The role of the liver in the cutaneous respiratory compensation of the frog (Rana esculenta)

Experiments on anaesthetized specimens of Rana esculenta (L.) demonstrate that at low temperatures (6°C) the liver of the frog-like the spleen of the newt-regulates cutaneous respiration, storing red blood cells when the animal is well oxygenated and releasing these into circulation under conditions of hypoxia. During the shift from the congested to decongested state, the liver is reduced to about two-thirds of its weight, while the erythrocyte concentration and correlated parameters in the bloodstream can increase by more than 50%. At higher temperatures (18°C) this compensatory mechanism becomes inefficient and the liver oversees the survival of the hypoxic organism for a fairly long period of time (about 5 hours) by releasing glucose for anaerobic glycolysis.     

Splenic respiratory compensation and blood volume in the newt

In the newt Triturus cristatus carnifex , reversible increase in size of the spleen is linked to the respiratory state of the animal: when the newt is exposed to the air, and thus well oxygenated, the spleen hoards erythrocytes; when immersed in still water, in an hypoxic state, the spleen releases erythrocytes into the bloodstream. In chlorobutanol-anaesthetized specimens exposed to the air, the maximum size reached by the spleen diminishes with a rise in temperature up to the disappearance of all congestion at 33°C. The blood volume of newts kept in humid air at 6°C and 18°C after anaesthesia varies from about 6–9 ml per 100 g of body weight, while the red blood cell count and haematocrit value remain stable. In anaesthetized specimens kept in still water at the same temperatures the blood volume is stable, at about 7 ml per 100 g, but the red cell count and haematocrit are notably higher. At 33°C, a critical temperature for the newts, the specimens in still water succumb while those in air present the same blood volume as at 18°C, but have a higher erythrocyte count and haematocrit value.     

Haematological changes by splenic respiratory compensation in the cave salamander, Hydromantes genei

Like other amphibians, the salamander Hydromantes genei , an exclusively terrestrial lungless plethodontid which lives in cold humid caves, possesses a haematological mechanism of respiratory compensation: the spleen can hoard erythrocytes, which are then released into the bloodstream when necessary, analogous to what happens in the Italian crested newt (which, however, is predominantly aquatic). The cave salamander, anaesthetized with chlorobutanol and kept in a humidity-saturated environment at a constant temperature long enough for its haematological conditions to stabilize, presents homogeneous and very low blood parameters at the extreme temperature ranges to which it is adapted (6 °C and 18 °C): about 16 10⁹ red blood cells/1, haematocrit value approximately 12, and haemoglobin concentration slightly below 3g/dl. The increase in temperature triggers the release of erythrocytes into the bloodstream from the spleen, which shrinks from 0.8% of the animal's body weight at 6 °C to 0.25% at 18 °C; however, a parallel increase in blood plasma maintains the blood composition unaltered. At 24 °C, a critical temperature for this species, the erythrocyte parameters increase by 50% owing to plasma loss, as happens in other amphibians in hypoxic conditions.     

Hepatic respiratory compensation and haematological changes in the cave cyprinid, Phreatichthys andruzzii

In several ectotherms, including all members of the Osteichthyes studied so far, the spleen is capable of storing and releasing erythrocytes according to the animal's respiratory needs. The tropical cave cyprinid Phreatichthys andruzzii uses its liver rather than the spleen as the site of accumulation in the respiratory compensation process, like the amphibian Rana esculenta. The reversible process of erythrocyte accumulation in the liver is very evident in animals anaesthetized with chlorobutanol; MS-222, an anaesthetic widely used in lower vertebrates alters all the haematological parameters and is not suitable for studies on blood and respiration. The hepatic respiratory compensation mechanism is as efficient as the splenic one: in animals kept at 18 °C for 24 h the mean liver weight percentage was 70% higher than in specimens kept at 28 °C (2.43% of the body weight compared to 1.39%, in groups of six specimens) while mean red blood cell counts fell from 2.49 to 1.60 · 10¹² per l, in agreement with the haematocrit value and haemoglobin concentration; mean corpuscular volume remained constant (at about 177 fl). Accepted: 15 April 1997     

Estimates of plasma, packed cell and total blood volume in tissues of the rainbow trout (Salmo gairdneri)

