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Bundle sheath proteins are more sensitive to oxidative damage than those of the mesophyll in maize leaves exposed to paraquat or low temperatures.

In maize leaves growth at low temperatures causes decreases in maximum catalytic activities of photosynthetic enzymes and reduced amounts of proteins, rather than effects on regulation or co-ordination of the photosynthetic processes. To test the hypothesis that differential localization of antioxidants between the different types of photosynthetic cell in maize leaves is a major determinant of the extreme sensitivity of maize leaves to chilling damage, oxidative damage to proteins, induced by incubation of maize leaves with paraquat, has been measured and compared with the effects incurred by growth at low temperatures. While the increase in protein carbonyl groups caused by paraquat treatment was much greater than that caused by low temperature growth conditions, most carbonyl groups were detected on bundle sheath proteins in both stress conditions. With one or two exceptions proteins located in the mesophyll tissues were free of protein carbonyl groups in both situations. Paraquat treatment caused a complete loss of the psaA gene products, modified the photosystem II reaction centre polypeptide, D1, and increased the number of peptides arising from breakdown of ribulose 1,5-bisphosphate carboxylase oxygenase (Rubisco). In contrast, growth at 15 degrees C increased the abundance (but not number) of Rubisco breakdown products and decreased that of the psaB gene product while the psaA gene product and PEP carboxylase were largely unaffected. Since bundle sheath proteins are more susceptible to oxidative damage than those located in the mesophyll cells, strategies for achieving a more balanced system of antioxidant defence may be effective in improving chilling tolerance in maize.

Adaptation, Physiological↗

Ionic mechanisms of action potential prolongation at low temperature in guinea-pig ventricular myocytes.

1. We studied the effects of low temperature on the action potentials and membrane currents of guinea-pig ventricular myocytes, using a tight-seal whole-cell clamp technique. 2. The action potential duration at 95% repolarization was prolonged from 146 +/- 33 ms (mean +/- S.D., n = 6) at 33-34 degrees C (control temperature) to 314 +/- 83 ms at 24-25 degrees C (low temperature). 3. In whole-cell clamp experiments, low temperature decreased the calcium current (ICa), the delayed rectifier potassium current (IK), and the inwardly rectifying potassium current (IK1) with 'apparent' Q10 (temperature coefficient) values of 2.3 +/- 0.6 for ICa, 4.4 +/- 1.2 for IK tail current and 1.5 +/- 0.3 for IK1 (n = 7). 4. The effect of low temperature on IK was further studied in the presence of 0.6 microM nicardipine to block ICa. The decay phase of the IK tail consisted of two exponential components. The fast but not the slow component was highly sensitive to the temperature change with an apparent Q10 of 4.5. 5. We found that a component of time-independent current is also sensitive to the temperature. The current had a linear I-V relationship and remained almost unchanged after inhibition of Na(+) -K+ pump in K(+)-free external solution. 6. Using our mathematical model of the ventricular action potential (a modification from the DiFrancesco-Noble model), we simulated the action potential at low temperature by modifying some of the membrane currents, namely IK, IK1, ICa and a component of background current. It was shown that simultaneous changes in these currents could reproduce approximately 75% of the action prolongation induced by low temperature.

Action Potentials↗

Effects of thyroidal, gonadal and adrenal hormones on tissue respiration of streaked frog, Rana limnocharis, at low temperature.

In vivo and in vitro effects of thyroidal, gonadal and adrenal hormones were studied on the rate of liver and skeletal muscle respiration in both the sexes of R. limnocharis during active and inactive phases of the annual activity cycle. Triiodothyronine (L-T3) and thyroxine (L-T4) did not stimulate tissue (liver and muscle) respiration in any of the experiments irrespective of season, sex and temperature. Testosterone, estradiol and corticosterone stimulated O2 uptake significantly irrespective of season, sex and temperature. Adrenaline and nor-adrenaline also stimulated tissue respiration significantly during the winter month. Since the ambient temperature was low even during the active phase (max. temperature 21 degrees C), it seems that the frog might have developed tissue sensitivity for gonadal and adrenal hormones at low temperatures when thyroid hormones are calorigenically ineffective.

