Effect of ageing of myosin on its ability to form synthetic filaments and on proteolysis of the LC2 light chain.
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Animals↗
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A general chain binomial model for infectious diseases.
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Common Cold↗
Effect of cold-temperature exposure and acclimation on amino acid pool changes and enzyme activities of rat brown adipose tissue.
The amino acid pool composition and its concentration ratios with respect to blood and plasma, as well as the activities of alanine, aspartate and branched chain amino acid transaminases, glutamine synthetase, adenylate deaminase and glutamate dehydrogenase have been studied in the interscapular brown adipose tissue of control, 12-h cold-exposed and 15-day cold-acclimated rats. Cold temperature affected the amino acid metabolism and pool composition more intensely after 15 days than after 12-h cold-exposure, even though the patterns of change were very similar in both groups. Cold temperatures induced a decrease in glutamine and an increase in glutamate concentration in the tissue. This probably increased the metabolism of branched chain amino acids and caused a decrease in adenylate deaminase activity. It also seemed to increase alanine utilization. We concluded that amino acid metabolism in brown adipose tissue is enhanced by cold temperature acclimation.
AMP Deaminase↗
Effects of diet and cold acclimation on lipid composition of rat interscapular brown adipose tissue.
The effects of diet and of cold acclimation on lipid composition of interscapular brown adipose tissue (BAT) were studied in rats. Three groups of animals were acclimated for 20 weeks to 28 degrees C (controls), 5 degrees C (CA) or to a daily fluctuating temperature (Cy). They received either normal fat (5 p. 100 lipids) [NF] or high fat (26 p. 100) [HF] diets. Cold acclimation increased the tissue weight and its protein, phospholipid, free fatty acid and cholesterol content. It decreased the total lipid proportion in the tissue. HF diet led to an increase in lipid content and to a decrease in the protein content only in control and Cy groups. The mean chain length of triglyceride or phospholipid fatty acids was increased by HF diet and cold acclimation. The proportion of unsaturated acids was not changed in cold acclimated rats, but the amount of arachidonic acid bound to phospholipid was significantly increased. It is concluded that the total amount of lipid in the diet acts on the synthesis of BAT lipids. The cold dependant changes in lipid composition which are similar but less important in cyclic cold acclimation than in constant cold adaptation seem to be proportional to the increase in the tissue energy metabolism.
Acclimatization↗
The Antarctic Psychrobacter sp. TAD1 has two cold-active glutamate dehydrogenases with different cofactor specificities. Characterisation of the NAD+-dependent enzyme.
Psychrobacter sp. TAD1 is a psychrotolerant bacterium from Antarctic frozen continental water that grows from 2 to 25 degrees C with optimal growth rate at 20 degrees C. The new isolate contains two glutamate dehydrogenases (GDH), differing in their cofactor specificities, subunit sizes and arrangements, and thermal properties. NADP+-dependent GDH is a hexamer of 47 kDa subunits and it is comparable to other hexameric GDHs of family-I from bacteria and lower eukaria. The NAD+-dependent enzyme, described in this communication, has a subunit weight of 160 kDa and belongs to the novel class of GDHs with large size subunits. The enzyme is a dimer; this oligomeric arrangement has not been reported previously for GDH. Both enzymes have an apparent optimum temperature for activity of approximately 20 degrees C, but their cold activities and thermal labilities are different. The NAD+-dependent enzyme is more cold active: at 10 C it retains 50% of its maximal activity, compared with 10% for the NADP+-dependent enzyme. The NADP+-dependent enzyme is more heat stable, losing only 10% activity after heating for 30 min, compared with 95% for the NAD+-dependent enzyme. It is concluded that in Psychrobacter sp. TAD1 not only does NAD+-dependent GDH have a novel subunit molecular weight and arrangement, but that its polypeptide chains are folded differently from those of NADP+-dependent GDH, providing different cold-active properties to the two enzymes.
Adaptation, Physiological↗
Expression of axolotl RNA-binding protein during development of the Mexican axolotl.
