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How do cells respond to their thermal environment?

Changes in growth temperature induce both activating and inactivating responses from cells, with the magnitude of the temperature change being among the factors that influence which type of response dominates. Aside from upregulated enzyme activity, induction of thermotolerance is the most widely studied and best understood activating response that cells exhibit following heat shock. Inactivating responses to heat shock that are of biomedical interest include heat radiosensitization and cytotoxicity. Interestingly, the activation energy for inducing thermotolerance, heat cytotoxicity, and radiosensitization all fall within a similar range of 120-146 kcal per mole. The relatively high activation energy for each of these responses suggests that they all involve a heat-induced molecular transition as a trigger, and several lines of research suggest strongly that protein denaturation is the common transition that triggers all three responses. Low levels of protein denaturation are sufficient to attract the 90 kDa heat shock protein (HSP90) such that it frees up heat shock factor 1, which then trimerizes to form an active transcription factor that upregulates expression of heat shock proteins. Upregulation of heat shock proteins and other heat-induced events result in the development of thermotolerance, which protects cells from subsequent exposure to heat shock and other stresses. A more severe heat shock increases protein denaturation proportionately and leads to aggregation of both denatured and native proteins. This results in inactivation of protein synthesis, cell cycle progression, and DNA repair processes such that cells either die or are sensitized to radiation and other cytotoxic events. The ultimate fate of cells following a heat shock depends upon the summation of the activation and inactivation events that are induced, which appears to be governed by the resultant magnitude of protein denaturation and aggregation. Treatments that stabilize cellular proteins against denaturation and aggregation reduce the magnitude of inactivating responses while increasing that of activating responses for a given heat shock (time at temperature), while treatments that sensitize proteins to denaturation and aggreation have the converse effect. These findings support the conclusion that the determinant of the cellular response to heat shock is the amount of heat-induced protein denaturation and aggregation and not the time at temperature.

Animals↗

Adaptation of microorganisms and their transport systems to high temperatures.

Growth of Bacteria and Archaea has been observed at temperatures up to 95 and 110 degrees C, respectively. These thermophiles are adapted to environments of high temperature by changes in the membrane lipid composition, higher thermostabilities of the (membrane) proteins, higher turnover rates of the energy transducing enzymes, and/or the (exclusive) use of sodium-ions rather than protons as coupling ion in energy transduction. The proton permeability of the cytoplasmic membrane of bacteria and archaea was observed to increase with the temperature. This increased proton permeability limits the maximum temperature of growth of bacteria. Higher growth temperatures can be reached by an increased proton pumping activity by using the less permeable sodium ions as coupling ions or by changing the lipid composition of the cytoplasmic membrane. The Na+/H+/glutamate transport proteins of the thermophiles Bacillus stearothermophilus (GltTBs) and Bacillus caldotenax (GltTBc) were studied extensively. These transportproteins have unique features. Transport of L-glutamate occurs in symport with 1 Na+ and 1 H+ when the transport proteins are expressed in their natural environment. The sodium ion dependency of the GltT transporters of these Bacillus strains was found to increase with temperature. However, when the GltT proteins are expressed in the mesophile Escherichia coli, electrogenic symport of L-glutamate occurs with > or = 2 H+. These observations suggest that the conformation of the transport proteins in the E. coli and the Bacillus membranes differs, and that the conformation influences the coupling ion selectivity. The Na+/H+/glutamate transport proteins of B. stearothermophilus (GltTBs) and B. caldotenax (GltTBc) are homologous to transport systems of glutamate and structurally related compounds from mesophilic organisms. Both sodium, as well as proton coupled transporters, belong to this family of carboxylate transporters (FCT).

ATP-Binding Cassette Transporters↗

Regulation of sleep and body temperature in response to exposure to cool and warm environments in neonates.

