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Interactions of energy and bovine somatotropin with heat stress.

Heat stress, resulting when dairy cows are exposed to hot or to hot and humid environmental conditions, reduces DMI and milk yield and increases maintenance costs of the cow. Decreased DMI may help to maintain homeothermy through reduced metabolic heat production. The high yielding cow has greater metabolic activity and produces more body heat than those with lower yields; thus, greater milk yield may increase heat stress if the cause of that stress is not mitigated. Use of bST increases milk yield, and, during hot or hot and humid environmental conditions, bST increased milk yield with or without noticeable effects on heat stress, depending on the study. The use of bST does not change maintenance requirements or partial efficiencies of milk yield. Thus, the greater heat stress that occurred in some studies probably was related to the increased metabolic activity and heat production associated with milk yield and to the difficulties of cows in dissipating the additional body heat from environmental conditions. Cows that are administered bST are subject to heat stress as are other high yielding cows if sufficient metabolic heat is not dissipated. Management to minimize effects on the cow of environments conducive to heat stress and strategies that improve DMI are necessary to sustain the high milk yield potential offered by technologies, including bST.

Animals↗

SB 203580, a mitogen-activated protein kinase inhibitor, abolishes resistance to myocardial infarction induced by heat stress.

Heat stress (HS) is known to confer protection against ischemia-reperfusion injury, including mechanical dysfunction and myocardial necrosis. However, the mechanisms involved in this cardioprotection are yet to be elucidated. Mitogen-activated protein (MAP) kinase cascades have been demonstrated to be involved in cellular response to different stresses. In particular, p38 MAP kinase is known to be activated by HS. Therefore, we investigated the implication of this kinase in HS-induced resistance to myocardial infarction, in the isolated rat heart model, using SB 203580 (SB) to selectively inhibit p38 MAP kinase. Rats were treated with SB (2.83 mg/kg, i.p.) or vehicle (1% DMSO in saline, i.p.) before they were either heat stressed (42 degrees C for 15 minutes) or sham anesthetized. Their hearts were isolated 24 hours later, retrogradely perfused, and subjected to a 35-minute occlusion of the left coronary artery followed by 120 minutes of reperfusion. The infarct-to-risk ratio was significantly reduced in HS (16.9 +/- 2.0%) compared with sham (41.6 +/- 2.5%) hearts. This reduction in infarct size was abolished in the SB 203580-treated group (37.8 +/- 1.9% in HS + SB vs. 42.0 +/- 1.9% in sham + SB). Risk zones were similar between experimental groups. Western blot analysis of the myocardial HSP72 showed an HS-induced increase of this protein, which was not modified by the p38 MAP kinase inhibitor, SB 203580. We conclude that activation of p38 MAP kinase appears to play a role in the functional cardioprotection associated with the heat stress response, which seems to be unrelated to the HSP72 level. Further investigations are required to elucidate the precise role of the p38 MAP kinase and heat stress proteins in this adaptative response.

Animals↗

Superoxide radical production in chicken skeletal muscle induced by acute heat stress.

Heat stress is of major concern for poultry, especially in the hot regions of the world because of the resulting poor growth performance, immunosuppression, and high mortality. To assess superoxide (O2*-) production in mitochondria isolated from skeletal muscle of chickens (n = 4 to 8) exposed to acute heat stress, electron spin resonance (ESR) spectroscopy using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin trap agent and lucigenin-derived chemiluminescence (LDCL) method were applied. ESR spectra of suspensions containing mitochondria from control and acute heat-treated meat-type chickens showed similar hyperfine coupling constants (aN = 1.44 mT, aHbeta = 0.12 mT, and aHbeta = 0.11 mT) to those of DMPO-O2*- adducts observed in a hypoxanthine-xanthine oxidase system. Heat exposure resulted in enhancement of the DMPO-O2*- signal. The results using LDCL showed significantly enhanced superoxide production in heat stress-treated skeletal muscle mitochondria of meat-type chickens, whereas no such increase was observed in laying chickens. The enhancement of superoxide production in the former case was associated with heat-induced increments in rectal and muscle temperatures, leading to significant body weight loss. In contrast, the latter case showed no increase in temperatures, although there was a slight decrease in body weight gain. Percentage increases of superoxide production in the presence of carboxyatractylate, a specific inhibitor of adenine nucleotide translocator (ANT), were the same for skeletal muscle mitochondria from meat- and laying-type chickens from the control or heat stress-treated group. This finding suggests the irrelevance of ANT in the regulation of reactive oxygen species flux under heat stress conditions. The study provides the first evidence of superoxide anion production in the skeletal muscle mitochondria of meat-type chickens in response to acute heat stress.

