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Cloning of two Hsp70 genes and association analysis between SNP haplotypes and high temperature tolerance trait in red swamp crayfish (Procambarus clarkii).

Aquaculture is suffering the challenge from high temperature climate. Two Hsp70 genes, PcHsp70-1 and PcHsp70-2, as key genes involved in the high temperature tolerance of red swamp crayfish (Procambarus clarkii) were identified and cloned in this study. Their molecular features and expression patterns were characterized, revealing the distinct tissue-specific upregulation expression under high temperature stress (33 °C). Two SNPs, PcHsp70-1 (SNP258) and PcHsp70-2 (SNP555) were examined to associate with high temperature tolerance in three populations (n = 675). The genotypes of PcHsp70-1-SNP258 (GA) and PcHsp70-2-SNP555 (TT) were significantly associated with stronger high temperature tolerance. Notably, individuals carrying the haplotype of Hap I (GG + TT) showed a survival rate exceeding 70% under high temperature stress, whereas, the Hap VIII (AA + CT) showed it at 5.2%. RNA interference of PcHsp70-1 resulted in a significant decrease expression of the gene GSH-Px and its encoding protein (glutathione peroxidase) activity, and damage in intestinal tissue under high temperature stress. The transcriptome result revealed that PcHsp70-1 participates in regulation of the pathways related to cytoskeletal construction, immune response, apoptosis, and antioxidant defense. These findings indicate that PcHsp70 genes are crucial for the cellular stress response under high temperature stress. The developed Kompetitive Allele Specific PCR (KASP) markers provide valuable tools for the marker-assisted selection of high temperature tolerant crayfish varieties, supporting the sustainable development of aquaculture under the challenge of global warming.

Animals↗

Similarity of the conformation of diphtheria toxin at high temperature to that in the membrane-penetrating low-pH state.

At high temperature, nicked free monomers of diphtheria toxin undergo a transition to a thermally denatured state, with a midpoint of 45-50 degrees C. In this report, the high-temperature (60 degrees C) conformation has been compared to the native (neutral pH) and low-pH (pH less than 5) conformations. The low-pH and high-temperature conformations are similar although not identical. As in the conformation at low pH, and unlike the toxin in its native conformation, the protein in its conformation at high temperature is hydrophobic, has low fluorescence intensity, and has increased exposure of tryptophan to aqueous solution. As at low pH, at high temperature the circular dichroism spectrum shows at most only partial unfolding of secondary structure. In contrast, the conformation of the toxin in guanidinium chloride is much closer to a random coil. The effects of high temperature and low pH interact in the sense that sensitivity of the native conformation to one is increased by the other. That is, the transition temperature between native and thermally denatured states is decreased as pH is decreased, and the transition pH between neutral-pH and low-pH states is increased as temperature is increased. This implies that there is some region on the protein where high temperature and low pH can disrupt conformation in a similar manner. Taken together, these results indicate that the low-pH and high-temperature conformations can both be defined as partially denatured states, even though unfolding may not be extensive at low pH. Similar behavior may occur in other proteins that undergo functionally important conformational disruption at low pH.

Chemical Phenomena↗

Analysis of phospholipids in cow's milk by high-temperature injection gas chromatography and high-performance liquid chromatography.

An investigation of the high-temperature injection (350 degrees C) gas chromatographic behaviour of standards of various classes of phospholipids has elucidated certain characteristic fragments ("tracers") in the pyrogram of each class. Under these conditions of pyrolysis of the phospholipids, a natural mixture of such substances (extracted and purified from cows' milk) has provided evidence for the same "tracers" as for the standards. Analogous results were obtained with a more representative specimen of cows' milk of various breeds grown in areas of different altitudes. The content of fatty acids in phospholipids, tested on each class (separated by means of radial compression high-performance liquid chromatography from polar lipids of milk) appeared to be relatively similar for all the phospholipid classes, and over half the content consisted of unsaturated fatty acids. The major component was delta 9-octadecenoid (oleic) acid.

