Physiological and Genomic Consequences of Intermittent Hypoxia: Selected Contribution: Role of spleen emptying in prolonging apneas in humans

This study addressed the interaction between short-termadaptation to apneas with face immersion and erythrocyte release from the spleen. Twenty healthy volunteers, including ten splenectomized subjects, participated. After prone rest, they performed five maximal-duration apneas with face immersion in 10°C water, with 2-minintervals. Cardiorespiratory parameters and venous blood samples werecollected. In subjects with spleens, hematocrit and hemoglobinconcentration increased by 6.4% and 3.3%, respectively, over theserial apneas and returned to baseline 10 min after the series. A delayof the physiological breaking point of apnea, by 30.5% (17 s), wasseen only in this group. These parameters did not change in thesplenectomized group. Plasma protein concentration, preapneic alveolarPCO₂, inspired lung volume, and divingbradycardia remained unchanged throughout the series in both groups.Serial apneas thus triggered the hematological changes that have been previously observed after long apneic diving shifts; they were rapidlyreversed and did not occur in splenectomized subjects. This suggeststhat splenic contraction occurs in humans as a part of the divingresponse and may prolong repeated apneas.

     

Cardiovascular and respiratory responses to apneas with and without face immersion in exercising humans.

The effect of the diving response on alveolar gas exchange was investigated in 15 subjects. During steady-state exercise (80 W) on a cycle ergometer, the subjects performed 40-s apneas in air and 40-s apneas with face immersion in cold (10 degrees C) water. Heart rate decreased and blood pressure increased during apneas, and the responses were augmented by face immersion. Oxygen uptake from the lungs decreased during apnea in air (-22% compared with eupneic control) and was further reduced during apnea with face immersion (-25% compared with eupneic control). The plasma lactate concentration increased from control (11%) after apnea in air and even more after apnea with face immersion (20%), suggesting an increased anaerobic metabolism during apneas. The lung oxygen store was depleted more slowly during apnea with face immersion because of the augmented diving response, probably including a decrease in cardiac output. Venous oxygen stores were probably reduced by the cardiovascular responses. The turnover times of these gas stores would have been prolonged, reducing their effect on the oxygen uptake in the lungs. Thus the human diving response has an oxygen-conserving effect.     

Diving response and arterial oxygen saturation during apnea and exercise in breath-hold divers.

This study addressed the effects of apnea in air and apnea with face immersion in cold water (10 degrees C) on the diving response and arterial oxygen saturation during dynamic exercise. Eight trained breath-hold divers performed steady-state exercise on a cycle ergometer at 100 W. During exercise, each subject performed 30-s apneas in air and 30-s apneas with face immersion. The heart rate and arterial oxygen saturation decreased and blood pressure increased during the apneas. Compared with apneas in air, apneas with face immersion augmented the heart rate reduction from 21 to 33% (P < 0.001) and the blood pressure increase from 34 to 42% (P < 0.05). The reduction in arterial oxygen saturation from eupneic control was 6.8% during apneas in air and 5.2% during apneas with face immersion (P < 0.05). The results indicate that augmentation of the diving response slows down the depletion of the lung oxygen store, possibly associated with a larger reduction in peripheral venous oxygen stores and increased anaerobiosis. This mechanism delays the fall in alveolar and arterial PO(2) and, thereby, the development of hypoxia in vital organs. Accordingly, we conclude that the human diving response has an oxygen-conserving effect during exercise.     

Increased pulmonary vascular resistance and reduced stroke volume in association with CO2 retention and inferior vena cava dilatation.

