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Pathogenicity in isolates of Salmonella enterica serotype Enteritidis PT4 which differ in RpoS expression: effects of growth phase and low temperature.

Experiments with 2 wild type isolates of Salmonella enterica serotype Enteritidis PT4, which differed in RpoS expression, tolerance to certain hostile environments and pathogenicity, found that changes in in vitro acid, heat, or peroxide tolerance had no effect on the ability of the isolates to multiply in the spleens of C57/BL7/J mice infected orally. Thus, with the pathogenic RpoS-positive isolate, the infectivity of log phase chilled cells, which are profoundly acid-sensitive, was the same as that of non-chilled stationary phase cells which are acid-tolerant. Similarity the infectivity of the RpoS-negative, sensitive isolate, was not enhanced by increases in any tolerance. The ability to survive on surfaces, like infectivity, was also largely unaffected by either growth phase or cold exposure. These two attributes may thus be related and, given that the pathogenic PT4 isolate is capable of prolonged survival and the nonpathogenic isolate survives poorly, survival could serve as a potential marker of pathogenicity. Although the pathogenicity of the two isolates was very different, they showed an almost identical increase in acid tolerance following culture at pH 4.0 for up to 60 min.

Acids↗

Discrimination between chilling-sensitive and chilling-resistant plants based on measurements of free fatty acid accumulation and inactivation of oxygen evolution in aged chloroplasts.

The effect of aging of isolated chloroplasts of two chilling-sensitive (CS) and three chilling-resistant (CR) plants on the inactivation of oxygen evolution and accumulation of free fatty acids (FFA) was studied at 30 degrees C, pH 5.5 or 7.0, in the absence or presence of either sorbitol or NaCl. Considerable accumulation of FFA in aged chloroplasts of CS plants: bean and maize line F7-RpIII was accompanied by a marked inactivation of oxygen evolution. This relation was not, however, found in chloroplasts of CR species: pea, wheat and maize line EP1-RpI, in which the accumulation of FFA upon aging was very low whereas the decline of the rate of oxygen evolution was pronounced. In contrast to changes observed at pH 5.5, the inactivation of oxygen evolution in chloroplasts of CR species aged at pH 7.0 was dependent on the composition of the medium, especially in wheat chloroplasts. Thus, for the evaluation of chilling sensitivity based on the measurements of oxygen evolution activity solely, either aging of chloroplasts at pH 5.5 or possibly at pH 7.0 with NaCl included into the incubation medium may be recommended. It is concluded that determination of both the extent of FFA accumulation and inactivation of oxygen evolution in aged chloroplasts might be applied as chilling tolerance indexes.

Chloroplasts↗

Genome-wide analysis of FATA associated with drought tolerance in tetraploid potato (Solanum tuberosum).

The cuticle represents the outer most protective barrier against biotic and abiotic stresses. It is composed of cutin and waxes and protects plants from desiccation, UV, cold, mechanical stresses, and pathogens. GWAS/BSAseq combined with SeqSNP analyses in an association panel of 34 potato cultivars had revealed that the acyl-ACP thioesterase FATA (Soltu.DM.06G033680.1) is significantly associated with drought tolerance in potato. Apart from three FATB genes, only one FATA gene is present in potato that has the highest homology to FATA2 in Arabidopsis. FATA is responsible for the export of C18:1 fatty acid from chloroplast into cytosol, which is necessary for the biosynthesis of cutin. A knockout mutant of AtFATA2 was analyzed with regard to the cuticle permeability and to drought tolerance as well as recovery. Loss of FATA function leads to higher sensibility to water deficit in Arabidopsis, but to no change in recovery. The increased permeability of the cuticle in the fata2 knockout mutant as shown indirectly by higher chlorophyll leaching might play a role in this. Haplotypes for FATA were identified for the two potato cultivars Albatros and Désirée. All Désirée haplotypes and Albatros haplotypes 1, 3 and 4 were also revealed by former potato pan genome studies, while Albatros haplotype 2 is unique and has not been described before. Protein models were developed to investigate the influence of different SNPs in the haplotypes on the predicted protein structure and especially the substrate cavity. In potato, protein modeling suggests that only the hypothetical isoform B of FATA might be able to process oleoyl-ACP, but not hypothetical isoform A. However, this hypothesis needs to be verified by enzyme activity assays.

