Given that the chemistry of lactate production disproves the existence of a lactic acidosis, there is a need to further reveal and explain the importance of the organic and computational chemistry of pH dependent competitive cation fractional (~) proton (H+) exchange (~H+e). An additional importance of this knowledge is that it could potentially contradict the assumption of the Stewart approach to the physico-chemical theory of acid-base balance. For example, Stewart proposed that chemical reaction and pH dependent H+ dissociation and association do not directly influence the pH of cellular and systemic body fluids. Yet at the time of Stewart's work, there were no data that quantified the H+ exchange during chemical reactions, or from pH dependent metabolite H+ association or dissociation. Consequently, the purpose of this review and commentary was three-fold; 1) to provide explanation of pH dependent competitive cation ~H+e exchange; 2) develop a model of and calculate new data of substrate flux in skeletal muscle during intense exercise; and 3) then combine substrate flux data with the now known ~H+e from chemical reactions of non-mitochondrial energy catabolism to quantify chemical reaction and metabolic pathway ~H+e. The results of purpose 3 were that ~H+ release for the totality of cytosolic energy catabolism = −187.2 mmol·L−1, where total glycolytic ~H+te = −85.0 mmol·L−1. ATP hydrolysis had a ~H+te = −43.1 mmol·L−1. Lactate production provided the largest metabolic ~H+ buffering with a ~H+te = 44.5 mmol·L−1. The total ~H+ release to La ratio = 4.25. The review content and research results of this manuscript should direct science towards new approaches to understanding the cause and source of H+e during metabolic acidosis and alkalosis.
Thermal tolerance limits of ectotherms may result from respiratory limitations. In response to declining oxygen availability, organisms have shown to exhibit oxyregulation by enhancing ventilation and heartbeat rates. In this study we examined how this regulatory response in mussels (Mytilus) changes with increasing temperature. Experimental mussels showed extensive oxyregulation at temperatures near to their habitat temperature, but increasingly lost this capacity towards higher temperatures. At breakpoint temperature no regulation took place and respiration rates changed proportional to oxygen availability. These results revealed how thermal limitations relate to respiratory capacity of mussels.
Respiration of bulky plant organs such as roots, tubers, stems, seeds, and fruit depends very much on oxygen (O2) availability and often follows a Michaelis-Menten-like response. A multiscale model is presented to calculate gas exchange in plants using the microscale geometry of the tissue, or vice versa, local concentrations in the cells from macroscopic gas concentration profiles. This approach provides a computationally feasible and accurate analysis of cell metabolism in any plant organ during hypoxia and anoxia. The predicted O2 and carbon dioxide (CO2) partial pressure profiles compared very well with experimental data, thereby validating the multiscale model. The important microscale geometrical features are the shape, size, and three-dimensional connectivity of cells and air spaces. It was demonstrated that the gas-exchange properties of the cell wall and cell membrane have little effect on the cellular gas exchange of apple (Malus×domestica) parenchyma tissue. The analysis clearly confirmed that cells are an additional route for CO2 transport, while for O2 the intercellular spaces are the main diffusion route. The simulation results also showed that the local gas concentration gradients were steeper in the cells than in the surrounding air spaces. Therefore, to analyze the cellular metabolism under hypoxic and anoxic conditions, the microscale model is required to calculate the correct intracellular concentrations. Understanding the O2 response of plants and plant organs thus not only requires knowledge of external conditions, dimensions, gas-exchange properties of the tissues, and cellular respiration kinetics but also of microstructure.
Fourteen acidic and neutral free amino acids (FAA) were investigated in soft tissue of Macoma balthica from different depth zones of the Gulf of Gdansk (Baltic Sea) over a full seasonal cycle. The dry weight of the bivalves and physico-chemical parameters of overlying bottom water and surface sediments were measured simultaneously at each site. In the brackish waters of the Baltic, the main pool of FAA is composed of Ala, Gln, Arg, Gly and Orn which represent approximately 80% of the total. Compared to the full saline environments, the composition of FAA in the clams from the Baltic differs substantially. The differences can be attributed to the lower salinity of the Baltic. In the Baltic, Gly appears to play a most important role in regulating intracellular osmolarity in the clams, a function performed primarily by Tau in Atlantic and North Sea populations. Spatio-temporal variations of the FAA are affected by biotic and environmental parameters; their respective influence differs with the amino acids. The concentration of Arg depends on its uptake from the external medium. However, its level might be temporarily modified by stress-induced metabolic transformation (e.g. hydrolysis to Orn) caused by changes in the ambient environment. The concentration of Ala increases with depth, probably because of physiological adaptations of the animal to diminishing oxygen concentration through anaerobic glucose catabolism. Biosynthesis of Ala, similarly to Gln, in the shallower zone is generally related to the physiological state of an organism. The concentration of Gly is most likely regulated by internal mechanisms driven by gonadal development and reproduction.
