Avian embryos in hypoxic environments.

Avian embryos at high altitude do not benefit of the maternal protection against hypoxia as in mammals. Nevertheless, avian embryos are known to hatch successfully at altitudes between 4,000 and 6,500 m. This review considers some of the processes that bring about the outstanding modifications in the pressure differences between the environment and mitochondria of avian embryos in hypoxic environments. Among species, some maintain normal levels of oxygen consumption ( VO2) have a high oxygen carrying capacity, lower the air cell-arterial pressure difference ( PAO2 - PaO2 ) with a constant pH. Other species decrease VO2, increase only slightly the oxygen carrying capacity, have a higher PAO2 - PaO2 difference than sea-level embryos and lower the PCO2 and pH. High altitude embryos, and those exposed to hypoxia have an accelerated decline of erythrocyte ATP levels during development and an earlier stimulation of 2,3-BPG synthesis. A higher Bohr effect may ensure high tissue PO2 in the presence of the high-affinity hemoglobin. Independently of the strategy used, they serve together to promote suitable rates of development and successful hatching of high altitude birds in hypoxic environments.

Effect of domperidone on ventilation and polycythemia after 5 weeks of chronic hypoxia in rats.

Chronically hypoxic humans and some mammals have attenuated ventilatory responses, which have been associated with high dopamine level in carotid bodies. Alveolar hypoventilation and blunted ventilatory response have been recognized to be at the basis of Chronic Mountain Sickness by generating arterial hypoxemia and polycythemia. To investigate whether dopamine antagonism could decrease the hemoglobin concentration by stimulating resting ventilation (VE) and/or hypoxic ventilatory response, 18 chronically hypoxic rats (5 weeks, PB=433 Torr) were studied with and without domperidone treatment (a peripheral dopamine antagonist). Acute and prolonged treatment significantly increased poikilocapnic ventilatory response to hypoxia (RVE ml/min/kg=VE at 0.1 FI(O(2))-VE at 0.21 FI(O(2))), from 506+/-36 to 697+/-48; and from 394+/-37 to 660+/-81, respectively. In addition, Domperidone treatment decreased hemoglobin concentration from 21.6+/-0.29 to 18.9+/-0.19 (P<0.01) in rats chronically exposed to hypobaric hypoxia. Our study suggests that the stimulant effect of D(2)-R blockade on ventilatory response to hypoxia seems to compensate the low hypoxic peripheral chemosensitivity after chronic exposure and the latter in turn decrease hemoglobin concentration.

Ventilatory and cardiovascular responses to hypoxia and exercise in Andean natives living at sea level.

This study was designed to determine in subjects born at high altitude who move to sea level (HA-SL: born at 3500 m or above; n = 25) whether their cardiorespiratory responses to hypoxia and exercise are similar to those of sea level natives (SL,n = 25). The average age (39 +/- 7.3 yr), weight (72 +/- 7.3 kg), and height (1.71 +/- 0.01 m) did not differ between the SL and HA-SL subjects. All subjects were studied at rest or during exercise (60 W on cycle ergometer) while breathing room air (F(IO2) = 0.21 and P(B) = 760) or hypoxia (F(IO2) = 0.115 and PB = 760) in the following order: (1) normoxia at rest (NX-Rs), (2) hypoxia at rest (HX-Rs, 11.5% O(2)), hypoxia at exercise (HX-Ex), and normoxia at exercise (NX-Ex). Each period lasted 5 min. In absolute values, HA-SL showed significantly higher ventilation (V(E), L/min) during exercise in both normoxia and hypoxia and higher oxygen saturation (Sa(O2), %) during hypoxia both at rest and in exercise. They also had lower end-tidal CO(2) values (P(ETCO2), torr) at rest in both normoxia and hypoxia, but a higher P(ETCO2) in hypoxic exercise. Heart rate (HR, beats/min) was lower at rest in both normoxia and hypoxia, but higher in exercise. With acute hypoxia, Sa(O2) decreased less in the HA-SL than in the SL at rest (HA-SL, 9.2 +/- 0.8; SL, 12.0 +/- 0.82) and during exercise (HA-SL, 18.3 +/- 1.1; SL, 21.2 +/- 1.2). In conclusion, this study shows that HA-SL natives have increased ventilation and heart rate during exercise once their lifelong hypoxia is relieved.

