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Starch-related alpha-glucan/water dikinase is involved in the cold-induced development of freezing tolerance in Arabidopsis.

Cold-induced soluble sugar accumulation enhances the degree of freezing tolerance in various cold-hardy plants including Arabidopsis (Arabidopsis thaliana), where soluble sugars accumulate in only a few hours at 2 degrees C. Hence, along with photosynthesis, starch degradation might play a significant role in cold-induced sugar accumulation and enhanced freezing tolerance. Starch-related alpha-glucan/water dikinase (EC 2.7.9.4), encoded by Arabidopsis STARCH EXCESS 1 (SEX1), is hypothesized to regulate starch degradation in plastids by phosphorylating starch, thereby ensuring better accessibility by starch-degrading enzymes. Here, we show that Arabidopsis sex1 mutants, when incubated at 2 degrees C for 1 d, were unable to accumulate maltooligosaccharides or normal glucose and fructose levels. In addition, they displayed impaired freezing tolerance. After 7 d at 2 degrees C, sex1 mutants did not show any of the above abnormal phenotypes but displayed slightly higher leaf starch contents. The impaired freezing tolerance of sex1 mutants was restored by overexpression of wild-type SEX1 cDNA using the cauliflower mosaic virus 35S promoter. The results demonstrate a genetic link between the SEX1 locus and plant freezing tolerance, and show that starch degradation is important for enhanced freezing tolerance during an early phase of cold acclimation. However, induction of starch degradation was not accompanied by significant changes in alpha-glucan/water dikinase activity in leaf extracts and preceded cold-induced augmentation of SEX1 transcripts. Therefore, we conclude that augmentation of SEX1 transcripts might be a homeostatic response to low temperature, and that starch degradation during an early phase of cold acclimation could be regulated by a component(s) of a starch degradation pathway(s) downstream of SEX1.

Adaptation, Physiological↗

Self-efficacy and perceived control: cognitive mediators of pain tolerance.

The cold-pressor task was used with 102 female undergraduates in 2 experiments to determine (a) whether self-efficacy has validity as a true causal determinant of behavior change or is a correlate of change that has already occurred and (b) how perceptions of control and self-efficacy interact to determine choice behavior, persistence, and the impact of an aversive stimulus. Results of Experiment 1 indicate that self-efficacy expectations affected performance beyond what would have been expected from past performance alone. Changes in self-efficacy expectations predicted changes in cold-pressor tolerance. These findings suggest that self-efficacy expectations can be causal determinants of behavior in an aversive situation. Results of Experiment 2 indicate that self-efficacy was separable from control and that performance was best if both high levels of perceived control and self-efficacy were present. These findings support the notion that self-efficacy expectations can mediate the desirability of providing control, in that those who benefit most from control are those who are most confident they can exercise it.

Adaptation, Psychological↗

Molecular cloning and relationship to freezing tolerance of cold-acclimation-specific genes of alfalfa.

Cold-acclimation-specific (CAS) gene expression has been examined by screening a cDNA library prepared from poly(A)(+) RNA of cold-acclimated seedlings of a freezing-tolerant variety of alfalfa (Medicago falcata cv Anik). Three CAS cDNA clones, pSM784, pSM2201, and pSM2358, representing different sequence species, have been used to investigate the relative abundance and time-course of accumulation of corresponding transcripts. Results obtained show that the expression of these CAS genes is regulated in a coordinated manner most likely at the level of transcription. The expression of genes, as measured by mRNA abundance corresponding to the three CAS cDNA clones, is not stimulated or induced by heat shock, water stress, abscisic acid, or wounding. A positive correlation is observed between the expression of these cloned sequences and the degree of freezing-tolerance in four alfalfa cultivars.

Journal Article↗

Cold-induced freezing tolerance in Arabidopsis.

Changes in the physiology of plant leaves are correlated with enhanced freezing tolerance and include accumulation of compatible solutes, changes in membrane composition and behavior, and altered gene expression. Some of these changes are required for enhanced freezing tolerance, whereas others are merely consequences of low temperature. In this study we demonstrated that a combination of cold and light is required for enhanced freezing tolerance in Arabidopsis leaves, and this combination is associated with the accumulation of soluble sugars and proline. Sugar accumulation was evident within 2 h after a shift to low temperature, which preceded measured changes in freezing tolerance. In contrast, significant freezing tolerance was attained before the accumulation of proline or major changes in the percentage of dry weight were detected. Many mRNAs also rapidly accumulated in response to low temperature. All of the cold-induced mRNAs that we examined accumulated at low temperature even in the absence of light, when there was no enhancement of freezing tolerance. Thus, the accumulation of these mRNAs is insufficient for cold-induced freezing tolerance.

Acclimatization↗

The Arabidopsis cold-responsive transcriptome and its regulation by ICE1.