In vivo and in vitro experiments demonstrated that the rectal caeca of the asteroid Asterias rubens selectively and actively excrete patent blue V (PBV) from the individual body cavity towards the gut lumen, a function already shown for another water-soluble dye (chlorophenol red, CPR) in several echinoderm species (including A. rubens). (These dyes are both known to be selectively and actively excreted by the vertebrate kidney.) Aquarium experiments were conducted in order to characterize in vivo CPR and PBV excretion kinetics in A. rubens. Dye eliminations were determined and were shown to be partly inhibited by para-aminohippuric acid (PAH), a competitive inhibitor of CPR transport in asteroid rectal caeca as well as in mammalian renal proximal tubules. The results show that the in vivo method is reliable and complements in vitro investigations. In particular, due to preservation of the organs’ integrity, the method preserves rectal caeca epithelium from rapid autolyse and allows the design of long-term experiments to study echinoderm excretion.
It is generally recognized that a decrease in carbohydrate availability can lead to the development of fatigue during prolonged exercise in humans. Administration of glucose or other carbohydrates before or during exercise has been shown to postpone fatigue, conserve muscle glycogen and improve performance. Carbohydrates can be categorised according to their ability to increase blood glucose concentration (known as glycaemic index) and by the extent they stimulate the release of insulin. The glycaemic index is reflected in the rate at which consumed carbohydrate is made available in the blood. Glucose is the only type of carbohydrate that can readily be oxidised by skeletal muscle for energy production. Gastric emptying is the primary factor limiting the rate of carbohydrate delivery to the blood and therefore influences the utilisation of exogenous carbohydrate ingested before or during exercise. Various methods have been used to assess the oxidation of exogenous carbohydrates during exercise. Peak rates of CHO oxidation during exercise have been reported between 0.4 and 1.0 g/min, and the rates of oxidation do not appear to be influenced to a major extent by the use of multiple drinking schedule in comparison with a single bolus schedule. Previous studies also suggest that the ingestion of fructose during exercise does not offer any additional benefits over ingestion of glucose or glucose polymer solutions of similar concentration. The hormones insulin, glucagon and adrenaline together with cortisol and growth hormone play key roles in the regulation of carbohydrate metabolism during exercise. Ingestion of moderately concentrated carbohydrate solutions (4-8%) enhances prolonged exercise performance and is appropriate for optimising energy and fluid delivery without causing adverse effects. The ergogenic effects of carbohydrate ingestion on performance during intermittent exercise such as competitive sports are less well established, although the evidence to date suggests diminished performance when carbohydrate are limiting.
The objective of this review is to summarize data on the interrelationships that exist between nutrition, the endocrine system and their modulation of plasma tumor necrosis factor-α responses to endotoxin in cattle. During stress, intake of nutrients often is compromised and a percentage of available nutrients are diverted away from growth processes to stabilize other physiological processes of a higher survival priority. Management practices that minimize the magnitude and duration of disease stress will aid in speeding the return to homeostatic equilibrium. However, the shift away from growth during stress is almost inevitable as a mechanism to survive. Some degree of control and management of the metabolic cost of disease stress involves understanding the integration of nutritional, endocrine and immune signals by cells and working with the natural homeostatic processes. Endocrine hormones and immune system cytokine signals participate in redirecting nutrient use during disease stress. In an intricate interplay, hormones and cytokines regulate, modify and modulate each other's production and tissue interactions to alter metabolic priorities. Levels of dietary protein and energy intake affect patterns of hormones and cytokines in the blood after endotoxin challenge and further modulate the biological actions of many of these regulatory effectors. In vivo, administration of growth hormone to young calves has significant effects to decrease the many specific physiological responses to endotoxemia. Many aspects of nutrition can attenuate or facilitate this effect.
We cloned chick cellular retinol-binding protein, type two (CRBP II) cDNA and compared it with those of some mammals. The deduced amino acid sequence showed that chick CRBP II was one amino acid greater in size than those of mammals, and the nucleotide sequence of chick CRBP II shared 72%–75% similarity with those of mammals. RNA blot hybridization analysis showed that CRBP II transcript of 0.7 kb was first detected in the duodenum of day-18 embryonic chick, and exhibited a rapid increase during 24 hr around the hatching. Northern blot hybridization also revealed that the transcripts of two types of retinoid X receptors (RXR α and RXR γ) and peroxisome proliferator-activated receptor (PPAR) were expressed in the chick duodenum at hatching. The organ culture of day 16 embryonic chick duodenum showed that the addition of 9-cis retinoic acid in the medium caused a significant increase in CRBP II mRNA levels. In addition, arachidonic acid, from which putative ligands for PPAR were supposed to be generated, was accumulated around hatching in the duodenum. The results may suggest that the abrupt increase of the CRBP II gene expression in the chick duodenum around hatching may be related with RXRs and/or PPAR.
