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PubMed1975

Determination of unfrozen water in winter cereals at subfreezing temperatures.

The freezing of water in acclimated and nonacclimated cereals was studied using pulsed nuclear magnetic resonance spectroscopy. The quantity of unfreezable water per unit dry matter was not strongly dependent on the degree of cold acclimation. In contrast, the fraction of water frozen which was tolerated by nonacclimated winter cereals and by an acclimated spring wheat (Triticum aestivum L.) was less than in acclimated hardy cereals. The freezing curves had the following form:L(T) = L(0)DeltaTm/T + KL(T) and L(0) are liquid water per unit dry matter at T and 0 C, respectively. DeltaTm is the melting point depression and K is the liquid water which does not freeze.

Journal Article↗
PubMed2004

Isolation of a dehydrin cDNA from orange and grapefruit citrus fruit that is specifically induced by the combination of heat followed by chilling temperatures.

Dehydrins (DHNs; late embryogenesis abundant D-11) are a family of plant proteins induced in response to environmental stresses such as water stress, salinity and freezing or which occur during the late stages of embryogenesis. Previously, it was reported that citrus contains a small gene family encoding a unique class of dehydrins that differs from most other plant dehydrins in various respects, such as having an unusual K-segment similar to that of gymnosperms. In the present study, we identified by cDNA differential display analysis a 'Navel' orange 202-bp polymerase chain reaction (PCR) fragment, which encoded the typical plant angiosperm-type K-segment consensus sequence, and of which the expression was down-regulated by exposure to low oxygen levels. The full-length cDNA sequence of the orange DHN, designated csDHN (for Citrus sinensis DHN), was further isolated by 5'-and 3'-RACE; it had a total length of 933 bp and encoded a predicted polypeptide of 235 amino acids. In addition, the same 202-bp 'Navel' dehydrin PCR fragment was used to screen a 'Star Ruby' grapefruit flavedo cDNA library, and its full-length grapefruit homologue, designated cpDHN (for C. paradisi DHN) was isolated and found to have a total length of 1024 bp and to encode a predicted polypeptide of 234 amino acids. The defined orange and grapefruit DHN proteins were completely identical in the 196 amino acids of their N-terminus but differed in their C-terminus region. Overall, the csDHN and cpDHN proteins share 84% identity and contain the conserved dehydrin serine cluster (S-segment) and a putative nuclear localization signal, but csDHN has one conserved dehydrin K-segment consensus sequence, whereas cpDHN contains two dehydrin K-segments. Both csDHN and cpDHN represent single copy genes, in 'Navel' orange and 'Star Ruby' grapefruit genomes, respectively. We found that the cpDHN gene was consistently expressed in the fruit peel tissue at harvest, but that its message levels dramatically decreased during storage at either ambient or low temperatures. However, a pre-storage hot water treatment, given to enhance fruit-chilling tolerance, increased cpDHN mRNA levels during the first 3 weeks of cold storage at 2 degrees C, and enabled the message levels to be retained for up to a further 8 weeks of cold storage at 2 degrees C. The hot water treatment by itself had no inductive effect on cpDHN gene expression when the fruits were held at non-chilling temperatures. Other stresses applied to the fruit, such as wounding, UV irradiation, water stress, low oxygen and exposure to the stress hormone ethylene decreased DHN mRNA levels, whereas abscisic acid had no effect at all.

Journal Article↗
PubMed1998

The nature of antioxidant defense mechanisms: a lesson from transgenic studies.

