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PubMed2002

Differential gene expression in the brain of channel catfish ( Ictalurus punctatus) in response to cold acclimation.

Aquatic ectotherms can adapt to a wide range of temperature changes, but the molecular mechanisms that underlie this adaptability are not well understood. We identified genes that are differentially expressed in the catfish ( Ictalurus punctatus) brain using a cDNA microarray approach to gain an initial understanding of adaptation to low temperature. Among 660 genes analyzed, 61 were differentially expressed when compared at 12 degrees C and 24 degrees C. Gene induction was rapid, occurring within 2 h of the temperature shift. The major categories of differentially expressed genes included (1) genes for chaperones such as Hsp70 and Hsp70/Hsp90 organizing protein; (2) genes for transcription factors and gene products involved in signal transduction pathways such as zinc-finger proteins, calmodulin kinase inhibitor, the nuclear autoantigen SG2NA, interferon regulatory factor 3, and inorganic pyrophosphatase; (3) genes involved in lipid metabolism such as TB2 and acyl CoA binding protein; and (4) genes involved in the translational machinery such as ribosomal proteins. Some genes were induced transiently, whereas others were induced in an enduring fashion. Several genes, primarily ribosomal protein genes, were down regulated, indicating reduced metabolic activities after extended incubation at the low temperature. Thus channel catfish respond to low temperature by adjusting expression of a large number of genes. The rapid induction of proteins involved in signal transductions and chaperones suggests that both de novo synthesis of cold-induced proteins and modification of existing proteins are required for adaptation and tolerance of catfish to low environmental temperature.

Acclimatization↗
PubMedDate not supplied

Sucrose metabolism of perennial ryegrass in relation to cold acclimation.

Sugar metabolism is one of the important factors involved in winter hardiness and since the discovery of sucrose biosynthesis, considerable advances have been made in understanding its regulation and crucial role. This investigation examined the changes in activities of sucrose metabolizing enzymes and sugar content during cold hardening of perennial ryegrass (Lolium perenne L.). Changes in acid invertase (AI), sucrose synthase (SS) and sucrose phosphate synthase (SPS) along with all the three soluble sugars glucose, fructose and sucrose were measured in leaves and stem base tissue during cold acclimation. Although fructans were the predominant carbohydrate the changes in glucose, fructose and sucrose were significant. All the three soluble sugars in both leaf and stem tissues started to decrease from the first day and continued up to day 7 and thereafter started to increase until day 28. AI in the soluble fraction showed a higher activity than that in the cell wall bound fraction. In both the leaf and stem bases soluble AI activity increased during the first week and after that it started to decrease gradually. On the other hand both the SS and SPS increased gradually throughout the acclimation period. Sucrose content was negatively correlated with AI and positively correlated with SS and SPS accounting well for the relation between the substrate and enzyme activity. These results suggest that AI, SS and SPS in ryegrass are regulated by cold acclimation and play an important role in sugar accumulation and acquisition of freezing tolerance.

Acclimatization↗
PubMed1998

Survival of Escherichia coli O157:H7 in synthetic gastric fluid after cold and acid habituation in apple juice or trypticase soy broth acidified with hydrochloric acid or organic acids.