Energetic metabolism in mouse cerebral cortex during chronic hypoxia.

We measured the activities of Na(+)K(+) ATPase and of enzymes of the glycolytic pathway, Krebs cycle, and the respiratory chain in cerebral cortex of mice exposed to chronic hypoxia for three weeks and compared their values with those of sea level controls. There were no differences in Na(+)K(+) ATPase activity or in the activity of glycolytic enzymes. In the Krebs cycle, a 66% increase of succinate dehydrogenase activity was found due to a lower Km. In contrast, respiratory chain cytochrome oxidase activity was reduced by 12% in mice exposed to hypoxia. This suggested that the metabolic demand would be satisfied despite the respiratory chain depression (cytochrome oxidase), probably due to anaerobic energy production within the mitochondria (succinate dehydrogenase).

Relationship of ovarian hormones to hypoxemia in women residents of 4,300 m.

Prevalence of excessive erythrocytosis, the main sign of chronic mountain sickness (CMS), is greater in postmenopausal Andean women than in premenopausal women. It is uncertain whether this greater prevalence is related to the decline in female hormones and ventilatory function after the occurrence of the menopause. To study this, we compared the physiological variables involved in the physiopathology of CMS [end-tidal CO(2) (PET(CO(2)), Torr) and end-tidal O(2) (PET(O(2)), Torr), arterial oxygen saturation (Sa(O(2)), %), and Hb concentration (g/dl)] and progesterone and estradiol levels between postmenopausal and premenopausal women, both in the luteal and follicular phases. Women residing in Cerro de Pasco (n = 33; 4,300 m) aged 26--62 yr were studied. Postmenopausal women compared with premenopausal women in the luteal phase had lower PET(O(2)) (48 +/- 4 vs. 53 +/- 2 Torr, P = 0.005) and Sa(O(2)) levels (82 +/- 12 vs. 88 +/- 12%, P < 0.005) and higher PET(CO(2)) (34 +/- 2 vs. 29 +/- 3 Torr, P = 0.005) and Hb concentration (19 +/- 1 vs. 14 +/- 2 g/dl, P < 0.005). In addition, plasma progesterone was negatively correlated with PET(CO(2)) and positively correlated with PET(O(2)) and Sa(O(2)). No clear relationship was found among the cycle phases between estradiol and the variables studied. In conclusion, our results reveal that, before menopause, there is better oxygenation and lower Hb levels in women long residing at altitude, and this is associated with higher levels of progesterone in the luteal phase of the cycle.

Hematological parameters in high altitude residents living at 4,355, 4,660, and 5,500 meters above sea level.

There have been a number of reports describing the hematological indicators of Andean residents living at altitudes above 4,000 m, but several confounding factors have made the published results difficult to interpret. To clear up the effect of hypoxia on hemoglobin concentration (Hb, g/dL), hematocrit (Hct, %) and red blood cell concentration (RBC, cells/microL), this publication describes and analyzes these variables in children, men, and women from three large and homogeneous populations living at 4,355 m (n = 151), 4,660 m (n = 400), and 5,500 m (n = 273) in the Southern Peruvian Andes. Hb, Hct, and RBC increase with age in men (p < 0.001), as well as in women (p < 0.001) at the three altitudes of the study. In children (boys and girls) living at 5,500, Hb increases 11% when compared with children living at 4,355 m, and in adults, Hb increases 9.6% when comparing the same altitudes. The maximum percentage increase in Hb with age was 5.6% at 5,500 m, in men and 3.2% at 4,355 m, in women. The average percentage of difference for the Hb concentration between adult men and women is 6.6% at 4,355 m, 9.8% at 4,660 m, and 11.6% at 5,500 m. The differences in Hb concentration between men and women can only be seen after puberty. Finally, Hb is higher in older than younger women, which confirms the role of menopause in the development of erythremia. The result of this analysis reinforces the notion that Hb and Hct seem to be stable and useful parameters for acclimatization only at moderate altitudes; with aging or with increasing altitude, they may become excessive and lose their efficiency to protect the venous oxygen pressure.