Animals↗

Influence of sire and sire breed (Gyr versus Holstein) on establishment of pregnancy and embryonic loss in lactating Holstein cows during summer heat stress.

Heat stress has negative effects on pregnancy rates of lactating dairy cattle. There are genetic differences in tolerance to heat stress; Bos taurus indicus (B. t. indicus) cattle and embryos are more thermotolerant than Bos taurus taurus (B. t. taurus). In the present study, the effects of sire and sire breed on conception and embryonic/fetal loss rates of lactating Holstein cows during the Brazilian summer were determined. In Experiment 1, cows (n=302) were AI after estrus detection or at a fixed-time with semen from one Gyr (B. t. indicus) or one Holstein sire (B. t. taurus). Pregnancy was diagnosed 80 days after AI. In Experiment 2, cows (n=811) were AI with semen from three Gyr and two Holstein sires. Pregnancy was diagnosed at 30-40 and at 60-80 days after AI. Cows diagnosed pregnant at the first examination but non-pregnant at the second were considered as having lost their embryo or fetus. Data were analyzed by logistic regression. The model considered the effect of sire within breed, sire breed, days postpartum, period of lactation, and AI type (AI after estrus versus fixed-time). There was no effect of the AI type, days postpartum or milk production on conception or embryonic loss rates. The use of Gyr bulls increased pregnancy rate when compared to Holstein bulls [9.1% (60/657) versus 5.0% (23/456), respectively, P=0.008; data from Experiments 1 and 2 combined]. Additionally, in Experiment 2, cows inseminated using semen from sire #4 (Gyr) had lower embryonic loss (10%) when compared with other B. t. indicus (35.3% and 40%) or B. t. taurus sires (18.2% and 38.5%, P=0.03). In conclusion, the use of B. t. indicus sires may result in higher conception rates in lactating Holstein cows during summer heat stress. Moreover, sire can affect embryonic loss and selection of bulls according to this criterion may result in higher parturition rates in lactating Holstein cows.

Abortion, Veterinary↗

Protein kinase C is involved in resistance to myocardial infarction induced by heat stress.

