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Induction of chilling resistance by water stress, and cDNA sequence analysis and expression of water stress-regulated genes in rice.

Exposure of seedlings of a chilling-sensitive variety of rice (Oryza sativa L. cv. Wasetoittu) to water stress (0.5 M mannitol, 30 min) at room temperature induced a degree of chilling resistance. No such resistance was induced by exogenous abscisic acid (ABA) application (10 microM, 60 min). Upon short-term water stress, new transcripts were expressed in both seedlings and suspension-cultured cells. We suggest that the genes induced by short-term water stress, and not those induced by ABA, are related to acquired chilling resistance in this chilling-sensitive rice variety. A total of nine different cDNA clones, specifically induced by short-term water stress, were isolated by differential hybridization and partial sequencing. Northern hybridization analysis using RNAs from the seedlings subjected to chilling after water stress treatment reveal three distinct groups of above mentioned nine cDNA clones: wsi (water stress-induced) 18, 76, and 724, representative of genes whose expression increases, decreases, and remains almost fixed during chilling, respectively. The nucleotide and deduced amino acid sequences of the three representative clones were determined. Characteristic features of wsi18 are the presence of one set of amino acid sequence repeats, a conserved amino acid sequence common to LEA-group genes in the N-terminal region, and an alanine- and lysine-rich tract in the C-terminal region.

Abscisic Acid↗

Coming to terms with stress: effects of stress location in sentence processing.

The purpose of this research was to determine the role of syllabic stress in language processing during the early on-line processing of speech and later in the representation of a sentence in memory. Experiment 1 used a syllable monitoring task while Experiment 3 used a probe task in which subjects heard a sentence and then were asked to determine whether a probe syllable had occurred in the sentence. In the monitoring task, stressed syllables were detected more rapidly in word-initial position, but unstressed syllables were detected more rapidly in word-final position. Stress facilitation in initial syllables was strongly related to high relative F0, but not to changes in perceived vowel quality as assessed in Experiment 2. This pattern is interpreted as evidence that lexical stress is used on-line to guide lexical access and/or lexical segmentation. The probe task of Experiment 3 showed stress facilitation in both positions, indicating that stress is independently retained in the postperceptual representation of a sentence.

Adolescent↗

Nitrate uptake by the halotolerant cyanobacterium Aphanothece halophytica grown under non-stress and salt-stress conditions.

We have compared the characteristics of nitrate uptake by Aphanothece halophytica grown under non-stress and salt-stress conditions. Both cell types showed essentially similar patterns of nitrate uptake toward ammonium, nitrite, and DL-glyceraldehyde. Although the affinities of nitrate to non-stress cells and salt-stress cells were not significantly different, i.e., Ks = 416 and 450 microM, respectively, the V(max) value for non-stress cells was about twofold of that for salt-stress cells (9.1 vs 5.3 micromol min(-1) mg(-1) Chl). Nitrate uptake by A. halophytica was found to be dependent on Na+. Ammonium inhibited nitrate uptake, and the presence of methionine sulfoximine could not release the inhibition by ammonium. Nitrite appeared to competitively inhibit nitrate uptake with a K(i) value of 84 microM. Both chloride and phosphate anions did not affect nitrate uptake. DL-Glyceraldehyde, an inhibitor of CO2 fixation, caused a reduction in the uptake of nitrate.

Chlorides↗

Blockade of the hypothalamic-pituitary-adrenal response to stress by intraventricular injection of dexamethasone: a method for studying the stress-induced peripheral effects of glucocorticoids.

Interest in the mechanisms whereby stressors can influence behavior and physiological functioning has involved the use of a variety of methods to prevent the stress-induced release of glucocorticoids, an important and commonly studied stress hormone. We examined the effect of intracerebral ventricular dexamethasone (ICV DEX) on the stress-induced release of adrenocorticotropic hormone (ACTH), corticosterone, plasma epinephrine (E), and plasma norepinephrine (NE). Male Sprague-Dawley rats were stereotaxically implanted with third ventricle ICV cannulae, administered DEX or vehicle, and exposed to 100 1.6-mA tail shocks. Stress hormones were assessed from blood taken during and after the cessation the shock. We report an ICV DEX injection protocol (10 microgram given four times) that results in blocking the stress-induced release of ACTH and corticosterone, and attenuating the stress-induced release of plasma E and NE. We hypothesize that ICV DEX reduces hypothalamic corticotropin releasing hormone (CRH) synthesis and/or release. This method would be especially useful for those studying the effect of pituitary-adrenal hormones on steroid sensitive peripheral targets, such as the immune system.