Carbonic anhydrase activity in the red blood cells of sea level and high altitude natives.

Red blood cell carbonic anhydrase (CA) activity has not been studied in high altitude natives. Because CA is an intraerythocytic enzyme and high altitude natives are polycythemic, it is important to know if the activity of CA per red cell volume is different from that of their sea level counterparts. Blood was collected from healthy subjects living in Lima (150m) and from twelve subjects from Cerro de Pasco (4330m), and hematocrit and carbonic anhydrase activity were measured. As expected, the high altitude natives had significantly higher hematocrits than the sea level controls (p = 0.0002). No difference in the CA activity per milliliter of red cells was found between the two populations. There was no correlation between the hematocrit and CA activity.

Reduced oxygen diffusion across the shell of Gray gull (Larus modestus) eggs.

Gray gulls, Larus modestus, nest 1500 m above sea level in northern Chile's Atacama Desert, one of the driest in the world. Their eggshell gas permeability, one third of that found in other Larus species, is an adaptation that reduces water loss, but at the expense of oxygen diffusion into the air cell with resultant hypoxia and reduced metabolic rate. This contrasts with characteristics found in birds nesting at very high altitudes where oxygen diffusion across the egg shell is maximized at the expense of water conservation. The oxygen consumption (MO2) of Larus modestus is 66% that of Larus argentatus; the oxygen conductance (GO2) is equivalent to 48% of that obtained in 5 other bird species. The oxygen partial pressure (PAO2) in the air chamber of Larus modestus (84 Torr) is lower than that of 10 other bird species whose average (PAO2) is 106 Torr. The CO2 partial pressure (PACO2) in the air chamber of Larus modestus is 68 Torr, a higher value than that found in 9 other bird species whose average (PACO2) is 39 Torr.

Reduced oxygen diffusion across the shell of Gray gull (Larus modestus) eggs

Gray gulls, Larus modestus, nest 1500 m above sea level in northern Chile's Atacama Desert, one of the driest in the world. Their eggshell gas permeability, one third of that found in other Larus species, is an adaptation that reduces water loss, but at the expense of oxygen diffusion into the air cell with resultant hypoxia and reduced metabolic rate. This contrasts with characteristics found in birds nesting at very high altitudes where oxygen diffusion across the egg shell is maximized at the expense of water conservation. The oxygen consumption (MO2) of Larus modestus is 66% that of Larus argentatus; the oxygen conductance (GO2) is equivalent to 48% of that obtained in 5 other bird species. The oxygen partial pressure (PAO2) in the air chamber of Larus modestus (84 Torr) is lower than that of 10 other bird species whose average (PAO2) is 106 Torr. The CO2 partial pressure (PACO2) in the air chamber of Larus modestus is 68 Torr, a higher value than that found in 9 other bird species whose average (PACO2) is 39 Torr.

Carbonic anhydrase activity in the red blood cells of sea level and high altitude natives

Red blood cell carbonic anhydrase (CA) activity has not been studied in high altitude natives. Because CA is an intraerythocytic enzyme and high altitude natives are polycythemic, it is important to know if the activity of CA per red cell volume is different from that of their sea level counterparts. Blood was collected from healthy subjects living in Lima (150m) and from twelve subjects from Cerro de Pasco (4330m), and hematocrit and carbonic anhydrase activity were measured. As expected, the high altitude natives had significantly higher hematocrits than the sea level controls (p = 0.0002). No difference in the CA activity per milliliter of red cells was found between the two populations. There was no correlation between the hematocrit and CA activity.

