Blockage of hemichannels alters gene expression in osteocytes in a high magneto-gravitational environment.

Osteocytes, the most abundant cells in bone, are highly responsive to external environmental changes. We tested how Cx43 hemichannels which mediate the exchange of small molecules between cells and extracellular environment impact genome wide gene expression under conditions of abnormal gravity and magnetic field. To this end, we subjected osteocytic MLO-Y4 cells to a high magneto-gravitational environment and used microarray to examine global gene expression and a specific blocking antibody was used to assess the role of Cx43 hemichannels. While 3 hr exposure to abnormal gravity and magnetic field had relatively minor effects on global gene expression, blocking hemichannels significantly impacted the expression of a number of genes which are involved in cell viability, apoptosis, mineral absorption, protein absorption and digestion, and focal adhesion. Also, blocking of hemichannels enriched genes in multiple signaling pathways which are enaged by TGF-beta, Jak-STAT and VEGF. These results show the role of connexin hemichannels in bone cells in high magneto-gravitational environments.

Cytokine secretion and latent herpes virus reactivation with 28 days of horizontal hypokinesia.

INTRODUCTION: Hypokinesia is associated with spaceflight and prolonged illnesses and may lead to secondary immune deficiency. METHODS: The distribution of immunocytes in whole blood, mitogen-induced cytokine secretion in vitro, Epstein-Barr virus (EBV) reactivation, and plasma cortisol levels were studied in 13 healthy volunteers subjected to a horizontal bed rest (BR) regime for 28 d. Samples were collected before the study, weekly during BR, and then 3-5 d after the regime ended. Additionally, subjects were treated with hydrocortisone on the 1st and 27th d of BR to simulate the hypercortisolemia that occurs during stress. RESULTS: The factors of 28-d BR regime accompanied by acute hypercortisolemia significantly decreased the relative and absolute number of total lymphocytes, CD3+ T-cells, T-helper subset, and monocytes, but increased the percentage of the CD8+ T-cells, and NK cells at the 4th wk compared with the baseline. A significant decrease in mitogen-activated secretion of IL-2, IFN-gamma, TNF-beta, IL-6, and IL-10 was registered at the same interval. Also, secretion of IL-2 and IFN-gamma declined at the 2nd week of the BR regime. Secretion of IL-4 was significantly higher at the 2nd and 3rd weeks compared with the baseline. A significant increase in the shedding of EBV DNA in saliva was observed as early as the 3rd wk of BR. CONCLUSIONS: Stress factors associated with BR significantly alter immune responsiveness in vitro and in vivo. Changes in the cytokine secretion and cytokine imbalance precede latent EBV reactivation. PHA/LPS-activated cytokine secretion in whole blood can be used as a test system for predicting latent virus activation.

Effect of estradiol and dihydrotestosterone on hypergravity-induced MAPK signaling and occludin expression in human umbilical vein endothelial cells.

Female astronauts have been reported to have a higher incidence of post-flight orthostatic intolerance (POI) compared with that of their male counterparts. POI may result from increased permeability of the endothelial cell (EC) layer in the vasculature. The goal of this study has been to determine whether estradiol (E(2)) and dihydrotesterone (DHT) alter human umbilical vein ECs (HUVECs) responses to short term (10 min) hypergravity (1-3 g) mimicking the g force experienced by astronauts during liftoff. E(2) and DHT rapidly (within 5 min) activated MAPK (mitogen-activated protein kinase) in HUVEC at 1 g in a receptor-dependent manner. Liftoff inhibited MAPK phosphorylation, and rapid E(2) and DHT activation of MAPK was blocked. Liftoff simulation or brief (5-90 min) treatment with E(2) or DHT at 1 g had no effect on the expression of the EC tight-junction protein occludin. However, 24-h pre-treatment of HUVECs with E(2) and DHT prior to liftoff simulation significantly increased occludin expression, and hypergravity exposure did not alter this increase. These data provide evidence for a possible protective effect of E(2) and DHT on EC function as indicated by increased occludin; this may help maintain the integrity of EC tight junction and could thus retard or reduce the incidence of POI.

[Outcome of the long-term isolation on the development of allergic reactions in humans]. / Vliianie dlitel'noi izoliatsii na formirovanie allergicheskikh reaktsii u cheloveka.