Amphibians occupy a central position in phylogeny between aquatic and terrestrial vertebrates and are widely used as model systems for studying vertebrate development. We have undertaken a comprehensive molecular approach to understand the early events related to embryonic development in the Mexican axolotl, Ambystoma mexicanum, which is an exquisite animal model for such explorations. Axolotl RBP is a RNA-binding protein which was isolated from the embryonic Mexican axolotl by subtraction hybridization and was found to show highest similarity with human, mouse, and Xenopus cold-inducible RNA-binding protein (CIRP). The reverse transcriptase polymerase chain reaction (RT-PCR) analysis suggests that it is expressed in most of the axolotl tissues except liver; the expression level appears to be highest in adult brain. We have also determined the temporal and spatial pattern of its expression at various stages of development. RT-PCR and in situ hybridization analyses indicate that expression of the AxRBP gene starts at stage 10-12 (gastrula), reaches a maxima around stage 15-20 (early tailbud), and then gradually declines through stage 40 (hatching). In situ hybridization suggests that the expression is at a maximum in neural plate and neural fold at stage 15 (neurula) of embryonic development.
Ambystoma↗
Uncoupling protein 3: its possible biological role and mode of regulation in rodents and humans.
The recently discovered uncoupling protein 3 (UCP3) is highly homologous to the mitochondrial inner membrane protein UCP1, which generates heat by uncoupling the respiratory chain from oxidative phosphorylation. The thermogenic function of UCP1 protects against cold and regulates the energy balance in rodents. We review in vitro studies investigating the uncoupling activity of UCP3 and in vivo studies, which address UCP3 gene expression in brown adipose tissue and skeletal muscle under various metabolic conditions. The data presented are, for the most, consistent with an uncoupling role for UCP3 in regulatory thermogenesis. We also discuss mediators of UCP3 regulation and propose a potential role for intracellular fatty acids in the mechanism of UCP3 modulation. Finally, we hypothesize a role for UCP3 in the metabolic adaptation of the mitochondria to the degradation of fatty acids.
Animals↗
Heterogeneity and specificity of human anti-insulin antibodies determined by isoelectric focusing.
Anti-insulin antibodies are present in the majority of insulin treated diabetics, and in some cases these antibodies have been found to be highly specific for limited epitopes on the molecule. To determine how the human response differs from that seen in inbred animals, we have examined the heterogeneity and specificity of human anti-insulin antibodies by isoelectric focusing (IEF). In addition, we have used human insulin to examine the extent of autoreactivity in the serum of subjects treated with animal insulins. The majority of diabetic sera exhibited complex IEF spectra that were composed of discrete bands and unresolved smears. Autoradiography using 125I-beef, pork, and human insulin revealed some affinity differences; however, the predominant antibodies were capable of binding all insulins, including human. These specificity studies were extended by comparing competitive inhibition with excess cold insulins, and sera with highly specific binding of the A chain loop of beef insulin were identified. The spectra by IEF of these highly specific sera were found to be variable. Our results indicate that the majority of insulin-treated diabetics develop a heterogeneous antibody response that is more complex than the response of inbred animals and includes reactivity with autologous insulin. Although infrequent, individuals having antibodies directed at limited regions of the molecule can be identified and will provide valuable tools for dissecting this complex response.
Animals↗
Suppression of fatty acids synthesis in brown adipose tissue of mice fed diets rich in long chain fatty acids.
The influence of dietary lipid on fatty acid synthesis in brown adipose tissue has been investigated by feeding different high-fat diets to cold-acclimated mice for a period of 2 weeks. Fatty acid synthesis was measured in vivo with 3H2O, and the fats used in the study were maize oil, beef tallow and medium chain triacylglycerol oil. In the mice fed the maize oil and the beef tallow diets fatty acids synthesis was inhibited in all tissue examined--interscapular brown adipose tissue, epididymal white adipose tissue, the liver and the carcass. Synthesis was more inhibited, however, in brown adipose tissue than in other tissues, and the inhibition was greater on the maize oil diet than on the beef tallow. The medium chain triacylglycerol oil had no inhibitory effect on fatty acid synthesis in any tissue, and hepatic synthesis was even elevated on this diet. It is concluded that fatty acid synthesis in brown adipose tissue, as in other lipogenic tissues, is subject to strong suppression by dietary long chain fatty acids, and particularly by linoleic acid.