Changes in cardiovascular parameters elicited during a maximal breath hold are well described. However, the impact of consecutive maximal breath holds on central hemodynamics in the postapneic period is unknown. Eight trained apnea divers and eight control subjects performed five successive maximal apneas, separated by a 2-min resting interval, with face immersion in cold water. Ultrasound examinations of inferior vena cava (IVC) and the heart were carried out at times 0, 10, 20, 40, and 60 min after the last apnea. The arterial oxygen saturation level and blood pressure, heart rate, and transcutaneous partial pressures of CO(2) and O(2) were monitored continuously. At 20 min after breath holds, IVC diameter increased (27.6 and 16.8% for apnea divers and controls, respectively). Subsequently, pulmonary vascular resistance increased and cardiac output decreased both in apnea divers (62.8 and 21.4%, respectively) and the control group (74.6 and 17.8%, respectively). Cardiac output decrements were due to reductions in stroke volumes in the presence of reduced end-diastolic ventricular volumes. Transcutaneous partial pressure of CO(2) increased in all participants during breath holding, returned to baseline between apneas, but remained slightly elevated during the postdive observation period (approximately 4.5%). Thus increased right ventricular afterload and decreased cardiac output were associated with CO(2) retention and signs of peripheralization of blood volume. These results indicate that repeated apneas may cause prolonged hemodynamic changes after resumption of normal breathing, which may suggest what happens in sleep apnea syndrome.     

Bronchoconstriction: a component of the 'diving response' in man

We have investigated the possibility that a bronchoconstriction accompanies the vagally-mediated bradycardia induced in man by immersion of the face in cold water. Forced expiratory flows (FEF) at 40% and 25% of vital capacity (VC) have been measured from partial flow-volume curves obtained during forced expirations starting at 70% VC. These were performed immediately after 15 s apnoea with or without face immersion, and compared with control expirations having the same volume history but without the preceding apnoea. Five of the 10 subjects showed evidence of a greater than 10% reduction in FEF, which averaged 17% (Fig. 2). Half the response was attributable to the apnoea alone and the other half, which was abolished by ipratropium, to cold face immersion (Fig. 3). This bronchoconstriction appears to be a new component of the 'diving response' in man, mediated, like the bradycardia, by vagal efferents.     

The human spleen as an erythrocyte reservoir in diving-related interventions.

Twelve subjects without and ten subjects with diving experience performed short diving-related interventions. After labeling of erythrocytes, scintigraphic measurements were continuously performed during these interventions. All interventions elicited a graduated and reproducible splenic contraction, depending on the type, severity, and duration of the interventions. The splenic contraction varied between approximately 10% for "apnea" (breath holding for 30 s) and "cold clothes" (cold and wet clothes applied on the face with no breath holding for 30 s) and approximately 30-40% for "simulated diving" (simulated breath-hold diving for 30 s), "maximal apnea" (breath holding for maximal duration), and "maximal simulated diving" (simulated breath-hold diving for maximal duration). The strongest interventions (simulated diving, maximal apnea, and maximal simulated diving) elicited modest but significant increases in hemoglobin concentration (0.1-0.3 mmol/l) and hematocrit (0.3-1%). By an indirect method, the splenic venous hematocrit was calculated to 79%. No major differences were observed between the two groups. The splenic contraction should, therefore, be included in the diving response on equal terms with bradycardia, decreased peripheral blood flow, and increased blood pressure.     

Repetitive apneas induce periodic hypertension in normal subjects through hypoxia.

Periodic increases in blood pressure (BP) can occur in the sleep apnea syndrome (SAS) during recurrent apneas. To investigate the mechanisms causing this periodic hypertension, we simulated SAS by imposing a matching breathing pattern on seven healthy awake male volunteers. Continuous finger arterial BP, electrocardiogram, arterial O2 saturation (SaO2), end-tidal CO2, and tidal volume were measured. The role of hypoxia was studied by comparing apneas during depletion of O2 in the spirometer with those during 100% O2 breathing. In all subjects, BP periodically reached values greater than 150/95 mmHg in the hypoxic series. During the hyperoxic apnea series, however, BP remained stable. End-apneic mean BP was shown to be inversely correlated to SaO2 in six subjects in the SaO2 range from 60 to 100%. Although the hypoxic BP pattern closely mimicked that in SAS, the heart rate pattern in four of our subjects remained distinct from that in patients. Atropine could not prevent large BP swings in the hypoxic series. We conclude that SaO2 is a major determinant of periodic hypertension in recurrent apneas. Its effect probably results from chemoreflex modulation of peripheral resistance.     