Animals↗

The acute toxicity evaluation of a low-temperature hydraulic fluid.

A low-temperature version of MIL-H-83282 (LT 83282) is a candidate hydraulic fluid to be used as a replacement for the current low-temperature fluid used on Strategic Air Command aircraft. A single neat dose of 0.1 mL LT 83282 into New Zealand White (NZW) rabbit eyes resulted in slight conjunctival irritation for up to 24 hr after treatment in two of nine rabbits. Rinsing the eyes after treatment appeared beneficial. A single treatment of 0.5 mL neat LT 83282 to rabbit skin produced no irritation. A total of 40% of the guinea pigs receiving repeated dermal application of the fluid demonstrated a positive sensitization response. A single oral dose of 5 g LT 83282/kg body weight given to five male and five female Fischer 344 (F-344) rats and a single dermal application of 2 g LT 83282/kg body weight applied to five male and five female NZW rabbits resulted in no deaths. Inhalation exposures to aerosol concentrations of LT 83282 resulted in an LC50 of 2.13 and 1.50 mg/L for male and female F-344 rats, respectively. No clinical signs of acute delayed neurotoxicity were observed in hens twice dosed at limit levels (5 g/kg) and observed for 21 days.

Administration, Inhalation↗

Temperature control in Lowicryl K4M and glycol methacrylate during polymerization: is there a low-temperature embedding method?

An apparatus for embedding tissues at resin temperatures down to 228 K is described. By placing thermocouples in the resin the temperature has been monitored during embedding at low temperature with glycol methacrylate (GMA) and Lowicryl K4M. Even in this apparatus with a liquid cooling bath the heat of polymerization is not dissipated and the resin temperature rises. This rise is directly proportional to the resin temperature at the onset of polymerization and is higher in Lowicryl K4M than GMA. The initial resin temperature also affects the time taken for polymerization. The time to the onset of the peak and its duration are both increased as the temperature is lowered. This effect is more pronounced with GMA than Lowicryl K4M and polymerization of GMA is inhibited at the lowest temperature used. When Lowicryl K4M, polymerized at low temperature, is warmed up to ambient a further exothermic reaction occurs, which causes the resin temperature to rise well above ambient. Both this temperature peak and that during polymerization are reduced, but not totally eliminated, by reducing the resin volume. Air-cooled systems are inefficient compared with the low-temperature apparatus used here and the resin temperature rise is consequently greater and, even with small resin volumes, it can be very high. It is therefore unlikely for published methods that the temperature specified has been maintained in the resin during polymerization. The implications of these findings are discussed in relation to enzyme and antigen survival. Recommendations include use of very small volumes of resin, refrigerated liquid-bath rather than air-cooled systems and contact with a heat sink when specimens are warmed up to ambient temperature. Examples of enzyme reaction, antigen survival and structural preservation obtained with the method are presented.

Acrylates↗

[Heat-capacity of DNA at low temperatures and varying humidities].

Adiabatic low temperature microcalorimeter working in a wide temperature range, including that of liquid helium, is described. Heat capacity of DNA at various water concentrations was measured at 4-400 degrees K. It is shown that the pattern of its change is determined by the hydration parameter n (number of water molecules per mole of base pairs). According to the experimental findings the DNA molecule can be modelled by a sufficiently rigid "thin rod" and the dynamic characteristics of the subject under study can be explained in terms of monomeric continuum model. When studying low temperature heat capacity of DNA, account should be taken of the double helic hydration depending on GC-content. Measurement of heat capacity of DNA in native and denaturated states in a wide temperature range (4-400 degrees K) taking into account specific hydration of double helix allows a correct description of its thermodynamic properties.

Calorimetry↗

Promoters from kin1 and cor6.6, two homologous Arabidopsis thaliana genes: transcriptional regulation and gene expression induced by low temperature, ABA, osmoticum and dehydration.