BACKGROUND AND OBJECTIVE: Thermoregulation is impaired during desynchronized sleep in animals and in adults. This can lead to a conflict between homeothermy and sleep in nonthermoneutral conditions. This study aimed to analyze thermoregulation during sleep, especially during desynchronized sleep (active sleep, AS) and to determine whether the conflict between thermoregulation and sleep might exist in the newborn sleeping in warm or cool conditions. METHODS: Esophageal and skin (cheek and abdomen) temperatures, local sweating rate (ventilated sweat collection capsule stuck on the abdomen), metabolism (indirect respiratory calorimetry), and sleep variables were recorded in 10 newborns exposed, in an incubator, to thermoneutral, warm, and cool environments. Body movements and apneas were also considered. Exposures were performed after a first habituation condition. RESULTS: Sleep structure was not modified by the first exposure nor by the warm environment. Exposure to cool temperatures increased AS duration (+13% of total sleep time) and the quantity of body movements during AS (+11.3% of AS duration), whereas these parameters were not modified during quiet sleep. The thermoregulatory response to warm and cool environments was not impaired during AS. During exposure to mild thermal load, analyses revealed large interindividual differences in the strategy for thermoregulation during AS. Depending on the newborn, the thermoregulatory response to cool temperatures could be described by an increase either in nonshivering thermogenesis or in frequency of body movement. In warm conditions, most newborns exhibited an increased sweating rate. The interindividual differences (lack of increase sweating in three newborns) seemed to be linked to changes in the sensitivity of the sweating response. CONCLUSION: Because thermoregulation is not impaired during AS, this sleep stage seems to be a well-protected one from a thermoregulatory point of view. This difference from adults and animals may be due to the important role of AS in newborn's nervous maturation.

Body Temperature Regulation↗

Disruption of protein-protein interaction in the Mgl-1 oncoprotein.

Mammalian homologues of the Lethal giant larvae (Lgl) tumor suppressor gene have been identified and these homologues can complement the yeast double mutant of Sop1 and Sop2, the yeast homologue of Lgl, as reported previously. In the absence of these genes in yeast, cellular viability is affected at restrictive temperature and salt environments. Members of this family contain five or more of the WD-40 repeat motifs, which is known to be involved in protein-protein interaction. In order to investigate the biochemical roles for conserved amino acids within the most conserved WD-40 repeat motif amongst these family members, we generated deletion mutants for five conserved amino acids (G450, H451, D453, W459 and D460) in mouse Lgl-1 (Mgl-1), located between 450-460 amino acids. We found that the deletion mutants of Mgl-1, DeltaG450 and DeltaD453, were not capable of complementing yeast mutants of Sop1 and Sop2 at restrictive temperature and high salt environments. These results indicate that the WD-40 repeat motif is important for cellular viability by regulating temperature-sensitivity and salt tolerance in yeast.

Amino Acid Motifs↗

Evidence for a hormonal tactic maximizing green turtle reproduction in response to a pervasive ecological stressor.

Mortality of breeding sea turtles due to excessive heat exposure after nesting activities is an unusual feature of the Raine Island green turtle rookery. Breeding turtles that fail to return to the ocean after oviposition can experience increasing body temperatures that exceed lethal limits (>39 degrees C) as ambient temperatures rise after sunrise. We investigated how acute increases in body temperature influenced plasma corticosterone (B) concentrations of individual turtles. Furthermore, interactions between progesterone (P) and testosterone (T) and increasing body temperature and the glucocorticoid corticosterone were examined for negative correlations. Breeding green turtles exhibited a 16-fold mean increase in plasma corticosterone concentration as body temperature (cloacal) rose from 28.2 to 40.7 degrees C in less than 6 h. However, the absolute increase in plasma B was small and much less than expected, despite the lethal stressor. Comparatively, the maximal B response to lethal heat stress was similar to plasma B concentrations obtained from breeding female turtles exposed to 8 h of capture stress. However, the maximal B response of breeding turtles exposed to heat and capture stressors was significantly less than the B response of nonbreeding adult female turtles subjected to an 8-h capture stressor. No negative correlations were observed between plasma T and plasma B, between plasma T and body temperature, between plasma P and plasma B, or between plasma P and body temperature. Our findings provide further evidence that reduced adrenocortical function operates in breeding green turtles in the presence of even the most pervasive of environmental stressors.

Adrenal Cortex↗

Adaptive and differential changes on beta- and alpha 2-adrenoreceptors mediated hyperthermia after chronic treatment with antidepressant drugs in the rat kept at high ambient temperature.

