV. For those subjects with an observable MEMC (n = 50), 48 subjects (96%) did not show evidence of an anticipatory response after conditioning, whereas only 2 subjects (4%) did. These findings reveal that MEMCs are not readily conditioned in most individuals, suggesting that anticipatory MEMCs are not prevalent within the general population. The prevalence of anticipatory MEMCs does not appear to be sufficient to justify inclusion as a protective mechanism in auditory injury risk assessments.

Spatial variation in signal and sensory precision both constrain auditory acuity at high frequencies.

Sensory performance is constrained by the information in the stimulus and the precision of the involved sensory system(s). Auditory spatial acuity is robust across a broad range of sound frequencies and source locations, but declines at eccentric lateral angles. The basis of such variation is not fully understood. Low-frequency auditory spatial acuity is mediated by sensitivity to interaural time difference (ITD) cues. While low-frequency spatial acuity varies across azimuth and some physiological models predict strong medial bias in the precision of ITD sensitivity, human psychophysical ITD sensitivity appears to vary only slightly with reference ITD magnitude. Correspondingly, recent analyses suggest that spatial variation in human low-frequency acuity is well-accounted for by acoustic factors alone. Here we examine the matter of high-frequency auditory acuity, which is mediated by sensitivity to interaural level difference (ILD) cues. Using two different psychophysical tasks in human subjects, we demonstrate decreasing ILD acuity with increasing ILD magnitude. We then demonstrate that the multiplicative combination of spatially variant sensory precision and physical cue information (local slope of the ILD cue) provides improved prediction of classic high-frequency spatial acuity data. Finally, we consider correlates of magnitude dependent acuity in neurons that are sensitive to ILDs.

Acclimation of C2C12 myoblasts to physiological glucose concentrations for in vitro diabetes research.

AIMS: The interplay between hyper-glycemia and -lipidemia in diabetes mellitus (DM) is important in simulating diabetic conditions. However, cell culture media typically contain supraphysiological levels of glucose to stimulate cellular growth, which also desensitizes cells to elevated glucose levels. Moreover, creating hyperlipidemic conditions in vitro requires specialized carriers because unbound lipids form micelles when introduced to liquid media. This study sought to develop a novel method for simulating DM conditions in vitro. MATERIALS AND METHODS: We acclimated the C2C12 mouse myoblasts to culture medium with 5.6 mM glucose, which mimics physiological levels, and created a bovine serum albumin-palmitic acid conjugate for lipid transport to explore the effects of hyperlipidemia. We simulated diabetic conditions in vitro by using both hyper-glycemic and -lipidemic conditions and compared the results to that of only hyperglycemic or hyperlipidemic conditions. KEY FINDINGS: Acclimated cells exposed to these hyper-glycemic (15 mM glucose) and/or -lipidemic (0.25 mM palmitate) conditions for 2 h showed increased mitochondrial fragmentation and membrane potential as well as elevated reactive oxygen species production compared to control cells. These findings suggest altered mitochondrial morphology and function, which have been confirmed using isolated rat flexor digitorum brevis myofibers. Hyper-glycemic and/or -lipidemic stimulations for 24 h significantly increased mitogen-activated protein kinase kinase MEK 1/2 protein expression, upregulated the early pro-apoptotic transcription factor C/EBP homologous protein (CHOP), and induced apoptosis. SIGNIFICANCE: Our results further support and confirm the utility of this method which will allow for subsequent investigations studying the effects of hyper-glycemia and/or -lipidemia in vitro.

Adrenoceptor blockade modifies regional cerebral blood flow responses to hyperbaric hyperoxia: protection against CNS oxygen toxicity.

Exposure to extreme hyperbaric oxygen (HBO2) >5-6 atmospheres absolute (ATA) produces baroreflex impairment, sympathetic hyperactivation, hypertension, tachycardia, and cerebral hyperemia, known as phase II, culminating in seizures. We hypothesized that attenuation of the effects of high sympathetic outflow would preserve regional cerebral blood flow (rCBF) and protect against HBO2-induced seizures. To explore this possibility, we tested four adrenoceptor antagonists in conscious and anesthetized rats exposed to HBO2 at 5 and 6 ATA, respectively: phentolamine (nonselective α1 and α2), prazosin (selective α1), propranolol (nonselective ß1 and ß2), and atenolol (selective ß1). In conscious rats, four drug doses were administered to rats before HBO2 exposures, and seizure latencies were recorded. Drug doses that provided similar protection against seizures were administered before HBO2 exposures in anesthetized rats to determine the effects of adrenoceptor blockade on mean arterial pressure, heart rate, rCBF, and EEG spikes. All four drugs modified cardiovascular and rCBF responses in HBO2 that aligned with epileptiform discharges, but only phentolamine and propranolol effectively increased EEG spike latencies by ~20 and 36 min, respectively. When phentolamine and propranolol were delivered during HBO2 at the onset of phase II, only propranolol led to sustained reductions in heart rate and rCBF, preventing the appearance of epileptiform discharges. The enhanced effectiveness of propranolol may extend beyond ß-adrenoceptor blockade, i.e., membrane stability and reduced metabolic activity. These results indicate that adrenoceptor drug pretreatment will minimize the effects of excessive sympathetic outflow on rCBF and extend HBO2 exposure time.NEW & NOTEWORTHY Blocking adrenergic receptors with phentolamine (nonselective α1 and α2), prazosin (selective α1), propranolol (nonselective ß1 and ß2), and atenolol (selective ß1) modified cardiovascular and regional cerebral blood flow (rCBF) responses in hyperbaric oxygen (HBO2) at 6 atmospheres absolute (ATA); however, only phentolamine and propranolol extended EEG spike latencies. When these two agents were delivered at the onset of sympathetic hyperactivation, only propranolol reduced heart rate and rCBF throughout the exposure and prevented epileptiform discharges. These data validate the strong role of adrenergic control of cardiovascular function and rCBF in extreme HBO2 and the potential use of antiadrenergic drugs to prevent seizures.