The Arabidopsis thaliana genes kin1 and cor6.6 belong to the same family and were expressed at higher levels following low temperature and ABA treatments. In an attempt to elucidate the mechanism of gene regulation by low temperature, the relationship between low-temperature- and abscisic acid (ABA)-induced gene expression and possible differential expression of the two genes, we have cloned a 5.3 kb genomic fragment harboring kin1 and cor6.6 and their respective 5' sequences. The putative promoters of both genes were fused to the beta-glucuronidase (GUS) coding sequence and GUS expression was analysed in transgenic tobacco and Arabidopsis plants. The cor6.6 promoter produced a higher basal level of expression than the kin1 promoter in transgenic tobacco. Enzyme assays of inducible GUS activity in transgenic Arabidopsis and tobacco plants showed that GUS activity directed by both kin1 and cor6.6 promoters was significantly induced by ABA, dehydration and osmoticum, but not by low temperature. Northern analysis revealed, in contrast, that GUS mRNA was significantly induced in these transgenic plants by low temperature. Further analysis showed that, at low temperature, GUS protein synthesis from the induced GUS mRNA was inhibited. Together these results reveal induction of kin1 and cor6.6 transcription by low temperature, exogenous ABA and dehydration. However, low-temperature expression is dramatically reduced at the translational level.

Abscisic Acid↗

Stability of lysosomal and cell membranes in haemocytes of the common mussel (Mytilus edulis): effect of low temperatures.

Expanding industrial activities in the Arctic require an urgent assessment of the toxicity of chemicals at low temperatures. Organisms acclimatized to low temperature exhibit specific adaptations. For example, the amount of unsaturated lipids is increased to maintain the fluidity of the cell membranes. It has been hypothesized that such temperature-induced alterations in membrane lipid composition may affect the stability of lysosomal and cell membranes in the common mussel, Mytilus edulis, an organism exposed to seasonal temperature extremes. As mussels may be exposed to petroleum compounds along industrialized coastlines, we tested the combined effects of exposure to low temperature and the petroleum compound, phenanthrene, on haemocyte membrane stability. Test animals, acclimated to either 0 or 10 degrees C, were exposed to phenanthrene (0 = control or 500 micrograms l-1) and haemocytes were examined using the neutral red retention assay (lysosomal stability) and a fluorescence assay (cell membrane stability). At 0 degree C, lysosomal and cell membranes from uncontaminated mussels were destabilized compared with 10 degrees C (P = 0.0005). No significant effects (P > 0.05) of phenanthrene were detected at either temperature. Possible mechanisms underlying membrane destabilization include a weaker physical resistance of the membrane due to a higher amount of unsaturated lipids, a potentially higher level of reactive oxygen radicals at low temperature and the higher susceptibility of unsaturated lipids to oxidative stress. More work is required to better understand the consequences of this membrane destabilization at low temperature on the susceptibility of the organism to pollutants.

Adaptation, Physiological↗

Alteration of [beta]-Tubulin Gene Expression during Low-Temperature Exposure in Leaves of Arabidopsis thaliana.

Responses of [beta]-tubulin gene expression to low-temperature exposure (4[deg]C) have been investigated in leaves of Arabidopsis thaliana. During low-temperature exposure, the patterns of both [alpha]- and [beta]-tubulin isoforms are altered; the effect is smaller for the [alpha]-tubulins than for the [beta]-tubulins, however. An examination of [beta]-tubulin gene expression revealed that during low-temperature exposure, transcript levels of TUB2, TUB3, TUB6, and TUB8 decrease, whereas those of TUB4, TUB5, and TUB7 remain constant, and the TUB9 transcript level increases. The changes in transcript levels of TUB6, TUB8, and TUB9 were detectable after 6 h of low-temperature treatment. As shown by transcription-blocking experiments, the in vivo decay rates at 25[deg]C are comparable to those at 4[deg]C for TUB5, TUB6, and TUB8 mRNAs, whereas TUB9 mRNA appears to be more stable at 4[deg]C than at 25[deg]C. Thus, decreases in transcript levels of TUB6 and TUB8 in response to low temperature appear to be regulated at the transcriptional level, and the increase in TUB9 transcript level that results from lowering the temperature from 25[deg]C to 4[deg]C may be due in part to its slower rate of decay at 4[deg]C. When a chimeric gene containing 1061 base pairs of TUB8 5[prime] flanking DNA fused to the [beta]-glucuronidase coding region was used to produce transgenic Arabidopsis plants, the chimeric gene expression was down-regulated in response to low temperature as assayed by histochemical localization and RNA gel blots. These results confirm that the alteration of transcript levels of TUB8 in response to low temperature is regulated at the transcriptional level.