Extreme acid tolerance of Escherichia coli O157:H7 has raised doubts about the safety of acidic foods. This study examined whether prior storage in acidic and/or cold conditions enhanced survival of E. coli O157:H7 in synthetic gastric fluid (SGF). Three E. coli O157:H7 strains were stored in trypticase soy broth (TSB; acidified with HCl, malic acid, citric acid, or lactic acid) or pH 3.5 and 6.5 (nonacidic control) apple juice at 4 and 21 degrees C for < or = 7 days and then were incubated in pH 2.5 SGF at 37 degrees C for 4 h. Cells survived better in apple juice than in TSB containing organic acids, suggesting that juice constituents other than organic acids protect E. coli O157:H7. Refrigeration combined with low pH best protected cells in apple juice and acidified TSB, but, compared to the nonacidic control, only acidified TSB enhanced subsequent survival in pH 2.5 SGF. Equal survival in SGF occurred after storage in pH 3.5 or 6.5 apple juice at 4 degrees C, suggesting that low temperature alone in apple juice enhanced acid tolerance. Two strains stored at 4 degrees C in TSB containing malic or citric acid subsequently survived better in SGF than cells stored in nonacidified TSB but poorer than cells stored in the presence of HCl. These differences reflect the higher pKa of these organic acids. However, subsequent survival of these strains in SGF was poorer after refrigerated storage in apple juice than in TSB containing citric or malic acids. Cells stored in lactic acid were most likely to be completely eliminated upon transfer to SGF. Differences in survival in storage media or SGF related to strain, storage conditions, or acidifier were consistent and often statistically significant (P < 0.05). Although the survival of E. coli O157:H7 in refrigerated acidic beverages may not be affected by the type of acidifier used, the subsequent survival in SGF of this pathogen may be critically dependent on this factor.

Beverages↗
PubMed2006

Freezing tolerance in grasses along an altitudinal gradient in the Venezuelan Andes.

The tropical high Andes experience greater daily temperature oscillations compared to seasonal ones as well as a high frequency of night frost occurrence year round. Survival of organisms, under such environmental conditions, has been determined by selective forces which have evolved into adaptations including avoidance or tolerance to freezing. These adaptations have been studied in different species of trees, shrubs and perennial herbs in páramo ecosystems, while they have not been considered in grasses, an important family of the páramo. In order to understand survival of Poaceae, resistance mechanisms were determined. The study was performed along an altitudinal gradient (2,500-4,200 m a.s.l.) in the páramo. Supercooling capacity and frost injury temperature were determined in nine species in order to establish cold resistance mechanisms. Grasses registered a very low supercooling capacity along the altitudinal gradient, with ice formation between -6 and -3 degrees C. On the other hand, frost injury temperature oscillated between -18 and -7 degrees C. Our results suggest that grasses exhibit freezing tolerance as their main cold resistance mechanism. Since grasses grow at ground level, where greatest heat loss takes place, tolerance may be related to this life form as reported for other small life forms.

Adaptation, Physiological↗
PubMed2001

Regulation of a wheat actin-depolymerizing factor during cold acclimation.

We have previously shown that the wheat (Triticum aestivum) TaADF gene expression level is correlated with the plants capacity to tolerate freezing. Sequence analysis revealed that this gene encodes a protein homologous to members of the actin-depolymerizing factor (ADF)/cofilin family. We report here on the characterization of the recombinant TaADF protein. Assays for ADF activity showed that TaADF is capable of sequestering actin, preventing nucleotide exchange, and inducing actin depolymerization. In vitro phosphorylation studies showed that TaADF is a substrate for a wheat 52-kD kinase. The activity of this kinase is modulated by low temperature during the acclimation period. Western-blot analyses revealed that TaADF is expressed only in cold-acclimated Gramineae species and that the accumulation level is much higher in the freezing-tolerant wheat cultivars compared with the less tolerant ones. This accumulation was found to be regulated by a factor(s) encoded by a gene(s) located on chromosome 5A, the chromosome most often found to be associated with cold hardiness. The induction of an active ADF during cold acclimation and the correlation with an increased freezing tolerance suggest that the protein may be required for the cytoskeletal rearrangements that may occur upon low temperature exposure. These remodelings might be important for the enhancement of freezing tolerance.

Acclimatization↗
PubMed2003

Cold exposure induces tissue-specific modulation of the insulin-signalling pathway in Rattus norvegicus.