FATA↗

[Clinical aspects of global ischemia of the heart].

The vast majority of modern surgical open-heart procedures require a period of global myocardial ischaemia due to the interruption of the coronary circulation, since a bloodless operative field is of great importance for delicate surgical techniques. Several cardiac protective measures have been applied in order to minimize ischaemic damage to the heart muscle: 1. Intermittent myocardial ischaemia, combined with mild to moderate hypothermia: ischaemic periods of 15-30 min at 28-32 degrees C are tolerated. Multiple ischaemic periods are possible after intermittent (3-5 min) reperfusion. 2. Deep hypothermia, combined with ischaemia: myocardial cooling to 18-23 degrees C protects the heart for ischaemic periods of up to 45-60 min. 3. Cardioplegic arrest, combined with hypothermia: a great variety of cardioplegic solutions are still used today in clinical practice. a. Interruption of electrical activity: most cardioplegic solutions use K+ (15-35 mmol/l); complete suppression of electrical processes is not always achieved. b. Interruption of electrical activity and buffering of glycolytic end products. St. Thomas-, Kirklin-Solution, blood cardioplegia and their multiple variations can result in an 3-5 fold tolerance to myocardial ischaemia. c. Interruption and blockade of electro-mechanical activity, combined with an effective buffering. Bretschneider-HTK cardioplegia equilibrates the extracellular space due to a 6-10 min infusion of the cold, crystalloid solution and allows uninterrupted ischaemic periods of more than 2 h under clinical conditions. In order to achieve optimal cardiac tolerance to global ischaemia a careful protocol of perioperative myocardial proection has to be followed. Most heart centers use a combination of hypothermia and effective cardioplegia.

Cardioplegic Solutions↗

Developmental traits affecting low-temperature tolerance response in near-isogenic lines for the Vernalization locus Vrn-A1 in wheat (Triticum aestivum L. em Thell).

Investigation of low-temperature (LT) tolerance in cereals has commonly led to the region of the vyn-A1 vernalization gene or its homologue in related genomes. Two cultivars, one a non-hardy spring wheat and one a very cold-hardy winter wheat, whose growth habits are determined by the Vrn-A1 (spring habit) and vrn-A1 (winter habit) alleles, were chosen to produce reciprocal near-isogenic lines (NILs). These lines were then used to determine the relationship between rate of phenological development and the degree and duration of LT tolerance gene expression. Each allele was isolated in the genetic backgrounds of the non-hardy spring wheat 'Manitou' and the very cold-hardy winter wheat 'Norstar'. The effects of each allele on phenological development and low-temperature tolerance (LT50) were determined at regular intervals over a 4 degrees C acclimation period of 0-98 d. The vegetative/reproductive transition, as determined by final leaf number (FLN), was found to be a major developmental factor influencing LT tolerance. Possession of a vernalization requirement increased both the length of the vegetative growth phase and LT tolerance. Similarly, increased FLN in spring Norstar and winter Manitou NILs delayed their vegetative/reproductive transition and increased their LT tolerance relative to Manitou. Although the winter Manitou NILs had a lower FLN than the spring Norstar NILs, they were able to extend their vegetative stage to a similar length by increasing the phyllochron (interval between the appearance of successive leaves). Cereal plants have four ways of increasing the length of the vegetative phase, all of which extend the time that low-temperature tolerance genes are more highly expressed: (1) vernalization; (2) photoperiod responses; (3) increased leaf number; and (4) increased length of the phyllochron.

Acclimatization↗

Effects of hypnosis on diffuse noxious inhibitory controls.