Studied were consequences of long-term isolation in airtight environment for development of the type-I (IgE-antibodies production) and type-IV (involvement of sensibilized T-lymphocytes) allergic reactions in humans. No significant changes in total IgE, specific IgE-antibodies for domestic, epidermal, fungal, grass pollen and food-borne allergens or serum IL-4 level were found in the period of the 240-d isolation with the microclimate and atmospheric parameters within their normal variations, and on completion of the experiment. Yet, after 3 mos. in isolation all subjects exhibited inhibited leukocyte migration in the presence of tuberculin. These findings bear witness to an activation of sensibilized lymphocytes known as effectors of delayed hypersensitivity.

Incidence of adverse reactions from 23,000 exposures to simulated terrestrial altitudes up to 8900 m.

INTRODUCTION: Adverse reactions during hypobaric chamber operations result from changes in barometric pressure per se and to the related reductions in the partial pressure of inspired oxygen. Previous studies have indicated that an adverse reaction may occur in approximately 6% of exposures in chambers used for flight training. The purpose of this study was to determine the rate of adverse reactions in a chamber used exclusively for terrestrial altitude research studies. METHODS: Data for incidence of ear block, sinus block, toothache, and other adverse reactions were analyzed using a master history file that contained information such as annual and total number of studies and human exposures, and onset and outcome of adverse reactions. Incidence for a given time period was calculated as (# of reactions x 100)/(# of exposures). RESULTS: In 33 yr, there have been 23,656 human exposures and 296 adverse reactions. The overall incidence was 1.25 reactions per 100 exposures (0.72 for research volunteers and 0.53 for staff). The majority (75% of all reactions) were ear blocks (0.93 reactions per 100 exposures). Staff members were more likely than research volunteers to have more than one reaction. DISCUSSION: The incidence of 1.25 reactions per 100 exposures for our chamber is considerably lower than that reported in the literature. This is due primarily to our use of lower altitudes and slower rates of ascent and descent compared with other facilities whose emphasis is on aircraft flight training.

[Dental caries resistance under conditions of a space flight]. / Ustoichivost' zubov k kariesu v usloviiakh kosmicheskogo poleta.

The maxillodental status of aeronauts attracts special interest of scientists and causes apprehensions of physicians and scientists of countries with aeronautic industry because of long duration of space missions. Some scientists consider that the realization of mission to Mars can lead to development of multiple dental caries in aeronauts. The aim of this study was to study the effects of space mission factors on the resistance of hard dental tissues and remineralizing activity of the saliva in a simulation experiment. The results demonstrated dynamic changes in the studied parameters under the effect of space mission factors; based on these results, we determined the criteria for selection of candidates for prolonged experiments and space missions.

Parasympathetic activity during parabolic flight, effect of LBNP during microgravity.

BACKGROUND/HYPOTHESIS: During parabolic flight, in the standing position, changes are partly due to an acute shift in fluid between the lower extremities, the head and the thorax (Vaïda P, et al. J Appl Physiol 1997; 82:1091-7; and Bailliart O, et al. J Appl Physiol 1998; 85:2100-5). We hypothesized that modifications of parasympathetic activity associated with changes in hydrostatic pressure gradients induced by changes in gravity could be detected by analysis of short time periods. METHODS: We assessed heart rate variability (HRV) in 11 healthy volunteers by indices of temporal analysis (NN, SDNN, RMSSD) and normalized indices such as coefficients of variation CV-SDNN and CV-RMSSD and ratio SDNN/RMSSD. A lower body negative pressure (LBNP) at -50 mm Hg was randomly applied during the microgravity phase (0 Gz) to counteract the lack of hydrostatic pressure in the lower part of the body. RESULTS: NN, CV-SDNN and CV-RMSSD decreased during hypergravity phases and increased during microgravity and during early normogravity (1 Gz) period at the end of parabolas. With LBNP changes are less pronounced at 0 Gz and in the 1 Gz post parabolic period. CONCLUSION: We concluded that parasympathetic nervous activity is recordable by temporal analysis of HRV during short periods of time. LBNP applied during 0 Gz phase reduced the parasympathetic activation at 0 Gz and post parabolic 1 Gz.

Dietary salt and urinary calcium excretion in a human bed rest spaceflight model.