We found previously that the beta-agonist clenbuterol and the alpha-agonist clonidine produced hyperthermia in rats kept at high ambient temperature, which effects were mediated by beta- and alpha 2-adrenoceptors, respectively. In the present paper this observation was used for testing the responsiveness of beta- or alpha 2-adrenoceptors, changed by a pharmacological manipulation, i.e., by chronic treatment with antidepressants. The animals were pretreated with desipramine, imipramine or amitriptyline twice a day for 1 or 2 weeks. All antidepressants significantly attenuated the clenbuterol-induced hyperthermia after 2 weeks of treatment. The effect of desipramine was stronger than that of the other antidepressants and appeared as little as 1 week after the treatment. The hyperthermic effect of clonidine was significantly reduced by repeated treatment with desipramine, increased after 2 weeks administration of imipramine, whereas amitriptyline produced no significant changes. In conclusion, these data suggest that, after repeated treatment, the antidepressants tested produce an adaptive decrease in function of beta-adrenoceptors while the same drugs exert differential effects on alpha 2-receptors. Moreover, clenbuterol induced hyperthermia may be a useful test for examining possible functional changes in beta-adrenoceptor sensitivity.

Amitriptyline↗

Cold and heat tolerance of drosophilid flies with reference to their latitudinal distributions.

The relation between thermal tolerance and latitudinal distribution was studied with 30 drosophilid species collected from the cool-temperate region (Sapporo), the warm-temperate region (Tokyo and Kyoto) and the subtropical region (Iriomote island) in Japan. In addition, intraspecific variation was examined for five species collected from two localities. The subtropical strains of Scaptodrosophila coracina, Drosophila bizonata and D. daruma were less tolerant to cold than their temperate strains. However, the difference of cold tolerance between these two geographic strains was much smaller than the difference between the species restricted to the subtropical region and those occurring in the temperate region. In D. auraria and D. suzukii, no difference was observed in thermal tolerance between their cool- and warm-temperate strains. Thus, geographic variation in thermal tolerance within species was low or negligible. Interspecific comparisons by phylogenetic independent contrasts revealed that species which had the northern boundaries of their distributions at higher latitudes were generally more tolerant to cold than those which had their boundaries at lower latitudes. However, the data for some species did not agree with this trend. The use of man-protected warm places for overwintering, competition or predation would also affect their distributions. It also appeared that species which had their southern boundaries at higher latitudes were generally more cold-tolerant. The acquisition of cold tolerance may lower a fly's capacity to compete, survive or reproduce in warmer climates. On the other hand, no relation was observed between heat tolerance and latitudinal distribution. Heat tolerance was higher in species inhabiting openlands or the forest canopy than in those inhabiting the forest understorey.

Adaptation, Physiological↗

Effects of salts and temperatures on post-irradiation growth of Penicillium exposed to ultraviolet.

The growth of Penicillium notatum colonies after UV irradiation of dried mycelium or spores was studied in relation to post-irradiation temperature and salt environment. Dried mycelium and spores behaved differently with respect to sensitivity to temperature, salts and UV, especially the latter. Threshold inhibitory doses for spores were modified markedly either at 4 degrees C or in magnesium and calcium chlorides. It is suggested that these temperature and salt effects are related to prevention of photochemical membrane damage.

Calcium Chloride↗

Hibernation triggers and cryogens: do they play a role in hibernation?

A survey of the literary evidence on cryogens and hibernation induction triggers is given and the results of experiments on the effect of hypothalamic or i.v. injections of opioids and plasma from hibernating European hamsters on body temperature control of rabbits are presented. Pharmacological doses of a delta opioid--DADLE (25 or 50 micrograms), when injected into the anterior hypothalamus, induce a small and short-lasting hypothermic effect in cold exposed rabbits, due to the downward shift of the temperature threshold for shivering. Lower doses (5 micrograms) are without effect, similarly as i.v. administrations (500 micrograms/kg) of this substance. Intrahypothalamic injections of met-enkephalin (0.1-1 microgram) induce a slight hyperthermia due to the shift of all thermoregulatory effectors to higher body temperatures. Intrahypothalamic injections of plasma from hibernating European hamsters do not influence the body temperature control in rabbits.

Animals↗

Characterization of tryptophan environments in glutamate dehydrogenases from temperature-dependent phosphorescence.

