Therapeutic ketosis with ketone ester delays central nervous system oxygen toxicity seizures in rats.

Central nervous system oxygen toxicity (CNS-OT) seizures occur with little or no warning, and no effective mitigation strategy has been identified. Ketogenic diets (KD) elevate blood ketones and have successfully treated drug-resistant epilepsy. We hypothesized that a ketone ester given orally as R,S-1,3-butanediol acetoacetate diester (BD-AcAc(2)) would delay CNS-OT seizures in rats breathing hyperbaric oxygen (HBO(2)). Adult male rats (n = 60) were implanted with radiotelemetry units to measure electroencephalogram (EEG). One week postsurgery, rats were administered a single oral dose of BD-AcAc(2), 1,3-butanediol (BD), or water 30 min before being placed into a hyperbaric chamber and pressurized to 5 atmospheres absolute (ATA) O2. Latency to seizure (LS) was measured from the time maximum pressure was reached until the onset of increased EEG activity and tonic-clonic contractions. Blood was drawn at room pressure from an arterial catheter in an additional 18 animals that were administered the same compounds, and levels of glucose, pH, Po(2), Pco(2), ß-hydroxybutyrate (BHB), acetoacetate (AcAc), and acetone were analyzed. BD-AcAc(2) caused a rapid (30 min) and sustained (>4 h) elevation of BHB (>3 mM) and AcAc (>3 mM), which exceeded values reported with a KD or starvation. BD-AcAc(2) increased LS by 574 ± 116% compared with control (water) and was due to the effect of AcAc and acetone but not BHB. BD produced ketosis in rats by elevating BHB (>5 mM), but AcAc and acetone remained low or undetectable. BD did not increase LS. In conclusion, acute oral administration of BD-AcAc(2) produced sustained ketosis and significantly delayed CNS-OT seizures by elevating AcAc and acetone.

Human Physiology in an Aquatic Environment.

Water covers over 70% of the earth, has varying depths and temperatures and contains much of the earth's resources. Head-out water immersion (HOWI) or submersion at various depths (diving) in water of thermoneutral (TN) temperature elicits profound cardiorespiratory, endocrine, and renal responses. The translocation of blood into the thorax and elevation of plasma volume by autotransfusion of fluid from cells to the vascular compartment lead to increased cardiac stroke volume and output and there is a hyperperfusion of some tissues. Pulmonary artery and capillary hydrostatic pressures increase causing a decline in vital capacity with the potential for pulmonary edema. Atrial stretch and increased arterial pressure cause reflex autonomic responses which result in endocrine changes that return plasma volume and arterial pressure to preimmersion levels. Plasma volume is regulated via a reflex diuresis and natriuresis. Hydrostatic pressure also leads to elastic loading of the chest, increasing work of breathing, energy cost, and thus blood flow to respiratory muscles. Decreases in water temperature in HOWI do not affect the cardiac output compared to TN; however, they influence heart rate and the distribution of muscle and fat blood flow. The reduced muscle blood flow results in a reduced maximal oxygen consumption. The properties of water determine the mechanical load and the physiological responses during exercise in water (e.g. swimming and water based activities). Increased hydrostatic pressure caused by submersion does not affect stroke volume; however, progressive bradycardia decreases cardiac output. During submersion, compressed gas must be breathed which introduces the potential for oxygen toxicity, narcosis due to nitrogen, and tissue and vascular gas bubbles during decompression and after may cause pain in joints and the nervous system.

The effects of respiratory muscle training on respiratory mechanics and energy cost.

Resistance respiratory muscle training (RRMT) increases respiratory muscle strength and can increase swimming endurance time by as much as 85%. The purpose of this study was to examine potential mechanisms by which RRMT improves exercise endurance. Eight healthy adult male scuba divers underwent experiments in a hyperbaric chamber at sea level (1 atmosphere absolute (ATA)), 2.7 ATA and 4.6 ATA, both dry and fully submersed. Subjects rested, exercised, and rested while mimicking their own exercise breathing (ISEV). Airway resistance (R(aw)), exhaled nitric oxide output (V˙(NO)), and respiratory duty cycle (T(I)/T(Tot)) were determined before and after four weeks of RRMT. RRMT decreased T(I)/T(Tot) (-10% at rest at 1 ATA), V˙(O2) (-17% at 2.7 ATA during submersed exercise), V˙(E) (-6% at 2.7 ATA during submersed exercise), and R(aw) (-34% inspiratory at 4.6 ATA submersed, -38% expiratory at 2.7 ATA dry), independent of changes in V˙(NO). Most importantly, respiratory muscle efficiency increased (+83% at 2.7 ATA submersed).

Female rats are more susceptible to central nervous system oxygen toxicity than male rats.