Adrenocorticotropic Hormone↗

Abiotic stress, the field environment and stress combination.

Farmers and breeders have long known that often it is the simultaneous occurrence of several abiotic stresses, rather than a particular stress condition, that is most lethal to crops. Surprisingly, the co-occurrence of different stresses is rarely addressed by molecular biologists that study plant acclimation. Recent studies have revealed that the response of plants to a combination of two different abiotic stresses is unique and cannot be directly extrapolated from the response of plants to each of the different stresses applied individually. Tolerance to a combination of different stress conditions, particularly those that mimic the field environment, should be the focus of future research programs aimed at developing transgenic crops and plants with enhanced tolerance to naturally occurring environmental conditions.

Crops, Agricultural↗

Hypoxia differentially regulates stress proteins in cultured cardiomyocytes: role of the p38 stress-activated kinase signaling cascade, and relation to cytoprotection.

OBJECTIVE: Stress proteins (heat shock proteins, HSPs) are molecular chaperones that have been shown to enhance the survival of cells exposed to environmental stress. We sought to investigate the effects of hypoxia on the levels of HSP27 and heme oxygenase-1 (HO-1 or HSP32) in an established model of rat neonatal cardiac myocytes in culture. METHODS: Myocytes were subjected to hypoxia (<0.5% O(2) for 16 h). Studies of cell viability and nuclear morphology showed no evidence of cell death under these conditions. RESULTS: Messenger RNA analysis demonstrated constitutive expression of HSP27 and low levels of HO-1. Hypoxia strongly induced HO-1 mRNA without affecting HSP27 mRNA. In parallel to mRNA levels, hypoxia increased HO-1 protein level without affecting HSP27. To further assess the signaling pathways implicated in HO-1 induction, we used inhibition experiments. The tyrosine kinase inhibitor tyrphostin and the mitogen-activated protein kinase inhibitor PD98059 did not prevent HO-1 induction, while the protein kinase C inhibitor chelerythrine partially blocked this response. The p38 stress-activated kinase inhibitor SB203580 was the most potent in suppressing hypoxia-induced HO-1. In vitro kinase assays, cell labeling and immunoprecipitation showed activation of signaling pathways downstream of p38 stress-activated kinase as revealed by an increase in phosphorylation of MAPKAPK-2/3 kinases and HSP27. CONCLUSIONS: These data show a differential pattern of hypoxia-induced HSP expression and implicate the stress kinase in HO-1 induction. Thus, selective regulation of HSP levels may play a role in the cardioprotective mechanisms that participate in the adaptive response to hypoxia-induced stress.

Alkaloids↗

Fluctuating shear stress effects on stress fiber architecture and energy metabolism of cultured renal cells.

The project investigates the relationship between the external shear force and the actin cytoskeleton along with the metabolic changes occurring inside the cells due to this force. Anchorage-dependent Madin Darby canine kidney (MDCK) cells were placed in spinner flasks with paddle-type stirrers agitated at 20 rpm, where they experienced shear stress fluctuations from 0.02 to 0.27 dyn/cm2 in magnitude. Following fixation, permeabilization, and staining with rhodamine-phalloidin, the relative amounts and distribution of F-actin stress fibers in the 1 micron basal layer of the cells were visualized by confocal microscopy. These structures disappeared after 12-15 h of exposure to shear stress. Previous results showed that the stress fibers disappear, leading to loss of epithelial attachment, after only 1 h of starvation-induced energy depletion. Therefore, in this study, the energy metabolism of the cells was established by measuring adenosine triphosphate (ATP) levels at different time intervals. No statistical difference in ATP content was found between the shear-stressed cells and the controls, showing that shear stresses cause cytoskeletal reorganization by a mechanism other than ATP depletion.

Actins↗

Life stress, the "kindling" hypothesis, and the recurrence of depression: considerations from a life stress perspective.

Major depression is frequently characterized by recurrent episodes over the life course. First lifetime episodes of depression, however, are typically more strongly associated with major life stress than are successive recurrences. A key theoretical issue involves how the role of major life stress changes from an initial episode over subsequent recurrences. The primary conceptual framework for research on life stress and recurrence of depression is the "kindling" hypothesis (R. M. Post, 1992). Despite the strengths of the kindling hypothesis, a review of the research literature reveals inconsistencies and confusion about life stress and its implications for the recurrence of depression. Adopting a life stress perspective, the authors introduce 3 major themes that resolve the inconsistencies in the current literature. They integrate these themes and extrapolate the ideas with available data to develop a preliminary framework for evaluating competing explanatory models and to guide research on life stress and the recurrence of depression.