Reactive oxygen species (ROS) have been implicated in the pathogenesis of many clinical disorders such as adult respiratory distress syndrome, ischemia-reperfusion injury, atherosclerosis, neurodegenerative diseases, and cancer. Genetically engineered animal models have been used as a tool for understanding the function of various antioxidant enzymes in cellular defense mechanisms against various types of oxidant tissue injury. Transgenic mice overexpressing three isoforms of superoxide dismutase, catalase, and the cellular glutathione peroxidase (GSHPx-1) in various tissues show an increased tolerance to ischemia-reperfusion heart and brain injury, hyperoxia, cold-induced brain edema, adriamycin, and paraquat toxicity. These results have provided for the first time direct evidence demonstrating the importance of each of these antioxidant enzymes in protecting the animals against the injury resulting from these insults, as well as the effect of an enhanced level of antioxidant in ameliorating the oxidant tissue injury. To evaluate further the nature of these enzymes in antioxidant defense, gene knockout mice deficient in copper-zinc superoxide dismutase (CuZnSOD) and GSHPx-1 have also been generated in our laboratory. These mice developed normally and showed no marked pathologic changes under normal physiologic conditions. In addition, a deficiency in these genes had no effects on animal survival under hyperoxida. However, these knockout mice exhibited a pronounced susceptibility to paraquat toxicity and myocardial ischemia-reperfusion injury. Furthermore, female mice lacking CuZnSOD also displayed a marked increase in postimplantation embryonic lethality. These animals should provide a useful model for uncovering the identity of ROS that participate in the pathogenesis of various clinical disorders and for defining the role of each antioxidant enzyme in cellular defense against oxidant-mediated tissue injury.

Animals↗
PubMedDate not supplied

Phytoremediation of pesticide wastes in soil.

Soils at agrochemical dealer sites often are contaminated with pesticide residues from decades of accidental and incidental spillage. We have determined that prairie grasses native to the Midwestern U.S. are suitable for phytoremediation because they are tolerant of most herbicides and of climatic extremes, such as heat, cold, drought, and flooding. A mixed stand of big bluestem, switch grass, and yellow indiangrass develops a rhizosphere with microflora that can readily detoxify pesticide residues. Specific atrazine-degrading bacteria or the free enzyme atrazine chlorohydrolase also can enhance the rate of biotransformation of atrazine in soil. Metolachlor degradation can be accelerated significantly by the prairie grass/rhizosphere effect. Several grasses used in filter strips have also been evaluated for their pesticide-degradation capabilities. The prairie grasses also have been demonstrated to reduce the rates of leaching of pesticides through intact soil columns, since less water leaches out of vegetated soil columns compared to non-vegetated soil columns. The evaluation of the degree of success of remediation has relied heavily on chemical residue analysis, but recent studies on biological endpoints have shown promise for providing more ecologically relevant indications of the potential exposure of organisms to pesticides in the soil. Earthworm 8-day bioaccumulation assays and root growth assays have shown the value of assessing the bioavailability of the residues. Mass balance experiments have utilized radiolabeled atrazine and metolachlor to ascertain the complete metabolism and binding profile of those two pesticides in phytoremediation studies.

Atrazine↗
PubMed1995

[Current trends in the preservation of frozen fruits, as markers of hygienic quality of food].

Spore germination and growth rate of pathogens can be greatly reduced by cooling right after harvest. Most tropical or subtropical crops, and even some temperate, fruits undergo from chilling injury (CI) at temperatures considerably above their freezing point. Several procedures have been designed to increase tolerance to CI. Prestorage temperature conditioning can reduce CI in subsequent cold storage. Intermittent warming interrupting the chilling period by periods at nonchilling temperatures may become a practical treatment to prolong the storage of chilling sensitive crops. Modified atmospheres extend the postharvest life of fruit by reducing their respiration rate. The maintenance of fruit in good physiological condition may result in considerable disease resistance. A direct effect against pathogens is also possible at concentrations of O2 lower than 1.8% or concentrations of CO2 higher than 10%.

Food-Processing Industry↗
PubMed1983

Lower temperature and orthophosphate control of alkaline phosphatase in a cold-requiring plant.

The elevated activity of alkaline phosphatase observed in roots and shoots of Verbascum rosettes grown in vitro at 15 degrees C may be the result of a reduced supply of orthophosphate. The rosette reveals greater activities of this enzyme under conditions of lower external phosphate or chilling temperature. Either of these treatments gives lower amounts of endogenous phosphate. The stimulation of phosphatase in tissues having lower levels of phosphate is thus a possible feature of other plant species which also display freezing tolerance.