BACKGROUND: Dietary salt is known to increase the excretion of urinary calcium (Ca). To determine the potential role of dietary sodium (Na) on the calciuria associated with a spaceflight simulation model, we evaluated urinary Ca in two groups of bed rest subjects fed either high or low normal amounts of salt. METHODS: We analyzed urinary Ca excretion expressed in terms of creatinine (UCa/Cr), fractional Ca excretion (FECa), and urinary cAMP (UCAMP) as an index of parathyroid function, in the urine of 30-50-yr-old male volunteers for 6 degrees head down tilt bed rest studies. Dietary Na was in the high normal range (190 mmol x d(-1)) in 8 men for 7 d (HiNa), and in the low normal range (114 mmol x d(-1)) in 11 men for 30 d (LoNa) bed rest. Dietary Ca averaged 20 mmol x d(-1) in both studies. RESULTS: Within the first 3 bed rest days, subjects in the HiNa study showed increases in UCa/Cr (0.1130 +/- 0.05 to 0.161 +/- 0.05, p < 0.002) and in FECa (1.95 +/- 0.70 to 3.19 +/- 0.93, p < 0.001); those in LoNa showed no change in UCa/Cr (0.125 +/- 0.06 to 0.121 +/- 0.07, NS) or FECa (1.93 +/- 0.75 to 2.22 +/- 0.63). After the 5th bed rest day UCa/Cr stabilized at similar levels in both dietary groups. UCAMP decreased 20% during the first week of bed rest with HiNa, but not until the third week with LoNa diets (p < 0.05). CONCLUSION: These findings implicate high salt diets in Ca excretion in a spaceflight model and suggest that low normal salt diets may reduce early calciuria associated with spaceflight.

Thermoregulation and heat exchange in a nonuniform thermal environment during simulated extended EVA. Extravehicular activities.

BACKGROUND: Nonuniform heating and cooling of the body, a possibility during extended duration extravehicular activities (EVA), was studied by means of a specially designed water circulating garment that independently heated or cooled the right and left sides of the body. The purpose was to assess whether there was a generalized reaction on the finger in extreme contradictory temperatures on the body surface, as a potential heat status controller. METHOD: Eight subjects, six men and two women, were studied while wearing a sagittally divided experimental garment with hands exposed in the following conditions: Stage 1 baseline--total body garment inlet water temperature at 33 degrees C; Stage 2--left side inlet water temperature heated to 45 degrees C; right side cooled to 8 degrees C; Stage 3--left side inlet water temperature cooled to 8 degrees C, right side heated to 45 degrees C. RESULTS: Temperatures on each side of the body surface as well as ear canal temperature (Tec) showed statistically significant Stage x Side interactions, demonstrating responsiveness to the thermal manipulations. Right and left finger temperatures (Tfing) were not significantly different across stages; their dynamic across time was similar. Rectal temperature (Tre) was not reactive to prevailing cold on the body surface, and therefore not informative. Subjective perception of heat and cold on the left and right sides of the body was consistent with actual temperature manipulations. CONCLUSIONS: Tec and Tre estimates of internal temperature do not provide accurate data for evaluating overall thermal status in nonuniform thermal conditions on the body surface. The use of Tfing has significant potential in providing more accurate information on thermal status and as a feedback method for more precise thermal regulation of the astronaut within the EVA space suit.

Rat growth, body composition, and renal function during 30 days increased ambient CO2 exposure.

BACKGROUND: In experiments using rodents onboard orbiting spacecraft, specimens may be exposed to an increase in ambient CO2. HYPOTHESIS: Many of the physiological changes reported in rats (and humans) for spaceflight are similar to those observed with increased CO2, raising the question whether the observed changes are due to spaceflight or more specifically, the elevated ambient CO2. METHODS: To evaluate the effects of increased CO2, at levels similar to those experienced during spaceflight, three groups of adult male rats (n = 10) were exposed to ambient CO2 concentrations of 0.3, 0.7 and 2.0% for 30 d. Control rats were exposed to atmospheric conditions (0.03% CO2) for each group. RESULTS: There were alterations in water turnover, food intake, and renal function with increased CO2. Blood pH, total CO2, and plasma concentrations of Na+, Ca2+, and corticosterone were significantly elevated at the 2.0% exposure, while plasma PO4(3-) was reduced. At the 0.3% and 0.7% CO2 exposures, many of these changes were not significant. Animals exposed to 0.3% CO2 showed a significant increase in total body Na+. Urinary Ca2+, K+, creatinine, corticosterone, and total CO2 excretion were higher at 2.0%, but only Ca2+ and CO2 excretion were significantly elevated at 0.7%, and there was no significant alteration in renal function at 0.3%. CONCLUSION: Chronic increased ambient CO2 levels, similar to those observed on the Space Shuttle and proposed for the International Space Station, elicit compensatory responses in rats which may affect interpretation of experiments designed to evaluate the effects of exposure to microgravity.