Journal Article↗

Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds.

Exposure of imbibed seeds to low temperature (typically 4 degrees C) is widely used to break seed dormancy and to improve the frequency of germination. However, the mechanism by which temperature accelerates germination is largely unknown. Using DNA microarray and gas chromatography-mass spectrometry analyses, we found that a subset of gibberellin (GA) biosynthesis genes were upregulated in response to low temperature, resulting in an increase in the level of bioactive GAs and transcript abundance of GA-inducible genes in imbibed Arabidopsis thaliana seeds. Using a loss-of-function mutant, the cold-inducible GA biosynthesis gene, AtGA3ox1, was shown to play an essential role in mediating the effect of low temperature. Besides temperature, AtGA3ox1 also is positively regulated by active phytochrome and negatively regulated by GA activity. We show that both red light and GA deficiency act in addition to low temperature to elevate the level of AtGA3ox1 transcript, indicating that multiple signals are integrated by the AtGA3ox1 gene to control seed germination. When induced by low temperature, AtGA3ox1 mRNA was detectable by in situ RNA hybridization in an additional set of cell types relative to that in red light-induced seeds. Our results illustrate that the GA biosynthesis and response pathways are activated during seed imbibition at low temperature and suggest that the cellular distribution of bioactive GAs may be altered under different light and temperature conditions.

Arabidopsis↗

Isolation of calcium-phosphate crystals of bone by non-aqueous methods at low temperature.

We have developed low temperature nonaqueous solution methods as well as low power plasma ashing for the degradation of the organic matrix of bone power which have permitted us to obtain bone crystals essentially free of organic matrix constituents without any significant change in their composition, overall structure, or internal short-range order. We have also been able to disperse the crystals, which has made it possible to examine the isolated crystals by X-ray diffraction and resolution-enhanced Fourier transform infrared (FTIR) spectroscopy and isolated single crystals by high resolution transmission electron microscopy (TEM) and electron diffraction. TEM of isolated single crystals of fish, chicken, mouse and bovine bone have clearly demonstrated that the crystals are very thin plates. No rod or needle-like crystals were observed in any of the bone samples in the four species studied including the earliest crystals deposited. Although there were variations in the size distribution of the crystals in the different species studied, in general the average crystal dimensions were very similar.

Animals↗

Low temperatures during early development influence subsequent maternal and reproductive function in adult female mice.

Challenging conditions early in development can have enduring effects into adulthood. The effects of low temperatures were examined on subsequent sex-specific morphology (anogenital distance [AGD]), maternal care, and reproductive performance in adult female mice. Dams (F0) were maintained either in (1) standard laboratory room temperatures (21 +/- 2 degrees C) or (2) low temperatures (10 +/- 2 degrees C) throughout gestation. Their progeny (F1) either remained in the temperature condition in which they were conceived or were switched to the other temperature condition at 2 days of age until weaning. Reproductive performance and maternal behaviors were assessed in adulthood. F0 dams that were maintained in low temperatures bore larger litters as compared to F0 animals housed in standard temperatures throughout their pregnancy. In contrast, mean litter size was reduced for all groups of F1 females that experienced low temperatures. Infant mortality was elevated in litters of F1 females that were exposed to low temperatures both before and after birth. Prenatal exposure to low temperatures was associated with reduced responsiveness towards the nursing young and decreased maternal aggression in F1 animals. Prenatally treated F1 females had longer, male-like AGDs on Day 2 following birth compared to animals not subjected to experimental manipulations. Our results indicate that exposure to low temperatures during early development impairs reproductive function and is associated with important fitness costs as evidenced by reduced offspring survival. Our findings also suggest that chronic low temperatures experienced only after birth may have less deleterious effects than exposure to a combination of pre- and postnatal or prenatal treatments alone.