Depression↗

Stress tolerance and metabolic response to stress in Drosophila melanogaster.

A potentially important physiological response to stress may be alteration in the gross regulation of energy metabolism. Different genotypes may respond differently to environmental stress, and the variation in these norms of reaction may be of key importance to the maintenance of genetic variation in metabolic traits. In the study reported here, a set of genetically defined lines of Drosophila melanogaster were exposed to four stresses (acetic acid, ethanol, starvation and thermal stress) in order to assess the magnitude of environmental effects and genotype x environment interactions. In addition to scoring metabolic traits, distributions of survival times under each stress were also quantified. Although both metabolic traits and survival times exhibited strong differences among genotypes, the correlations between enzyme traits and survival were generally weak. Many of the genetic correlations exhibit significant heterogeneity across environments. The results suggest that transient environmental stress may play an important role in the evolution of this highly intercorrelated set of metabolic traits.

Acetic Acid↗

Stress induction of the Bacillus subtilis clpP gene encoding a homologue of the proteolytic component of the Clp protease and the involvement of ClpP and ClpX in stress tolerance.

The Bacillus subtilis clpP gene, encoding the proteolytic component of the Clp or Ti protease, was cloned and sequenced. The amount of clpP-specific mRNA increased after heat shock, salt and ethanol stress, as well as after treatment with puromycin. Two transcriptional start sites upstream of the clpP structural gene were identified, preceded by sequences resembling the consensus sequences of promoters recognized by sigmaA and sigmaB transcriptional factors of the B. subtilis RNA polymerase respectively. Transcription initiation occurred predominantly at the putative sigmaA-dependent promoter in exponentially growing cells and was induced under stress conditions. After exposure to stress, initiation of transcription also increased at the sigmaB-dependent promoter, but to a lesser extent, indicating that clpP belongs to a double promoter-controlled subgroup of class III general stress genes in B. subtilis. In a sigB mutant strain, clpP remained heat and stress inducible at the sigmaA-dependent promoter. BgaB-reporter gene fusions, carrying either the sigmaA- or the sigmaB-dependent promoter, showed a higher bgaB induction at the sigmaA-dependent promoter, whereas a significantly lower level of induction was measured at the sigmaB-dependent promoter. The sigmaA-dependent promoter appeared to be crucial for the heat-inducible transcription of clpP. A CIRCE (controlling inverted repeat of chaperone expression) element, the characteristic regulation target of class I heat shock genes such as dnaK and groESL, was not found between the transcriptional and translational start sites. Mutants lacking either the proteolytic component ClpP or the regulatory ATPase component ClpX were phenotypically distinct from the wild type. Both mutants produced chains of elongated cells and exhibited severely impaired growth under stress conditions and starvation. Comparison of two-dimensional protein gels from wild-type cells with those from clpP and clpX mutant cells revealed several changes in the protein pattern. Several proteins, such as GroEL, PpiB, PykA, SucD, YhfP, YqkF, YugJ and YvyD, which were found preferentially in higher amounts in both clpP and clpX mutants, might be potential substrates for the ClpXP protease.

ATPases Associated with Diverse Cellular Activitie↗

Transient expression and heat-stress-induced co-aggregation of endogenous and heterologous small heat-stress proteins in tobacco protoplasts.

Heat-stress granules (HSG) are highly ordered, cytoplasmic chaperone complexes found in all heat-stressed plant cells. We have developed an experimental system involving expression of cytosolic class I and class II small heat-stress proteins (Hsps) of pea, Arabidopsis and tomato in tobacco protoplasts to study the structural prerequisites for the assembly of HSG or HSG-like complexes. Class I and class II small Hsps formed class-specific dodecamers of 210-280 kDa, which, upon heat stress, were incorporated into HSG complexes. Interestingly, class II dodecamers alone could form HSG-like complexes (auto-aggregation), whereas class I dodecamers could do so only in the presence of class II proteins (recruitment). By analysing C-terminal deletion forms of Hsp17 class II, we obtained evidence that the intact C-terminus is critical for the oligomerization state, for the heat-stress-induced auto-aggregation and for recruitment of class I proteins. The class-specific formation of dimers as a prerequisite for oligomerization was analysed by the yeast two-hybrid system. In the presence of the endogenous (tobacco) set of heat-stress-induced proteins, all heterologous class I and class II proteins were incorporated into HSG complexes, whose ultrastructure was different from that of complexes formed by class I and class II proteins alone. Although other, more distantly related, members of the Hsp20 family, i.e. the plastidic pea Hsp21, the Drosophila Hsp23 and the mouse Hsp25, were well expressed in tobacco protoplasts and formed homo-oligomers of 200-700 kDa, none of them could be recruited to HSG complexes.

