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.

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.

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.

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.

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.

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.

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.

Secondary PDZ domain-binding site on class B plexins enhances the affinity for PDZ-RhoGEF.

PDZ domains are abundant protein interaction modules and typically recognize a short motif at the C terminus of their ligands, with a few residues in the motif endowing the binding specificity. The sequence-based rules, however, cannot fully account for the specificity between the vast number of PDZ domains and ligands in the cell. Plexins are transmembrane receptors that regulate processes such as axon guidance and angiogenesis. Two related guanine nucleotide exchange factors (GEFs), PDZ-RhoGEF and leukemia-associated RhoGEF (LARG), use their PDZ domains to bind class B plexins and play critical roles in signaling. Here, we present the crystal structure of the full-length cytoplasmic region of PlexinB2 in complex with the PDZ domain of PDZ-RhoGEF. The structure reveals that, in addition to the canonical C-terminal motif/PDZ interaction, the 3D domain of PlexinB2 forms a secondary interface with the PDZ domain. Our biophysical and cell-based assays show that the secondary interface contributes to the specific interaction between plexin and PDZ-RhoGEF and to signaling by plexin in the cell. Formation of secondary interfaces may be a general mechanism for increasing affinity and specificity of modular domain-mediated interactions.

Mixed stimulation rates to improve sensitivity of interaural timing differences in bilateral cochlear implant listeners.

Normal hearing listeners extract small interaural time differences (ITDs) and interaural level differences (ILDs) to locate sounds and segregate targets from noise. Bilateral cochlear implant listeners show poor sensitivity to ITDs when using clinical processors. This is because common clinical stimulation approaches use high rates [∼1000 pulses per-second (pps)] for each electrode in order to provide good speech representation, but sensitivity to ITDs is best at low rates of stimulation (∼100-300 pps). Mixing rates of stimulation across the array is a potential solution. Here, ITD sensitivity for a number of mixed-rate configurations that were designed to preserve speech envelope cues using high-rate stimulation and spatial hearing using low rate stimulation was examined. Results showed that ITD sensitivity in mixed-rate configurations when only one low rate electrode was included generally yielded ITD thresholds comparable to a configuration with low rates only. Low rate stimulation at basal or middle regions on the electrode array yielded the best sensitivity to ITDs. This work provides critical evidence that supports the use of mixed-rate strategies for improving ITD sensitivity in bilateral cochlear implant users.

Effects of oral sodium nitrate on forearm blood flow, oxygenation and exercise performance during acute exposure to hypobaric hypoxia (4300 m).

A reduction in oxygen transport contributes to impaired exercise capacity at high altitude. Since blood flow is mediated, in part, by nitric oxide (NO), we hypothesized that sodium nitrate provided before forearm grip exercise performed at a simulated altitude of 4300 m (hypobaric hypoxia (HH)) would increase forearm blood flow and oxygenation, and decrease the decrement in grip performance. In a double-blind, randomized crossover study, 10 healthy subjects (9 males and 1 female) performed continuous (CGrip) and repeated rhythmic (RGrip) isometric forearm exercise until task failure in normobaric normoxia (NN), 2.5 h following consumption of placebo and sodium nitrate (15 mmol) in HH, and then again post-HH at sea-level pressure. Measurements included forearm blood flow (FBF) and anterior forearm tissue oxygenation (StO2), mean arterial blood pressure (MAP), arterial blood O2 saturation (SpO2), plasma NO reaction products (NOx) and nitrite, and exhaled NO (PENO). Compared to baseline testing in NN, performing CGrip and RGrip exercise in HH resulted in significant reductions in forearm blood flow, SaO2 and StO2, responses that were accompanied by significant performance decrements (∼10%) in both CGrip and RGrip exercise. In spite of a 10-fold increase in plasma NOx levels and a significant decrease in MAP during CGrip exercise following nitrate consumption, there were no significant main effects of treatment (placebo vs. sodium nitrate) for forearm blood flow, SpO2, StO2, or grip performance. PENO remained unchanged between NN, HH and post-HH conditions with placebo, but increased (∼24%) following nitrate supplementation in HH and post-HH. These data do not support a benefit in consuming a single dose of supplemental nitrate on forearm blood flow and isometric exercise in healthy adults at a simulated altitude of 4300 m.

The Effect of Microphone Placement on Interaural Level Differences and Sound Localization Across the Horizontal Plane in Bilateral Cochlear Implant Users.