Scuba diving enhances endogenous antioxidant defenses in lymphocytes and neutrophils

The aim was to study the effects of a scuba diving session on the lymphocyte antioxidant system, NO synthesis, the capability to produce reactive oxygen species and the antioxidant response in neutrophils. For that purpose seven male divers performed an immersion at a depth of 40 m for 25 min. The same parameters were measured after an hyperbaric oxygen (HBO) treatment at resting conditions in a hyperbaric chamber. Lymphocyte H₂O₂ production rose after diving and after HBO treatment. Glutathione peroxidase (GPx) and catalase activities increased after diving in lymphocytes, while after HBO exposure only increased GPx activity. Lymphocyte HO-1 mRNA expression increased after diving and after HBO exposure, while iNOS levels and nitrite levels significantly increased after diving. The hyperoxia associated to scuba diving leads to a condition of oxidative stress with increased lymphocyte H₂O₂ production, HO-1 expression, NO synthesis and antioxidant enzyme adaptations in order to avoid oxidative damage.     

Body cooling and the diving capabilities of muskrats (Ondatra zibethicus): a test of the adaptive hypothermia hypothesis

We tested the hypothesis that immersion hypothermia enhances the diving capabilities of adult and juvenile muskrats by reducing rates of oxygen consumption (V O2). Declines in abdominal body temperature (T(b)) comparable to those observed in nature (0.5-3.5 degrees C) were induced by pre-chilling animals in 6 degrees C water. Pre-chilling did not reduce diving V O2 of any animal tested in 10 degrees C or 30 degrees C water, irrespective of the nature of the dive. Most behavioural indices of dive performance, including average and cumulative dive times, were unaffected by T(b) reduction in adults, but depressed in hypothermic juveniles (200-400 g). Hypothermia reduced diving heart rate only on short (<25s) dives (16% reduction, P=0.01), but did not affect the temporal onset of diving bradycardia. Post-immersion V O2 was higher for pre-chilled than for normothermic muskrats, but the difference became insignificant on longer (>90 s) dives. Our findings suggest that the mild hypothermia experienced by muskrats in nature has minimal effect on diving and post-immersion metabolic costs, and thus has little impact on the dive performance of this northern semi-aquatic mammal.     

Changes in heart rate during breathing interrupted by recurrent apneas in humans

Heart rate varies with breathing patterns, especially in sleep apnea. To assess the effects on heart rate of recurrent apneas interrupting tidal breathing, we studied five normal awake male subjects. These subjects voluntarily changed their breathing pattern from regular tidal breathing to tidal breathing interrupted by breath holding at end expiration. This recurrent apneic breathing pattern did not change mean heart rate but increased its variance significantly. In addition, the variations in heart rate formed a cyclic pattern of oscillation with a mean cycle length identical to both arterial O2 saturation (SaO2) (R = 0.95; P less than 0.01) and ventilation (R = 0.92; P less than 0.01). Cyclic changes in either SaO2 or ventilation reproduced the oscillatory patterns of heart rate seen with tidal breathing interrupted by multiple apneas, but the amplitude of the variance in heart rate was smaller. Finally, preventing the cyclic declines in SaO2 with supplemental O2 did not significantly alter the heart rate changes seen in tidal breathing interrupted by apneas.     

