Changes in energy system contributions to the Wingate anaerobic test in climbers after a high altitude expedition.

PURPOSE: The Wingate anaerobic test measures the maximum anaerobic capacity of the lower limbs. The energy sources of Wingate test are dominated by anaerobic metabolism (~ 80%). Chronic high altitude exposure induces adaptations on skeletal muscle function and metabolism. Therefore, the study aim was to investigate possible changes in the energy system contribution to Wingate test before and after a high-altitude sojourn. METHODS: Seven male climbers performed a Wingate test before and after a 43-day expedition in the Himalaya (23 days above 5.000 m). Mechanical parameters included: peak power (PP), average power (AP), minimum power (MP) and fatigue index (FI). The metabolic equivalents were calculated as aerobic contribution from O2 uptake during the 30-s exercise phase (WVO2), lactic and alactic anaerobic energy sources were determined from net lactate production (WLa) and the fast component of the kinetics of post-exercise oxygen uptake (WPCr), respectively. The total metabolic work (WTOT) was calculated as the sum of the three energy sources. RESULTS: PP and AP decreased from 7.3 ± 1.1 to 6.7 ± 1.1 W/kg and from 5.9 ± 0.7 to 5.4 ± 0.8 W/kg, respectively, while FI was unchanged. WTOT declined from 103.9 ± 28.7 to 83.8 ± 17.8 kJ. Relative aerobic contribution remained unchanged (19.9 ± 4.8% vs 18.3 ± 2.3%), while anaerobic lactic and alactic contributions decreased from 48.3 ± 11.7 to 43.1 ± 8.9% and increased from 31.8 ± 14.5 to 38.6 ± 7.4%, respectively. CONCLUSION: Chronic high altitude exposure induced a reduction in both mechanical and metabolic parameters of Wingate test. The anaerobic alactic relative contribution increased while the anaerobic lactic decreased, leaving unaffected the overall relative anaerobic contribution to Wingate test.

Responses of peripheral blood mononuclear cells to moderate exercise and hypoxia.

The purpose of this study was to analyze the physiological features of peripheral blood mononuclear cells (PBMCs) isolated from healthy female trekkers before and after physical activity carried out under both normoxia (low altitude, < 2000 m a.s.l.) and hypobaric hypoxia (high altitude, > 3700 m a.s.l.). The experimental design was to differentiate effects induced by exercise and those related to external environmental conditions. PBMCs were isolated from seven female subjects before and after each training period. The PBMCs were phenotypically and functionally characterized using fluorimetric and densitometric analyses, to determine cellular activation, and their intracellular Ca(2+) levels and oxidative status. After a period of normoxic physical exercise, the PBMCs showed an increase in fully activated T lymphocytes (CD3(+) CD69(+) ) and a reduction in intracellular Ca(2+) levels. On the other hand, with physical exercise performed under hypobaric hypoxia, there was a reduction in T lymphocytes and an increase in nonactivated B lymphocytes, accompanied by a reduction in O2 (-) levels in the mitochondria. These outcomes reveal that in women, low- to moderate-intensity aerobic trekking induces CD69 T cell activation and promotes anti-stress effects on the high-altitude-induced impairment of the immune responses and the oxidative balance.

Gokyo Khumbu/Ama Dablam Trek 2012: effects of physical training and high-altitude exposure on oxidative metabolism, muscle composition, and metabolic cost of walking in women.

PURPOSE: We investigated the effects of moderate-intensity training at low and high altitude on VO2 and QaO2 kinetics and on myosin heavy-chain expression (MyHC) in seven women (36.3 yy ± 7.1; 65.8 kg ± 11.7; 165 cm ± 8) who participated in two 12- to 14-day trekking expeditions at low (598 m) and high altitude (4132 m) separated by 4 months of recovery. METHODS: Breath-by-breath VO2 and beat-by-beat QaO2 at the onset of moderate-intensity cycling exercise and energy cost of walking (Cw) were assessed before and after trekking. MyHC expression of vastus lateralis was evaluated before and after low-altitude and after high-altitude trekking; muscle fiber high-resolution respirography was performed at the beginning of the study and after high-altitude trekking. RESULTS: Mean response time of VO2 kinetics was faster (P = 0.002 and P = 0.001) and oxygen deficit was smaller (P = 0.001 and P = 0.0004) after low- and high-altitude trekking, whereas ˙ QaO2 kinetics and Cw did not change. Percentages of slow and fast isoforms of MyHC and mitochondrial mass were not affected by low- and high-altitude training. After training altitude, muscle fiber ADP-stimulated mitochondrial respiration was decreased as compared with the control condition (P = 0.016), whereas leak respiration was increased (P = 0.031), leading to a significant increase in the respiratory control ratio (P = 0.016). CONCLUSIONS: Although training did not significantly modify muscle phenotype, it induced beneficial adaptations of the oxygen transport-utilization systems witnessed by faster VO2 kinetics at exercise onset.

