Introduction to the special issue: Thermodynamic aspects of cryobiology

This brief paper introduces the subject of thermodynamics and the papers of the special issue on thermodynamic aspects of cryobiology. Thermodynamic terminology is defined for the non-specialist.     

Proteomic analysis of hypothalamic injury in heatstroke rats

Ischemic and oxidative damage to the hypothalamus may be associated with decreased heat tolerance as well as heatstroke formation. The present study explores the hypothalamic proteome mechanisms associated with heatstroke‐mediated hypothalamic ischemia, and oxidative damage. Heatstroke rats had hypotension, hypothalamic ischemia, and lethality. In addition, they had hyperthermia and hypothalamic blood–brain–barrier disruption, oxidative stress, activated inflammation, and neuronal apoptosis and degeneration. 2DE combined LC‐MS/MS revealed that heatstroke‐induced ischemic injury and apoptosis were associated with upregulation of L‐lactate dehydrogenase but downregulation of both dihydropyriminase‐related protein and 14‐3‐3 Zeta isoform protein. Heat‐induced blood–brain–barrier disruption might be related to upregulation of glial fibrillary acidic protein. Oxidative stress caused by heatstroke might be related to upregulation of cytosolic dehydrogenase‐1. Also, heat‐induced overproduction of proinflammatory cytokines might be associated with downregulation of stathmin 1. Heat‐induced hypothalamic ischemia, apoptosis, injury (or upregulation of L‐lactate dehydrogenase), blood–brain–barrier disruption (or upregulation of glial fibrillary acidic protein), oxidative stress (or upregulation of cytosolic dehydrogenase‐1), and activated inflammation (or downregulation of stathmin 1) were all significantly reversed by whole body cooling. Our data indicate that cooling therapy improves outcomes of heatstroke by modulating hypothalamic proteome mechanisms.     

Recovery of plants from “Near-Lethal” stress

This study reports on the dieback and recovery of red-osier dogwood, Cornus sericea L. plants from near-lethal (NL, sublethal) stress after varying lengths of post-stress environment (PSE). Intact dormant stems were subjected to 47° C for one hour during either October, November or December, and then placed into either constant 0° C or 23° C (dark condition) or kept under natural conditions at Corvallis, OR. Plants exposed to NL-heat stress in October died prior to 9 weeks of 0° C PSE, while none of the plants from other PSE treatments showed signs of injury. For plants exposed to NL-heat stress during November and December, stemdieback occurred at 0° C after 12 and 15 weeks, respectively. None of the plants from the other PSE treatments were injured. Post-stress temperatures of 0° or 5° C following NL-heat in October were lethal while temperatures above 10° C allowed recovery. Post-stress exposure to 0° C injured excised stems within 48 h, whereas irreversible damage to whole plants occurred by two weeks. Dormant plants exposed in October to other stresses, e.g., freezing temperature and hydrogen cyanamide, at NL dosages showed that these stresses also caused plant dieback at 0° C and little or no dieback at 23° C PSE.Abbreviations NL Near-Lethal - PSE Post-Stress Environment     

Behavioral and mortality responses of the bivalves Scrobicularia plana and Cerastoderma edule to temperature,as indicator of climate change's potential impacts

Temperature is one of the most important abiotic factors affected by climate change. It determines physiological processes, ecological patterns and establishes the limits of geographic distribution of species. The induced thermal stress frequently results in physiological and behavioral responses and, in extreme cases, may lead to mortality episodes. Scrobicularia plana and Cerastoderma edule behavioral and mortality responses to temperature were evaluated. Specimens were sampled in the Mondego estuary (Portugal), acclimated and exposed to different temperature treatments (5–35 °C). Individual activity and mortality were registered during 120 h laboratory assays. Both species showed a thermal optimum for their activity (S. plana: 15–23 °C; C. edule: 20–23 °C), and survival was mainly affected by high temperature (S. plana: LC50_120 h = 28.86 °C; C. edule: LC50_120 h = 28.01 °C), with 100% mortality above critical values (≥32 °C). Results further indicated that both species are more affected the higher the temperature and the longer the exposure time. This study indicates that the occurrence of extreme climatic events, especially heat waves, may be particularly impairing for these species.     

The induction of stress proteins in three marine Vibrio during carbon starvation

Abstract The induction of DnaK and GroEL homologous proteins by heat-shock and long-term carbon starvation was studied in Vibrio vulnificus, Vibrio sp. strain S14, and Vibrio sp. strain DW1. In each Vibrio strain one protein (60 kDa) reacted with antibodies against Escherichia coli -GroEL and two proteins, DnaK (69 kDa) and Sis1 (62-60 kDa), reacted with antibodies against E. coli -Dnak. The carbon starvation elicited induction of the stress proteins was strain-specific, suggesting that the induction of stress proteins like DnaK and GroEL in marine Vibrios might not be a uniform starvation response. It appears as of these proteins, only DnaK in Vibrio sp. strain S14 remains induced after long-term carbon starvation in the three marine bacterial strains that were tested.     