Alkaline Phosphatase↗
PubMed1998

A study of the interactive effects of oral contraceptive use and dietary fat intake on blood pressure, cardiovascular reactivity and glucose tolerance in normotensive women.

OBJECTIVE: To investigate the interactive effects of oral contraceptive pill use and dietary fat intake on cardiovascular haemodynamics and metabolic parameters in young normotensive women. DESIGN: Thirty-two women participated, of whom 16 were taking oral contraceptive pills (ethinyl-oestradiol plus levonorgestrel) and 16 were age-matched and weight-matched controls not taking such pills. Subjects consumed either a high-fat or a low-fat diet for 2 weeks in an open, randomized, crossover study lasting 6 weeks. Investigations were performed at the end of each diet during the luteal phase of the menstrual cycle. METHODS: Blood pressure was measured by 24 h ambulatory recording; cardiovascular reactivity was determined by examining blood pressure responses to systemic infusions of noradrenaline and angiotensin II and to the cold pressor test; and carbohydrate metabolism was investigated by an intravenous glucose-tolerance test. RESULTS: Plasma triglyceride levels were significantly higher in women taking oral contraceptive pills compared with non-users on both diets; however, responses of lipoprotein levels to the two diets did not differ between study groups (total and low-density lipoprotein cholesterol levels decreased by 15 and 17% in oral contraceptive pill users and by 14% each in non-users, on the low-fat compared with the high-fat diet). Fasting plasma insulin levels, the insulin-production response to administration of glucose (insulin area under the curve) and resting clinic and night-time systolic blood pressures were all significantly reduced on the low-fat diet, but only in non-users. Blood pressure responses to noradrenaline and maximal heart rate response to cold were significantly attenuated during the low-fat diet in oral contraceptive pill users. During the low-fat diet, resting systolic, 24 h systolic and diastolic blood pressures and insulin area under the curve were all significantly higher for women taking the oral contraceptive pills. Users of these pills also exhibited a greater systolic sensitivity to administration both of noradrenaline and of angiotensin II and had a higher plasma renin activity irrespective of dietary phase. CONCLUSIONS: These results confirm that oral contraceptive pills have the potential to cause adverse effects on blood pressure, cardiovascular reactivity and the insulin-production response to administration of glucose and suggest that some of the beneficial effects of a low-fat diet on these parameters may be negated in women taking oral contraceptive pills.

Adult↗
PubMed2001

Acid stress, starvation, and cold stress affect poststress behavior of Escherichia coli O157:H7 and nonpathogenic Escherichia coli.

The effects of acid shock, acid adaptation, starvation, and cold stress of Escherichia coli O157:H7 (ATCC 43895), an rpoS mutant (FRIK 816-3), and nonpathogenic E. coli (ATCC 25922) on poststress heat resistance and freeze-thaw resistance were investigated. Following stress, heat tolerance at 56 degrees C and freeze-thaw resistance at -20 to 21 degrees C were determined. Heat and freeze-thaw resistance of E. coli O157:H7 and nonpathogenic E. coli was enhanced after acid adaptation and starvation. Following cold stress, heat resistance of E. coli O157:H7 and nonpathogenic E. coli was decreased, while freeze-thaw resistance was increased. Heat and freeze-thaw resistance of the rpoS mutant was enhanced only after acid adaptation. Increased or decreased tolerance of acid-adapted, starved, or cold-stressed E. coli O157:H7 cells to heat or freeze-thaw processes should be considered when processing minimally processed or extended shelf-life foods.

Adaptation, Physiological↗
PubMed2006

Correlating gene expression to physiological parameters and environmental conditions during cold acclimation of Pinus sylvestris, identification of molecular markers using cDNA microarrays.