Cold exposure provides a reproducible model of improved glucose turnover accompanied by reduced steady state and glucose-induced insulin levels. In the present report we performed immunoprecipitation and immunoblot studies to evaluate the initial and intermediate steps of the insulin-signalling pathway in white and brown adipose tissues, liver and skeletal muscle of rats exposed to cold. Basal and glucose-induced insulin secretion were significantly impaired, while glucose clearance rates during a glucose tolerance test and the constant for glucose decay during a 15 min insulin tolerance test were increased, indicating a significantly improved glucose turnover and insulin sensitivity in rats exposed to cold. Evaluation of protein levels and insulin-induced tyrosine (insulin receptor, insulin receptor substrates (IRS)-1 and -2, ERK (extracellular signal-related kinase)) or serine (Akt; protein kinase B) phosphorylation of proteins of the insulin signalling cascade revealed a tissue-specific pattern of regulation of the molecular events triggered by insulin such that in white adipose tissue and skeletal muscle an impaired molecular response to insulin was detected, while in brown adipose tissue an enhanced response to insulin was evident. In muscle and white and brown adipose tissues, increased 2-deoxy-D-glucose (2-DG) uptake was detected. Thus, during cold exposure there is a tissue-specific regulation of the insulin-signalling pathway, which seems to favour heat-producing brown adipose tissue. Nevertheless, muscle and white adipose tissue are able to take up large amounts of glucose, even in the face of an apparent molecular resistance to insulin.

Adaptation, Physiological↗
PubMed1999

Context-dependence of amino acid residue pairing in antiparallel beta-sheets.

In an effort to understand the driving forces behind antiparallel beta-sheet assembly, we have investigated the mutational tolerance of four pairs of residues in CspA, the major cold shock protein of E. coli. Two buried pairs and two exposed pairs of neighboring amino acids were separately randomized and the corresponding effects on protein stability were assessed using a protein expression screen. The thermal denaturation of a subset of the recovered proteins was measured by circular dichroism spectroscopy in order to determine the range of stabilities sampled by the expressed mutants. As anticipated, buried sites are substantially less tolerant of substitutions than exposed sites with more than half of the exposed residue combinations giving rise to stably folded proteins. The two exposed residue pairs, however, display different degrees of tolerance to substitution and accept different residue pair combinations. Except for the prohibition of proline from interior strand positions, no obvious correlations of mutant stability with any single parameter such as beta-sheet propensity or hydrophobicity can be detected. Mutant combinations recovered in both orientations (e.g. XY and YX) at a given exposed pair site often show markedly different stabilities, indicating that the local environment plays a substantial role in modulating the pairing preferences of residues in beta-sheets.

Amino Acids↗
PubMed1997

The induction of kin genes in cold-acclimating Arabidopsis thaliana. Evidence of a role for calcium.

The involvement of calcium signaling during cold-induction of the kin genes of Arabidopsis thaliana (L.) Heynh. was examined. Treatments with chemicals which either chelate extracellular calcium (EGTA) or block the plasma-membrane calcium channels (La3+, Gd3+) inhibited cold acclimation as well as kin gene expression. Ruthenium red, an inhibitor of calcium release from intracellular stores partially inhibited kin gene expression and development of freezing tolerance. An inhibitor of calcium-dependent protein kinases (CDPKs) and calmodulin prevented cold acclimation as well as the cold induction of kin genes. Using restriction fragment length polymorphism-coupled domain-directed differential display, five CDPK clones were identified which showed differential regulation by cold. The amplified fragments showed homology to known plant CDPKs. The involvement of calcium and calcium-binding proteins in cold acclimation of A. thaliana is discussed.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1995

Expression of a Low-Temperature-Induced Protein in Brassica napus.

BN28 is a low-temperature-induced, boiling-soluble protein in Brassica napus. We used antibodies raised against a recombinant BN28 to examine the expression of this protein in cold-acclimating plants and to investigate its relationship to plant freezing tolerance. Changes in the steady-state levels of BN28 protein appear to lag several days behind those of the mRNA. BN28 is first detected on immunoblots after approximately 8 d of exposure to low temperature, and thereafter levels remain stable while plants are maintained at 4[deg]C. Radiolabeling studies indicate that BN28 is synthesized at a relatively low rate. A decline in protein levels is observed soon after returning plants to control temperatures, and little or no protein can be detected after 7 d of deacclimation. The disappearance of the protein precedes a loss in freezing tolerance, suggesting that BN28 is not involved in maintaining plasma membrane integrity. Expression of BN28 is observed primarily in leaves and appears to be low-temperature specific. Quantitative analysis indicated that BN28 accumulates to approximately 82.7 pmol mg-1 total protein in cold-acclimated leaves. This concentration is similar to that reported for two group 2 late-embryogenesis-abundant-like proteins.