Sperm forward motility is negatively affected by short-term exposure to altitude hypoxia.

Human exposure to altitude is a model to study the role of oxygen in different areas of physiology and pathophysiology. The aim of this study was to evaluate whether a short exposure to hypoxia (5 days) combined with exercise, at altitude ranging from 900 m above sea level to 5895 m above sea level (Kilimanjaro Expedition) can modify seminal and reproductive hormonal parameter levels in human beings. During the ascent, blood oxygen saturation at 3.848 m above sea level was found to be decreased when compared to sea level (P < 0.02). The sperm forward motility at sea level after the expedition showed a significant reduction ​​(P < 0.02). There were no changes in other seminal parameters among those compared. Determination of the hormonal plasma concentrations showed that baseline values of follicle-stimulating hormone, total testosterone, prolactin and oestradiol were unchanged at sea level after the hypoxic experience, with respect to baseline values at sea level. On the other hand, luteinising hormone levels after altitudes trekking significantly increased compared to levels before the expedition (P < 0.05). Because of the short-term exposure, we can assume that the reduced forward motility described here may result from the effects of the acute altitude hypoxia on spermatozoa during the epididymal transit where they mature acquiring their motility.

Adaptation of olfactory threshold at high altitude.

The aim of this study was to investigate the effects of the extreme environment of high altitude hypoxia on olfactory threshold. The study was conducted before, during, and after a scientific expedition to Mera Peak (5,800 m). The n-butanol test was used for the assessment of the magnitude of the olfactory threshold. The finding was that the olfactory threshold dramatically increased at high altitude. We conclude that there is a physiological adaptation of olfaction due to altitude-hypoxia.

Proteomic analysis of the carotid body: a preliminary study.

We present a proteomic analysis of the rat carotid body (CB) preparation by comparison between normoxia and hypoxia. Proteomic investigation would be helpful to identify the stress-induced protein during hypoxia and to know what O(2) species are being sensed by CB cells. Adult Wistar rats were used, one group was kept in room air (21% O(2)) as control, and the other was kept in a Plexiglas chamber for 12 days in chronic hypoxia (10-11% inspired oxygen). A total protein extract for each lysated tissue was separated using a broad pH range no-linear IPG strip (3-10) and the second dimension was performed on a 9-16% polyacrylamide gel. Exposure to hypoxia for 12 days produced significant changes in protein expression, providing an initial insight into the mechanism underlying differences in susceptibility to hypoxia. Further investigation is needed to have an overview of the specific set of proteins present in the CB and the functions of such proteins in signal transduction and adaptation during hypoxia.

Physiological analysis of 8-ISO-PGF2 alpha: a homeostatic agent in superficial bladder cancer.

Recent studies underscore the importance of oxygen supply in bladder cancer. Tumour growth stimulates the production of vasoactive factors to increase oxygen delivery to tissues by vasodilatation. Any vasoconstrictor mediator could impair this vasodilatation reducing the oxygen supply. 8-Iso-PGF2 alpha is a potent vasoconstrictor agent. The aim of this work is to determine 8-Iso-PGF2 alpha release in healthy bladder mucosa and in superficial bladder cancer in order to investigate a pathophysiological vasoconstrictor answer of the superficial bladder cancer. The study was conducted on a sample of 12 patients; for every subject studied 8-Iso-PGF2 alpha release was assayed in healthy bladder mucosa and in superficial bladder tumour. 8-Iso-PGF2 alpha release was significantly reduced (p less than 0.001) in superficial bladder cancer compared with healthy bladder mucosa. The inhibition of the production of a powerful vasoconstrictor such as 8-Iso-PGF2 alpha in the vascular homeostatic mechanism of bladder cancer can represent a response of the tumor tending to contrast an antagonist effect of vasodilatation and the necessary to support the oxygen supply.