Similar hypoxic ventilatory responses in sea-level natives and high-altitude Andean natives living at sea level.

High-altitude (HA) natives have blunted ventilatory sensitivities to hypoxia, and it is uncertain whether this blunting is reversible on migration to sea level (SL). To study this, the ventilatory sensitivities to hypoxia of HA natives residing near SL were compared with those of SL natives. Two studies were performed. In study A, 24 HA subjects who had lived above 3,000 m for an average of 14 yr and had been resident at SL for an average of 23 yr were compared with 23 SL controls. In study B, 25 HA subjects who had lived above 3,500 m for at least 20 yr and had been resident at SL for no more than 5 yr were compared with 25 SL controls. Hypoxic sensitivities were assessed by breathing seven progressively more hypoxic gas mixtures that contained progressively more CO2 in an attempt to maintain isocapnia throughout. The ventilatory sensitivities to hypoxia (l . min-1 . %-1 . m-2) did not differ significantly (by analysis of variance) between HA and SL natives in either study A (-0.51 +/- 0.25, mean +/- SD) or study B (-0.34 +/- 0. 15), but the ventilatory sensitivities did differ significantly between the two studies for reasons which are not entirely clear. We conclude that HA natives residing at SL, even if previously at HA for >20 yr, do not maintain the severely blunted hypoxic responses that have been reported in such individuals.

Pentoxifylline and enalapril and its effects on polycythemia induced by hypobaric hypoxia in mice

Se estudió 70 ratones Swiss macho de 3-4 meses de edad, para evaluar los efectos profilácticos y terapéuticos del analapril y la pentoxifilina en la policitemia inducida por hipoxia. Se dividió a los animales en dos grupos: grupo pentoxifilina (n=39) y grupo enalapril (n=31). Cada uno fue adicionalmente dividido en un grupo profiláctico que recibió el medicamento antes de la exposición a hipoxia hipobárica intermitente (IHH) y en un grupo terapéutico que recibió el medicamento luego de la exposición a IHH. Cada Subgrupo tuvo su respectivo control. La exposición a IHH fue realizada a través de una cámara hipobárica que simulaba una altura equivalente a 4500m, 22 horas por día. Se midió semanalmente peso y hematocrito. La evolución del peso corporal en el tiempo no mostró diferencias sustanciales entre los animales tratados y los controles en los grupos profilácticos y terapéuticos, tanto en los grupos pentoxifilina como enalapril. Hubo una disminución significativa del hematocrito a los 36 y 47 días del inicio de la profilaxis en el grupo pentoxifilina. En el grupo profiláctico enapril los hematocritos fueron significativamente menores en los animales tratados. Concluimos que ambas drogas son efectivas cuando son usadas profilácticamente antes de la exposición a IHH. Se sugiere que las drogas podrían ejercer su efecto a través de un bloqueo parcial de la producción de eritropoyetina (EPO).

Efecto de la menopausia en la relación entre las presiones alveolares de O2 y CO2 y el mal de montaña crónico / Effect of the menopause in the relation among the end tidal O2 y CO2 pressures and chronic mountain sicknessm