Heat stress (HS) is known to protect against mechanical dysfunction and myocardial necrosis in myocardial ischemia-reperfusion models both in vivo and in vitro. However, the mechanisms involved in this form of cardioprotection remain unclear. Protein kinase C (PKC) and tyrosine kinase activation have both been shown to be involved in the delayed phase of protection following ischemic preconditioning, a phenomenon which appears to be analogous to HS-induced protection. Therefore, we investigated the role of PKC and tyrosine kinase in HS-induced resistance to myocardial infarction, in the isolated rat heart. The selective inhibitors chelerythrine (Che) and genistein (Gen) were used to inhibit PKC and tyrosine kinase, respectively. Rats were treated with Che (5 mg/kg, i.p.) or Gen (5 mg/kg, i.p.) or vehicle before they were either heat stressed (42 degrees C for 15 min) or sham anesthetized. Twenty-four h later their hearts were isolated, retrogradely perfused, and subjected to 35-min occlusion of the left coronary artery followed by 120-min of reperfusion. Infarct-to-risk ratio was significantly reduced in HS (19.9+/-1.1%) compared to sham (43.1+/-1.1%) hearts. This reduction in infarct size was abolished in chelerythrine-treated groups (43.8+/-1.9% in HS+Che v 44.9+/-2.0% in sham+Che), but was conserved in genistein-treated groups (17.7+/-0.9% in HS+Gen v 36.4+/-2.8% in sham+Gen). In order to confirm that genistein at this dose was effectively inhibiting tyrosine kinase activity, we observed the ability of the agent to prevent the hypoglycemic responses to insulin in a separate group of anesthetised rats receiving an i.v. insulin infusion. Western blot analysis of the myocardial hsp72 showed a HS-induced increase of this protein, which was modified by neither the PKC inhibitor, chelerythrine, nor the tyrosine kinase inhibitor, genistein. We conclude that the activation of PKC, but not of tyrosine kinase, appears to play a role in the functional cardioprotection associated with the heat stress response. Although protection appears to be dissociated from induction of hsp72, further work is required to explore the importance of hsp72 phosphorylation to cytoprotective activity of the protein.

Alkaloids↗

Heat stress- and heat shock transcription factor-dependent expression and activity of ascorbate peroxidase in Arabidopsis.

To find evidence for a connection between heat stress response, oxidative stress, and common stress tolerance, we studied the effects of elevated growth temperatures and heat stress on the activity and expression of ascorbate peroxidase (APX). We compared wild-type Arabidopsis with transgenic plants overexpressing heat shock transcription factor 3 (HSF3), which synthesize heat shock proteins and are improved in basal thermotolerance. Following heat stress, APX activity was positively affected in transgenic plants and correlated with a new thermostable isoform, APX(S). This enzyme was present in addition to thermolabile cytosolic APX1, the prevalent isoform in unstressed cells. In HSF3-transgenic plants, APX(S) activity was detectable at normal temperature and persisted after severe heat stress at 44 degrees C. In nontransgenic plants, APX(S) was undetectable at normal temperature, but could be induced by moderate heat stress. The mRNA expression profiles of known and three new Apx genes were determined using real-time PCR. Apx1 and Apx2 genes encoding cytosolic APX were heat stress and HSF dependently expressed, but only the representations of Apx2 mRNA met the criteria that suggest identity between APX(S) and APX2: not expressed at normal temperature in wild type, strong induction by heat stress, and HSF3-dependent expression in transgenic plants. Our data suggest that Apx2 is a novel heat shock gene and that the enzymatic activity of APX2/APX(S) is required to compensate heat stress-dependent decline of APX1 activity in the cytosol. The functional roles of modulations of APX expression and the interdependence of heat stress and oxidative stress response and signaling mechanisms are discussed.

Arabidopsis↗

Preliminary studies on the relationship between autoantibodies to heat stress proteins and heat injury of pilots during acute heat stress.

Comparison in the heart rate, oral temperature and lymphocyte DNA damage during heat stress was made in pilots with negative antibodies to heat stress proteins (HSPs) and those with positive antibodies in the man-made climate room with Western blot and comet assay. Our results showed that the increase in oral temperature, heart rate and lymphocyte DNA damage in pilots with the positive antibodies to HSPs were higher than those in pilots with the negative antibodies during heat stress. These results indicated that the presence of autoantibodies in plasma of pilots might reflect heat damage and high sensitivity to heat.

Adult↗

[Dynamics of chaperone complex Hdj1-Hsp70-Bag1 as a response of erythroleukemia K562 cells to heat stress].