二价金属离子在分离大豆种子尿囊素酶中的作用

大豆 (Glycine max L.cv.Keyu 10 )种子和叶片提取液中的尿囊素酶 (ALNase)是热稳定的 ,但在Ca2 、Mg2 或 Mn2 存在的条件下对热有不同的反应。 75℃热处理 5 min,种子提取液中的酶除 Mg2 尚存有 1/ 3酶活性外 ,其余全部丧失 ,56%以上的蛋白质变性 ;叶片提取液中的酶除 Mn2 使活性下降 36%外其余仍然稳定 ,热变性蛋白也不因 Ca2 、Mg2 和 Mn2 的存在有明显增加。在室温下二价金属离子可以清除大豆种子提取液中 50 %以上非酶蛋白 ,但不影响酶活性。二价金属离子这种作用和其浓度无关 ,增加浓度反而有减弱的趋势 ,也不受 Na Cl的影响。两种 ALNase提取液对不同的难溶性钙盐也有不同的反应 ,种子提取液经 5% Ca SO4 处理后非酶蛋白减少近 50 % ,酶活性受影响不大 ;叶片提取液对Ca3 (PO4 ) 2 敏感 ,经处理后酶活性损失 50 %左右 ,蛋白损失近 70 %。 Mn2 对种子提取液中 ALNase有激活作用 ,但这种激活作用随着酶的纯化而消失。乙二胺四乙酸钠 (EDTA)对两种提取液中 ALNase活性没有影响 ,但外加二价金属离子浓度接近 DETA浓度时酶活性很快丢失     

Genetic and environmental factors in the resistance of Drosophila subobscura adults to high temperature shock

Summary Survival time following a high temperature shock of Drosophila subobscura adults in dry air has shown great variability. This experimental condition involved desiccation as the first cause of death. Here survival is studied under saturation humidity, so that the mortality may be imputed only to thermic stress. We analyze the influence of culture temperature and crowding on resistance for different sex and age of the adults. The results show strong influences of these environmental factors on heat shock resistance and show interactions with the age and sex of the adults. We suggest that these facts could be due to acclimatization and/or to adaptation. The acclimatization would occur during development and would affect physiological processes related to aging of the flies. The adaptation would take place for selection, acting through differential mortality before the heat shock. Of course, other processes could be significant. Whatever the causal explanation, it will be necessary in any future research related with heat shock resistance to take these factors into account.     

Environmental stress responses in Lactobacillus: a review

Environmental stress responses in Lactobacillus, which have been investigated mainly by proteomics approaches, are reviewed. The physiological and molecular mechanisms of responses to heat, cold, acid, osmotic, oxygen, high pressure and starvation stresses are described. Specific examples of the repercussions of these effects in food processing are given. Molecular mechanisms of stress responses in lactobacilli and other bacteria are compared.     

Heat stress-induced H2O2 is required for effective expression of heat shock genes in Arabidopsis

The mechanisms of sensing and signalling of heat and oxidative stresses are not well understood. The central question of this paper is whether in plant cells oxidative stress, in particular H₂O₂, is required for heat stress- and heat shock factor (HSF)-dependent expression of genes. Heat stress increases intracellular accumulation of H₂O₂ in Arabidopsis cell culture. The accumulation was greatly diminished using ascorbate as a scavenger or respectively diphenyleneiodonium chloride (DPI) as an inhibitor of reactive oxygen species production. The mRNA of heat shock protein (HSP) genes, exemplified by Hsp17.6, Hsp18.2, and the two cytosolic ascorbate peroxidase genes Apx1, Apx2, reached similar levels by moderate heat stress (37°C) or by treatment with H₂O₂, butylperoxide and diamide at room temperature. The heat-induced expression levels were significantly reduced in the presence of ascorbate or DPI indicating that H₂O₂ is an essential component in the heat stress signalling pathway. Rapid (15 min) formation of heat shock promoter element (HSE) protein-binding complex of high molecular weight in extracts of heat-stressed or H₂O₂-treated cells and the inability to form this complex after ascorbate treatment suggests that oxidative stress affects gene expression via HSF activation and conversely, that H₂O₂ is involved in HSF activation during the early phase of heat stress. The heat stress induction of a high mobility HSE-binding complex, characteristic for later phase of heat shock response, was blocked by ascorbate and DPI. H₂O₂ was unable to induce this complex suggesting that H₂O₂ is involved only in the early stages of HSF activation. Significant induction of the genes tested after diamid treatment and moderate expression of the sHSP genes in the presence of 50 mM ascorbate at 37°C occurred without activation of HSF, indicating that other mechanisms may be involved in stress signalling. Electronic Supplementary Material Supplementary material is available for this article at http//dx.doi.org/10.1007/s11103-006-0045-4 Roman A. Volkov and Irina I. Panchuk contributed equally     

Thermosensitivity of the <Emphasis Type="Italic">Saccharomyces cerevisiae gpp1gpp2</Emphasis> double deletion strain can be reduced by overexpression of genes involved in cell wall maintenance

A Saccharomyces cerevisiae strain in which the GPP1 and GPP2 genes, both encoding glycerol-3-phosphate phosphatase isoforms, are deleted, displays both osmo- and thermosensitive (ts) phenotypes. We isolated genes involved in cell wall maintenance as multicopy suppressors of the gpp1gpp2 ts phenotype. We found that the gpp1gpp2 strain is hypersensitive to cell wall stress such as treatment with β-1,3-glucanase containing cocktail Zymolyase and chitin-binding dye Calcofluor-white (CFW). Sensitivity to Zymolyase was rescued by overexpression of SSD1, while CFW sensitivity was rescued by SSD1, FLO8 and WSC3—genes isolated as multicopy suppressors of the gpp1gpp2 ts phenotype. Some of the isolated suppressor genes (SSD1, FLO8) also rescued the lytic phenotype of slt2 deletion strain. Additionally, the sensitivity to CFW was reduced when the cells were supplied with glycerol. Both growth on glycerol-based medium and overexpression of SSD1, FLO8 or WSC3 had additive suppressing effect on CFW sensitivity of the gpp1gpp2 mutant strain. We also confirmed that the internal glycerol level changed in cells exposed to cell wall perturbation.