Animals↗

Body temperature of acclimated broilers during exposure to high temperature.

Short-term acclimation to high temperatures increased resistance of broilers to heat exposure. Two trials were conducted to determined the effect of acclimation on body temperature (Tb) during heat exposure. Broilers, 46-days old, were maintained at control or acclimating temperatures for 4 days. The control temperature was a constant 21 C with a 10 C dewpoint, and the acclimating temperature was a diurnal cycle of 24-35-24 C with a constant 21 C dewpoint. During the 4th day, broilers of both groups were exposed to temperatures of 41 C and 23 C dewpoint for 210 min. Body temperature was determined at the beginning of the heat exposure and at 30-min intervals until the end of the exposure. The Tb of both groups increased throughout heat exposure, but acclimated broilers had significantly lower Tb than control broilers from 90 through 210 min. Acclimated broilers had significantly lower heat stress mortality. The data show that acclimated broiler's Tb rises upon exposure to heat but that acclimation gives the capacity to stabilize Tb above normal Tb. The data also suggest Tb is preferable to mortality as a measure of acclimation in broilers.

Acclimatization↗

Dry Matter Production and Rate of Change of Harvest Index at High Temperature in Peanut.

The concept of a linear increase in harvest index, dHI/dt, has proven very useful for crop simulation modeling. The effect of high temperature on the response of dHI/dt of pods and seeds of peanut (Arachis hypogaea L.) has not been described. The objectives of this work were to determine (i) whether dHI/dt was linear at high temperature, (ii) whether high temperature affected dHI/dt and/or the timing of the linear phase of increase in HI, and (iii) whether there was genotypic variation in the response of dHI/dt to high temperature. Four peanut genotypes varying in heat tolerance were grown in pots at either 28/22 or 38/22 degrees C from 21 to 90 d after planting (DAP). Plants were harvested on 10 occasions starting 27 DAP and total dry matter accumulation and partitioning measured. High temperature reduced total dry weight by 20 to 35%, seed HI by 0 to 65%, and seed dry weight by 23 to 78%. At 28/22 degrees C, dHI/dt for pods and seeds was linear and varied from 0.0058 to 0.0109 d(-1). At 38/22 degrees C, dHI/dt of pods and seeds was also linear and varied from 0.0028 to 0.0089 d(-1). There were genotypic differences in response to temperature. High temperature had no effect on dHI/dt in moderately tolerant genotypes 796 and 47-16. In susceptible genotypes ICGV 86016 and ICGV 87282, however, the start of pod and seed filling was delayed by 5 to 9 d and dHI/dt reduced by 20 to 65% at 38/22 degrees C. Reductions in pod and seed dry weight at 38/22 degrees C were therefore due to reductions in total dry matter and dHI/dt, depending on the heat tolerance of the genotype. Crop models need to account for genotypic differences in the response of timing and rate of dHI/dt to high temperature to successfully simulate yields in warmer environments.

Journal Article↗

Short metal capillary columns packed with polymer-coated fibrous materials in high-temperature gas chromatography.

The high-temperature gas chromatographic (GC) separation of several semivolatile compounds is studied with a short metal capillary column packed with fibrous material, having a polydimethylsiloxane coating thereon. Taking advantage of the excellent heat-resistance of the fiber and also the combination of the surface-deactivated metal capillary, a temperature-programmed separation up to 450 degrees C is successfully demonstrated for the separation of polymer standard samples. The average molecular weight of the commercially-available polymer standard samples for size exclusion chromatography (SEC) is estimated by high-temperature GC analysis and compared with the nominal value determined by a conventional SEC method. Although a slight deviation for the number-average molecular weight is observed between the GC and SEC analysis, the data for the weight-average molecular weight shows a good agreement in these methods. The results also suggest the future possibility of the fiber-packed metal capillary as a miniaturized GC column with an increased sample loading capacity.