Effect of head-out immersion on plasma atrial natriuretic factor in man

This study was conducted to examine the role of atrial natriuretic factor (ANF) in the development of diuresis and natriuresis in response to the head-out immersion in 35 degrees C water. Six male subjects were hydrated (0.5% body wt), sat for 1 hr in air (preimmersion), were immersed in water to the neck for 3 hr, and then sat for 1 hr in air (postimmersion). In another series they were similarly hydrated and then sat for 5 hr in air for the time control. Urine and venous blood samples were collected hourly for creatinine and electrolyte measurements. In addition, the concentration of ANF was determined in unextracted plasma by a radioimmunoassay. The pattern of electrolyte excretion was evaluated on the basis of fractional excretion of filtered load. In the time control series, urine flow and fractional excretion of Na and K remained low throughout the 5-hr experimental period. On the other hand, urine flow increased significantly from the preimmersion level of approximately 2 to approximately 7 ml/min during the first hour of immersion (P less than 0.05), after which it decreased to approximately 5 ml/min during the second hour of immersion (P less than 0.05) and to approximately 2 ml/min during the third hour of immersion. Fractional excretion of Na increased continuously from preimmersion level of approximately 1.0 to approximately 1.8% during the second and third hours of immersion (P less than 0.05) and then decreased to 1.2% during the 1-hr postimmersion period. The plasma ANF remained low (approximately 75 pg/ml) during the 5-hr time control period. In the immersion series, plasma ANF increased significantly from the preimmersion level of approximately 80 to approximately 120 pg/ml during the entire 3-hr immersion period and then returned to the preimmersion level during 1 hr postimmersion. These results indicate that the immersion diuresis and natriuresis are indeed associated with the increased ANF release. However, it can not be ascertained from the present study if the increased ANF contributes directly to these renal responses to immersion or in concert with other mediators.     

Scuba diving enhances endogenous antioxidant defenses in lymphocytes and neutrophils

The aim was to study the effects of a scuba diving session on the lymphocyte antioxidant system, NO synthesis, the capability to produce reactive oxygen species and the antioxidant response in neutrophils. For that purpose seven male divers performed an immersion at a depth of 40 m for 25 min. The same parameters were measured after an hyperbaric oxygen (HBO) treatment at resting conditions in a hyperbaric chamber. Lymphocyte H₂O₂ production rose after diving and after HBO treatment. Glutathione peroxidase (GPx) and catalase activities increased after diving in lymphocytes, while after HBO exposure only increased GPx activity. Lymphocyte HO-1 mRNA expression increased after diving and after HBO exposure, while iNOS levels and nitrite levels significantly increased after diving. The hyperoxia associated to scuba diving leads to a condition of oxidative stress with increased lymphocyte H₂O₂ production, HO-1 expression, NO synthesis and antioxidant enzyme adaptations in order to avoid oxidative damage.     

Changes in the jugular haematocrit of sheep during feeding.

Intact and splenectomized sheep with and without a rumen fistula were used to investigate changes in the jugular blood haematocrit and plasma osmolality during hourly and once-daily feeding regimes. Osmolality was also estimated in the ruminal fluid of fistulated sheep with spleens. Haematocrit decreased in sheep with spleens before they were given a once-daily feed; it increased when these sheep started to feed, reaching a maximum increase of 13% after 30 min of feeding; it decreased during the remaining 45 min of feeding time and usually continued to decrease after feeding stopped. These changes were not due to diurnal influences. Splenectomized sheep fed once daily showed only small decreases in haematocrit before they were fed. Increases occurred with the onset of eating but they were smaller (7%) than in intact sheep and were of shorter duration. In hourly fed sheep with spleens, haematocrit decreased in the early stages of sampling in a manner similar to that for sheep fed once daily. The changes in haematocrit that did occur were not related in any obvious manner to the feeding regime. The haematocrit in splenectomized sheep fed hourly was stable throughout feeding. Variations in the haematocrit in splenectomized sheep, equivalent to a range of 13% in one of them, were observed in a series of blood samples obtained during a 5-h period remote from the feeding time. Large increases occurred in osmolality of ruminal fluid when sheep were fed daily and this was abolished by hourly feeding. Plasma osmolality in sheep fed once daily increased slowly. Maxima occurred after 100 min from the start of eating and were 7% greater than prefeeding values. Only minor changes were observed when these sheep were fed hourly.     

Plasma fibronectin (Fn) study in homozygous beta-thalassemia: relation to splenectomy and transfusion

The concentration of plasma Fn was determined in non-splenectomized and splenectomized patients with homozygous beta-thalassemia, before and 7-10 days after blood transfusion. The mean Fn concentration of non-splenectomized patients before transfusion did not differ from that of matched normal controls but appeared significantly decreased following blood transfusion. On the other hand, in splenectomized thalassemics, Fn levels were increased but were unrelated to transfusion. It is concluded that Fn plays some homeostatic function when RES activity of thalassemic patients is altered either as a result of splenectomy or blood transfusion.     