Journal Article↗
PubMed2026

Rhodoglobus galactosidasius sp. nov., a psychrophilic Actinomycetota producing a cold-active &#x3b2;-galactosidase isolated from Cape Hallett, Antarctica.

A novel psychrophilic member of the phylum Actinomycetota, designated strain CPHL_1T, was isolated from the shoreline of Cape Hallett, Antarctica. Cells were Gram-positive, aerobic, non-motile, non-spore-forming rods (0.7-1.0&#x2009;&#xb5;m in length) with orange to yellow pigmentation. Optimal growth occurred at 15&#x2009;&#xb0;C, pH 8.0, with NaCl tolerance up to 3% (w/v). Strain CPHL_1T exhibited cytosolic cold-active &#x3b2;-galactosidase activity, with an optimum at 15&#x2009;&#xb0;C and pH 7.0 and formed non-adherent biofilms. Phylogenetic analysis based on 16S rRNA gene sequence placed the strain within the genus Rhodoglobus, showing high similarity to Rhodoglobus vestalii LV3T (99.4%) and Rhodoglobus aureus CMS 81yT (97.6%). Chemotaxonomic characteristics, including the predominance of anteiso-C15&#x200a;:&#x200a;0, anteiso-C17&#x200a;:&#x200a;0 and iso-C16&#x200a;:&#x200a;0 fatty acids, the presence of MK-11, MK-10 and MK-12 as the major respiratory quinones, and genomic G+C content of 60%, further supported its assignment to the genus Rhodoglobus. Whole-genome comparisons revealed clear taxonomic distinction, with strain CPHL_1T sharing 88.9% average nucleotide identity (ANI) and 38.3% digital DNA-DNA hybridization (dDDH) values with R. aureus, and 81.7% ANI and 25.1% dDDH with R. vestalii, confirming its status as a novel species. Phylogenetic analysis revealed that the &#x3b2;-galactosidases from these Rhodoglobus strains form a well-supported clade with high sequence similarity. Based on these data, a novel species is proposed: Rhodoglobus galactosidasius sp. nov. (type strain CPHL_1T =ITEM 19986T =KCTC 59592T). Genomic analyses also highlighted its biosynthetic potential for specialized metabolites, consistent with the recognized metabolic diversity of the genus.

Phylogeny↗
PubMed2006

A perennial ryegrass CBF gene cluster is located in a region predicted by conserved synteny between Poaceae species.

CBF/DREB1 proteins are the most important regulators of the cold temperature signaling pathway in many plants. CBF genes are candidates for low-temperature tolerance QTL in wheat and barley. Ten novel putative CBF cDNAs of perennial ryegrass (Lolium perenne L.) have been isolated from cold-treated leaf tissue. Their primary structures contain some conserved motifs, characteristic of the gene class. Phylogenetic analysis revealed that LpCBF genes were attributable to the HvCBF3-, and HvCBF4-subgroups following the previously proposed classification of barley CBF genes. RT-PCR analysis revealed that the expression of LpCBF genes was rapidly induced in response to low temperature and that the expression pattern under the low-temperature conditions for a long period was different between the various LpCBF genes. Five of the ten LpCBF genes were assigned to the genetic linkage map using the p150/112 reference mapping population. LpCBFIb, LpCBFII, LpCBFIIIb and LpCBFIIIc were mapped on LG5 forming a cluster within 2.2 cM, while LpCBFVb was located on LG1. Based on comparative genetic studies, conserved synteny for CBF gene family was observed between the Triticeae cereals and perennial ryegrass. Information on the perennial ryegrass CBF genes at both the molecular and genetic level obtained in this study would be useful for the further study on the role of CBF genes and low-temperature tolerance in grasses.