Cafeteria diet increases prostaglandin E2 levels in rat prostate, kidney and testis.

Nutrient composition, particularly the omega-6/omega-3 polyunsaturated fatty acids ratio, may differently affect inflammatory mediators production in tissues, which could be causally related to increased cancer incidence in obesity. We evaluated prostaglandin E(2) levels in male Wistar rat prostate, kidney and testicle tissues after 15 days of either a high fat, cafeteria-style diet (5.50 Kcal/g, 30 percent calories from fat, omega-6/omega-3 ratio 2.33) or a standard laboratory chow diet (3.35 Kcal/g, 3 percent calories from fat, omega-6/omega-3 ratio 0.56). In the cafeteria diet compared to standard laboratory diet rats, we found both an increase in weight gain and increased prostaglandin E(2) (PGE(2)) levels in prostate, kidney and testicle tissues. The increased levels of PGE(2) induced by the cafeteria diet could drive an inflammatory process leading to increased incidence of prostate, kidney and testicular cancer in overweight patients.

Reduced pulmonary function is age-dependent in the rat lung in normoxia.

BACKGROUND: Oxygen transport is optimized at the cellular level, since oxygen serves as the terminal electron acceptor in mitochondrial oxidative phosphorylation and several enzymatic processes require molecular oxygen as substrate. During development and aging, redundant cells and exhausted cells are eliminated, respectively, whereas others can adapt to the stressful environment and survive. OBJECTIVE: The study investigated the molecular mechanisms activated in the lung during normal aging, through the expression of hypoxia inducible factor (HIF), vascular endothelial growth factor (VEGF), p53, p66⊃Shc, putative cysteine protease (CPP32) and kinaseB-α phosphorylation (pIkB-α). MATERIAL AND METHODS: Twelve male Wistar rats divided into two age-groups, each consisting of 6 animals, 3 and 24 months old, were used. The rats were anesthetized with Nembutal (40 mg/kg, ip) and the lungs were excised from each rat and processed for TUNEL and Western blotting analyses. RESULTS: The expressions of p53, p66⊃Shc and CPP32 were significantly increased in the old normoxic rat lung specimens, when compared with the young ones. In parallel, expressions of VEGF and pIkBα were increased in old rather than young rats. CONCLUSIONS: Aging leads to increased expressions of p53, p66⊃Shc and CPP32, suggesting that apoptosis is in progress. At the same time, the lung tries to counteract apoptosis through the production of VEGF and pIkB-α to adapt itself to a stressful situation. The aging lung creates a life-support system in order to counteract the apoptotic process.

Neuroimmune biology of physical exercise.

Physical inactivity plays a negative role in several aspects of health status. A sedentary lifestyle represents a predisposing base for the development of obesity, type 2 diabetes, cardiovascular disease, osteoporosis, depression and cancer. On the other hand, an active life is distinguished by physical exercise and the beneficial effects it brings to health, reducing the risk of contracting these diseases. The general state of exercise-related psycho-physical health is regulated by a variety of mechanisms that affect the highly integrated and synergistic responses of the central nervous system and efficiency of the overall immune system. There is positive evidence concerning the influence of neuro-immune systems on the status of health, during moderate and regular physical exercise.

Does hypoxia cause sarcopenia? Prevention of hypoxia could reduce sarcopenia.

Sarcopenia is the physiological age-related reduction of muscle mass and strength. Considering that life span is correlated with metabolic rate and mitochondria are the site of oxygen consumption, muscle mitochondria volume densities were determined by morphometric analysis. We found a tight correlation between aging and hypoxia with decrease in muscle total mitochondria volume. Therefore, hypoxia and aging seem to share some common pathways, allowing hypoxic models to be used for the study of the aging processes. Additional research will be required to fully elucidate the correlations among aging, sarcopenia and hypoxia, but these findings provide a starting point for such investigations.