Heat shock protein Hsp70 is known to play an important role in cell protection against a variety of harmful factors. This property, at least in part, is due to Hsp70 ability to restore the native conformation of newly synthetized or damaged proteins. In this activity Hsp70 is accompanied by two proteins, Hdj1 and Bag1, that enable Hsp70 to peform cycles of binding-release of target proteins. The aim of this study was to investigate interactions of Hdj1 and Bag1 co-chaperones with Hsp70 in vivo. The accumulation of Hsp70 was stimulated by heat stress, and later, at certain periods following the stress, cell probes were collected for biochemical and microscopic analysis. The data of Western blotting showed that within 24 h after heat shock amounts of Hsp70 and Hdj1 raised to remain at the elevated level for nearly 48 h. Several time points within this period were chosen for analysis of the complexes between Hsp70 and co-chaperones. The data of reciprocal immunoprecipitation/immunoblotting and confocal microscopy showed that Hsp70-Hdj1 complexes were detected primarily at early stage after heat shock, then Hsp70 was preferably bound to Bag1. The dynamics of chaperone complex formation and changes in their intracellular localization are discussed in terms of cell reaction to stress.

Chaperonins↗

[Distribution of 70kDa heat shock protein in rabbit brains after heat stress and heat stroke].

OBJECTIVE: Evaluation of the relationship between the induction of 70kDa heat shock protein in rabbit brains and heat stress. METHODS: HSP70 was detected using monoclonal antibody by ABC method in rabbit hypothalamus, hippocampus and cerberal cortex. RESULTS: Intense HSP70 staining was displayed in rabbit brains of the heat stroke group (rectal temperature 43 degrees C to death). Positive cells were distributed mainly in the CA1, CA2 regions of the hippocampus; granular cell layer I and pyramidal layer (II) of the cerebral cortex; and the periventricular area of hypothalamus. HSP70-psoitive substances were localized in the cytoplasm and neuronal processes, a few neurons exhibited dark staining nucle. Hosever, the rabbit brains of the general heat stress group (rectal temperature 42.0 degrees C, 30 minutes) had much weaker staining. CONCLUSION: Hyperthermia causes neuronal expression of HSP70, particularly under strong heat stress, and may be sustained till death.

Animals↗

Occupational heat stress.

Heat stress is an occupational health problem affecting Man in diverse, hostile environments ranging from deep mining to outer space. Healthy, young military recruits and industrial workers exposed to hot, humid environments are typically at risk. The International Classification of Diseases classifies heat disorders into 10 separate entities, of which the most clinically significant are heat cramps, heat exhaustion and heat stroke. The latter is a medical emergency of the first order. Clinical management hinges upon early treatment based on a high index of suspicion. Effective preventive measures and adequate education of those at risk form the basis for its control.

Emergencies↗

Identification of sites of injury in Lactobacillus bulgaricus during heat stress.

Heat resistance of Lactobacillus bulgaricus in skimmed milk at 62 degrees, 64 degrees, 65 degrees and 66 degrees C was studied. The response to increasing temperatures in this range was not linear, with temperatures at 65 degrees C and above giving a lower survival rate than would be predicted from experiments at lower temperatures. To identify sites of injury at these temperatures, chemical markers were used. Heating at 64 degrees C and below resulted in damage to the cytoplasmic membrane. At temperatures of 65 degrees C and above chemical markers also indicated damage in the cell wall and proteins. Using differential scanning calorimetry analysis of whole cells of Lact. bulgaricus seven main peaks were observed (1-51, m1-61, m2-73, n-80, p-89, q-100, r-112 degrees C). Three of these peaks (l(r), m(r) and p(r)) were the result of reversible reactions. Analysis of cell fractions identified the cell structure involved in giving rise to each of the three reversible peaks; l(r), cell membrane lipids, m(r), ribosomes, and p(r), DNA. The evidence presented in this paper shows that irreversible reactions in the cell ribosomes are a critical site of damage in Lact. bulgaricus during heat stress in liquid media at 65 degrees C and above.

Animals↗

CHIP interacts with heat shock factor 1 during heat stress.

