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.
This article reviews briefly some general aspects of the enteric nervous system (ENS). Furthermore, the ENS control of epithelial transport is exemplified by a description of the enteric nervous reflexes activated by cholera toxin.
The cellular signalling pathways participating in physiological color change are reviewed, particularly in crustaceans, teleosts, amphibians, and reptiles. This review is an attempt to summarize what is known and to raise some hypotheses about basic questions still to be elucidated. The first picture that emerges from the literature is that the transduction pathways are identical in the various types of chromatophores of a single species, except for the iridophore. The cAMP-dependent pathway has been well conserved throughout evolution: cAMP increase is the pigment dispersion signal whereas the nucleotide decrease leads to granule aggregation. On the other hand, the Ca(-2)-dependent pathways evoke pigment aggregation in teleosts and crustaceans, and dispersion in amphibians and probably reptiles as well. Another interesting point is the ultimate convergence of the signalling pathways of different agonists inducing the same response in one chromatophore type. A hypothesis is raised about why different chromatophores behave differently in the absence of agonists, that is, why some are punctate, whereas others are stellate.
Cl− secretion in the rat colonic crypt base cell (bc) requires the coordinated (a) opening of Cl− channels in the luminal membrane; (b) activation of the Na+2Cl−K+ cotransporter; (c) enhanced conductive K+ exit from the cell; and (d) increased pumping by the (Na+ + K+)-ATPase. In this study we focus on the importance of conductive K+ exit. After stimulation with the cholinergic agonist carbachol (CCH, 0.1–10 μmol/l) bc respond with a marked increase in whole cell (wc) conductance and a hyperpolarization of the membrane voltage (Vm). This is paralleled by a marked increase in the (Cl− secretory) short-circuit current (Isc) in Ussing chamber studies of the intact distal colon. Current evidence favors the view that CCH, via IP3, enhances cytosolic Ca2+ activity, and that Ca2+ increases the open probability of Cl− channels indirectly and that of K+ channels directly. After stimulation with PGE2 bc also enhance the wc conductance, but this is paralleled by a marked depolarization of Vm. Again these effects correspond to a marked increase in (Cl− secretory) Isc. The depolarization and enhanced wc conductance is partly due to the activation of Cl− channels. However, current evidence suggests that these effects on Cl− channels are paralleled by an activation of K+ channels. The chromanol 293B, by inhibiting these K+ channels specifically, abolishes PGE2-induced Cl− secretion completely, but has no effect on basal K+ conductance or on CCH-induced Cl− secretion. CCH apparently activates a Ca2+-dependent K+ channel with a conductance of 10–20 pS, whilst PGE2 (or cAMP) activate a much smaller K+ channel. Only the latter K+ channel can be inhibited by 293B in excised patches. Noise analysis suggests that this K+ channel has a conductance of <3 pS and fast kinetics. The complete 293B induced inhibition of Cl− secretion caused by PGE2 can be explained by the fact that PGE2 induces a marked depolarization and that this depolarization reduces the basal K+ conductance. Current evidence suggests that this inhibition of the basal K+ conductance is caused by a depolarization induced inhibition of Ca2+ entry.
The intestinal uptake of water-soluble vitamins, nicotinamide, riboflavin, biotin and folic acid, was studied in isolated everted intestinal sleeves of the cold-water teleost rainbow trout (Onchorhynchus mykiss). The presence of a carrier-mediated transport mechanism was determined by competitive inhibition and by Michaelis-Menten kinetics. The uptake of riboflavin, biotin or folic acid was not only subject to competitive inhibition but also a saturable function of increasing vitamin concentration in the incubation medium. The kinetic constants of the saturable mechanism were for riboflavin: K(m), 2.32 +/- 0.76 microM; Vmax, 0.26 +/- 0.04 pmol/mg min; for biotin: K(m), 9.70 +/- 3.76 microM; Vmax, 0.31 +/- 0.07 pmol/mg min; and for folic acid: K(m), 32.9 +/- 21.2 microM; Vmax, 3.63 +/- 0.99 pmol/mg min. In contrast, the uptake of nicotinamide was not subject to competitive inhibition and was a linear function of concentration (Kd, 0.140 +/- 0.012 pmol/mg min microM). Folic acid was absorbed more rapidly than and was not inhibited by its derivative, 5-methyl-tetrahydrofolate. Thus, the intestinal uptake of riboflavin, biotin and folic acid is carrier-mediated while that of nicotinamide occurs by simple diffusion. These mechanisms are similar to those found in the channel catfish for the same vitamins, except for folic acid, which is absorbed by diffusion in this warm-water omnivorous species.
The purposes of these studies were: 1) to determine the effects of various doses of propranolol, a nonspecific beta-adrenergic antagonist, on ventilation, oxygen consumption, and body temperature in hamsters, and 2) to test the hypothesis that in hamsters the stimulatory effects of naloxone, an opioid receptor antagonist, on ventilation and oxygen consumption occur, at least in part, through the release of catecholamines that act via beta-adrenergic receptors. Propranolol, a non-specific beta adrenergic receptor antagonist, at a 20 mg/kg depressed body temperature, oxygen consumption, tidal volume, and ventilation relative to saline. The lower dose of 10 mg/kg had only transitory effects on tidal volume at 60 min and ventilation at 30 min post-injection-Naloxone (1 mg/kg) relative to saline stimulated ventilation and oxygen consumption. These effects were blocked by propranolol pretreatment. The results of these experiments demonstrate that in the hamster, 1) body temperature, oxygen consumption, and ventilation appear to be modulated by beta-adrenergic receptors, and 2) the stimulatory effects of naloxone on oxygen consumption and ventilation may occur through the interaction of endogenous opioids and beta-adrenergic receptor systems.
Embryos of Atlantic herring (Clupea harengus L.) from the Buchan (Northern North Sea) stock were incubated from fertilisation until hatching at temperatures of 5, 8, 12, and 15 degrees C. The relative timing of development of the Kolmer-Agduhr (KA) neurons, the posterior lateral line nerve, the motor neurons, and myotubes were determined with respect to somite stage of the embryo. Development of the KA neurons, the lateral line nerve, and the myotubes was similar at all temperatures. In contrast, timing of outgrowth of the motor neuron axons with respect to somite stage was earlier at higher (> or = 12 degrees C) than at lower temperatures (< or = 8 degrees C) although it reached a similar point at all temperatures by the 58-somite stage. Our hypothesis to explain these observations is that delayed motor axon outgrowth in the lower temperature groups is probably due to a delay in a signalling interaction between motor neurons and the somite.