Neuroglobin in aging carotid bodies.

Neuroglobin (Ngb) is a member of the vertebrate globin family expressed particularly in the brain and in the retina. Ngb is concentrated in the mitochondria-containing areas of neurons, and its distribution is correlated with oxygen consumption rates. Previously we have shown that Ngb is expressed in carotid body (CB) tissues. Considering that hypoxia and aging may be linked through a series of adaptive and protective mechanisms (e.g. reduction in mitochondrial numbers), we investigate the role of Ngb during aging and hypoxia. Two groups of six rats (age-matched 3 and 24 months old) were kept in room air as a control groups, the others two groups were kept in a Plexiglas chamber for 12 days in chronic hypoxia (10-12% inspired oxygen). The presence of Ngb in the CB tissue was detected by immunohistochemistry using a polyclonal antibody. Ngb immunoreactivity was significantly higher in CB tissues from young rats exposed to chronic intermittent hypoxia, whereas CB tissues from old rats did not show any significant increase in Ngb levels after hypoxia. Similar to hemoglobin, Ngb may act as a respiratory protein by reversibly binding gaseous ligands NO and O(2) and could act as a NO scavenger and participate in detoxification of Reactive Oxygen Species (ROS) generated under hypoxic conditions.

Physiological carotid body denervation during aging.

Aging is characterized by a lower homeostatic capacity and the carotid body (CB) plays an important role during aging. Here, we sought to elucidate whether the aging effects on the oxygen-sensitive mechanisms in CB cells occur through a reduction of the contact surfaces in the synaptic junctions. The hypothesis was that the CB would undergo a "physiological denervation" in old age. Two groups of male Wistar rats, young (2-3 months old) and senescent (22 months old) were used. CBs were rapidly dissected and the specimens were subjected to a routine transmission electron microscopic procedure. Expressions of HIF-1 proportional, variant, VEGF and NOS-1 were evaluated by immunohistochemical analysis. Our results show that in the old CB, HIF-1 proportional, variant, VEGF and NOS-1 expressions decrease. The cell volume, the number of mitochondria and that of dense-cored vesicles were reduced, and the nucleus shrank. There also was an accumulation of lipofuscin and a proliferation of extracellular matrix. Most importantly, there were fewer synaptic connections between chemoreceptor cells. The total number of synapses observed in all electronograms decreased from 125 in the young to 28 in the old CB. These results suggest the aging CB undergoes a "physiological denervation" leading to a reduction in homeostatic capacity. The age-related reduction of synaptic junctions may be a self-protective mechanism through which cells buffer themselves against reactive oxygen species accumulation during aging.

Pampiniform plexus and oxidative stress during chronic hypoxia and hyperoxia.

The aim of the present study is to investigate oxidative stress produced by experimental hypoxia and hyperoxia in young and old pampiniform plexus rats, in order to evaluate the oxidative role of oxygen. Oxidative stress causing molecular and cellular dysfunction increases in hypertension and can therefore be considered a state of oxidative stress. This consideration makes us reflect on the responsibility of oxidative stress in the veins of the pampiniform plexus, notoriously under high hydrostatic pressure. After experimental hypoxia and hyperoxia we studied the 8-iso-PGF2alpha release (a specific index of cellular oxidative stress) in young and old left pampiniform plexus rats. The basal 8-iso-PGF2alpha release showed a statistically significant difference P=0.0067 between young and old rats PP. After hypoxia and hyperoxia, the release was higher in young rats as compared to normoxia, respectively P=0.0001 and P=0.0002. After hypoxia the release was not modified in old rats P=0.544 while after hyperoxia the release was increased in old rats as compared to control P less than 0.0001. The results show how chronic hypoxia and hyperoxia represent two important causes of oxidative stress and lipid peroxidation in pampiniform plexus rats. In young rats an increase of oxidative stress suggests that pampiniform plexus is sensitive to variations of oxygen supply. In old rats the pampiniform plexus is liable to a reduction of oxygen-sensing mechanisms and it is possible that the missing oxidative answer to the hypoxia in old rats is attributable in all likelihood to adaptation to a hypoxic condition typical of aging.

The role of hypoxia in erectile dysfunction mechanisms.