1. Total blood volume and relative blood volumes in selected tissues were determined in non-anesthetized, confined rainbow trout by using 51Cr-labelled trout erythrocytes as a vascular space marker. 2. Mean total blood volume was estimated to be 4.09 +/- 0.55 ml/100 g, or about 75% of that estimated with the commonly used plasma space marker Evans blue dye. 3. Relative tissue blood volumes were greatest in highly perfused tissues such as kidney, gills, brain and liver and least in mosaic muscle. 4. Estimates of tissue vascular spaces, made using radiolabelled erythrocytes, were only 25-50% of those based on plasma space markers. 5. The consistently smaller vascular volumes obtained with labelled erythrocytes could be explained by assuming that commonly used plasma space markers diffuse from the vascular compartment.     

Effects of temperature, oxygen and carbon dioxide on osmotic fragility of carp, Cyprinus carpio L., erythrocytes

The in vitro effects of gases and temperature on the osmotic fragility of carp erythrocytes were studied. At the three different temperatures analyzed (5, 11 and 20°C) there was no noticeable modification in erythrocyte membrane osmotic resistance. Osmotic fragility of red blood cells was altered by CO₂ and air treatment, as compared to the standard procedure. This suggests the need to take into account a possible moderate hypoxia that develops in the routine procedure of nucleated erythrocyte osmotic fragility tests.     

Change in the Respiratory Rate of Sea Urchin Spermatozoa following Sperm-egg Interaction in the Absence of Mg2+

Spermatozoa of the sea urchin, Hemicentrotus pulcherrimus (10⁸ cells/ml), preincubated with unfertilized eggs deprived of jelly coats (more than l0⁵ cells/ml) at 20°C for 20min in Mg²⁺ free artificial sea water containing 1 mM Ca²⁺ (MFASW), exhibited very low respiration, which was enhanced by 2, 4 dinitrophenol (DNP). The fertilization rate in MFASW was usually less than 5% and was about 25% at most. Preincubation with fertilized eggs (with and without a fertilization membrane) in MFASW did not reduced the respiratory rate of spermatozoa. The rate of sperm respiration was lower in MFASW than in artificial sea water (ASW), but was higher than the respiratory rate of spermatozoa preincubated in MFASW with unfertilized eggs. Sperm respiration in MFASW or in ASW was not stimulated by 2, 4 dinitrophenol. Almost complete inhibition of sperm respiration was obtained with unfertilized eggs fixed with glutaraldehyde at concentrations of above 10⁵ cells/ml in MFASW and of about l0⁴ cells/ml in ASW. The respiratory rate of spermatozoa treated with fixed eggs was enhanced by DNP. It is concluded that the respiratory rate of the spermatozoa is reduced by their interaction with unfertilized eggs before their penetration into the eggs.     

Hemoglobin functions in the blood of Bufo marinus

The effects of three physical-chemical factors, temperature, hydrogen ion concentration, and partial pressure of oxygen, on the respiratory functions of blood of the toad (Bufo marinus) have been studied. Measurements of oxygen affinity of hemoglobin in whole blood were measured tonometrically by a method devised for small quantities of blood. At pH 7.40 and 25°C blood was found to be 50% saturated with oxygen at a partial pressure of 44 mm Hg of oxygen. The Bohr effect was measured at various temperatures and found to be about one-half that found for mammalian blood. Carbon dioxide content of toad blood changes only slightly in the oxygenated and reduced states. Thus the “Haldane” effect parallels the small Bohr effect. Toad blood was found to have average hematocrit values of 37% for erythrocytes and average hemoglobin values of 11 gm/100 ml per cubic millimeter of blood. The respiratory functions of the blood of the toad conform to the pattern of respiratory mechanisms available for gas exchange between the environment and tissues of the organism.     

Dynamics of the changes in homogenate respiratory activity after rat whole liver storage at 4 degrees C

Homogenate respiratory activity was studied after different storage terms of the whole rat liver at 4 degrees C in sucrose-based solution and following normothermic reperfusion. Preservation of homogenate respiratory activity in all metabolic states after normothermic reperfusion of the control liver (60 min, 4 degrees C) is shown. Further storage (6 and 24 hrs) of isolated liver under the mentioned above conditions strengthens the substrate respiration of homogenate both after storage and after normothermic reperfusion. At the same time oxidative phosphorylation does not practically change. No change was noted in respiratory activity in the states 3, 4ADP and 3DNP after 24 hrs of liver storage in respect of a previous term. Following normothermic liver reperfusion contributes to a statistically true reduction of mentioned parameters of respiration, that correlates with a decrease in the degree of respiration and phosphorylation coupled of the studied system.     