Tryptophan room temperature phosphorescence in solution was detected in glutamic dehydrogenase from bovine liver and Escherichia coli with lifetimes of 1.2 and 0.65 s, respectively. Although these enzymes possess three and five tryptophanyl residues per polypeptide chain, respectively, the temperature dependence of the phosphorescence quantum yield estimates that the room temperature emission is due, in either case, to a single residue. Long triplet-state lifetimes and very small rates of O2 quenching indicate that these tryptophanyl side chains are embedded in a highly inflexible internal region of the macromolecule. Aided by sequence homology with dehydrogenases of known structure and theoretical predictions of secondary structure [Wootton, J.C. (1974) Nature (London) 252, 542-546; Brett, M., Chambers, G.K., Holder, A. A., Fincham, J.R.S., & Wootton, J.C. (1976) J. Mol. Biol. 106, 1-22], the phosphorescing tryptophans have been tentatively placed in the catalytic coenzyme binding domain of each enzyme. The particular sensitivity of the triplet-state lifetime in probing local changes in conformation provides a strong indication that within the time window of phosphorescence measurements the six subunits in the hexameric enzymes are equivalent. Furthermore, while in the bovine enzyme this parameter is markedly affected by the interaction with ligands which have a functional role, the constancy of the phosphorescence lifetime at various degrees of polymerization suggests that the association process is not accompanied by important conformational changes in the macromolecule.

Alcohol Dehydrogenase↗

Novel method using a temperature-sensitive polymer (methylcellulose) to thermally gel aqueous alginate as a pH-sensitive hydrogel.

A novel method using a temperature-sensitive polymer (methylcellulose) to thermally gel aqueous alginate blended with distinct salts (CaCl2, Na2HPO4, or NaCl), as a pH-sensitive hydrogel was developed for protein drug delivery. It was noted that the salts blended in hydrogels may affect the structures of an entangled network of methylcellulose and alginate and have an effect on their swelling characteristics. The methylcellulose/alginate hydrogel blended with 0.7 M NaCl (with a gelation temperature of 32 degrees C) demonstrated excellent pH sensitivity and was selected for the study of release profiles of a model protein drug (bovine serum albumin, BSA). In the preparation of drug-loaded hydrogels, BSA was well-mixed to the dissolved aqueous methylcellulose/alginate blended with salts at 4 degrees C and then gelled by elevating the temperature to 37 degrees C. This drug-loading procedure in aqueous environment at low temperature may minimize degradation of the protein drug while achieving a high loading efficiency (95-98%). The amount of BSA released from test hydrogels was a function of the amount of alginate used in the hydrogels. The amount of BSA released at pH 1.2 from the test hydrogel with 2.5% alginate was relatively low (20%), while that released at pH 7.4 increased significantly (86%). In conclusion, the methylcellulose/alginate hydrogel blended with NaCl could be a suitable carrier for site-specific protein drug delivery in the intestine.

Alginates↗

Molecular recognition in cyclodextrin complexes of amino acid derivatives: the effects of kinetic energy on the molecular recognition of a pseudopeptide in a nonconstraining host environment as revealed by a temperature-dependent crystallographic study.

The crystal structure of a triclinic 2:2 inclusion complex of beta-cyclodextrin with N-acetyl-L-phenylalanine methyl ester has been determined at several temperatures between 298 and 20 K to further study molecular recognition using solid-state supramolecular beta-cyclodextrin complexes. The study reveals kinetic energy dependent changes in guest molecule conformations, orientations, and positions in the binding pocket presented by the crystal lattice. Accompanying these changes are observable differences in guest-guest interactions and hydrogen-bonding interactions in the binding pocket that involve guest molecules, water of hydration molecules, and beta-cyclodextrin molecules. On the basis of the differences observed in the crystal structures, we present a solid-state example of a system that displays the properties of both classical and quantum chemical models. At higher temperatures, the structure conforms to a classical mechanical model with dynamic disorder. At lower temperatures, the observations conform to examples in which there is static disorder representative of models in which quantum states differing in conformation, position, and orientation of components in the crystal structure are occupied. Ab initio theoretical calculations on the different guest molecule conformations have been carried out. Superpositions of theoretical electrostatic surface potential diagrams on the observed molecular positions in the complexes provide confidence that the deconvolution of the guest molecule disorder is acceptable. Temperature-dependent solid-state magic angle spinning deuteron NMR measurements provide evidence for large-amplitude, diffusive motion on a microsecond time scale in the complex.

Amino Acids↗