Heat shock factor 1 (HSF1) is a major transactivator of heat shock genes in response to stress and mediates cell protection against various harmful conditions. In this study, we identified the interaction of CHIP (carboxyl terminus of the heat shock cognate protein 70-interacting protein) with the N-terminus of HSF1. Using GST full-down assay, we found that CHIP directly interacts with C-terminal deleted HSF1 (a.a. 1-290) but not with full-length HSF1 under non-stressed conditions. Interestingly, interaction of CHIP with full-length HSF1 was induced by heat shock treatment. The structural change of HSF1 was observed under heat stressed conditions by CD spectra. These observations demonstrate the direct interaction between HSF1 and CHIP and this interaction requires conformational change of HSF1 by heat stress.

Binding Sites↗

Polo-like kinase 1 phosphorylates heat shock transcription factor 1 and mediates its nuclear translocation during heat stress.

Heat shock transcription factor 1 (HSF1) is activated by pathophysiologic stresses and activation leads to an increased cellular level of heat shock proteins (Hsp(s)). Although the activation of HSF1 occurs via multiple stress-induced processes such as hyperphosphorylation, the exact cellular mechanism of HSF1 activation is still unclear. Here we show polo-like kinase 1 (PLK1) and HSF1 interact in vivo using the tandem affinity purification system. Although the interaction between HSF1 and PLK1 is increased by thermal stress, overexpression of PLK1 did not affect HSF1 trimerization or DNA binding activity. This interaction results in the phosphorylation of HSF1 on serine 419 by PLK1. Interestingly, mutation of serine 419 to alanine inhibited heat-stress induced HSF1 nuclear translocation. Our results suggest that the phosphorylation of HSF1 by PLK1 is an essential step for HSF1 nuclear translocation by heat stress.

Active Transport, Cell Nucleus↗

Mitochondrial respiratory electron carriers are involved in oxidative stress during heat stress in Saccharomyces cerevisiae.

In the present study we sought to determine the source of heat-induced oxidative stress. We investigated the involvement of mitochondrial respiratory electron transport in post-diauxic-phase cells under conditions of lethal heat shock. Petite cells were thermosensitive, had increased nuclear mutation frequencies, and experienced elevated levels of oxidation of an intracellular probe following exposure to a temperature of 50 degrees C. Cells with a deletion in COQ7 leading to a deficiency in coenzyme Q had a much more severe thermosensitivity phenotype for these oxidative endpoints following heat stress compared to that of petite cells. In contrast, deletion of the external NADH dehydrogenases NDE1 and NDE2, which feed electrons from NADH into the electron transport chain, abrogated the levels of heat-induced intracellular fluorescence and nuclear mutation frequency. Mitochondria isolated from COQ7-deficient cells secreted more than 30 times as much H(2)O(2) at 42 as at 30 degrees C, while mitochondria isolated from cells simultaneously deficient in NDE1 and NDE2 secreted no H(2)O(2). We conclude that heat stress causes nuclear mutations via oxidative stress originating from the respiratory electron transport chains of mitochondria.

Cell Nucleus↗

The regulatory domain of human heat shock factor 1 is sufficient to sense heat stress.

Heat shock factor (HSF) activates transcription in response to cellular stress. Human HSF1 has a central regulatory domain which can repress the activity of its activation domains at the control temperature and render them heat shock inducible. To determine whether the regulatory domain works in tandem with specific features of the HSF1 transcriptional activation domains, we first used deletion and point mutagenesis to define these activation domains. One of the activation domains can be reduced to just 20 amino acids. A GAL4 fusion protein containing the HSF 1 regulatory domain and this 20-amino-acid activation domain is repressed at the control temperature but potently activates transcription in response to heat shock. No specific amino acids in this activation domain are required for response to the regulatory domain; in particular, none of the potentially phosphorylated serine and threonine residues are required for heat induction, implying that heat-induced phosphorylation of the transcriptional activation domains is not required for induction. The regulatory domain is able to confer heat responsiveness to an otherwise completely heterologous chimeric activator that contains a portion of the VP16 activation domain, suggesting that the regulatory domain can sense heat in the absence of other portions of HSF1.