Amino Acid Sequence↗
PubMed2004

QTL mapping of chromosomal regions conferring reproductive frost tolerance in barley ( Hordeum vulgare L.).

Spring radiation frost is a major abiotic stress in southern Australia, reducing yield potential and grain quality of barley by damaging sensitive reproductive organs in the latter stages of development. Field-based screening methods were developed, and genetic variation for reproductive frost tolerance was identified. Mapping populations that were segregating for reproductive frost tolerance were screened and significant QTL identified. QTL on chromosome 2HL were identified for frost-induced floret sterility in two different populations at the same genomic location. This QTL was not associated with previously reported developmental or stress-response loci. QTL on chromosome 5HL were identified for frost-induced floret sterility and frost-induced grain damage in all three of the populations studied. The locations of QTL were coincident with previously reported vegetative frost tolerance loci close to the vrn- H1 locus. This locus on chromosome 5HL has now been associated with response to cold stress at both vegetative and reproductive developmental stages in barley. This study will allow reproductive frost tolerance to be seriously pursued as a breeding objective by facilitating a change from difficult phenotypic selection to high-throughput genotypic selection.

Acclimatization↗
PubMed2005

Carbohydrate partitioning between upper and lower regions of the crown in oat and rye during cold acclimation and freezing.

Carbohydrates have long been recognized as an important aspect of freezing tolerance in plants but the association between these two factors is often ambiguous. To help clarify the relationship, the allocation of carbohydrates between specific tissues within the over wintering organ (crown) of winter cereals was measured. A winter-hardy and non-winter-hardy oat (Avena sativa L.), and a rye (Secale cereale L.) cultivar were grown and frozen under controlled conditions. Crown tissue was fractionated into an upper portion, called the apical region, and a lower portion, called the lower crown. These tissues were ground in liquid N and extracted with water. Extracts were analyzed by HPLC for the simple sugars, sucrose, glucose, fructose, and for fructan of various size classes. After 3 weeks of cold acclimation at 3 degrees C, carbohydrates accounted for approximately 40% of the dry weight of oats and 60% of the dry weight of rye. The apical region, which is the tissue within the crown that acclimates to the greatest extent, was generally 10% higher in total carbohydrates than the lower crown. During a mild freeze, various carbohydrates were allocated differently between specific tissues in the three genotypes. When frozen, fructan generally decreased to a greater extent in the lower crown than in the apical region but sugars increased more in the apical region than in the lower crown. Results suggest that to understand how carbohydrates relate to freezing tolerance, regions of the crown that endure freezing stress differently should be compared.

Acclimatization↗
PubMed1986

Detrimental effects of four hours of cold storage on porcine pancreaticoduodenal transplantation.

Transplantation of the pancreaticoduodenal allograft (PDA) has recently been advocated as a technique that is superior to the use of the segmental allograft. However, the effect of simple cold storage preservation on the PDA has not been studied. We investigated the effect of 24 and 4 hours of cold storage in Eurocollins solution on porcine PDA function after transplantation in pancreatectomized pigs. A regimen of cyclosporine and prednisone was used, which prevented rejection for at least 28 days after transplantation. Cold storage preservation for 24 hours uniformly resulted in PDA failure. Compared with recipients of immediately transplanted PDA, recipients of PDA cold stored for 4 hours had marked plasma hyperamylasemia (10,000 U/L versus 1932 U/L), relative glucose intolerance (K value -2.15 versus -2.66), hypoinsulinemia (peak immunoreactive insulin 11.0 microU/ml versus 34.7 microU/ml), and an abnormal pattern of insulin secretion as demonstrated with intravenous glucose tolerance testing. There was also a higher incidence of technical complications in the group transplanted with cold-stored PDAs. Our results suggest that there is a detrimental effect on porcine PDA function after only 4 hours of cold storage in Eurocollins solution.