Effects of gender on the autonomic modulation of the cardiovascular responses to lower body negative pressure.

BACKGROUND: The cardiovascular responses to submaximal lower body negative pressure (LBNP) appear to differ between genders, but the underlying mechanisms are uncertain. HYPOTHESIS: These differences are due to differences in the autonomic modulation of the cardiovascular system. METHODS: There were 14 women and 13 men who underwent LBNP to -50 mmHg in 10 mmHg increments of 6 min each. Heart rate (HR), stroke volume (SV), BP, forearm blood flow and R-R interval data were acquired. Spectral analysis of the R-R interval data was used to assess autonomic modulation with the low frequency component (LF) set at 0.04 to 0.15 Hz and the high frequency component (HF) at 0.15 to 0.4 Hz. RESULTS: The responses to LBNP to -40 mmHg did not differ between groups. LBNP of -50 mmHg evoked greater HR increases in the women than the men (7.2 +/- 1.0 vs. 3.8 +/- 1.1 bpm; p < 0.05), while SV, cardiac output and total peripheral conductance decreased more (-15 +/- 2 vs. -8 +/- 2 ml x beat(-1); -0.668 +/- 0.131 vs. -0.1778 +/- 0.124 L x min(-1); -0.009 +/- 0.002 vs. -0.004 +/- 0.001 units; p < 0.05). Normalized HF, an indicator of the vagal influence on HR variability, declined below rest at -40 mmHg while the LF/HF ratio, an indicator of sympathetic neural modulation of HR variability, increased above rest at -40 mmHg. These responses did not differ significantly between groups. CONCLUSIONS: These results suggest that gender differences in the cardiovascular responses to LBNP are not due to gross differences in modulation of the autonomic nervous systems.

Antiorthostatic suspension for 14 days does not diminish the oxidative response of neutrophils in mice.

The effects of long-term spaceflight on inflammatory responses have not been well-studied in either humans or animals. It is thus important to determine if the functions of immune and inflammatory cells are altered in models of spaceflight. One such animal model is antiorthostatic suspension (AOS), in which the experimental animal is subjected to a head-down tilt that mimics both the stress and the cephalad fluid shift experienced in spaceflight. A previous study reported that the peritoneal neutrophils from mice experiencing AOS generated less superoxide than unsuspended controls. We expanded on this study using several different stimuli and measuring the oxidative response of murine neutrophils in a variety of ways. These responses included the rate, lag period, and dose/response characteristics for superoxide generation, FACS analysis with dihydrodichlorofluorescein as a substrate, and a chemiluminescence response with luminol as a substrate. We also examined phagocytosis of three different microorganisms. While some effects of orthostatic suspension (attributable to the stress of the apparatus) were observed, no clear effects of AOS on oxidative function of the peritoneal neutrophils were seen.

Hormonal changes during a 20-week confinement.

BACKGROUND: When the European Space Agency planned the EUROMIR'95 long-duration flight with a European astronaut on board the Russian orbital MIR station, it organized simultaneously a ground simulation, called the Human Behaviour Study, of this manned space mission. The ground simulation was a confinement experiment, and this paper describes the changes in volume-regulating hormones that occurred during and after 20 weeks of confinement. METHODS: In a normobaric diving chamber, 3 subjects were confined for 135 d. Arterial pressure, plasma concentrations of blood volume-regulating hormones (active renin and arginine-vasopressin), and urinary variables (aldosterone, arginine-vasopressin, and metabolites of catecholamines) were measured before, during, and after confinement. RESULTS: Arterial pressure was increased from week 1 until week 15 of confinement, while heart rate was elevated from week 6 until the end of the simulation. Plasma active renin was elevated throughout the confinement (after week 6). Urine volume increased transitively on the first 2 d of confinement. CONCLUSIONS: The results obtained during this long-term confinement experiment have major importance regarding concerns about spaceflight and bed rest data, because we observed hormonal changes during the experiment that normally are assigned to the fluid shift that occurs in weightlessness or in the head-down tilt position (i.e., an increase of renin, an increase of urinary volume during the first two days, and a decreased urinary cyclic guanosine monophosphate.

Cardiovascular and hormonal changes induced by a simulation of a lunar mission.