Effect of the adrenergic beta receptor blocker propranolol on the dilatation of cerebrocortical vessels evoked by arterial hypoxia

In order to elucidate the importance of adrenergic beta receptors in the regulation of cerebral microcirculation during arterial hypoxia, chloralose anaesthetized cats were treated with propranolol hydrochloride. Arterial hypoxia, lasting for approximately 4 min, was induced by respiring the animals with a gas mixture containing 7% oxygen balanced in nitrogen gas. Arterial hypoxia was induced in the same animals before and during continuous infusion of propranolol (0.05 mg/kg/min into the lingual artery). Cerebrocortical vascular volume ( CVV ) and NADH fluorescence were measured through a cranial window with a microscope fluororeflectometer . Control arterial hypoxia (no treatment) increased CVV and NADH reduction by 22.2 +/- 2% and 20.4 +/- 2.1%, respectively. Following 1 mg/kg propranolol treatment arterial hypoxia of the same severity resulted in only approximately 2/3 of the CVV response obtained during the control arterial hypoxia. Since arterial hypoxia induced similar changes in arterial blood gases, arterial blood pressure, and intracranial pressure in both cases, our results indicate that the cortical vasodilatation occurring during arterial hypoxia is due, at least in part, to the activation of adrenergic beta receptors.     

Oxidative phosphorylation of liver mitochondria from mice acclimatized to hypobaric hypoxia

Mice exposed to intermittent hypobaric hypoxia for 20 hours a day, 6 days a week, develop extracellular adaptive responses similar to those found in humans exposed to oxygen tension equivalent to that found at an altitude of 4500 m. Isolated liver mitochondria from these animals show no significant differences in rates of substrate-stimulated respiration, ADP-stimulated respiration and the respiratory control ratio (RCR), when compared with sea level controls. Undetectable or negligible differences in these parameters are also noted when sea level animals are exposed for one hour to severe hypoxia (7% O₂). We therefore conclude that the oxidative phosphorylation capacity of the isolated mouse liver mitochondria remains unaltered in both acute and chronic hypoxia. However thein vivo oxygen consumption by mice at this degree of hypoxia was markedly reduced. Lack of observable changes in oxidative phosphorylation could be accounted for by extracellular adaptations in mitochondria isolated from acclimatized animals. This explanation, however, is not consistent with the lack of changes on oxidative phosphorylation in mitochondria isolated from mice undergoing acute hypoxia at sea level. It is then suggested that isolated mitochondrial preparations are of limited value for investigating biochemical mechanisms underlying the variation of cellular respiration occurringin vivo.     

Augmentation of hypoxic respiration after brief hyperoxia in the anesthetized cat: Putative function of GABA<Subscript>A</Subscript> neurotransmission

In this study, we attempted to determine the role of GABA neurotransmission in augmentation of hypoxic respiration by antecedent hyperoxic breathing. The experiments were performed in anesthetized, paralyzed and vagotomized cats divided into control and bicuculline (a GABA_A receptor blocker)-injected groups. The experimental protocol consisted of exposing the animals to successive hypoxic-hyperoxic-hypoxic conditions. Respiration was assessed using phrenic electroneurograms, from which the peak phrenic height, a surrogate of the tidal volume component, and respiratory rate were obtained, and their product, the respiratory minute output, was calculated. We found that prior hyperoxic ventilation increased the subsequent respiratory response to hypoxia by an average of 23.5%, compared with the preoxygen response. This increase was driven by volume respiration. The biphasic character of the hypoxic respiratory response, consisting of stimulatory and depressant phases, was sustained. Bicuculline abolished the augmentative effect on hypoxic respiration of prior hyperoxia, which suggests that oxygenation induces GABA_A-mediated hyperexcitability of respiratory neurons, possibly by the liberation of reactive oxygen species. We concluded that GABA neurotransmission is pertinent to the effect of hyperoxia on hypoxic respiratory reactivity.     