Abstract Tonic-clonic seizures typify central nervous system oxygen toxicity (CNS-OT) in humans and animals exposed to high levels of oxygen, as are encountered during scuba diving. We previously demonstrated that high doses of pseudoephedrine (PSE) decrease the latency to seizure (LS) for CNS-OT in young male rats. This study investigated whether female rats respond similarly to PSE and hyperbaric oxygen (HBO). We implanted 60 virgin stock (VS) and 54 former breeder (FB) female rats with radio-telemetry devices that measured brain electrical activity. One week later, rats were gavaged with saline or PSE in saline (40, 80, 120, 160, or 320 mg/kg) before diving to five atmospheres absolute in 100% oxygen. The time between reaching maximum pressure and exhibiting seizure was LS. Vaginal smears identified estrus cycle phase. PSE did not decrease LS for VS or FB, primarily because they exhibited low LS for all conditions tested. VS had shorter LS than males at 0, 40, and 80 mg/kg (-42, -49, and -57%, respectively). FB also had shorter LS than males at 0, 40, and 80 mg/kg (-60, -86, and -73%, respectively). FB were older than VS (286 ± 10 days vs. 128 ± 5 days) and weighed more than VS (299 ± 2.7 g vs. 272 ± 2.1 g). Males tested were younger (88 ± 2 days), heavier (340 ± 4.5 g), and gained more weight postoperatively (7.2 ± 1.6 g) than either VS (-0.4 ± 1.5 g) or FB (-1.6 ± 1.5 g); however, LS correlated poorly with age, body mass, change in body mass, and estrus cycle phase. We hypothesize that differences in sex hormones underlie females' higher susceptibility to CNS-OT than males.

Metabolic therapy with Deanna Protocol supplementation delays disease progression and extends survival in amyotrophic lateral sclerosis (ALS) mouse model.

Amyotrophic Lateral Sclerosis (ALS), also known as Lou Gehrig's disease, is a neurodegenerative disorder of motor neurons causing progressive muscle weakness, paralysis, and eventual death from respiratory failure. There is currently no cure or effective treatment for ALS. Besides motor neuron degeneration, ALS is associated with impaired energy metabolism, which is pathophysiologically linked to mitochondrial dysfunction and glutamate excitotoxicity. The Deanna Protocol (DP) is a metabolic therapy that has been reported to alleviate symptoms in patients with ALS. In this study we hypothesized that alternative fuels in the form of TCA cycle intermediates, specifically arginine-alpha-ketoglutarate (AAKG), the main ingredient of the DP, and the ketogenic diet (KD), would increase motor function and survival in a mouse model of ALS (SOD1-G93A). ALS mice were fed standard rodent diet (SD), KD, or either diets containing a metabolic therapy of the primary ingredients of the DP consisting of AAKG, gamma-aminobutyric acid, Coenzyme Q10, and medium chain triglyceride high in caprylic triglyceride. Assessment of ALS-like pathology was performed using a pre-defined criteria for neurological score, accelerated rotarod test, paw grip endurance test, and grip strength test. Blood glucose, blood beta-hydroxybutyrate, and body weight were also monitored. SD+DP-fed mice exhibited improved neurological score from age 116 to 136 days compared to control mice. KD-fed mice exhibited better motor performance on all motor function tests at 15 and 16 weeks of age compared to controls. SD+DP and KD+DP therapies significantly extended survival time of SOD1-G93A mice by 7.5% (p = 0.001) and 4.2% (p = 0.006), respectively. Sixty-three percent of mice in the KD+DP and 72.7% of the SD+DP group lived past 125 days, while only 9% of the control animals survived past that point. Targeting energy metabolism with metabolic therapy produces a therapeutic effect in ALS mice which may prolong survival and quality of life in ALS patients.

Relative effects of submersion and increased pressure on respiratory mechanics, work, and energy cost of breathing.

Submersion and increased pressure (depth) characterize the diving environment and may independently increase demand on the respiratory system. To quantify changes in respiratory mechanics, this study employed a unique protocol and techniques to measure, in a hyperbaric chamber, inspiratory and expiratory alveolar pressures (interrupter technique), inspiratory and expiratory resistance in the airways (RawI and RawE, esophageal balloon technique), nitric oxide elimination (thought to correlate with Raw), inspiratory and expiratory mechanical power of breathing, and the total energy cost of ventilation. Eight healthy adult men underwent experiments at 1, 2.7, and 4.6 atmospheres absolute (ATA) in dry and fully submersed conditions. Subjects rested, cycled on an ergometer at 100 W, and rested while voluntarily matching their ventilation to their own exercise hyperpnea (isocapnic simulated exercise ventilation). During isocapnic simulated exercise ventilation, increased O2 uptake (above rest values) resulted from increased expired ventilation. RawI decreased with submersion (mean 43% during rest and 20% during exercise) but increased from 1 to 4.6 ATA (19% during rest and 75% during exercise), as did RawE (53% decrease with submersion during rest and 10% during exercise; 9% increase from 1 to 4.6 ATA during rest and 66% during exercise). Nitric oxide elimination did not correlate with Raw. Depth increased inspiratory mechanical power of breathing during rest (40%) and exercise (20%). Expiratory mechanical power of breathing was largely unchanged. These results suggest that the diving environment affects ventilatory mechanics primarily by increasing Raw, secondary to increased gas density. This necessitates increased alveolar pressure and increases the work and energy cost of breathing as the diver descends. These findings can inform physician assessment of diver fitness and the pulmonary risks of hyperbaric O2 therapy.