Acclimatization↗
Influence of early postnatal cold exposure on myofiber maturation in pig skeletal muscle.
Early after birth, piglets rely almost exclusively on muscular shivering thermogenesis to produce heat in the cold and this can possibly modulate skeletal muscle development. An experiment involving 10 individually housed piglets was conducted to determine the influence of cold (24-15 degrees C, D5C group) vs. thermoneutrality (34-30 degrees C, D5TN group) between birth and 5 days on myosin heavy chain (MyHC) polymorphism and metabolic characteristics of longissimus lumborum (LL) and rhomboideus (RH) muscles. Five additional piglets were sacrificed at birth. Piglets exposed to cold received 43% more artificial milk on a liveweight basis in order to achieve similar growth rates. D5C piglets produced 93% more heat and exhibited intense shivering during the whole experiment. Contractile and metabolic characteristics of muscles were determined by immunocytochemistry, electrophoresis and enzyme activities. At least eight MyHC isoforms were detected, including atypical expressions of the alpha-cardiac and extraocular isoforms. Dramatic changes in MyHC composition, myofiber cross-sectional area (CSA) and energy metabolism occurred between birth and 5 days. Cold exposure did not affect either the total number of fibers or the CSA, but it did influence muscle maturation. In particular, it increased the expression of alpha-cardiac and type I MyHC, and decreased that of fetal MyHC, confirming an acceleration in the rate of postnatal maturation. An increase in oxidative enzyme activities was observed in both muscles in the cold, whereas the activity of a glycolytic enzyme, lactate dehydrogenase, remained unchanged. Cold exposure also induced an increase in T3 plasma levels. The extent to which these changes are the result of sustained shivering or are due to the action of hormonal factors, such as thyroid hormones, are discussed.
Age Factors↗
A branched-chain amino acid aminotransferase gene isolated from Hordeum vulgare is differentially regulated by drought stress.
Differential display was used to isolate cDNA clones showing differential expression in response to ABA, drought and cold in barley seedling shoots. One drought-regulated cDNA clone (DD12) was further analyzed and found to encode a branched-chain amino acid aminotransferase (HvBCAT-1). A genomic clone was isolated by probing the Morex BAC library with the cDNA clone DD12 and the structure of Hvbcat-1 was elucidated. The coding region is interrupted by six introns and contains a predicted mitochondrial transit peptide. Hvbcat1 was mapped to chromosome 4H. A comparison was made to rice and Arabidopsis genes to identify conserved structural patterns. Complementation of a yeast (Saccharomyces cerevisiae) double knockout strain revealed that HvBCAT-1 can function as the mitochondrial (catabolic) BCATs in vivo. Transcript levels of Hvbcat-1, increased in response to drought stress. As the first enzyme in the branched-chain amino acid (BCAA) catabolic pathway, HvBCAT-1 might have a role in the degradation of BCAA. Degradation of BCAA could serve as a detoxification mechanism that maintains the pool of free branched-chain amino acids at low and non toxic levels, under drought stress conditions.
Abscisic Acid↗
Functional and compositional studies of arteries stored in University of Wisconsin solution compared with Krebs-Bülbring buffer.
Endothelium-dependent and -independent vasorelaxation in ring segments of rabbit thoracic aorta is reduced and noradrenaline-induced vasoconstriction unaltered after prolonged storage in University of Wisconsin solution (UW) compared to arteries stored in extracellular-type solutions such as Krebs-Bülbring buffer (KBB). The aims of the present study were to determine whether angiotensin-II-induced vasoconstriction, alterations in myosin light chains, protein synthetic capacity, and subcellular structures are altered after 8 days of UW storage at 4 degrees C. The present study showed reduced contractility to angiotensin II, following 8 days of cold storage in UW, that was not reversed in the presence of a nitric oxide synthase inhibitor, N(G)-nitro-l-arginine methyl ester (100 microM). Measurements of contractile protein ratios in the same tissues after cold storage in UW or KBB did not show any significant alterations in smooth muscle myosin light chains or protein synthetic capacity (reflected by total RNA). It is concluded that reductions in vasoconstriction in UW-stored tissue are unlikely to be due to increased release of nitric oxide nor reduced availability of myosin light chains for phosphorylation and vasoconstriction.