Journal Article↗

Development of a high-temperature air-blown gasification system.

Current status of high-temperature air-blown gasification technology development is reviewed. This advanced gasification system utilizes preheated air to convert coal and waste-derived fuels into synthetic fuel gas and value-added byproducts. A series of demonstrated, independent technologies are combined to form the core of this gasification system. A high-temperature, rapid devolatilization process is used to enhance the volatile yields from the fuel and to improve the gasification efficiency. A high-temperature pebble bed filter is used to remove to the slag and particulates from the synthetic fuel gas. Finally, a novel regenerative heater is used to supply the high-temperature air for the gasifier. Component development tests have shown that higher gasification efficiencies can be obtained at more fuel-rich operating conditions when high-temperature air is used as the gasification agent. Test results also demonstrated the flex-fuel capabilities of the gasifier design. Potential uses of this technology range from large-scale integrated gasification power plants to small-scale waste-to-energy applications.

Biomass↗

[Changes in the biochemical and hematological indices of chickens raised under high temperature and humidity].

Biochemical and hematological investigations were carried out with broilers raised at extreme temperature and humidity regimes-high temperature in combination with high relative humidity. Under the effect of high temperature (30-35 degrees C) and high relative humidity of the air (85-95 per cent) there was noted a rise of the lymphocytes and monocytes and a drop of the basophile, eosinophile, and pseudoeosinophile leukocytes in the white blood picture. After vaccination of the birds of the test groups with the vaccinal La Sota strain the blood serum showed negative titers. It was found that the Ca and P content in the blood serum of such broilers was higher as compared with that of the control group.

Animals↗

High-temperature repetitive nerve stimulation in myasthenia gravis.

High temperature enhances the decrement on repetitive nerve stimulation (RNS) in patients with myasthenia gravis (MG). However, the limit of this phenomenon at high temperature is unknown. Three-hertz ulnar RNS was performed in 7 patients with MG at a skin temperature of 32 degrees C and then with the hand in a 44 degrees C water bath. At 32 degrees C, the mean decrement preactivation was 5% (range, 0-24%); after 1 min of exercise, the mean decrement reached a maximum of 11% (range, 1-34%) 2 min postactivation. At a hand temperature of 42 degrees C, the mean decrement preactivation was 17% (range, 0-65%); after exercise, the mean decrement reached a maximum of 29% (range, 5-74%) 1 min postactivation. In 3 subjects, RNS was normal at 32 degrees C, but a definite decrement developed with heating. These findings demonstrate that very high temperature can improve the sensitivity of ulnar RNS for postsynaptic neuromuscular transmission defects.

Action Potentials↗

Arabidopsis hot mutants define multiple functions required for acclimation to high temperatures.

Plants acquire thermotolerance to lethal high temperatures if first exposed to moderately high temperature or if temperature is increased gradually to an otherwise lethal temperature. We have taken a genetic approach to dissecting acquired thermotolerance by characterizing loss-of-function thermotolerance mutants in Arabidopsis. In previous work, we identified single recessive alleles of four loci required for thermotolerance of hypocotyl elongation, hot1-1, hot2-1, hot3-1, and hot4-1. Completed screening of M2 progeny from approximately 2500 M1 plants has now identified new alleles of three of these original loci, along with three new loci. The low mutant frequency suggests that a relatively small number of genes make a major contribution to this phenotype or that other thermotolerance genes encode essential or redundant functions. Further analysis of the original four loci was performed to define the nature of their thermotolerance defects. Although the HOT1 locus was shown previously to encode a major heat shock protein (Hsp), Hsp101, chromosomal map positions indicate that HOT2, 3, and 4 do not correspond to major Hsp or heat shock transcription factor genes. Measurement of thermotolerance at different growth stages reveals that the mutants have growth stage-specific heat sensitivity. Analysis of Hsp accumulation shows that hot2 and hot4 produce normal levels of Hsps, whereas hot3 shows reduced accumulation. Thermotolerance of luciferase activity and of ion leakage also varies in the mutants. These data provide the first direct genetic evidence, to our knowledge, that distinct functions, independent of Hsp synthesis, are required for thermotolerance, including protection of membrane integrity and recovery of protein activity/synthesis.