To understand the gene network controlling tolerance to cold stress, we performed an Arabidopsis thaliana genome transcript expression profile using Affymetrix GeneChips that contain approximately 24,000 genes. We statistically determined 939 cold-regulated genes with 655 upregulated and 284 downregulated. A large number of early cold-responsive genes encode transcription factors that likely control late-responsive genes, suggesting a multitude of transcriptional cascades. In addition, many genes involved in chromatin level and posttranscriptional regulation were also cold regulated, suggesting their involvement in cold-responsive gene regulation. A number of genes important for the biosynthesis or signaling of plant hormones, such as abscisic acid, gibberellic acid, and auxin, are regulated by cold stress, which is of potential importance in coordinating cold tolerance with growth and development. We compared the cold-responsive transcriptomes of the wild type and inducer of CBF expression 1 (ice1), a mutant defective in an upstream transcription factor required for chilling and freezing tolerance. The transcript levels of many cold-responsive genes were altered in the ice1 mutant not only during cold stress but also before cold treatments. Our study provides a global picture of the Arabidopsis cold-responsive transcriptome and its control by ICE1 and will be valuable for understanding gene regulation under cold stress and the molecular mechanisms of cold tolerance.

Adaptation, Physiological↗

PLANT COLD ACCLIMATION: Freezing Tolerance Genes and Regulatory Mechanisms.

Many plants increase in freezing tolerance upon exposure to low nonfreezing temperatures, a phenomenon known as cold acclimation. In this review, recent advances in determining the nature and function of genes with roles in freezing tolerance and the mechanisms involved in low temperature gene regulation and signal transduction are described. One of the important conclusions to emerge from these studies is that cold acclimation includes the expression of certain cold-induced genes that function to stabilize membranes against freeze-induced injury. In addition, a family of Arabidopsis transcription factors, the CBF/DREB1 proteins, have been identified that control the expression of a regulon of cold-induced genes that increase plant freezing tolerance. These results along with many of the others summarized here further our understanding of the basic mechanisms that plants have evolved to survive freezing temperatures. In addition, the findings have potential practical applications as freezing temperatures are a major factor limiting the geographical locations suitable for growing crop and horticultural plants and periodically account for significant losses in plant productivity.

Journal Article↗

Age-related differences in response to acute cold challenge (-10 degrees C) in male F344 rats.

Male F344 rats, 3, 12, and 24 months of age, were acclimated to either 5 degrees C (CA) or 23 degrees C (non-CA). At the end of acclimation, all CA rats exhibited higher basal oxygen uptake than the non-CA when measured at 28 degrees C. When challenged by a 3-hour exposure to -10 degrees C, significant difference in tolerance to cold was observed not only between the CA and non-CA rats but among the different age groups. The 3-month-olds tolerated cold better than 12-month-olds, and the 12-month-olds did better than the 24-month-olds. The ranking order for cold tolerance in -10 degrees C was: 3-month CA greater than 12-month CA greater than 3-month non-CA = 12-month non-CA = 24-month CA greater than 24-month non-CA. Total 3-hour O2 uptake decreased progressively with advancing age. Heat productions were doubled in -10 degrees C in all age groups, and all CA rats had higher heat production than non-CAs. The increase of heat production in -10 degrees C was insufficient to maintain body temperature in all animals. All animals had loss of body heat content, the loss becoming greater with advancing age. Non-CA rats had greater loss of heat content or faster decline of colonic temperature than the CAs. Significant strain-related difference in cold tolerance was noted between the F344 and Sprague-Dawley (S-D) rats, because 24-month-old non-CA male F344 rats still exhibited a better tolerance to -10 degrees C than 3-month-old CA S-D rats.

Aging↗

Catalase in the heat-induced chilling tolerance of cold-stored hybrid Fortune mandarin fruits.

Hybrid Fortune mandarins developed chilling injury (CI) upon cold storage, unless the fruits were conditioned at 37 degrees C for 3 days before they were held at low temperature. This heat treatment induced 2.5-, 1.2-, and 1.4-fold increases in the activities of catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD), respectively, and reduced the activity of glutathione reductase (GR). The differences in the activities afforded by the heat treatment were, in general, maintained during cold storage. However, SOD levels in nonconditioned Fortune fruits exhibiting CI were similar to those of conditioned fruits stored for 0 or 6 weeks at 2 degrees C. No difference between APX activity in the conditioned and nonconditioned fruits stored for 6 weeks at 2 degrees C was found. The data indicate that CAT may be a major antioxidant enzyme operating in the heat-induced chilling tolerance of cold-stored Fortune mandarin fruits.

Ascorbate Peroxidases↗

Preference for safflower oil in rats exposed to a cold environment under free-feeding conditions.