Chronic hypoxia is related to many pathological conditions: aging, heart and respiratory failure, sleep apneas, smoke, chronic obstructive pulmonary disease (COPD), diabetes, hypertension and arteriosclerosis, all characterized by reductions of sleep-related erections (SREs) and by erectile dysfunction (ED). Sleep-related erections occur naturally during rapid eye movement (REM) sleep in sexually potent men. Hypoxia is also a physiological condition at altitude. The level of inspired oxygen decreases progressively with the increase of altitude; for this reason, this study was performed to evaluate the relationship of SREs with hypoxic environment. SREs have been recorded by an erectometer (RigiScan) on three mountain climbers (mean age: 32.5) during a 26-day stay at an altitude ranging from 2000 to 5600 m above sea level. Twenty-four records have been made at progressively increasing altitudes. A data analysis was carried out on a statistical mean of the three values of each variable and an analysis of variance (ANOVA) and Newman-Keuls test were carried out for multiple comparison among groups. At altitudes over 4450 m, we found lack of rigidity at 80-100% and 60-79%. Mean % of rigidity and rigidity time of 80-100% (tip and base) decreased progressively with altitude. No significant reductions were shown in rigidity time at 0-19% and at 20-39% (tip and base), of total number, of total and mean duration of SREs. Pathological rigidometric records at high altitude in sexually potent men at sea level clarify the primary role of hypoxia in physiopathological ED pathway.

Improved VO2 uptake kinetics and shift in muscle fiber type in high-altitude trekkers.

The study investigated the effect of prolonged hypoxia on central [i.e., cardiovascular oxygen delivery (Q(a)O(2))] and peripheral (i.e., O(2) utilization) determinants of oxidative metabolism response during exercise in humans. To this aim, seven male mountaineers were examined before and immediately after the Himalayan Expedition Interamnia 8000-Manaslu 2008, lasting 43 days, among which, 23 days were above 5,000 m. The subjects showed a decrease in body weight (P < 0.05) and of power output during a Wingate Anaerobic test (P < 0.05) and an increase of thigh cross-sectional area (P < 0.05). Absolute maximal O(2) uptake (VO(2max)) did not change. The mean response time of VO(2) kinetics at the onset of step submaximal cycling exercise was reduced significantly from 53.8 s ± 10.9 to 39.8 s ± 10.9 (P < 0.05), whereas that of Q(a)O(2) was not. Analysis of single fibers dissected from vastus lateralis biopsies revealed that the expression of slow isoforms of both heavy and light myosin subunits increased, whereas that of fast isoforms decreased. Unloaded shortening velocity of fibers was decreased significantly. In summary, independent findings converge in indicating that adaptation to chronic hypoxia brings about a fast-to-slow transition of muscle fibers, resulting in a faster activation of the mitochondrial oxidative metabolism. These results indicate that a prolonged and active sojourn in hypoxia may induce muscular ultrastructural and functional changes similar to those observed after aerobic training.

Aging and expression of heme oxygenase-1 and endothelin-1 in the rat carotid body after chronic hypoxia.

Hypoxia transiently increases transcription of the gene encoding heme oxygenase-1 (HO-1) and potently activates production of endothelin-1 (ET-1), the latter of which plays a central role in cellular adaptation to hypoxia. The ventilatory response to hypoxia attenuates with aging, and decreased responsiveness to hypoxia is seen in the aged vs. young rats, suggesting that the functionality of the oxygen-sensitive mechanism is age-dependent. In the present study, we examined the effects of aging on the expression of HO-1 and ET-1 in the carotid body, which is a small cluster of chemoreceptors and supporting cells that measure changes in the composition of arterial blood flowing through it. Our results revealed that HO-1 and ET-1 were expressed in carotid bodies of both young and old rats, although less so in the old ones. Exposure to chronic intermittent hypoxia significantly increased both HO-1 and ET-1 immunoreactivity in both young and old carotid body tissues, with the persisting age-dependent inequality to the disadvantage of old age. Considering that ET-1 is capable of enhancing intermittent hypoxia-induced chemosensory responses by the carotid body, our results suggest that decreased induction of ET-1 and HO-1 during aging could form the basis for age-related reductions in chemosensory discharge.