Aggression↗

Temperature-dependent neutron powder diffraction study of the Ba(OD)(2) polymorphs: a new low-temperature phase.

The structural behaviour of both low-temperature beta- and high-temperature alpha-Ba(OD)(2), barium dihydroxide-d, was investigated at temperatures between 10 and 552 K by neutron powder diffraction. While the beta phase (P2(1)/n) remains stable to the lowest temperature investigated, the quenchable alpha phase (Pnma) shows a reversible orthorhombic-to-monoclinic phase transition between 100 and 150 K. The structure of the new alpha(m) phase (P2(1)/n) is quite similar to that of the alpha phase. This behaviour is unusual as a metastable phase transforms to another metastable phase. The Pnma <--> P2(1)/n low-temperature phase transition is driven by an order-disorder mechanism, mainly caused by one of the D atoms, which is disordered on positions off the mirror plane, mimicking a special position on the mirror plane in the orthorhombic phase. Refinements of the alpha phase above the phase transition indicate this disorder across the mirror plane through a conspicuously high isotropic displacement parameter if compared to the other D atoms. At low temperature the energy of the vibration is lowered and the D atom is frozen at a general position in a correlated way, thus violating the mirror plane and reducing the space-group symmetry.

Journal Article↗

Low-temperature modification of the inhibitory effects of volatile anesthetics on airway smooth muscle contraction in dogs.

BACKGROUND: Because exposure to low temperature can modify the effect of volatile anesthetics on airway smooth muscle contraction, this study was conducted to investigate low-temperature modifications of the inhibitory effects of isoflurane and sevoflurane on canine tracheal smooth muscle tone by simultaneously measuring the muscle tension and intracellular concentration of Ca2+ ([Ca2+]i) and by measuring voltage-dependent Ca2+ channel activity. METHODS: [Ca2+]i was monitored by the 500-nm light emission ratio of preloaded fura-2, a Ca2+ indicator. Isometric tension was measured simultaneously. Whole cell patch clamp recording techniques were used to observe voltage-dependent Ca2+ channel activity in dispersed muscle cells. Isoflurane (0-3.0%) or sevoflurane (0-3%) was introduced to a bath solution at various temperatures (37, 34, or 31 degrees C). RESULTS: Low temperature (34 or 31 degrees C) reduced high-K+-induced (72.7 mm) muscle contraction and increased [Ca2+]i, but it enhanced carbachol-induced (1 microm) muscle contraction with a decrease in [Ca2+]i. The volatile anesthetics tested showed significant inhibition of both high-K+-induced and carbachol-induced airway smooth muscle contraction, with a concomitant decrease in [Ca2+]i. The inhibition of the carbachol-induced muscle contraction by volatile anesthetics was abolished partially by exposure to low temperature. Volatile anesthetics and low-temperature exposure significantly inhibited voltage-dependent Ca2+ channel activity of the smooth muscle. CONCLUSIONS: Exposure of airway smooth muscle to low temperature leads to an increase in agonist-induced muscle contractility, with a decrease in [Ca2+]i. The inhibition of voltage-dependent Ca2+ channel activity by exposure to low temperature and by volatile anesthetics cam be attributed, at least in part, to the decrease in [Ca2+]i.

Anesthetics, Inhalation↗

Titanium casting: the surface reaction layer of castings obtained using ultra-low-temperature molds.