There are several benefits to a high-fat diet for animals exposed to cold, including improved tolerance to severe cold conditions and increased survival rates in cold environments. It is therefore of interest to examine whether animals exposed to cold will selectively consume lipids. We examined the intake of safflower oil (SO) by rats exposed to cold (4 +/- 2 degrees C) under a feeding condition in which the rats were given free access to SO. Rats exposed to cold consumed more SO than those housed at 25 +/- 2 degrees C. This finding suggests that rats prefer SO in a cold environment. There was no significant difference in the ratio of calories of SO ingested to that of matter (standard laboratory chow plus SO) ingested between rats exposed to cold and those at 25 +/- 2 degrees C. The high SO intake also affected cold tolerance and metabolite kinetics in the rats. Factors that affected the SO intake of rats exposed to cold are also discussed.

Animal Feed↗

Mapping regulatory genes as candidates for cold and drought stress tolerance in barley.

Cereal crop yield is greatly affected in many growing areas by abiotic stresses, mainly low temperature and drought. In order to find candidates for the tolerance genes for these stresses, 13 genes encoding for transcription factors and upstream regulators were screened by amplification and SSCP on six parental genotypes of three barley mapping populations ('Nure' x 'Tremois', 'Proctor' x 'Nudinka', and 'Steptoe' x 'Morex'), and mapped as newly developed STS, SNP, and SSCP markers. A new consensus function map was then drawn using the three maps above, including 16 regulatory candidate genes (CGs). The positions of barley cold and drought tolerance quantitative trait loci (QTLs) presently described in the literature were added to the consensus map to find positional candidates from among the mapped genes. A cluster of six HvCBF genes co-mapped with the Fr-H2 cold tolerance QTL, while no QTLs for the same trait were positioned on chromosome 7H, where two putative barley regulators of CBF expression, ICE1 and FRY1, found by homology search, were mapped in this work. These observations suggest that CBF gene(s) themselves, rather than their two regulators, are at present the best candidates for cold tolerance. Four out of 12 drought tolerance QTLs of the consensus map are associated with regulatory CGs, on chromosomes 2H, 5H, and 7H, and two QTLs with effector genes, on chromosomes 5H and 6H. The results obtained could be used to guide MAS applications, allowing introduction into an ideal genotype of favourable alleles of tolerance QTLs.

Chromosome Mapping↗

The leaf-order-dependent enhancement of freezing tolerance in cold-acclimated Arabidopsis rosettes is not correlated with the transcript levels of the cold-inducible transcription factors of CBF/DREB1.

The central part of cold-acclimated rosettes of Arabidopsis thaliana L. (ecotype Columbia) survived freezing at lower temperatures better than did those at the rosette periphery. Electrolyte-leakage tests with detached leaves verified that freezing tolerance in central (or young) leaves increased faster and to a greater extent than in peripheral (or aged and mature) leaves at 2 degrees C. Cold-induced accumulation of sugars could partly account for the leaf-order-dependent enhancement of freezing tolerance after 1 d at 2 degrees C, whereas the role of proline remains to be determined. Cold-induced accumulation of the transcripts of stress-inducible CBF/DREB1 transcription factors apparently disagreed with the observed difference in the freezing tolerance in different leaf orders. However, the levels of COR78/RD29A transcripts were almost the same between different leaf orders after 1-3 d at 2 degrees C, and COR78/RD29A content per total leaf protein was similar between different leaf orders after 7 d at 2 degrees C. Thus, cold-induced accumulation of COR78/RD29A does not seem to account for the observed difference in freezing tolerance in different leaf orders. Although further studies are required for comprehensive understanding of the phenomenon, the present work does provide an important and interesting physiological aspect in our understanding of the freezing tolerance in plants.

Adaptation, Physiological↗

Transgenic B lymphocytes expressing a human cold agglutinin escape tolerance following experimental infection of mice by Mycoplasma pulmonis.

Several microbial infections, including Mycoplasma pneumoniae respiratory infection, are capable, in man, of transiently inducing the expression of anti-red blood cell autoantibody called cold agglutinins (CA). To analyze the mechanisms by which immune tolerance is broken following a mycoplasma infection, we used transgenic mice expressing a pathogenic human CA, designated CA-GAS, specific for sialylated carbohydrates. In these mice peripheral deletion of autoreactive B lymphocytes and receptor editing, prevent the development of autoimmune hemolytic anemia. Experimental infections of transgenic mice with Mycoplasma pulmonis resulted in a high anti-mycoplasma antibody response (despite a severe B cell depletion at the onset of infection), and an important induction of serum CA concentrations, reaching in some mice pathological titers. Whereas in naïve mice, only a small percentage of CA-expressing cells could be detected, in infected mice, a majority of circulating B lymphocytes were large B220(-) cells, which expressed the transgenic immunoglobulin. Immunization of the transgenic mice with keyhole limpet hemocyanin and Freund's adjuvant, to nonspecifically stimulate the expression of the passenger transgenes, only moderately increased the CA titers. These results indicate that M. pulmonis infection is capable of breaking immune tolerance in the CA-transgenic mice, in part through specific activation of CA-expressing B lymphocytes. This experimental infection mimics the induction of CA in humans and provide an animal model for studying the genesis of the autoimmune hemolytic anemia.