Amino Acid Sequence↗

Psychological hardiness, workplace stress and related stress reduction strategies.

Contending with stressful situations in the workplace is a common occurrence for all health care providers. Stress has numerous devastating effects on the workplace environment, as well as upon individuals who become victims of stress. However, enhancing psychological hardiness may facilitate an individual's ability to deal with workplace stress. Psychological hardiness, a personality style consisting of commitment, control and challenge, encourages human survival and the enrichment of life through development. This article provides an overview of psychological hardiness and workplace stress, and proposes strategies that one can use to enhance commitment, control and challenge in an effort to reduce the impact of stress.

Adaptation, Psychological↗

Stress nuclear myocardial perfusion imaging versus stress echocardiography: prognostic comparisons.

The use of noninvasive stress cardiac imaging for stratifying risk in patients with known or suspected coronary artery disease is growing as a tool for identification of the subgroup most likely to benefit from the expense and risk of more invasive procedures, including cardiac catheterization and coronary revascularization. In this setting, it is especially important that a test be able to identify patients with sufficiently low risk that clinicians are comfortable in deferring such interventions, especially in those with other markers of increased risk. Previous data have shown that cardiac risk is most closely related to the presence and extent of jeopardized viable myocardium on noninvasive stress cardiac imaging. Although stress echocardiography may have comparable ability to detect coronary artery disease, current data suggest that stress echocardiography detects significantly less jeopardized viable myocardium than stress nuclear myocardial perfusion imaging and consequently fewer patients at risk for cardiac events. Stress nuclear myocardial perfusion imaging may therefore have important advantages for risk stratification and the direction of future care of patients with known or suspected coronary artery disease.

Echocardiography↗

Herp is dually regulated by both the endoplasmic reticulum stress-specific branch of the unfolded protein response and a branch that is shared with other cellular stress pathways.

The mammalian unfolded protein response (UPR) includes two major branches: one(s) specific to ER stress (Ire1/XBP-1 and ATF6-dependent), and one(s) shared by other cellular stresses (PERK/eIF-2alpha phosphorylation-dependent). Here, we demonstrate that the ER-localized protein Herp represents a second target, in addition to CHOP, that is dually regulated by both the shared and the ER stress-specific branches during UPR activation. For the first time, we are able to assess the contribution of each branch of the UPR in the induction of these targets. We demonstrate that activation of the shared branch of the UPR alone was sufficient to induce Herp and CHOP. ATF4 was not required during ER stress when both branches were used but did contribute significantly to their induction. Conversely, stresses that activated only the shared branch of the UPR were completely dependent on ATF4 for CHOP and Herp induction. Thus, the shared and the ER stress-specific branches of the UPR diverge to regulate two groups of targets, one that is ATF6 and Ire1/XBP-1-dependent, which includes BiP and XBP-1, and another that is eIF-2alpha kinase-dependent, which includes ATF4 and GADD34. The two branches also converge to maximally up-regulate targets like Herp and CHOP. Finally, our studies reveal that a PERK-dependent target other than ATF4 is contributing to the cross-talk between the two branches of the UPR that has previously been demonstrated.

Activating Transcription Factor 4↗

Comparison of the stress response to cryopreservation in monolayer and three-dimensional human fibroblast cultures: stress proteins, MAP kinases, and growth factor gene expression.

Stress responses induced in fibroblasts by cryopreservation were compared in suspension or three-dimensional cultures at various times up to 5 days of recovery. Cryopreservation caused an 86% inhibition in [(35)S]methionine incorporation, with recovery over 2 days to 45% &plusmn: 14% of its original value. Stress proteins, including heat shock protein (hsp) and glucose-regulated proteins (GRP), detected by immunoblotting, responded with transient increases in cellular content (hsp27 and hsp90 in suspension and three-dimensional culture, and hsp70 only in three-dimensional culture), decreases at 24 h (hsp56, hsp70, hsp90, and GRP78 in three-dimensional culture and hsp90 in suspension), or little change (hsp70 in suspension). Polyacrylamide gel electrophoresis of [(35)S]methionine-labeled proteins showed transient induction of hsp47 within 4 h, and increased synthesis of hsp90 and GRP78 and other unidentified proteins at 24 h, but no change in hsp70. The mitogen-activated protein (MAP) kinase, p38, showed a transient increase after thawing, followed by a peak in extracellular signal-regulated kinase at 24 h. The stress-activated protein kinase (JNK) was not activated. In both stress protein and MAP kinase responses, the three-dimensional cultures showed a more intense response than fibroblasts in suspension. Although some responses were related to osmotic and cold stress during freezing, others were unique. Cryopreservation induced mRNA for selected growth factors, including vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) A chain, which increased 5- to 20- fold at 48 h returning to basal levels by 120 h. Our results indicate the novel finding that cryopreservation of fibroblasts grown in three-dimensional culture induced a specific cellular stress response including growth factors.