The neurophysiological mechanisms of hypnotic analgesia are still under debate. It is known that pain occurring in one part of the body (counterstimulation) decreases pain in the rest of the body by activating the diffuse noxious inhibitory controls (DNICs). The aim of this study was to explore the effects of hypnosis on both pain perception and heterotopic nociceptive stimulation. The A forms of both the Harward Group Scale of Hypnotic Susceptibility and the Stanford Hypnotic Susceptibility Scale were administered to 50 healthy students. Twenty subjects were selected and assigned to two groups: group A, consisting of 10 subjects with high hypnotic susceptibility; and group B, consisting of 10 subjects with low hypnotic susceptibility. The subjects were then randomly assigned first to either a control session or a session of hypnotic analgesia. The nociceptive flexion reflex (RIII) was recorded from the biceps femoris muscle in response to stimulation of the sural nerve. The subjective pain threshold, the RIII reflex threshold, and the mean area with suprathreshold stimulation were determined. Heterotopic nociceptive stimulation was investigated by the cold-pressor test (CPT). During and immediately after the CPT, the subjective pain threshold, pain tolerance, and mean RIII area were determined again. The same examinations were repeated during hypnosis. Hypnosis significantly reduced the subjective pain perception and the nociceptive flexion reflex. It also increased pain tolerance and reduced pain perception and the nociceptive reflex during the CPT. These effects were found only in highly susceptible subjects. However, the DNIC's activity was less evident during hypnosis than during the CPT effects without hypnosis. Both hypnosis and DNICs were able to modify the perception of pain. It seems likely that DNICs and hypnosis use the same descending inhibitory pathways for the control of pain. The susceptibility of the subject is a critical factor in hypnotically induced analgesia.

Adult↗

Short-day potentiation of low temperature-induced gene expression of a C-repeat-binding factor-controlled gene during cold acclimation in silver birch.

Development of winter hardiness in trees is a two-stage process involving sequential perception of distinct environmental cues, short-day (SD) photoperiod and low temperature (LT). We have shown that both SD and LT are recognized by leaves of silver birch (Betula pendula cv Roth) leading to increased freezing tolerance, and thus leaves can be used as an experimental model to study the physiological and molecular events taking place during cold acclimation. To obtain a molecular marker for the acclimation process in birch we cloned a gene, designated Bplti36, encoding a 36-kD acidic SK2 type of dehydrin. The gene was responsive to LT, drought, salt, and exogenous abscisic acid. This responsiveness to abiotic stresses and abscisic acid was retained when Bplti36 was introduced to Arabidopsis (Arabidopsis thaliana). The LT induction of the gene appeared to be under the control of the C-repeat-binding factor pathway as suggested by the presence of several C-repeat/dehydration-responsive element/LT-responsive elements in the Bplti36 promoter and its constitutive expression in C-repeat-binding factor overproducing Arabidopsis. In birch SD photoperiod at normal-growth temperature did not result in significant induction of Bplti36. However, preexposure to SD followed by LT treatment resulted in a remarkable increase in Bplti36 transcript accumulation as compared to LT-treated plants grown at long-day photoperiod. This suggests that SD photoperiod potentiates the LT response by conditioning the leaf tissue to be more responsive to the LT stimulus.

Acclimatization↗

H-2 restriction of virus-specific cytotoxicity across the H-2 barrier. Separate effector T-cell specificities are associated with self-H-2 and with the tolerated allogeneic H-2 in chimeras.

During infection with lymphocytic choriomeningitis or vaccinia virus, F1 irradiation chimeras reconstituted with bone marrow cells from or both parents generate cytotoxic T cells which can lyse targets across the H-2 barrier. However, activity of chimera T cells is H-2 restricted as shown by cold target competition experiments and selective restimulation of a secondary response in vitro; T cells of H-2k specificity which lyse tolerated infected H-2d target cells do not lyse infected H-2k or unrelated target cells and vice versa. Therefore, H-2 restriction of virus-specific cytotoxic T cells probably does not reflect need for like-like self-interactions for lysis to occur. The specificity of virus immune T cells is thus determined by the H-2K and H-2D specificities present in the infected animal and which are probably recognized unidirectionally by T cells. The results are compatible with the idea the T cells are specific for "altered alloantigen", i.e., a complex of cell surface marker and viral antigen. Alternatively, explained with a dual recognition model, T cells may possess two independently, clonally expressed receptors, a self-recognizer which is expressed for one of the syngeneic or tolerated allogeneic K or D "self" markers, and an immunologically specific receptor for viral antigen.