Angiotensin II↗
The primary structure of the Fab fragment of protein KAU, a monoclonal immunoglobulin M cold agglutinin.
The complete amino acid sequence of the Fab fragment of protein KAU, a human monoclonal cold agglutinin (IgMk) with anti-I activity, was determined. The light chain (L-chain) consists of 215 residues; the variable (V)L region belongs to the Hum/Kv325/kIIIb sub-subgroup that is preferentially selected in human IgM autoimmune response. The joining (J) region is encoded by the Jk4 gene, and the constant region (C)L domain expresses the km3 allotypic marker. The Fd fragment contains 232 amino acids, and 120 of them comprise the variable domain. The VH region corresponds to the VHIV subgroup and is closely related to the VHIV 2.1 gene isolated from genomic DNA expressed in peripheral blood of a healthy Caucasian. The complementary-determining region 1 has a unique amino acid (Asp) at position 31, and the complementary-determining region 3 codified by the diversity segment (D) gene, shows poor homology with other known D sequences. The joining segment with two unusual substitutions at the D-J junction is encoded by the JH4 gene. Thus, cold agglutinin KAU is an IgM, VkIIIb-Jk4-km3; VHIV-JH4-C mu.
Agglutinins↗
Circulatory disturbances of the foot in vibration syndrome.
Circulatory disturbances of the foot in patients with vibration syndrome were studied by measuring the skin temperature of both index fingers and great toes through a 3-min immersion of the right foot in cold water at 10 degrees C. Subjects included 11 patients with vibration-induced white finger (VWF) [VWF(+) group], 12 patients without VWF [VWF(-) group], and 20 healthy referents not exposed to vibration. Patients were all male chain saw operators who had scarcely been exposed to vibration of the foot. The prevalence of coldness felt in the upper and lower extremities was greater than 90% in the VWF(+) group, about 60% in the VWF(-) group, and less than 10% in the referents. The extent of the coldness was greatest in the VWF(+) group. The skin temperature of both fingers and toes was lowest in the VWF(+) group, somewhat higher in the VWF(-) group, and highest in the referents both before and after immersion. These findings indicate that patients with vibration syndrome, especially those with VWF, have circulatory disturbances in the foot as well as in the hand. The disturbances in the foot may be related to long-term repeated vasoconstriction in the foot induced by hand-arm vibration through the sympathetic nervous system.
Aged↗
Adaptation of fatty acid composition to temperature--a study on carp (Cyprinus carpio L.) liver slices.
Liver slices obtained from warm-, and cold-adapted carp were incubated in the presence of [1-14C]sodium acetate, -stearate, -linoleate, and -linolenate at various temperatures and the distribution of radioactivity among different phospholipid fatty acids was determined. Relative labelling of saturated fatty acids is reduced, while that of long chain polyunsaturated fatty acids, especially of docosahexanoate, is increased with decreasing temperatures. Liver slices of cold adapted carp produced a fatty acid population at 25 degrees C indistinguishable from that produced by warm adapted ones at the same temperature. Liver slices obtained from cold-, and warm-adapted animals start to reorganise the pattern of labelling immediately after the exposure to the opposite temperatures as evident from pulse-chase labelling experiments. Desaturation of saturated and various unsaturated fatty acids is initiated immediately after down-shift of the temperature. This cold induced increase in desaturase activity is prevented by cycloheximide in the incubation medium. It is concluded that phospholipid fatty acid composition is continuously adjusted to the temperature and is governed partly by temperature coefficient of fatty acid synthetase and partly by induction or deactivation of desaturases in cold and warm, respectively.
Acclimatization↗
Drought acclimation confers cold tolerance in the soil collembolan Folsomia candida.