BACKGROUND: This is the first simulation of a 14-d lunar mission including 6 d on the Moon. HYPOTHESIS: We hypothesized that a lunar gravity simulation in the middle of a head-down tilt (HDT) might result in some reversal of body fluid/hormonal responses, and influence cardiovascular deconditioning. METHODS: Six men (28 +/- 2.5 yr) were placed in bed rest (BR): in (HDT) (-6 degrees) to simulate microgravity during the travel (two 4-d periods), and in head-up tilt (HUT) (+10 degrees) (6-d period) to simulate lunar gravity (1/6 g). Muscular exercise was performed during the HUT period to simulate 6 h of lunar EVA. Heart rate variability (HRV) and hormonal responses were studied. RESULTS: An orthostatic arterial hypotension was observed after the BR (tilt test) in 4 of the 6 subjects. Plasma volume measured at D14 decreased by -11.1% (vs. D-3, sitting position). A decrease in atrial natriuretic peptide (26 +/- 3.5 pg.ml-1 (D14) vs. 37.9 +/- 3.5 pg.ml-1 (D-3, sitting) and an increase in plasma renin activity (198 +/- 9.2 mg.L-1.min-1 (D14) vs. 71 +/- 9.2 mg.L-1.min-1 (D-3, sitting) were observed during the BR, more pronounced in HUT at 7:00 p.m. Sympathetic-parasympathetic balance (HRV) at rest showed a decrease in parasympathetic indicator and an increase in sympathetic indicator in BR (p < 0.05), without differences within HDT and HUT periods. CONCLUSION: These changes were mostly similar to those reported in spaceflights, and HDT. Although the exposure to 1/6 g with exercise modified some hormonal and body fluid responses, this partial gravity simulation was not sufficient to prevent the decrease in orthostatic tolerance observed here as well as after Apollo lunar missions.

Effects of exercise during head-down bed rest on postural control.

BACKGROUND: The effects of exercise on postural control during walking were evaluated following exposure to head-down bed rest (BR). METHODS: Two groups of male subjects (N = 18, mean age = 21.4 yr +/- SE 1.0) were exposed to 5 d of 6 degrees head-down bed rest. The experimental group (E) exercised 90 min.d-1 (n = 12 subjects) during the BR while the control group received no intervention (n = 6). The exercise treatment consisted of a combination of isotonic and isokinetic lower extremity exercise training. Electromyographic (EMG) activity in the tibialis anterior, gastrocnemius, vastus lateralis, and biceps femoris was measured during walking before and after BR. RESULTS: Following BR, EMG activity increased in both test subject groups for all muscles (p < 0.05). However, there were no significant differences in EMG activity between the C and E groups either before or after BR. CONCLUSIONS: These data suggest that the ability of the postural control system to adjust to the gravitational environment was compromised as a result of BR. In addition, we conclude that the exercise protocol used was not an effective countermeasure to the alterations in the postural control system.

Changes in the immune system during and after spaceflight.

The results of immunological analyses before, during and after spaceflight, have established the fact that spaceflight can result in a blunting of the immune mechanisms of human crew members and animal test species. There is some evidence that the immune function changes in short-term flights resemble those occurring after acute stress, while the changes during long-term flights resemble those caused by chronic stress. In addition, this blunting of the immune function occurs concomitant with a relative increase in potentially infectious microorganisms in the space cabin environment. This combination of events results in an increased probability of inflight infectious events. The realization of this probability has been shown to be partially negated by the judicious use of a preflight health stabilization program and other operational countermeasures. The continuation of these countermeasures, as well as microbial and immunological monitoring, are recommended for continued spaceflight safety.

Psychosocial value of space simulation for extended spaceflight.