Animals↗

The association among gene expression responses to nine abiotic stress treatments in Arabidopsis thaliana.

The identification and analysis of genes exhibiting large expression responses to several different types of stress may provide insights into the functional basis of multiple stress tolerance in plant species. This study considered whole-genome transcriptional profiles from Arabidopsis thaliana root and shoot organs under nine abiotic stress conditions (cold, osmotic stress, salt, drought, genotoxic stress, ultraviolet light, oxidative stress, wounding, and high temperature) and at six different time points of stress exposure (0.5, 1, 3, 6, 12, and 24 hr). In roots, genomewide correlations between transcriptional responses to different stress treatments peaked following 1 hr of stress exposure, while in shoots, correlations tended to increase following 6 hr of stress exposure. The generality of stress responses at the transcriptional level was therefore time and organ dependent. A total of 67 genes were identified as exhibiting a statistically significant pattern of gene expression characterized by large transcriptional responses to all nine stress treatments. Most genes were identified from early to middle (1-6 hr) time points of stress exposure. Analysis of this gene set indicated that cell rescue/defense/virulence, energy, and metabolism functional classes were overrepresented, providing novel insight into the functional basis of multiple stress tolerance in Arabidopsis.

Arabidopsis↗

Effects of freezing on plant mesophyll cells.

Freezing and thawing of leaves of herbaceous plants leads to damage when the freezing temperature falls below a certain tolerance limit, which depends on the plant species and state of acclimation. Such damage is expressed as an irreversible inhibition of photosynthesis observed after thawing. In frost-damaged leaves the capacity of photosynthetic reactions of the thylakoid membranes is impaired. Particularly, the water-oxidation system, photosystems II and I are inhibited. However, it appears that CO2 assimilation is more readily affected by freezing stress than the activity of the thylakoids. The inhibition of CO2 fixation seen in initial stages of damage seems to be independent of thylakoid inactivation. This can be shown by chlorophyll fluorescence analysis made simultaneously with measurement of CO2 assimilation. Fluorescence emission by leaves is strongly influenced by carbon assimilation activity, namely via the redox state of the photosystem II electron acceptor QA (QA-dependent quenching) and via energization of the thylakoid membranes depending on the transthylakoid proton gradient (energy-dependent quenching). Resolution of these components of fluorescence changes provides insight into alterations of the CO2 fixing capacity of the chloroplasts and properties of the thylakoids. The effects of freezing and thawing were studied in detail with isolated mesophyll protoplasts prepared from both non-hardened and cold-acclimated plants of Valerianella locusta L. Freezing damage was characterized by various parameters such as plasma membrane integrity, photosynthetic CO2 assimilation, chlorophyll fluorescence emission and activities of thylakoids isolated from the protoplasts. All tests indicated a substantially increased frost tolerance of protoplasts obtained from cold-acclimated as compared to non-hardened leaves. CO2 assimilation and related fluorescence changes were the most freezing-sensitive parameters in both types of protoplasts. Inactivation of CO2 assimilation was correlated neither to the disintegration of the plasma membrane nor to inactivation of the thylakoids. Experimental data indicate that freeze-thaw treatment affected the light-regulated enzymes of the carbon reduction cycle, such as fructose-1,6-bisphosphatase, sedoheptulose-1,7-bisphosphatase and ribulose-1,5-bisphosphate carboxylase. Inhibition of light-activation of these enzymes may be based on altered properties of the chloroplast envelope.

Acclimatization↗

Mechanisms of the antidiabetic effects of the beta 3-adrenergic agonist CL-316243 in obese Zucker-ZDF rats.