Blunted respiratory responses to hypoxia in mutant mice deficient in nitric oxide synthase-3.

In the present study, the role of nitric oxide (NO) generated by endothelial nitric oxide synthase (NOS-3) in the control of respiration during hypoxia and hypercapnia was assessed using mutant mice deficient in NOS-3. Experiments were performed on awake and anesthetized mutant and wild-type (WT) control mice. Respiratory responses to 100, 21, and 12% O(2) and 3 and 5% CO(2)-balance O(2) were analyzed. In awake animals, respiration was monitored by body plethysmography along with O(2) consumption (VO(2)) and CO(2) production (VCO(2)). In anesthetized, spontaneously breathing mice, integrated efferent phrenic nerve activity was monitored as an index of neural respiration along with arterial blood pressure and blood gases. Under both experimental conditions, WT mice responded with greater increases in respiration during 12% O(2) than mutant mice. Respiratory responses to hyperoxic hypercapnia were comparable between both groups of mice. Arterial blood gases, changes in blood pressure, VO(2), and VCO(2) during hypoxia were comparable between both groups of mice. Respiratory responses to cyanide and brief hyperoxia were attenuated in mutant compared with WT mice, indicating reduced peripheral chemoreceptor sensitivity. cGMP levels in the brain stem during 12% O(2), taken as an index of NO production, were greater in mutant compared with WT mice. These observations demonstrate that NOS-3 mutant mice exhibit selective blunting of the respiratory responses to hypoxia but not to hypercapnia, which in part is due to reduced peripheral chemosensitivity. These results support the idea that NO generated by NOS-3 is an important physiological modulator of respiration during hypoxia.     

Use of a membrane oxygenator with small volumes of perfusion for correction of functional changes in respiration disorders

Experiments on 11 dogs under hypoventilation hypoxia (a decrease in the respiratory minute volume by 40-50%) were made to study the efficacy of membrane oxygenation using a membrane Sever-OMP oxygenator of the blood under the conditions of minor perfusion (14-17% of the minute volume of circulation). The animals of the main series (7 dogs) with a veno-venous connection of the membrane oxygenator (MO) tolerated hypoxia quite well for 2 hours. The control animals died. The conclusion is made that membrane oxygenation with small volumes of perfusion (with the MO connected according to Seldinger) can be used in conjunction with artificial ventilation where the latter one is not effective enough.     

Perfluoro compounds as artificial erythrocytes.

Some liquid perfluoro compounds dissolve relatively large amounts of oxygen and can be used in dispersed form as substitutes for erythrocytes. The commonly used perfluoro compounds contain about the same amount of oxygen as do equal volumes of erythrocytes when equilibrated with 100% oxygen. However, when equilibrated with alveolar air, the perfluoro compounds contain much less oxygen than erythrocytes. The dispersed fluorochemicals are adequate substitutes for perfusion of isolated preparations of mammalian brain, heart kidney, lung and liver. However, when put into the circulation of the intact animal, the dispersed fluorochemicals tends to produce lesions of the lungs, dilation of the right heart, and ultimately fatal hypoxia. It is suggested that the course of events following intravenous injection of dispersed fluorochemical is initiated by an interaction of the perfluoro particles with blood platelets or blood clotting factors. The ensuing intravascular clotting could then cause the changes in the lungs which lead to a marked increase in pulmonary artery pressure and dilation of the right heart. These events would terminate in fatal hypoxia due to pulmonary pathology and heart failure.     

The blood oxygen binding properties of hypoxicSalmo gairdneri

Summary The blood oxygen binding properties of rainbow trout responded to environmental hypoxia (the oxygen saturation of water 30% at 11°C) in three ways. The quickest response was a moderate acidosis, leading to slightly lowered blood oxygen loading due to the Bohr effect. The second response, an increase of blood oxygen carrying capacity, was completed with 6 h from the onset of hypoxia. The speed of the response suggests that the formation of new haemoglobin played no practical role, the increase being caused either by a decrease of plasma volume or the liberation of erythrocytes from a storage organ. The slowest response, a 25% increase of the blood oxygen affinity within a week of hypoxia, was probably caused by the concurrent decrease of the erythrocyte ATP concentration from 4.45 to 2.51 mol/ml erythrocytes.     