Earlier studies identified two main defense strategies against hypoxia in hypoxia tolerant animals: (1) reduction in energy turnover, and (2) improved energetic efficiency of those metabolic processes that remain. We used two model systems from the highly anoxia-tolerant aquatic turtle: (1) tissue slices of brain cortex (to probe cell level electrophysiological responses to oxygen limitation), and (2) isolated liver hepatocytes (to probe signalling and defense). In the latter, a cascade of processes underpinning hypoxia defense begins with an oxygen sensor that is probably a heme protein and a signal transduction pathway that leads to the specific activation of some genes (increased expression of several proteins) and to specific down-regulation of other genes (decreased expression of several other proteins). The pathway seems to have characteristics in common with oxygen-regulated control elements in other cells. The probable roles of the oxygen sensing and signal transduction system include coordinate down-regulation of energy demand and energy supply pathways in metabolism. Because of this coordination, hypoxia tolerant cells stay in energy balance even as they down-regulate to extremely low levels of ATP turnover. The main ATP-demanding processes in normoxia (protein synthesis, protein degradation, glucose synthesis, urea synthesis and maintenance of electrochemical gradients) are all turned down to variable degrees during anoxia or extreme hypoxia. Most striking is the observation that ion pumping is the main energy sink in anoxia—despite reductions in cell membrane permeability (“channel arrest”). Neurons also show a much lower permeability than do homologous mammalian cells but, in this case under acute anoxia, there is no further change in cell membrane conductivity. We consider that, through this recent work, it is becoming evident how normoxic maintenance ATP turnover rates can be down-regulated by an order of magnitude or more—to a new hypometabolic steady state that is prerequisite for surviving prolonged hypoxia or anoxia. The implications of these developments extend to many facets of biology and medicine.
Hearts from rats fed a copper-deficient (Cu−) diet have decreased levels of nuclear-encoded peptides of cytochrome c oxidase (CCO). Studies were conducted to determine whether iron deficiency would lead to a similar finding, whether mRNA transcripts and the chaperonin heat shock proteins (HSP) 60 and 70 from hearts of Cu− rats were decreased as compared with copper-adequate controls and whether synthesis of mitochondrial and nuclear encoded peptides differed as affected by diet copper. In study 1, weanling rats were assigned to one of three groups (n = 6 in each group): (1) control copper and iron adequate fed rats; (2) Cu− rats and (3) iron-deficient (Fe−) rats. Western blotting of nonmyofibrillar cardiac proteins revealed that the nuclear encoded peptides of CCO from the Cu− rats were markedly decreased as compared with control and Fe− rats. Mitochondrial encoded subunits did not appear to differ by treatment groups. Iron-deficient rats had similar nuclear encoded peptide levels as those of controls. In study 2, mRNA transcripts from Cu− (n = 4) and control copper adequate (n = 4) rats did not appear to differ for subunits II and IV, which correspond to mitochondrial and nuclear encoded subunits, respectively. In study 3, levels of HSP 60 and 70 from hearts of Cu− rats (n = 3) did not differ from Cu+ rats (n = 3). In study 4, infusion of 3H-(4,5)-leucine into the hearts of Cu+ and Cu− rats suggested there was no difference in synthesis of the nuclear encoded peptides by copper status and some indication there was enhanced breakdown of the nuclear encoded peptides among the Cu− rats. As expected, more isotope was incorporated into the mitochondria of Cu− rats than Cu+ rats. These results demonstrate an independent effect of copper upon the apparent decrease in the nuclear encoded subunits of CCO, the effect of copper upon the CCO subunits is probably post-transcriptional and that translocation of the nuclear encoded subunits from the ribosomes to the mitochondria via the chaperonin proteins is not a primary defect in explaining these observations in hearts from Cu− rats and synthesis of the nuclear encoded subunits of CCO in not impaired in copper deficiency.