There have been over 60 studies of Earth-bound activities that can be viewed as simulations of manned spaceflight. These analogs have involved Antarctic and Arctic expeditions, submarines and submersible simulators, land-based simulators, and hypodynamia environments. None of these analogs has accounted for all the variables related to extended spaceflight (e.g., microgravity, long-duration, heterogeneous crews), and some of the stimulation conditions have been found to be more representative of space conditions than others. A number of psychosocial factors have emerged from the simulation literature that correspond to important issues that have been reported from space. Psychological factors include sleep disorders, alterations in time sense, transcendent experiences, demographic issues, career motivation, homesickness, and increased perceptual sensitivities. Psychiatric factors include anxiety, depression, psychosis, psychosomatic symptoms, emotional reactions related to mission stage, asthenia, and postflight personality, and marital problems. Finally, interpersonal factors include tension resulting from crew heterogeneity, decreased cohesion over time, need for privacy, and issues involving leadership roles and lines of authority. Since future space missions will usually involve heterogeneous crews working on complicated objectives over long periods of time, these features require further study. Socio-cultural factors affecting confined crews (e.g., language and dialect, cultural differences, gender biases) should be explored in order to minimize tension and sustain performance. Career motivation also needs to be examined for the purpose of improving crew cohesion and preventing subgrouping, scapegoating, and territorial behavior. Periods of monotony and reduced activity should be addressed in order to maintain morale, provide meaningful use of leisure time, and prevent negative consequences of low stimulation, such as asthenia and crew member withdrawal. Leadership roles and lines of authority need to be studied further to understand the factors leading to status leveling, leadership competition, and role confusion. Finally, the relationship between crews and ground personnel should be characterized in order to minimize the displacement of anger and tension to the outside, to counter the effects of inter-group miscommunications, and to develop support strategies that can help to counter in-group/out-group conflicts. Ground-based space simulations still have a role to play in terms of understanding the impact of these factors and ways of dealing with them. In particular, issues involving language, cultural differences, gender biases, career motivation, monotonous conditions, use of free time, leadership, lines of authority, and the relationship between crews and outside monitoring personnel need to be further characterized and examined under controlled conditions. Until such time as these factors can be studied directly in space, simulations provide an opportunity to learn more about these psychosocial issues and to plan ways of minimizing their negative consequences during actual space missions.

Frog experiment onboard space station Mir.

Japanese tree frogs (Hyla japonica) showed unique postures and behavior during an 8-day flight to the Russian space station Mir. When floating in the air, the animals arched their back and extended their four limbs. This posture resembles that observed during jumping or parachuting of the animals on the ground. Frog sitting on a surface bent their neck backward sharply, did not fold their hind limbs completely, and pressed their abdomen against the substrate. They walked backwards in this posture. The typical posture resembles that adopted during the emetic behavior process on the ground, although the posture in space lasts much longer. The possible mechanism of induction of this unique posture in orbit is discussed. Frogs in this posture might be in an emetic state, possibly due to motion sickness. Response behavior to some stimuli was observed in orbit. Body color change in response to the background color appeared to be delayed or slowed down. Response behavior to other stimuli showed little change as long as the animal maintained contact with a substrate. Once it left the surface, the floating frog could not control its movements so as to provide coordinated motility for locomotion and orientation. Adaptation to microgravity was observed in the landing behavior after jumping. Readaptation of the frogs to the Earth environment took place within a few hours after return. Postflight histological and biochemical analysis of organs and tissues showed some changes after the 8-day spaceflight. Weakening and density loss in vertebrae was noted. The beta-adrenoreceptor activity of the gastrocnemius was natriuretic decreased. Skin collagen and liver protein synthesis were lowered. The distribution of the atrial factor-like peptides in the brain was changed.

Incidentes fisiológicos durante entrenamiento en cámara hipobárica entre los años 1983-1992 / Physiological training in altitude chamber in the chilean airforce: years 1983-1992

En el presente trabajo, se ha analizado el Programa de Entrenamiento Fisiológico en el Departamento de Medicina Aeroespacial de la Dirección de Sanidad de la Fuerza Aérea de Chile, en lo que respecta a incidentes producidos durante el desarrollo de dicho programa y al que fueron sometidos tanto pilotos alumnos como instructores de vuelo de cámara hipobárica, durante los años 1983-1992. En este período se entrenó a 2.118 alumnos en vuelos de hipoxia y vuelos de descompresión rápida no registrándose ningún accidente, teniendo presente además, que los incidentes que se produjeron fueron sin consecuencias graves, tanto para alumnos como instructores. En el período analizado se produjeron un total de 274 incidentes en los alumnos correspondiendo a barotitis, barotitis tardía y barosinusitis, los porcentajes más altos. Asimismo, en los instructores el número total de incidentes fue de 56, siendo barotitis, barotitis tardía y barosinusitis los más frecuentes. Otros hechos importantes son la realización de 23 vuelos médicos y de 24 vuelos de prueba, todos sin consecuencia y con resultados altamente positivos. Cabe destacar que durante este período los incidentes han sido superados efectiva y oportunamente gracias a la preparación, capacidad y a las medidas de seguridad que en cada circunstancia específica se aplicaron