Previous studies have demonstrated that chronic cold exposure activates the sympathetic nervous system, increases energy expenditure, improves glucose uptake in peripheral tissues [brown and white adipose tissues (BAT and WAT) and muscles] of normal rats. The goal of the present studies was to test whether the selective beta 3-adrenergic agonist CL-316243 (CL) would mimic the beneficial beneficial effects of cold exposure in lean and obese ZDF/Gmi-fa male (ZDF) rats, a new model of type II diabetes. In obese ZDF rats, chronic infusion of CL (1 mg.kg-1.day-1 for 14 days) significantly decreased body weight gain, food intake, and WAT weight. It also increased total tissue cytochrome oxidase activity, not only in BAT (15 times), but also in WAT (2-4) times, suggesting that it progressively enhanced mitochondriogenesis in adipose tissues. CL treatment normalized hyperglycemia and reduced hyperinsulinemia and circulating free fatty acid (FFA) levels. It also improved glucose tolerance and reduced insulin response during an intravenous glucose tolerance test. In general, the beneficial effects of CL were more pronounced in obese than in lean rats. Hyperinsulinemic-euglycemic glucose clamps combined with the [2-3H]deoxyglucose method revealed that CL markedly improved insulin responsiveness in obese rats (3-4 times) and increased glucose uptake in BAT (21 times), WAT (3 times), skeletal muscles (2-3 times), and in the diaphragm (2.8 times), but not in the heart. It is concluded that chronic CL treatment improves glucose tolerance and insulin responsiveness in obese ZDF rats by a mechanism similar to that induced by chronic cold exposure, i.e., by stimulating facultative thermaogenesis, mitochondriogenesis, and glucose utilization in BAT and WAT. In addition to this mechanism, the reduction in plasma FFA levels induced by chronic CL treatment may further contribute to enhance glucose uptake in skeletal muscles (a tissue that does not express typical beta 3-adrenoceptors) via the "glucose-fatty acid" cycle. The antiobesity and antidiabetic properties of CL suggest that selective beta 3-adrenergic agonists may represent useful agents for the treatment of type II diabetes.

Adipose Tissue, Brown↗

Regulation of stress-induced phospholipid hydroperoxide glutathione peroxidase expression in citrus

Recent findings in our laboratory showed that in citrus cells, salt treatment induced the accumulation of mRNA and a protein corresponding to phospholipid hydroperoxide glutathione peroxidase (PHGPX), an enzyme active in the cellular antioxidant system. The protein and its encoding gene, csa, were isolated and characterized, and the expected enzymatic activity was demonstrated (G. Ben-Hayyim et al., 1993, Plant Sci. 88: 129-140; D. Holland et al., 1993, Plant Mol. Biol. 21: 923-927; D. Holland et al., 1994, FEBS Lett. 337: 52-55; T. Beeor-Tzahar et al., 1995, FEBS Lett. 366: 151-155). In an attempt to find out how salt induces the expression of an antioxidant enzyme, the regulation of PHGPX in citrus cells was studied at both the mRNA transcript and the protein levels. A high and transient response at the csa mRNA level was observed after 4-7 h of exposing salt-sensitive cells to NaCl, or abscisic acid, whereas no response could be detected in the salt-tolerant cells under the same conditions. tert-Butylhydroperoxide, a substrate of PHGPX, induced csa mRNA transcripts after only 2 h, and abolished the differential response between salt-sensitive and salt-tolerant cells. On the basis of these results and those obtained under heat and cold stresses, it is suggested that csa is directly induced by the substrate of its encoded enzyme PHGPX, and that salt induction occurs mainly via the production of reactive oxygen species and hydroperoxides.

Journal Article↗

A dose-response study of the efficacy and safety of ipratropium bromide nasal spray in the treatment of the common cold.

Rhinorrhea is an annoying symptom of the common cold for which effective therapy is not currently available. Ipratropium bromide (IB) is an anticholinergic drug that has been shown to decrease glandular secretion when applied topically to the nasal mucosa. The purpose of this study was to compare the efficacy and safety of three doses of IB nasal spray versus either vehicle or no treatment in relieving rhinorrhea in patients with naturally acquired colds. Rhinorrhea severity was measured objectively by determining nasal discharge weights and subjectively by means of visual analog scale scores. Compared with either vehicle or no treatment, IB nasal spray produced a significant decrease in the severity of rhinorrhea. A dose of 84 micrograms (two sprays of a 0.06% solution in buffered saline solution) in each nostril was more efficacious than a 42 microgram per nostril dose and only marginally less efficacious than a 168 micrograms per nostril dose. The 84 micrograms per nostril dose also was associated with fewer adverse events than was the higher dose. None of the adverse events related to intranasal IB therapy was of a serious nature. The use of IB nasal spray appears to be a rational and safe approach to relieving rhinorrhea associated with the common cold.