Interleukin-6 (IL-6) is a multifunctional cytokine produced by a variety of cell types in tissues of both the immune and endocrine systems. Among the major functions described for IL-6 are its role in the maturation of B cells to high-output antibody-producing cells and its contribution to the acute physiological responses to infection and inflammation, notably production of hepatic acute phase proteins and activation of the hypothalamic-pituitary-adrenal axis. In addition to these better known functions, IL-6 recently has been found within the pituitary of laboratory rats and also in the human pituitary. In rats, pituitary IL-6 mRNA is upregulated by peripheral exposure to bacterial endotoxin. However, the role of anterior pituitary IL-6 in host responses to infection and inflammation remains uncertain, although it may regulate pituitary hormone secretion. The following brief review summarizes the information available concerning cytokine production within the anterior pituitary of species of domestic livestock. To our knowledge, experiments conducted in our laboratory evaluating regulation of IL-6 mRNA expression and secretion from the porcine anterior pituitary provide most of the data in domestic species confirming the presence of IL-6 in the pituitary. Our data indicate that IL-6 mRNA is present in cultured porcine anterior pituitary cells and that the pituitary directly responds to stimulation with bacterial endotoxin by increasing secretion of IL-6. Furthermore, endotoxin-induced upregulation of IL-6 mRNA expression and secretion appears to be dependent upon production of cyclooxygenase products of arachidonic acid metabolism.
We have studied pH regulation in both intracellular and extracellular compartments of mouse colonic crypts, using distal colonic mucosa with intact epithelial architecture. In this work, we question how transepithelial SCFA gradients affect intracellular pH (pHi) and examine interactions between extracellular pH (pHo) and pHi regulation in crypts of distal colonic epithelium from mouse. We studied pH regulation in three adjacent compartments of distal colonic epithelium (crypt lumen, crypt epithelial cell cytosol, and lamina propria) with SNARF-1 (a pH sensitive fluorescent dye), digital imaging microscopy (for pHi), and confocal microscopy (for pHo). Combining results from the three compartments allows us to find how pHi and pHo are regulated and related under the influence of physiological transepithelial SCFA gradients, and develop a better understanding of pH regulation mechanisms in colonic crypts. Results suggest a complex interdependency between SCFA fluxes and pHo values, which can directly affect how strongly SCFAs acidify colonocytes.
Acidophilic microorganisms grow optimally at pH values between 1–4. They have adapted to the acid condition by maintaining their cytoplasmic pH at a value close to neutrality. Hence, only those (macro)-molecules, which face the acid medium, have had to adapt to this extreme condition. Literature data show that several exoproteins from thermoacidophilic prokaryotes are characterized by a low charge density. It is proposed that this property contributes to the stability of these proteins both below and above the pKa-values of their glutamate and aspartate residues. As an example of an acidophilic protein, the α-amylase from the Gram-positive Alicyclobacillus acidocaldarius ATCC27009 was studied. The enzyme is thermoacidophilic, with optima of temperature and pH of 75°C and pH 3, respectively. The nucleotide sequence of the cloned gene (8) indicates that the α-amylase belongs to a large family of starch-degrading enzymes with a characteristic catalytic (βα)8-domain. Three essential and probably catalytic acidic residues have been conserved, suggesting that the acidophilic α-amylase degrades starch with essentially the same mechanism as do its neutrophilic relatives. Still, the acidophilic protein contains three exchanges in residues uniformally or almost uniformally conserved among all members of the enzyme family. In order to test whether these exchanges contribute to the acidic pH optimum, the α-amylase gene was expressed in Escherichia coli. Sonication of the enzyme-producing cells released α-amylase activity associated with a 140 kDa protein. The optima of temperature and pH for the protein produced in E. coli were similar to those of the native enzyme. Experiments are underway in which it is tested which residues contribute to the acid pH optimum of the α-amylase.
This paper reports the effects of drugs affecting the homeostasis of cytosolic-free calcium on in vitro yeast phagocytosis by hemocytes of the colonial ascidian Botryllus schlosseri. Significant inhibition of phagocytosis is observed after exposure of hemocytes to 10 microM or higher concentrations of thimerosal, which is known to deplete intracellular calcium stores in mammalian cells. The two calcium channel blockers nifedipine and verapamil significantly decrease the phagocytic index, the minimum effective concentrations being 10 and 50 microM, respectively. As these substances have no effects at lower concentrations, they probably act through the inhibition of Ca(2+)-ATPase activity, required to restock intracellular calcium stores, due to their interaction with calmodulin. Analogously, pimozide, which suppresses ATPase activity by interacting with calmodulin, and thapsigargin, which inhibits Ca(2+)-ATPase activity, significantly reduce the phagocytic index. Moreover, nifedipine, by altering cytosolic calcium homeostasis, also lowers the production of superoxide anion associated with phagocytosis. Results indicate that in ascidians, as in mammals, a rise in intracellular calcium is required for phagocyte activation and induction of the respiratory burst.