OBJECTIVE: This study examined the effect of microphone placement on the interaural level differences (ILDs) available to bilateral cochlear implant (BiCI) users, and the subsequent effects on horizontal-plane sound localization. DESIGN: Virtual acoustic stimuli for sound localization testing were created individually for eight BiCI users by making acoustic transfer function measurements for microphones placed in the ear (ITE), behind the ear (BTE), and on the shoulders (SHD). The ILDs across source locations were calculated for each placement to analyze their effect on sound localization performance. Sound localization was tested using a repeated-measures, within-participant design for the three microphone placements. RESULTS: The ITE microphone placement provided significantly larger ILDs compared to BTE and SHD placements, which correlated with overall localization errors. However, differences in localization errors across the microphone conditions were small. CONCLUSIONS: The BTE microphones worn by many BiCI users in everyday life do not capture the full range of acoustic ILDs available, and also reduce the change in cue magnitudes for sound sources across the horizontal plane. Acute testing with an ITE placement reduced sound localization errors along the horizontal plane compared to the other placements in some patients. Larger improvements may be observed if patients had more experience with the new ILD cues provided by an ITE placement.

Lateralization of interaural timing differences with multi-electrode stimulation in bilateral cochlear-implant users.

Bilateral cochlear implant (BiCI) users have shown variability in interaural time difference (ITD) sensitivity at different places along the cochlea. This paper investigates perception of multi-electrode binaural stimulation to determine if auditory object formation (AOF) and lateralization are affected by variability in ITD sensitivity when a complex sound is encoded with multi-channel processing. AOF and ITD lateralization were compared between single- and multi-electrode configurations. Most (7/8) BiCI users perceived a single auditory object with multi-electrode stimulation, and the range of lateralization was comparable to single-electrode stimulation, suggesting that variability in single-electrode ITD sensitivity does not compromise AOF with multi-electrode stimulation.

Sound frequency-invariant neural coding of a frequency-dependent cue to sound source location.

The century-old duplex theory of sound localization posits that low- and high-frequency sounds are localized with two different acoustical cues, interaural time and level differences (ITDs and ILDs), respectively. While behavioral studies in humans and behavioral and neurophysiological studies in a variety of animal models have largely supported the duplex theory, behavioral sensitivity to ILD is curiously invariant across the audible spectrum. Here we demonstrate that auditory midbrain neurons in the chinchilla (Chinchilla lanigera) also encode ILDs in a frequency-invariant manner, efficiently representing the full range of acoustical ILDs experienced as a joint function of sound source frequency, azimuth, and distance. We further show, using Fisher information, that nominal "low-frequency" and "high-frequency" ILD-sensitive neural populations can discriminate ILD with similar acuity, yielding neural ILD discrimination thresholds for near-midline sources comparable to behavioral discrimination thresholds estimated for chinchillas. These findings thus suggest a revision to the duplex theory and reinforce ecological and efficiency principles that hold that neural systems have evolved to encode the spectrum of biologically relevant sensory signals to which they are naturally exposed.

Evidence for a neural source of the precedence effect in sound localization.

Normal-hearing human listeners and a variety of studied animal species localize sound sources accurately in reverberant environments by responding to the directional cues carried by the first-arriving sound rather than spurious cues carried by later-arriving reflections, which are not perceived discretely. This phenomenon is known as the precedence effect (PE) in sound localization. Despite decades of study, the biological basis of the PE remains unclear. Though the PE was once widely attributed to central processes such as synaptic inhibition in the auditory midbrain, a more recent hypothesis holds that the PE may arise essentially as a by-product of normal cochlear function. Here we evaluated the PE in a unique human patient population with demonstrated sensitivity to binaural information but without functional cochleae. Users of bilateral cochlear implants (CIs) were tested in a psychophysical task that assessed the number and location(s) of auditory images perceived for simulated source-echo (lead-lag) stimuli. A parallel experiment was conducted in a group of normal-hearing (NH) listeners. Key findings were as follows: 1) Subjects in both groups exhibited lead-lag fusion. 2) Fusion was marginally weaker in CI users than in NH listeners but could be augmented by systematically attenuating the amplitude of the lag stimulus to coarsely simulate adaptation observed in acoustically stimulated auditory nerve fibers. 3) Dominance of the lead in localization varied substantially among both NH and CI subjects but was evident in both groups. Taken together, data suggest that aspects of the PE can be elicited in CI users, who lack functional cochleae, thus suggesting that neural mechanisms are sufficient to produce the PE.

Effect of multi-electrode configuration on sensitivity to interaural timing differences in bilateral cochlear-implant users.