Scots pine (Pinus sylvestris L.) seedlings were grown under different conditions (three field locations, two seasons and two climate room regimes), and then analyzed for freezing tolerance of shoots and roots and for transcript abundance in apical buds based on a cDNA microarray containing about 1500 expressed sequence tags (ESTs) from buds of cold-treated Scots pine seedlings. In a climate room providing long daily photoperiods and high temperatures, seedlings did not develop freezing tolerance, whereas seedlings in a climate room set to provide declining temperatures and day lengths developed moderate freezing tolerance. Control seedlings grown outside under field conditions developed full freezing tolerance. Differences in physiological behavior of the different seedling groups, combined with molecular analysis, allowed identification of a large group of genes, expression of which changed during the development of freezing tolerance. Transcript abundance of several of these genes was highly correlated with freezing tolerance in seedlings differing in provenance, field location or age, making them excellent candidate marker genes for molecular tests for freezing tolerance.

Acclimatization↗
PubMed1988

Small particle aerosols of enviroxime-containing liposomes.

Enviroxime inhibits the replication of all rhinoviruses tested in vitro at very low concentrations (10-100 ng/ml), but evaluations in humans have not consistently shown efficacy. Lack of an appropriate method for administering this water-insoluble drug may have contributed to the latter result. The present report describes the characteristics and utilization of small particle aerosols to continuously deliver enviroxime-containing liposomes (LE) throughout the respiratory tract. The enviroxime content of liposomes and biological fluids of exposed individuals was quantified by high performance liquid chromatography using C18 resin, a mobile phase of 60:40 acetonitrile:water, and monitoring at 215 nm. Small particle aerosols of LE generated by Puritan-Bennett nebulizers had mass median diameters ranging from 2.4 to 3.1 microns. The concentration of enviroxime in aerosol particles was proportional to the reservoir concentration; during the first hour of operation, the mean concentration was 20 micrograms of enviroxime/l of aerosol. Liposome particles in the reservoir, although initially heterogeneous in size (less than 0.1 to greater than 1 micron), were processed by passage through the nebulizer to smaller, more homogeneous particles; the majority were less than 0.2 micron. In a preliminary study to evaluate short term tolerance and toxicity, five volunteers were exposed to small particle aerosol of LE for 1 h. At 1 h post-treatment, large amounts of enviroxime were still present in the nasal wash as determined both by HPLC and biological assay. Enviroxime was not detected in any urine sample and was detected in only 1 of 5 serum samples. No side effects were noted. This data suggest that liposome aerosols offer a method for the delivery of hydrophobic compounds for the treatment of respiratory diseases.

Adult↗
PubMedDate not supplied

Cold hardiness in the boreal adder, Vipera berus.

In three freezing experiments we examined the freeze tolerance in newborn adders, Vipera berus, from East-central Sweden. After a two to three hours exposure to freezing, ten out of eleven fast frozen snakes survived and recovered completely after being exposed to -3.1 degrees C on average. In the other two experiments with fast and slow freezing followed by long exposures lasting for 22-30 hours, none of the snakes survived average exposures at -4.8 degrees C or -3.8 degrees C, respectively. The glucose content of blood from frozen snakes was significantly higher than in unfrozen ones. The increase was small and its contribution to freeze tolerance doubtful. Compared to other freeze tolerant reptiles, the adder was categorised as virtually non freeze tolerant capable of surviving only a short exposure not colder than approximately -4 degrees C. Supercooling could play a role in winter survival but their precise choice of hibernation site is probably the most important.

Adaptation, Physiological↗
PubMed1997

Changes in ABA and gene expression in cold-acclimated sugar maple.

To determine if cold acclimation of sugar maple (Acer saccharum Marsh.) is associated with specific changes in gene expression under natural hardening conditions, we compared bud and root translatable mRNAs of potted maple seedlings after cold acclimation under natural conditions and following spring dehardening. Cold-hardened roots and buds were sampled in January when tissues reached their maximum hardiness. Freezing tolerance, expressed as the lethal temperature for 50% of the tissues (LT(50)), was estimated at -17 degrees C for roots, and at lower than -36 degrees C for buds. Approximately ten transcripts were specifically synthesized in cold-acclimated buds, or were more abundant in cold-acclimated buds than in unhardened buds. Cold hardening was also associated with changes in translation. At least five translation products were more abundant in cold-acclimated buds and roots compared with unhardened tissues. Abscisic acid (ABA) concentration increased approximately tenfold in the xylem sap following winter acclimation, and the maximum concentration was reached just before maximal acclimation. We discuss the potential involvement of ABA in the observed modification of gene expression during cold hardening.