Splenic contraction during breath-hold diving in the Korean ama

Major increases of hemoglobin concentration and hematocrit, possibly secondary to splenic contraction, have been noted during diving in the Weddell seal. We sought to learn whether this component of the diving response could be present in professional human breath-hold divers. Splenic size was measured ultrasonically before and after repetitive breath-hold dives to approximately 6-m depth in ten Korean ama (diving women) and in three Japanese male divers who did not routinely practice breath-hold diving. Venous hemoglobin concentration and hematocrit were measured in nine of the ama and all Japanese divers. In the ama, splenic length and width were reduced after diving (P = 0.0007 and 0.0005, respectively) and calculated splenic volume decreased 19.5 +/- 8.7% (mean +/- SD, P = 0.0002). Hemoglobin concentration and hematocrit increased 9.5 +/- 5.9% (P = 0.0009) and 10.5 +/- 4% (P = 0.0001), respectively. In Japanese male divers, splenic size and hematocrit were unaffected by repetitive breath-hold diving and hemoglobin concentration increased only slightly over baseline (3.0 +/- 0.6%, P = 0.0198). Splenic contraction and increased hematocrit occur during breath-hold diving in the Korean ama.     

Changes in arterial blood pressure, heart rate and haematocrit during acute hyperkalaemia in conscious sheep.

The systolic, diastolic and mean arterial blood pressures, heart rate and haematocrit were measured at 15 minute intervals before, during and after 2 hour infusions of 0-4 mol.l-1 NaCl at 2-2 ml min-1 into conscious intact sheep and 0-4 mol. l-1 KCl at 2-2 ml. min-1 into conscious sheep which were either intact or adrenalectomized. The haemotocrit was also measured in splenectomized sheep receiving 0-4 mol. l-1 KCl. The NaCl infusion had no significant effect on blood pressure(BP), heart rate and haematocrit. Both intact and adrenalectomized sheep were able to withstand an increase in plasma potassium concentration in excess of 50% of the preinfusion concentration before any substantial fall in BP occurred. In intact and adrenalectomized sheep, heart rate and haematocrit increased rapidly and progressively throughout the potassium infusions and at maximum plasma potassium concentration the mean increments in these parameters for both groups of sheep were 21-6+/-2-69 beats/min and 7-5+/-0-47% respectively. Heart rate and haematocrit were more closely correlated with the plasma potassium concentration than with any other variable measured in these experiments. Adrenalectomy did not reduce the ability of the sheep to maintain their BP or to increase their heart rate and haematocrit. As the mean increase in haematocrit during potassium infusion into splenectomized sheep was 1-3+/-0-45% most of the increase in haematocrit observed in the potassium-infused intact and adrenalectomized sheep was caused by ejection of red cells from the spleen into the circulation.     

Peripheral vascular resistance increases after termination of obstructive apneas.

The mechanisms by which obstructive apneas produce intermittent surges in arterial pressure remain poorly defined. To determine whether termination of obstructive apneas produce peripheral vasoconstriction, we assessed forearm blood flow during and after obstructive events in sleeping patients experiencing spontaneous upper airway obstructions. In all subjects, heart rate was monitored with an electrocardiogram and blood pressure was monitored continuously with digital plethysmography. In 10 patients (protocol 1), we used forearm plethysmography to assess forearm blood flow, from which we calculated forearm vascular resistance by performing venous occlusions during and after obstructive episodes. In an additional four subjects, we used simultaneous Doppler and B-mode images of the brachial artery to measure blood velocity and arterial diameter, from which we calculated brachial flow continuously during spontaneous apneas (protocol 2). In protocol 1, forearm vascular resistance increased 71% after apnea termination (29.3 +/- 15.4 to 49.8 +/- 26.5 resistance units, P < 0.05) with all patients showing an increase in resistance. In protocol 2, brachial resistance increased at apnea termination in all subjects (219.8 +/- 22.2 to 358.3 +/- 46.1 mmHg x l(-1) x min; P = 0.01). We conclude that termination of obstructive apneas is associated with peripheral vasoconstriction.     