It has been noted that both summer drought and sub-zero winter temperatures induce the synthesis of sugars and polyols in invertebrate tissues. This has led several authors to suggest that many of the adaptations, previously viewed as a response to cold, might be part of a more universal desiccation tolerance mechanism. Here we show that acclimation of the soil dwelling collembolan Folsomia candida to a sublethal desiccation stress confers tolerance to cold shock and a significant increase in the molar percent of membrane fatty acids with a mid-chain double bond. These changes in membrane fatty acids are interpreted as conferring a significant reduction in the transition temperature of cell membranes, as would be expected in acclimation to cold, and these changes are therefore interpreted as contributing to the cross-tolerance. Drought acclimation was also shown to trigger the synthesis of the 70kDa family of heat-shock proteins (Hsp70). This group of heat shock proteins is implicated in the reestablishment of the normal three-dimensional structure of partially unfolded proteins and therefore are also likely to contribute to the observed cross-tolerance. This study provides evidence that the stresses exerted by desiccation and cold at the cellular level have sufficient similarities to induce overlapping adaptations.
Journal Article↗
[Thyroid hormones, obesity and brown adipose tissue thermogenesis].
Brown adipose tissue (BAT) is the main site for hormone-dependent (non-shivering) thermogenesis in response to cold in lower mammals. The hypothalamus controls the cold-induced BAT activation by stimulating the sympathetic nerves and the secretion of norepinephrine (NE) in BAT. Mediated by beta-3 noradrenergic receptor and in the presence of triiodothyronine (T3), NE promotes the synthesis of the uncoupling protein 1 (UCP1). UCP1 is a 32 kDa protein located in the inner membrane of BAT mitochondria, where it dissipates the proton gradient created by oxidations in the mitochondria. UCP1 functions as a proton translocator, substituting for another translocator, the ATP synthetase. The uncoupling of oxidations and phosphorylations and the inhibition of ATP synthesis lead to dissipation as heat of all energy produced in the respiratory chain. The supply of adequate amounts of T3 is ensured by the cold-induced enhancement of the enzyme 5'-deiodinase type II activity, which deiodinates thyroxine (T4) to T3. The absence of T3 blocks UCP1 synthesis, leading to hypothermia. BAT has a limited significance in humans, except in the newborn, where it serves for a rapid acclimation to ambient temperature. The study of BAT physiology will provide more insight into the mechanisms regulating energy balance and body weight in humans, thus contributing to prevent and treat human obesity.
Adipose Tissue, Brown↗
Cold denaturation-induced conformational changes in phosphoglycerate kinase from yeast.
The temperature-dependent conformational equilibrium of 3-phosphoglycerate kinase has been studied in the temperature range from 1 to 30 degrees C by means of dynamic light scattering, small-angle X-ray scattering, differential scanning calorimetry, circular dichroism spectroscopy, and fluorescence spectroscopy. At 28 degrees C and in the presence of 0.7 M guanidine hydrochloride (GuHCl), the radius of gyration (RG) and the Stokes radius (RS) are 2.44 and 3.09 nm, respectively. Decreasing the temperature effects unfolding of the molecule, a process that involves two stages. The two stages correspond to the successive unfolding of the N-terminal and C-terminal domains. The peak maxima of the excess heat capacity, determined from differential calorimetric scans, extrapolated to 0 scan rate, are positioned at 16.5 degrees C for the N-terminal domain and at 6.3 degrees C for the C-terminal domain. At 4.5 degrees C, the radius of gyration and the Stokes radius increase to 7.8 and 4.8 nm, respectively. The persistence length and the length of the statistical chain segment of the unfolded polypeptide chain are 1.74 and 3.48 nm, corresponding to five and ten amino acids, respectively. At 1 degrees C, the dimensions of the unfolded chain nearly agree with the predicted dimensions under theta conditions. Thus, the conformational changes upon cold denaturation can be described by a transition from a compactly folded molecule to a random coil. The conformation-dependent ratio rho = RGRS-1 increases from rho = 0.79 to rho = 1.63. The volume of the unfolded chain is 30 times larger than that of the folded chain in the native state.(ABSTRACT TRUNCATED AT 250 WORDS)
Calorimetry, Differential Scanning↗