Este estudio se llevó a cabo con el fin de determinar el rol que tiene la llegada de la menopausia en la relación entre las presiones alveolares de O2 y CO2 y el mal de montaña crónico (MMC) o enfermedad de Monge. El estudio se realizó en una muestra poblacional femenina residente de Cerro de Pasco (4300 m) entre 26-62 años de edad (n=41). Se midieron, entre otras, las variables fisiológicas involucradas en la secuencia fisiopatológica del MMC: presiones tidales de O2 y CO2 (PO2T y PCO2T, torr), saturación de oxígenos (SaO2, por ciento), y concentración de hemoglobina (Hb, g/dl). Los resultados mostraron que, en las mujeres de altura después de la menopausia, se observa una disminución de la PO2T (48.4 ñ 3.3 vs 52.7 ñ 3.1 torr, prevalencia < 0.0001) y de la SaO2 (81.9 ñ 4.12 vs 84.7 ñ 3.14 por ciento, prevalencia 0.001), así como un aumento de la PCO2T (34 ñ 2.2 vs 30.4 ñ 3.3 torr, prevalencia < 0.0001) y de la Hb (18.6 ñ 1.7 vs 15.9 ñ 2.3 g/dl; prevalencia < 0.001). En las poblaciones masculinas de altura se observa un aumento de la prevalencia de eritrocitosis excesiva en forma continua con la edad, en el caso de las mujeres de altura, postulamos, en base a nuestros resultados, que un factor contribuyente adicional para la aparición abrupta (y no continua) del MMC sería la disminución de la ventilación, como lo indica el aumento de la PCO2T. Esta, al disminuir, traería como consecuencia hipoxemia y eritrocitosis excesiva, el signo preponderante del MMC.

Alveolar Pco2 and Po2 of high-altitude natives living at sea level.

This study was designed to determine whether subjects born at high altitude (HA; 2,000 m or above) who subsequently move to near sea level (SL) develop end-tidal PCO2 (PETCO2) and PO2 (PETO2) values that equal those of SL natives living near SL. A total of 108 male HA natives living near SL were identified by survey of a district in Lima, Peru, and a further 108 male SL natives from the same district were identified as control subjects. Of these subjects, satisfactory data for inclusion in the study were obtained from 93 HA and 82 SL subjects. Mean PETCO2 and PETO2 values were 37.7 +/- 2.5 (SD) and 104.7 +/- 3.2 Torr, respectively, in HA subjects and 37.7 +/- 2.2 and 104.8 +/- 3.0 Torr, respectively, in SL subjects. The average difference between SL natives and HA natives for PETCO2 was 0.07 Torr (-0.64 to 0.78; 95% confidence interval) and for PETO2 was 0.05 Torr (-0.89 to 0.99, 95% confidence interval). The average age and weight of the SL and HA subjects did not differ, but the HA subjects were shorter and tended to have larger vital capacities, consistent with their origin at HA. We conclude that the PETCO2 and PETO2 near SL of SL natives and HA natives do not differ.

Time-course of the polycythemic response in normoxic and hypoxic mice with high blood oxygen affinity induced by cyanate administration.

The Hb-O2 affinity and the erythropoietic response as a function of time were studied in mice treated with sodium cyanate for up to 2 months. Cyanate increased the Hb-O2 affinity in normoxic mice more than in chronically hypoxic mice. The hemoglobin concentration rose as a function of time both in normoxic and hypoxic conditions but reached higher levels in hypoxia. After 42 days of study (21 days of hypoxia) hemoglobin reached maximum levels and thereafter showed a plateau in both cyanate and control animals. It is concluded that a chronic left-shifted oxygen dissociation curve does not avoid the development of hypoxic polycythemia in mice. Moreover, prolonged cyanate administration potentiates the erythropoietic response to chronic hypoxia. Since polycythemia is an index of tissue hypoxia, the results show that the high hemoglobin affinity did not prevent tissue hypoxia in low PO2 conditions. Results showing beneficial effects of high hemoglobin oxygen affinity induced by cyanate based on acute hypoxic expositions should be cautiously interpreted with regard to their adaptive value in animals chronically exposed to natural or simulated hypoxia.

Pathophysiology and epidemiology of chronic mountain sickness.