Amylases↗
PubMed2007

Can a sexually dimorphic index of prenatal hormonal exposure be used to examine cold pressor pain perception in men and women?

There is considerable evidence to suggest that important differences exist between men and women in their experience of pain. Research has now turned to determine what the mechanisms of such differences actually are. One potential explanation is the effect of sex hormones, especially those typically found in greater concentration within women, e.g., estrogen, progesterone. However, it is also possible that other hormones, such as testosterone may be important. The current study employed a non-invasive sexually dimorphic index (digit ratio) that is believed to reflect prenatal exposure to testosterone, and related this to the cold pressor pain experiences of 23 men and 27 healthy women. As expected, females had greater symmetry between the second and fourth digits, and also reported lower pain tolerance levels. Although some significant relationships were found between digit ratio/digit length and cold pressor pain reports they were relatively inconsistent. Furthermore, the main finding, that pain thresholds were positively related to digit ratio in women but not men, is somewhat inconsistent with predictions. The results are discussed in light of methods for investigating the effect of prenatal hormonal exposure on pain sensitivity in men and women.

Adult↗
PubMed2001

Antisense inhibition of protein phosphatase 2C accelerates cold acclimation in Arabidopsis thaliana.

Two related protein phosphatases 2C, ABI1 and AtPP2CA have been implicated as negative regulators of ABA signalling. In this study we characterized the role of AtPP2CA in cold acclimation. The pattern of expression of AtPP2CA and ABI1 was studied in different tissues and in response to abiotic stresses. The expression of both AtPP2CA and ABI1 was induced by low temperature, drought, high salt and ABA. The cold and drought-induced expression of these genes was ABA-dependent, but divergent in various ABA signalling mutants. In addition, the two PP2C genes exhibited differences in their tissue-specific expression as well as in temporal induction in response to low temperature. To elucidate the function of AtPP2CA in cold acclimation further, the corresponding gene was silenced by antisense inhibition. Transgenic antisense plants exhibited clearly accelerated development of freezing tolerance. Both exposure to low temperature and application of ABA resulted in enhanced freezing tolerance in antisense plants. These plants displayed increased sensitivity to ABA both during development of frost tolerance and during seed germination, but not in their drought responses. Furthermore, the expression of cold-and ABA-induced genes was enhanced in transgenic antisense plants. Our results suggest that AtPP2CA is a negative regulator of ABA responses during cold acclimation.

Abscisic Acid↗
PubMed1999

Improved high altitude hypoxic tolerance and amelioration of anorexia and hypophagia in rats on oral glutamate supplementation.

BACKGROUND: The objective of this study is to evaluate the efficacy of oral glutamic acid supplementation in promoting hypoxic tolerance. METHODS: The experiments were conducted in albino rats by exposing them to three levels of hypoxia in a simulated environment for varying periods of time. The parameters studied include: gasping time at 35,000 ft (10,668 m), food and water intake, and heart to body weight (b.w.) ratio at 25,000 ft (7620 m), tolerance to composite stress at 15,000 ft (4572 m) and biodistribution of glutamate (glu). RESULTS: Supplementation of Glu (27 mg x kg(-1) b.w.) as glutamic acid dissolved in normal saline resulted in 4.8 times enhanced hypoxic tolerance (time taken for appearance of first gasp), 23% body weight gain and 24% increase in food consumption over control during hypoxia. When animals were subjected to composite stress of cold, hypoxia and restraint (CHR), the Glu fed animals showed higher resistance to fall in rectal temperature than the control group. Hypoxia significantly enhanced heart to body weight ratio compared with control, and Glu supplementation reduced and brought it down to that of control. CONCLUSION: The study reveals that Glu in optimal doses may be a conditionally essential amino acid resulting in enhanced tolerance to hypoxia and cold.

Administration, Oral↗