Agglutinins↗

Thermoregulatory responses of the inbred heat-tolerant FOK rat to cold.

The responses of inbred heat-tolerant FOK rats to cold were compared with those of Wistar King A/H (WKAH) and Std:Wistar (WSTR) strains. The fall of colonic temperature during cold exposure was unexpectedly smaller in FOK than in other groups, but the onset of shivering was delayed in FOK. Norepinephrine (NE)-induced in vivo oxygen consumption and the mitochondrial uncoupling protein 1 level of brown adipose tissue (BAT) were not different among the groups, but the cold-induced increases in in vivo oxygen consumption as well as plasma glycerol and free fatty acids were higher in FOK than in other groups. In vitro NE-induced oxygen consumption of BAT was less in FOK than WSTR, but not WKAH. The magnitude of the NE-induced increase in blood flow through BAT was higher in FOK than in other groups. These results suggest that FOK paradoxically have a high capacity for nonshivering thermogenesis in spite of their high capacity for heat tolerance, probably due to an increased lipid utilization and improved circulation of BAT.

Adaptation, Physiological↗

Safety and tolerability of cold-adapted influenza virus vaccine in children and adolescents with asthma.

BACKGROUND: Influenza infections can cause severe respiratory disease in high risk persons such as those with asthma, but immunization rates for high risk groups remain suboptimal. An investigational influenza virus vaccine, trivalent, types A and B, live, cold-adapted (CAIV-T) administered by intranasal spray was shown previously to be effective in healthy adults and healthy children. PURPOSE: To assess the safety and tolerability of CAIV-T in subjects 9 years of age and older with moderate to severe asthma. METHODS: In this randomized, double blind, placebo-controlled study, spirometry was performed twice before vaccination to establish a baseline forced expiratory volume at 1 s (FEV1) and once 2 to 5 days thereafter. The primary outcome index was the percent change in percent predicted FEV1 before and after vaccination. Peak flows, clinical asthma symptom scores and nighttime awakening scores were measured daily from 7 days pre- to 28 days postvaccination. RESULTS: The primary outcome index (percentage change in percent predicted FEV1) was not different between the two groups (0.2% vs. 0.4% for the treatment and placebo groups, respectively; P = 0.78). Secondary outcomes did not differ between the two groups; these included the number of subjects with a decrease in FEV1 > or =15% from baseline, reductions in peak flows > or =15%, > or =30% or > or =2 sd below baseline, use of beta-adrenergic rescue medications, asthma exacerbations and clinical asthma symptom scores before and after vaccination. The same proportion of subjects in each group experienced postvaccination symptoms within 10 days (92% and 91%, respectively; P = 1.0). No serious adverse event occurred. CONCLUSION: CAIV-T was generally safe and well-tolerated in children and adolescents with moderate to severe asthma.

Administration, Intranasal↗

Reduced cold pressor pain tolerance in non-recovered whiplash patients: a 1-year prospective study.

UNLABELLED: Whiplash injury and chronic whiplash syndrome represent major health problems in certain western communities, pain being the main symptom. Sensitization of the nociceptive system may play a role for non-recovery after whiplash injury. AIMS: This study examined if tolerance to endure pain stimuli may predict outcome in whiplash injury. In a prospective fashion, 141 acute whiplash patients exposed to rear-end car collision (WAD grade 1-3) and 40 ankle-injured controls were followed and exposed to a cold pressor test, respectively, 1 week, 1, 3, 6 and 12 months after the injury. VAS score of pain and discomfort was obtained before, during and after immersion of the dominant hand into cold water for 2 min. The McGill Pain Questionnaire showed that ankle-injured controls had higher initial pain scores than the corresponding whiplash group, while whiplash-injured subjects had higher scores at 6 months; pain scores being similar at other time points. No difference was found in cold pressor pain between recovered whiplash patients and ankle-injured subjects. Non-recovery was only encountered in whiplash injury. Eleven non-recovered whiplash patients (defined as: handicap after 1 year) showed reduced time to peak pain from 1 week to 3 months (P<0.001), 6 months (P<0.01), but not 12 months after the injury. A larger pain area was seen in non-recovered vs. recovered whiplash-injured subjects during the entire observation period (P<0.001). Non-recovery after whiplash was associated with initially reduced cold pressor pain endurance and increased peak pain, suggesting that dysfunction of central pain modulating control systems plays a role in chronic pain after acute whiplash injury.

Accidents, Traffic↗