To examine whether the surface reaction layer of titanium castings can be reduced by lowering the mold temperature during casting, we cast titanium at three mold temperatures, including an ultra-low temperature produced by cooling the mold with liquid nitrogen, then measured the tensile strength and elongation of the castings. The titanium was cast using a centrifugal casting machine, and the molds were incinerated according to the manufacturers' instructions. Castings were then made with the molds at 200 degrees C, 600 degrees C, and an ultra-low temperature (-196 degrees C). The castability of titanium cast in the mold at the ultra-low temperature was good. The Vickers hardness near the surface layer of castings decreased as the mold temperature decreased.

Cold Temperature↗

[Studies on medico-legal diagnosis in cold district. 2. Cadaveric phenomena in low temperature surroundings (author's transl)].

The authors have investigated 14 kinds of findings of 15 cadavers in low temperature surroundings which had remained at known postmortem interval and had been subjected to medico-legal autopsies in our laboratory, comparing with those at normal temperature. In low temperature surroundings, the onset and progress of the findings due to autolysis or putrefaction are retarded in a marked degree but the turbidity of the cornea which is considered to be due to physical or chemico-physical phenomenon is retarded merely in a slight degree. Hence, an erroneous determination of the postmortem interval can be avoided by taking care of the peculiarity. Even the cadavers at low temperature display in 3 to 5 months after death the findings whose degrees correspond to those of the cadavers at normal temperature in 4 to 7 days after death. Some fly-larvae pass the winter on cadavers demonstrating that the time of death was towards the end of autumn of the preceding year.

Adolescent↗

Moritella cold-active dihydrofolate reductase: are there natural limits to optimization of catalytic efficiency at low temperature?

Adapting metabolic enzymes of microorganisms to low temperature environments may require a difficult compromise between velocity and affinity. We have investigated catalytic efficiency in a key metabolic enzyme (dihydrofolate reductase) of Moritella profunda sp. nov., a strictly psychrophilic bacterium with a maximal growth rate at 2 degrees C or less. The enzyme is monomeric (Mr=18,291), 55% identical to its Escherichia coli counterpart, and displays Tm and denaturation enthalpy changes much lower than E. coli and Thermotoga maritima homologues. Its stability curve indicates a maximum stability above the temperature range of the organism, and predicts cold denaturation below 0 degrees C. At mesophilic temperatures the apparent Km value for dihydrofolate is 50- to 80-fold higher than for E. coli, Lactobacillus casei, and T. maritima dihydrofolate reductases, whereas the apparent Km value for NADPH, though higher, remains in the same order of magnitude. At 5 degrees C these values are not significantly modified. The enzyme is also much less sensitive than its E. coli counterpart to the inhibitors methotrexate and trimethoprim. The catalytic efficiency (kcat/Km) with respect to dihydrofolate is thus much lower than in the other three bacteria. The higher affinity for NADPH could have been maintained by selection since NADPH assists the release of the product tetrahydrofolate. Dihydrofolate reductase adaptation to low temperature thus appears to have entailed a pronounced trade-off between affinity and catalytic velocity. The kinetic features of this psychrophilic protein suggest that enzyme adaptation to low temperature may be constrained by natural limits to optimization of catalytic efficiency.

Amino Acid Sequence↗

Reactions at very low temperatures: gas kinetics at a new frontier.

Advances in experimental techniques, especially the development of the CRESU (Cinétique de Réaction en Ecoulement Supersonique Uniforme) method, allow many gas-phase molecular processes to be studied at very low temperatures. This Review focuses on the reactions of molecular and atomic radicals with neutral molecules. Rate constants for almost 50 such reactions have been measured at temperatures as low as 13 K by using the CRESU method. The surprising demonstration that so many reactions between electrically neutral species can be extremely rapid at these very low temperatures has excited interest both from theoreticians and from those seeking to understand the chemistry that gives rise to the 135 or so molecules that are present in low-temperature molecular clouds in the interstellar medium. Theoretical treatments of these reactions are based on the idea that a reaction occurs when the long-range potential between the reagent species brings them into close contact. The astrochemical context, theoretical studies, and the determination of the rate constants of these low-temperature reactions are critically discussed.

Journal Article↗