Adolescent↗

Reactivity and vulnerability to stress-associated risk for upper respiratory illness.

OBJECTIVE: We tested the hypothesis that the greater a person's laboratory stress-elicited elevation in cortisol, the greater the life stress-related risk for upper respiratory infection (URI). We also tested the prediction that the greater the laboratory stress-elicited rise in natural killer cell (NK) cytotoxicity, the smaller the life stress-related URI risk. Finally, we explored whether sympathetic nervous system (SNS) and enumerative immune reactivities to laboratory stress moderate the relation between life stress and URI. METHODS: At baseline, 115 healthy subjects were administered a negative stressful life events checklist and were tested to assess their SNS (blood pressure, heart rate, and catecholamines), HPA (cortisol), and immune (NK cell cytotoxicity and lymphocyte subsets) reactivities to laboratory speech tasks administered 2 weeks apart. Responses were averaged across the two laboratory assessments to create reactivity scores. After these assessments were completed, participants were followed weekly for 12 consecutive weeks. At each follow-up they completed a measure of perceived stress experienced over the last week. They were also instructed to contact the study coordinator if they had a cold or flu at any time during follow-up. A health care worker verified reported illnesses. RESULTS: In a traditional prospective analysis, high cortisol reactors with high levels of life events had a greater incidence of verified URI than did high reactors with low levels of life events and low reactors irrespective of their life event scores. Using hierarchical linear modeling, CD8(+) number, Natural Killer (NK) cell number, and NK cell cytotoxicity, each interacted with weekly perceived stress levels in predicting concurrent occurrences of self-reported URIs. For these outcomes, low immune reactors were more likely to experience an URI during high stress than low stress weeks. High immune reactors did not exhibit differences in weekly URIs as a function of weekly stress level. The SNS reactivity markers did not moderate the association of stress and URI incidence in either analysis. CONCLUSIONS: Acute HPA and immune responses to laboratory stressors are markers of how vulnerable people are to the increased risk for URI associated with stressors in the natural environment.

Adolescent↗

Short-term exercise training can improve myocardial tolerance to I/R without elevation in heat shock proteins.

We examined the effects of 3 days of exercise in a cold environment on the expression of left ventricular (LV) heat shock proteins (HSPs) and contractile performance during in vivo ischemia-reperfusion (I/R). Sprague-Dawley rats were divided into the following three groups (n = 12/group): 1) control, 2) exercise (60 min/day) at 4 degrees C (E-Cold), and 3) exercise (60 min/day) at 25 degrees C (E-Warm). Left anterior descending coronary occlusion was maintained for 20 min, followed by 30 min of reperfusion. Compared with the control group, both the E-Cold and E-Warm groups maintained higher (P < 0.05) LV developed pressure, first derivative of pressure development over time (+dP/dt), and pressure relaxation over time (-dP/dt) throughout I/R. Relative levels of HSP90, HSP72, and HSP40 were higher (P < 0.05) in E-Warm animals compared with both control and E-Cold. HSP10, HSP60, and HSP73 did not differ between groups. Exercise increased manganese superoxide dismutase (MnSOD) activity in both E-Warm and E-Cold hearts (P < 0.05). Protection against I/R-induced lipid peroxidation in the LV paralleled the increase in MnSOD activity whereas lower levels of lipid peroxidation were observed in both E-Warm and E-Cold groups compared with control. We conclude that exercise-induced myocardial protection against a moderate duration I/R insult is not dependent on increases in myocardial HSPs. We postulate that exercise-associated cardioprotection may depend, in part, on increases in myocardial antioxidant defenses.

Animals↗