Journal Article↗
PubMed2001

Gene transfer to the desiccation-tolerant cyanobacterium Chroococcidiopsis.

The coccoid cyanobacterium Chroococcidiopsis dominates microbial communities in the most extreme arid hot and cold deserts. These communities withstand constraints that result from multiple cycles of drying and wetting and/or prolonged desiccation, through mechanisms which remain poorly understood. Here we describe the first system for genetic manipulation of Chroococcidiopsis. Plasmids pDUCA7 and pRL489, based on the pDU1 replicon of Nostoc sp. strain PCC 7524, were transferred to different isolates of Chroococcidiopsis via conjugation and electroporation. This report provides the first evidence that pDU1 replicons can be maintained in cyanobacteria other than Nostoc and Anabaena. Following conjugation, both plasmids replicated in Chroococcidiopsis sp. strains 029, 057, and 123 but not in strains 171 and 584. Both plasmids were electroporated into strains 029 and 123 but not into strains 057, 171, and 584. Expression of P(psbA)-luxAB on pRL489 was visualized through in vivo luminescence. Efficiencies of conjugative transfer for pDUCA7 and pRL489 into Chroococcidiopsis sp. strain 029 were approximately 10(-2) and 10(-4) transconjugants per recipient cell, respectively. Conjugative transfer occurred with a lower efficiency into strains 057 and 123. Electrotransformation efficiencies of about 10(-4) electrotransformants per recipient cell were achieved with strains 029 and 123, using either pDUCA7 or pRL489. Extracellular deoxyribonucleases were associated with each of the five strains. Phylogenetic analysis, based upon the V6 to V8 variable regions of 16S rRNA, suggests that desert strains 057, 123, 171, and 029 are distinct from the type species strain Chroococcidiopsis thermalis PCC 7203. The high efficiency of conjugative transfer of Chroococcidiopsis sp. strain 029, from the Negev Desert, Israel, makes this a suitable experimental strain for genetic studies on desiccation tolerance.

Base Sequence↗
PubMed2005

Development of a novel microbial sensor with baker's yeast cells for monitoring temperature control during cold food chain.

A novel microbial sensor containing a commercial baker's yeast with a high freeze tolerance was developed for visibly detecting inappropriate temperature control of food. When the yeast cells fermented glucose, the resulting gas production triggered the microbial sensor. The biosensor was a simple, small bag containing a solution of yeast cells, yeast extract, glucose, and glycerol sealed up with multilayer transparent film with barriers against oxygen and humidity. Fine adjustment of gas productivity in the biosensor at low temperatures was achieved by changing either or both concentrations of glucose and yeast cells. Moreover, the amount of time that food was exposed to inappropriate temperatures could be deduced by the amount of gas produced in the biosensor. The biosensor was stable without any functional loss for up to 1 week in frozen storage. The biosensor could offer a useful tool for securing food safety by maintaining low-temperature control in every stage from farm to fork, including during transportation, in the store, and at home.

Biosensing Techniques↗
PubMed1989

[Study of bronchial reactivity by means of cold air inhalation].

In 24 children with bronchial asthma and 16 children of a control group provocation by three-minute inhalation of cold air was applied under eucapnic condition on a RHES apparatus (E. Jaeger, GFR). After hyperventilation changes in the resistance of the airways were investigated by the occlusion method and repeatedly parameters of ventilation were examined by the flow-volume loop method. In the group of asthmatic children a significantly increased resistance was recorded after provocation with an onset already during the first minute after completed hyperventilation, and it persisted to the 9th minute. In the investigated parameters of the flow-volume loop (FEV 1, MEF 25/75) the maximum drop was recorded between the 3rd and 5th minute after provocation. In the control group no significant changes developed after provocation. Provocation by eucapnic hyperventilation of cold air is in particular in paediatric pneumological and allergological diagnosis a perspective method which does not cause discomfort to the patient and is well tolerated.