Chronic mountain sickness, which affects permanent residents of high altitudes, is the outcome of a progressive loss of ventilatory rate which naturally occurs with age and resulting in excessive hypoxemia and polycythemia. A theoretical model predicts the progressive failure of homeostatic control of the hemoglobin concentration when the values increase above those found at sea level. This is confirmed by lack of feedback mechanism between high altitude erythrocytosis and serum erythropoietin. The results of epidemiological studies are in agreement with the physiological findings. In a male population living at 4,300 m, an increase with age of the prevalences of excessive erythrocytosis (Hb > 213 g/l), blood oxygen saturation < 83%, headaches and a high score of symptoms of chronic mountain sickness has been found. The studies suggest the possibility that in addition to an accentuated hypoxemia, the excessive erythrocytosis may also result from an overreaction of the bone marrow to a fixed level of hypoxemia in ageing individuals.

Serum immunoreactive erythropoietin in high altitude natives with and without excessive erythrocytosis.

We report the estimation of blood hemoglobin (Hb), arterial blood oxygen saturation (SaO2), and serum immunoreactive erythropoietin (siEPO) in a group of Peruvian workers residing in Cerro de Pasco at 4300 m showing "excessive erythrocytosis" (EE, Monge's disease, chronic mountain sickness). These estimates were compared with those of humans residing either in Cerro de Pasco and showing "normal erythrocytosis" (NE) or in Lima (sea level, SL) to determine whether Hb and SaO2 are related to siEPO in high altitude (HA) natives with NE or EE. The three parameters showed statistically significant differences between HA and SL groups--the values in SL being lower. Significant differences were also found between NE and EE groups in Hb and SaO2. There was no statistical difference in siEPo between the two groups. The results indicate, therefore, that HA residents who develop EE are not distinguishable from residents who develop NE on the basis of estimates of siEPO. As a result, siEPO and Hb do not show a dose-response relationship in HA residents, and variation in EPO does not explain the striking variation in Hb at high altitudes.

Different hematologic responses to hypoxia in Sherpas and Quechua Indians.

Previous studies of the erythropoietic response to hypoxia in high-altitude natives suggest that the hematocrit and hemoglobin values in Himalayan natives (Sherpas) are lower than expected for the altitude, perhaps because of a genetic adaptation. However, differences in sampling techniques and experimental methods make comparisons difficult. Our studies were carried out to compare the erythropoietic response with the same altitude in age-matched natives of the Himalayas and Andes by the same experimental techniques. Healthy male subjects were selected in Ollagüe, Chile (n = 29, 27.3 +/- 5.9 yr) and in Khunde, Nepal (n = 30, 24.7 +/- 3.8 yr). Both of these villages are located at 3,700 m above sea level. Hematologic measurements confirmed lower hematocrit values in Nepal (48.4 +/- 4.5%) than in Chile (52.2 +/- 4.6%) (P less than 0.003). When subjects were matched for hematocrit, erythropoietin concentrations in Chile were higher than in Nepal (P less than 0.01). Detailed measurements of blood O2 affinity in Nepal showed no differences in shape or position of the O2 equilibrium curve between Sherpas and Western sojourners. Our results indicate that although Quechua Indians have higher hematocrits than Sherpas living at the same altitude, nevertheless they may be functionally anemic.

Effects of hemodilution on O2 transport in high-altitude polycythemia.

A native of the Peruvian Andes (4,250 m) was studied before and after isovolemic hemodilution of the hematocrit from 62 to 42%. O2 transport was studied with newly developed catheters in the radial and pulmonary arteries. These catheters allowed continuous measurement of arteriovenous O2 content and intermittent cardiac output by thermodilution. During exercise tests, breath-by-breath gas exchange measurements also allowed cardiac output to be calculated by the O2-Fick technique. A complex series of interrelated physiological changes occurred in response to hemodilution. These included increased ventilation, increased arterial and mixed venous PO2, increased cardiac output (both heart rate and stroke volume), and improved ventilation-flow match. The general improvement in symptoms that followed hemodilution correlated well with increased anaerobic threshold and mixed venous PO2 during exercise.