Plasmodium berghei: The spleen in sporozoite-induced immunity to mouse malaria

A/J mice were splenectomized (Splx) or sham-splenectomized (SSplx) prior to administration of a single injection of irradiated sporozoites. Following challenge 7 days after immunization, it was found that none of the splenectomized mice were protected whereas 50% of the sham-splenectomized and intact animals were resistant to challenge. In another series of experiments similar groups, along with mice splenectomized just prior to challenge, were injected with 1.5 × 10⁵ irradiated sporozoites over a 5 week period. This resulted in protection of (1) 60–100% of the animals splenectomized before immunization, and (2) 90–100% protection of the animals splenectomized prior to challenge, as well as the intact and sham-splenectomized mice. Serum levels of antisporozoite antibodies (CSP and SNA) increased during immunization of the intact animals. Only 15–20% of the animals splenectomized prior to immunization presented positive CSP reactions and little if any sporozoite neutralizing activity (SNA) was detected. Serum from intact animals immunized and found resistant to sporozoite challenge was used for passive transfer studies. Immune serum recipients were challenged with small numbers of sporozoites. Only one out of 18 recipients was protected against sporozoite challenge.     

Stress-induced hormonal and mood responses in scuba divers: a field study

The majority of injuries in scuba-divers are attributable to inappropriate behavior under stressful diving conditions, predominantly involving panic reactions emerging from elevated levels of anxiety. Divers with an elevated level of anxiety and poor coping are at higher risk of developing panic reactions than those possessing more adequate stress-coping-mechanisms. In the comparison of two extreme groups of seven divers each with opposite stress coping strategies, prolactin was found to be a hormonal marker with a significant increase in the sub-group of the stress-controllers. This hormonal response was observed in a recreational and a stressful dive, and in the latter with a more distinct elevation. Along with the self-reported emotional conditions under immersion, these data suggest that an increased prolactin level reflects a state of elevated physical and mental activation and vigilance. Facing a stressful situation subjects with more emotional concern and the tendency to surrender react by "blunted responses" and show significantly lower elevations of the prolactin levels in contrast to subjects with the very opposite psychological features. The other observed somatic parameters (epinephrine, norepinephrine) showed significant increases during and after dives (with the exception of saliva cortisol), however without any significant group difference.     

The influence of support withdrawal on presaccadic EEG potentials in subjects with different asymmetry profiles

Characteristics of saccades and parameters of slow presaccadic potentials were studied in 12 volunteers, including seven subjects with a leading right eye (the RE group) and five subjects with a leading left eye (the LE group) before and on the sixth day of dry immersion. For visual stimulation, three light-emitting diodes were used; one of them was located in the center of the visual field, and two other, 10° away from the horizontal axis to the right and left of the first one. The subjects performed an anti-saccade test, which included making saccades at the spot that was symmetrically located in the visual field opposite to the stimulus. The EEG (19 standard unipolar derivations) and electrooculogram were recorded. To obtain slow presaccadic potentials (PSPs), backward averaging triggered by switching on a peripheral stimulus was performed. Before the immersion, there were no significant differences in the characteristics of saccades in both groups of subjects. At the same time, the amplitude of presaccadic negativity (PSN) in the LE group was decreased, especially in the frontal region, and had considerable asymmetry during the analysis. During the immersion, the latent periods of the saccades and the percentage of incorrect reactions did not change in RE subjects and were increased in LE subjects. Both groups demonstrated a decrease in the PSN amplitude and its shift to the right hemisphere; intergroup differences decreased in immersion conditions. The characteristic feature of the RE group was a significant decrease in PSN in frontal leas at immersion, apparently caused by sensory disintegration and a decrease in the tonic afferent input. In the LE group, the maximal amplitude of PSN was observed in the central region.