Cell Culture Techniques↗

Dictyostelium stress-activated protein kinase alpha, a novel stress-activated mitogen-activated protein kinase kinase kinase-like kinase, is important for the proper regulation of the cytoskeleton.

Mitogen-activated protein kinase cascades regulate various cellular functions, including growth, cell differentiation, development, and stress responses. We have identified a new Dictyostelium kinase (stress-activated protein kinase [SAPK]alpha), which is related to members of the mixed lineage kinase class of mitogen-activated protein kinase kinases. SAPKalpha is activated by osmotic stress, heat shock, and detachment from the substratum and by a membrane-permeable cGMP analog, a known regulator of stress responses in Dictyostelium. SAPKalpha is important for cellular resistance to stresses, because SAPKalpha null cells exhibit reduced viability in response to osmotic stress. We found that SAPKalpha mutants affect cellular processes requiring proper regulation of the actin cytoskeleton, including cell motility, morphogenesis, cytokinesis, and cell adhesion. Overexpression of SAPKalpha results in highly elevated basal and chemoattractant-stimulated F-actin levels and strong aggregation and developmental defects, including a failure to polarize and chemotax, and abnormal morphogenesis. These phenotypes require a kinase-active SAPKalpha. SAPKalpha null cells exhibit reduced chemoattractant-stimulated F-actin levels, cytokinesis, developmental and adhesion defects, and a motility defect that is less severe than that exhibited by SAPKalpha-overexpressing cells. SAPKalpha colocalizes with F-actin in F-actin-enriched structures, including membrane ruffles and pseudopodia during chemotaxis. Although SAPKalpha is required for these F-actin-mediated processes, it is not detectably activated in response to chemoattractant stimulation.

Actins↗

Osmotic stress in barley regulates expression of a different set of genes than salt stress does.

Under high salt conditions, plant growth is severely inhibited due to both osmotic and ionic stresses. In an effort to dissect genes and pathways that respond to changes in osmotic potential under salt stress, the expression patterns were compared of 460 non-redundant salt-responsive genes in barley during the initial phase under osmotic versus salt stress using cDNA microarrays with northern blot and real-time RT-PCR analyses. Out of 52 genes that were differentially expressed under osmotic stress, 11, such as the up-regulated genes for pyrroline-5-carboxylate synthetase, betaine aldehyde dehydrogenase 2, plasma membrane protein 3, and the down-regulated genes for water channel 2, heat shock protein 70, and phospholipase C, were regulated in a virtually identical manner under salt stress. These genes were involved in a wide range of metabolic and signalling pathways suggesting that, during the initial phase under salt stress, several of the cellular responses are mediated by changes in osmotic potential.

Enzymes↗

Trehalose metabolism in Escherichia coli: stress protection and stress regulation of gene expression.

Endogenously synthesized trehalose is a stress protectant in Escherichia coli. Externally supplied trehalose does not serve as a stress protectant, but it can be utilized as the sole source of carbon and energy. Mutants defective in trehalose synthesis display an impaired osmotic tolerance in minimal growth media without glycine betaine, and an impaired stationary-phase-induced heat tolerance. Mechanisms for stress protection by trehalose are discussed. The genes for trehalose-6-phosphate synthase (otsA) and anabolic trehalose-6-phosphate phosphatase (otsB) constitute an operon. Their expression is induced both by osmotic stress and by growth into the stationary phase and depend on the sigma factor encoded by rpoS (katF). rpoS is amber-mutated in E. coli K-12 and its DNA sequence varies among K-12 strains. For trehalose catabolism under osmotic stress E. coli depends on the osmotically inducible periplasmic trehalase (TreA). In the absence of osmotic stress, trehalose induces the formation of an enzyme IITre (TreB) of the group translocation system, a catabolic trehalose-6-phosphate phosphatase (TreE), and an amylotrehalase (TreC) which converts trehalose to free glucose and a glucose polymer.

Base Sequence↗