Oxygen sensing is a determinant function of mammals, especially humans, to maintain their activity under acute or chronic exposure to hypoxia. True O2 sensors (chemoreceptors, erythropoietin secreting cells) are involved in regulation loops, which aim to restore O2 availability to the cells. Pseudo O2 sensors are cells activated by the lack of oxygen but not clearly involved in regulation processes. Potassium channels in the carotid bodies have been suspected to be O2 sensitive and could mediate the chemosensitivity to hypoxia. Na,K-ATPase related ion transport in alveolar pneumocytes could be sensitive to O2 availability and regulate the flux of water and sodium in the alveolar space. Signal transduction in G-protein–dependent receptor systems is modified in hypoxia, such as in cardiac β-receptors and adenosinergic and muscarinic receptors. Recent studies have provided some evidence to the possible role of hypoxia inducible factors (HIF-1) in the regulation of protein synthesis at the transcriptional level. Similarities between O2-sensing mechanisms in erythropoiesis and in the synthesis of vascular endothelial growth factor were recently evidenced. Both genes are upregulated in hypoxia. However, the precise structure (heme-linked enzyme?) of all these O2-sensitive sites is not known, either in the erythropoietic system or in the chemoreceptor function. An adequate balance between hypoxia-induced upregulation and downregulation processes is necessary for optimal survival in a hypoxic environment.
We hypothetized that the Chilean mouse-opossum Thylamys elegans needs micronutrients that are in fruits and seeds, and the unsaturated fatty acids are such micronutrients that may allow individuals of this species to experience longer torpor bouts, lower body temperatures during torpor and higher energy savings during wintertime. To test this hypothesis, we studied: 1) wintertime preferences by artificial diets rich in saturated fatty acids, unsaturated fatty acids and control diets, and 2) the effect of acclimation to dietary fatty acids on the energetics and torpor patterns in this species. When individuals where allowed to choose between the experimental diets they always selected the unsaturated fatty acid diet. After 4 weeks of dietary acclimation, the average daily metabolic rate was not significantly different among treatments, neither was minimum metabolic rate during torpor significantly different, in spite of a tendency to lower values that was detected under unsaturated fatty acid treatment. A similar pattern was observed when body temperature during torpor was compared among treatments. Two explanations are proposed: 1) Acclimation time was not sufficient to obtain statistical significance, but physiological differences and 2) metabolic rate during torpor are not affected by dietary lipids in this species.
Histological studies have detected nitric oxide (NO) synthase in the central nervous system of all vertebrates examined, from lampreys to mamals. However, there are still very few comparative physiological studies on the function of NO synthase in the brain of non-mammalian vertebrates. So far, we know that acetylcholine can cause an NO-dependent increase in brain blood flow in turtles and some fish species (crucian carp and rainbow trout), whereas some other fishes appear to lack such a mechanism. Hypercapnia can induce NO-dependent cerebral vasodilation in mammals, but such a mechanism appears to be lacking in the ectothermic vertebrates examined. The number of species studied needs to be expanded before we can draw any firm conclusions about the origin of NO-dependent brain blood flow regulation: if it has evolved more than once or if it has been occasionally lost during evolution. We conclude that NO synthase may be present in all vertebrate brains but that its functions can vary, as judged from its role in cerebral blood flow regulation. The diversity of functions that NO has proven to have within the mammalian brain is likely to be paralleled by the same degree of diversity of function between vertebrate groups.
CF mice, i.e., mice without functional CFTR (cystic fibrosis transmembrane conductance regulator) exhibit a very low basal I-sc in all regions of the intestinal tract. The low basal I-sc in the intestinal epithelia of the CF mice appears to be a result of lack of spontaneous Cl- secretion (and possibly HCO3- secretion) mediated by neurotransmitter release from the enteric nervous system.In contrast to intestinal epithelia from normal mice, the intestinal epithelia of CF mice do not secrete Cl- in response to agents that increase cAMP (forskolin). Furthermore, as in human CF patients, agents that increase intracellular Ca2+ (bethanacol, ionomycin) failed to elicit Cl- secretion in the intestinal epithelia of CF mice.There was no difference in the electrogenic Na+-coupled glucose absorption in the CF murine jejuna compared to jejuna from normal mice. However, further studies are warranted to determine whether amiloride-sensitive Na+ absorption is upregulated in the murine CF colon.It was concluded that the intestinal epithelium of the CF mouse model exhibits some striking similarities to its human counterpart, and therefore should be very useful in further characterizing the ion transport defects in this disease. (C) 1997 Elsevier Science Inc.