Amino Acid Sequence↗

Effect of acute heat stress on heat shock protein 70 messenger RNA and on heat shock protein expression in the liver of broilers.

1. The synthesis of heat shock protein 70 (Hsp70) mRNA and the expression of Hsp70 in the liver of broiler chickens submitted to acute heat stress (35 degrees C for 5 h) was investigated. 2. Hsp70 expression was detected by SDS-PAGE and Western blot analysis using a polyclonal antiserum against Hsp70 of Blastocladiella emersonii. The specific signal of Hsp70 mRNA was analysed by Northern blot using as probe a Hsp70 cDNA of B. emersonii. 3. An increase in the amount of Hsp70 was detected from the first up to the fifth hour of acute heat exposure. This increase in the amount of Hsp70 was accompanied by an increase in Hsp70 mRNA which peaked at 3 h. 4. This study shows that the heat induced increase in Hsp70 mRNA and protein in broiler liver, in vivo, are time dependent, similar to that in mammals.

Animals↗

Whole-blood concentrations of glutathione in cattle exposed to heat stress or a combination of heat stress and endophyte-infected tall fescue toxins in controlled environmental conditions.

OBJECTIVE: To determine whether cattle exposed to heat stress alone or heat stress while consuming endophyte-infected fescue (EIF) have lower whole-blood (WB) concentrations of glutathione (GSH). ANIMALS: 10 Simmental cows. PROCEDURE: Cows were sequentially exposed to thermoneutral (TN; 2 weeks; 18 C, 50% relative humidity [RH]), heat stress (HS; 2 weeks; alternating 4-hour intervals at 26 and 33 C; 50% RH), and heat stress while consuming EIF (10 microg of ergovaline/kg/d; 2 weeks, HS + EIF). Blood samples were collected after each period and tested for GSH and oxidized glutathione (GSSG) concentrations. RESULTS: Feed consumption was similar when data were analyzed for time points at which WB concentrations of GSH or GSSG were determined. However, significant effects of treatment, cow, days exposed to heat, cow-by-treatment interaction, and treatment-by-days exposed to heat interaction were detected when data were considered simultaneously. Mean +/- SD hematocrit for TN, HS, and HS + EIF were 35.3+/-3, 33.3+/-2, and 37.1+/-3%, respectively. Mean WBGSH concentrations for TN, HS, and HS + EIF were 3.2+/-0.65, 2.7+/-0.62, and 2.4+/-0.56 mmol/L of RBC, respectively. Reduced WBGSH concentrations were associated with reduced feed intake during the later part of each heat period. CONCLUSIONS AND CLINICAL RELEVANCE: Decreased GSH and increased GSSG concentrations were evident during heat stress, especially when cattle consumed EIF These were associated with reduced feed intake during heat stress. Heat stress, reductions in feed intake, and thermoregulatory effects of EIF may induce oxidative stress in cattle.

Animals↗

Interaction of the fluorescent dye 1-N-phenylnaphthylamine with Escherichia coli cells during heat stress and recovery from heat stress.

The fluorescent dye 1-N-phenylnaphthylamine permeated Escherichia coli cells after exposure to a heat stress at 55 degrees C in Tris/Mg2+ buffer, pH 8.0. The rate of dye permeation increased with time during heat treatment and decreased gradually during subsequent incubation at 37 degrees C in a minimal medium. The initial level of rapid adsorption of the dye also increased with heating time, although it remained roughly constant during post-heating incubation. The results obtained suggest that the permeability barrier to the dye in the outer membrane was damaged by heat stress and was repaired after sublethal heating. RNA, protein and lipid syntheses, as well as an energy-yielding process, appeared to be necessary for the repair of impermeability to the dye.

1-Naphthylamine↗