Asthma↗
PubMed1997

Preservation of pig liver allografts after warm ischemia: normothermic perfusion versus cold storage.

Warm ischemia is known to induce substantial damage to the liver parenchyma. With respect to clinical liver transplantation, the tolerance of the liver to warm ischemia and the preservation of these organs have not been studied in detail. In isolated reperfused pig livers we proceeded according to the following concept: Livers were subjected to 1 or 3 h of warm ischemia. Subsequently, these organs were preserved by either normothermic perfusion or cold storage (histidine-tryptophan-alpha-ketoglutarate, HTK) for 3 h each. After storage, liver function was assessed in a reperfusion circuit for another 3 h. Parameters under evaluation were bile flow, perfusion flow, oxygen consumption, enzyme release into the perfusate (creatine kinase, glutamic oxaloacetic transaminase (GOT), lactic dehydrogenase, and glutamic pyruvic transaminase), and histomorphology. Damage to the liver was lowest after warm ischemia of 1 h. The results after cold storage were superior to those after normothermic perfusion (GOT: 3.2 +/- 0.3 and 2.6 +/- 0.2 U/g liver; cumulative bile production: 14.7 +/- 2.1 and 9.4 +/- 1 ml, respectively; P < 0.05). In contrast, we found substantial damage at the end of reperfusion in livers undergoing 3 h of warm ischemia under both preservation techniques with severe hepatocellular pyknoses and essentially altered nonparenchymal cells. The results suggest that pig livers undergoing 1 h of warm ischemia and cold storage for 3 h with HTK solution may lead to functioning after transplantation.

Alanine Transaminase↗
PubMed2001

Developmental and stress regulation of RCI2A and RCI2B, two cold-inducible genes of arabidopsis encoding highly conserved hydrophobic proteins.

The capability of most higher plants to tolerate environmental conditions strongly depends on their developmental stage. In addition, environmental factors have pleiotropic effects on many developmental processes. The interaction between plant development and environmental conditions implies that some genes must be regulated by both environmental factors and developmental cues. To understand their developmental regulation and obtain possible clues on their functions, we have isolated genomic clones for RCI2A and RCI2B, two genes from Arabidopsis ecotype Columbia (Col), whose expression is induced in response to low temperature, dehydration, salt stress, and abscisic acid. The promoters of RCI2A and RCI2B were fused to the uidA (GUS)-coding sequence and the resulting constructs used to transform Arabidopsis. GUS activity was analyzed in transgenic plants during development under both stressed and unstressed conditions. Transgenic plants with either the RCI2A or RCI2B promoter showed strong GUS expression during the first stages of seed development and germination, in vascular bundles, pollen, and most interestingly in guard cells. When transgenic plants were exposed to low temperature, dehydration, salt stress, or abscisic acid, reporter gene expression was induced in most tissues. These results indicate that RCI2A and RCI2B are regulated at transcriptional level during plant development and in response to different environmental stimuli and treatments. The potential role of RCI2A and RCI2B in plant development and stress response is discussed.

Abscisic Acid↗
PubMed2001

Chill-coma tolerance, a major climatic adaptation among Drosophila species.

Most drosophilid species can be classified either as temperate or tropical. Adults of species were submitted to a cold treatment (0 degrees C) and then brought back to ambient temperature. They generally exhibited a chill coma and the time needed to recover was measured. We found in a set of 26 temperate species that recovery was rapid (average 1.8 min, range 0.15-4.9). In contrast, a long recovery time (average 56 min, range 24-120) was observed for 48 tropical species. A few species, like Drosophila melanogaster, are cosmopolitan and can proliferate under temperate and tropical climates. In 9 of 10 such species, slight genetic differences were found: a shorter recovery in temperate than in tropical populations. Comparing physiological data to phylogeny suggests that chill-coma tolerance has been a recurrent adaptation that is selected for in cold climates but tends to disappear under a permanently warm environment. This major climatic adaptation, evidenced in drosophilids, seems to occur in other insect groups also.

Acclimatization↗