2-3 diphosphoglycerate and tissue oxygenation in the cirrhotic

Increased 2-3 Diphosphoglycerate levels in cirrhotic patients have been reported. Previous studies did not show significant changes in 2-3 DPG in anaemic cirrhotic patients when compared to non anaemic cirrhotic patients, but the role played by alkalosis and/or hypoxia has not been investigated. To study this question, haematic 2-3 DPG was measured in 8 male patients with liver cirrhosis (histologically diagnosed) together with PO2, PCO2, pH and Hct. 2-3 DPG was also measured in 6 healthy male volunteers. We found a significant increase in blood 2-3 DPG of cirrhotic patients compared to control subjects (5,55 +/- 0,4 vs 2,18 +/- 0,3 mmol/l erythrocytes respectively, p less than 0,001) in agreement with previous studies. PO2 levels and Hct value did not show important changes, whereas PCO2 and pH resulted to be very altered when compared to normal values, even though we could not correlate these values with blood 2-3 DPG. We conclude that the genesis of 2-3 DPG increase is multifactorial, however an alteration in acid-base equilibrium seems to play a more important role than hypoxia.     

Respiratory changes with stratification of pear seeds

Experiments on the stratification of seeds of pear ( Pyrus communis cv. Bartlett) indicate that there is a progressive increase in respiratory capacity, with about a three-fold increase developing during a 30-day stratification period. Evidence is presented suggesting this increase in respiratory capacity occurs in the cytochrome oxidase pathway and not in the alternative pathway of respiration. The increase is much less pronounced if the seeds are held at 25°C rather than at 6°C. The seed coat serves as a barrier to the expression of the increased respiratory capacity.     

Gill ventilation and respiratory efficiency of Sacramento blackfish, Orthodon microlepidotus Ayres, in hypoxic environments

Respiratory and gill ventilatory responses of Sacramento blackfish to three environmental temperatures (12, 20 and 28°C) and four environmental dissolved oxygen concentrations (130, 90, 65 and 40 torr P_O2) were examined to determine physiological strategies of survival in eutrophic lakes and suitibility for culture conditions. Situated in van Dam-type respirometers, experimental blackfish showed increased gill ventilatory flows from increased ventilatory frequencies and stroke volumes to meet higher respiratory oxygen demands at increased temperatures. Ventilation volumes also increased at reduced environmental dissolved oxygen levels by increased ventilatory stroke volumes alone, except at 28°C where frequency increases were also measured. Oxygen consumption rates remained essentially constant with declining dissolved oxygen, except at 28°C where excitement elevated respiratory metabolism at 65 and 40 torr. Percentage utilization of oxygen increased with temperature from 12°C, but levels at 20 and 28°C were insignificantly different. Contrary to most studies on other species, there was no change in percentage utilization under hypoxic conditions even with 4.7-fold increases in ventilation volume in excited fish at 28°C. The ability of blackfish to survive in hypoxic waters is quantitatively compared with other species by calculation of a respiratory efficiency index (I), which includes the relationship between ventilation volume and percentage utilization of oxygen under normoxic and hypoxic conditions as well as the half-saturation value (P₅₀) of the species' blood with oxygen.     

Functional reaction of the human body to different ways of respiration regulation

Reactions of the body to different ways of regulation of respiration, i.e., through the elastic resistance to respiratory movements and external dead space volume, as well as voluntary hyper- and hypoventilation, were studied. An increased elastic resistance to respiratory movements was established to create conditions for intense loading of respiratory musculature and to determine an increased in oxygen consumption. An increase in the external dead space produces a marked complex reaction of the body to the hypoxichypercapnic content of the inhaled gas mixture, causing hyperventilation and heightened work of the respiratory musculature and stimulating metabolism. Voluntary hyperventilation during muscular effort leads to a state of relative hypocapnia, gross loss of efficiency, and economic external respiration and gas exchange. Voluntary hypoventilation in the course of muscular effort brings about marked shifts in gas homeostasis towards alveolar hypoxia and hypercapnia. A concurrent increase in the efficiency and economy of external respiration and gas exchange is observed.