Recent psychophysical studies in bilateral cochlear implant users have shown that interaural timing difference (ITD) sensitivity with electrical stimulation varies depending on the place of stimulation along the cochlear array. While these studies have measured ITD sensitivity at single electrode places separately, it is important to understand how ITD sensitivity is affected when multiple electrodes are stimulated together because multi-electrode stimulation is required for representation of complex sounds. Multi-electrode stimulation may lead to poorer overall performance due to interference from places with poor ITD sensitivity, or from channel interaction due to electrical current spread. Alternatively, multi-electrode stimulation might result in overall good sensitivity if listeners can extract the most reliable ITD cues available. ITD just noticeable differences (JNDs) were measured for different multi-electrode configurations. Results showed that multi-electrode ITD JNDs were poorer than ITD JNDs for the best single-electrode pair. However, presenting ITD information along the whole array appeared to produce better sensitivity compared with restricting stimulation to the ends of the array, where ITD JNDs were comparable to the poorest single-electrode pair. These findings suggest that presenting ITDs in one cochlear region only may not be optimal for maximizing ITD sensitivity in multi-electrode stimulation.

Research report: Charcoal type used for hookah smoking influences CO production.

A hookah smoker who was treated for severe carbon monoxide poisoning with hyperbaric oxygen reported using a different type of charcoal prior to hospital admission, i.e., quick-light charcoal. This finding led to a study aimed at determining whether CO production differs between charcoals commonly used for hookah smoking, natural and quick-light. Our hypothesis was that quick-light charcoal produces significantly more CO than natural charcoal. A medium-sized hookah, activated charcoal filter, calibrated syringe, CO gas analyzer and infrared thermometer were assembled in series. A single 9-10 g briquette of either natural or quick-light charcoal was placed atop the hookah bowl and ignited. CO output (ppm) and temperature (degrees C) were measured in three-minute intervals over 90 minutes. The mean CO levels produced by quick-light charcoal over 90 minutes was significantly higher (3728 ± 2028) compared to natural charcoal (1730 ± 501 ppm, p = 0.016). However, the temperature was significantly greater when burning natural charcoal (292 ± 87) compared to quick-light charcoal (247 ± 92 degrees C, p = 0.013). The high levels of CO produced when using quick-light charcoals may be contributing to the increase in reported hospital admissions for severe CO poisoning.

The efficacy of vitamin D supplementation during a prolonged submarine patrol.

Submariners spend prolonged periods submerged without sunlight exposure and may benefit from vitamin D supplementation to maintain vitamin D status. The primary objective of this study was to determine the efficacy of daily vitamin D supplementation on maintenance of 25-hydroxyvitamin D (25(OH)D) during a 3-month submarine patrol. Submariners were randomly divided into three groups: placebo (n = 16), 1,000 IU/day (n = 20), or 2,000 IU/day (n = 17). Anthropometrics, self-reported dietary calcium and vitamin D intake, serum markers of vitamin D and bone metabolism, and peripheral quantitative computed tomography (pQCT) parameters of the tibia were determined before and after the patrol. Prior to departure, 49 % of the subjects were vitamin D insufficient (<50 nmol/L). Following the patrol, 25(OH)D increased in all groups (p < 0.001): 3.3 ± 13.1 (placebo), 4.6 ± 11.3 (1,000 IU/day), and 13 ± 14 nmol/L (2,000 IU/day). The changes in 25(OH)D levels were dependent upon the baseline concentration of 25(OH)D and body mass (p < 0.001). Osteocalcin increased by 38 % (p < 0.01), and pQCT analyses revealed small, yet significant increases in indices of tibial structure and strength (p < 0.05) that were independent of supplementation. These data suggest that vitamin D status was low prior to the patrol, and the subsequent changes in vitamin D status were dependent on the baseline 25(OH)D levels and body mass. Furthermore, short-term skeletal health does not appear to be negatively affected by 3 months of submergence in spite of a suboptimal response to vitamin D supplementation.

Biophysics of directional hearing in the American alligator (Alligator mississippiensis).

Physiological and anatomical studies have suggested that alligators have unique adaptations for spatial hearing. Sound localization cues are primarily generated by the filtering of sound waves by the head. Different vertebrate lineages have evolved external and/or internal anatomical adaptations to enhance these cues, such as pinnae and interaural canals. It has been hypothesized that in alligators, directionality may be enhanced via the acoustic coupling of middle ear cavities, resulting in a pressure difference receiver (PDR) mechanism. The experiments reported here support a role for a PDR mechanism in alligator sound localization by demonstrating that (1) acoustic space cues generated by the external morphology of the animal are not sufficient to generate location cues that match physiological sensitivity, (2) continuous pathways between the middle ears are present to provide an anatomical basis for coupling, (3) the auditory brainstem response shows some directionality, and (4) eardrum movement is directionally sensitive. Together, these data support the role of a PDR mechanism in crocodilians and further suggest this mechanism is a shared archosaur trait, most likely found also in the extinct dinosaurs.