Acclimatization↗

Synthesis of Hsp72 induced by exercise in high temperature.

An investigation was undertaken involving the physiological effects of exercise in high temperature and its effect on the induction of heat shock proteins synthesis in peripheral lymphocytes and gastrocnemius. Male Long-Evans rats were employed. Environmental temperature was designed as three conditions: room temperature (26-27 degrees C), middle temperature (30-31 degrees C), and high temperature (36-37 degrees C). Rats were divided into one of three groups and exercised passively in an automatic round treadmill. Body temperature, biochemical changes were determined and the synthesis of heat shock protein 72 were analyzed by SDS-PAGE and immunochemical stain with monoclonal anti-Hsp72 antibody. Those results indicated that the body temperature elevated faster and higher in rats of high-temperature exercise group than those of a house temperature significantly (p < 0.05). Hsp72 could be detected only in lymphocytes from rats exercising in high temperature more than 60 minutes whose body temperatures were elevated above 41 degrees C. The induced Hsp72 appeared in lymphocytes 2 hours after exercise and reached maximally at 16-24 hours and then disappeared gradually. In gastrocnemius, Hsp72 could be detected even before the passive exercise, and its amount changed as in lymphocytes did in all three groups. No difference was found in blood sugar, hematocrit and CK among the three groups of rats. On the basis of above results, we can conclude that Hsp72 is synthesized in the muscle of intact rats, and exercise is a sufficient inducer. For lymphocytes, the presence of Hsp72 can be viewed as a heat stress in exercise.

Animals↗

[Study on induced mutagenesis interaction of the high temperature and/or cigarette smoke].

The study aim is to research the genotoxicity and interaction of the High temperature and/or cigarette smoke. The micronucleus of human lymphocytes and bone marrow in mice were observed following heat stress and/or watersoluble cigarette smoke substances. The results indicated that the high temperature (40 degrees C, 41 degrees C and 42 degrees C for 5 min, respectively) significantly induced micronuclei in human lymphocytes in vitro (P < 0.01). A significant synergistic effect of the high temperature and cigarette was found. When high temperature combining with high concentration cigarette smoke (1.0-2.0 cigarette/20 g/d, for 10 d) was used, a significant induction and interaction (P < 0.01) on micronucleus of the bone marrow were also observed. The results indicated that high temperature substanance aberrations would be enhanced when the high temperature combines with cigarette smoke.

Animals↗

Performance of capillary columns for high-temperature gas chromatography

The developments in stationary-phase synthesis and capillary column technology have opened new perspectives in the analysis of high-molecular-weight compounds (600 daltons) and thermolabile organic compounds by high-temperature-high-resolution gas chromatography. This branch of high-resolution gas chromatography deals with analysis performed up to 390 degrees C oven temperature (with some applications going up to 420 degrees C and even a few applications to 450 degrees C maximum). The technique has been applied to many different fields of science (e.g., organic geochemistry, environmental chemistry, archeology, and natural product research). Apolar and medium-polar gum phases can now be operated at temperatures from 400 to 480 degrees C, but these higher temperatures are seldom used because of the thermostability of the material used to make the capillary tubing. This paper shows the performance of nine commercial high-temperature columns when used in routine applications.

Journal Article↗

In situ high-temperature visible microspectroscopy for volcanic materials.

In situ high-temperature visible microspectroscopy has been developed in order to study color change kinetics of volcanic materials. Olivine thin sections put on a synthetic alumina plate are heated on a heating stage at 600-800 degrees C under a visible microspectroscope. Changes in visible absorption spectra are monitored every 60 s for 5 hours. The obtained high-temperature visible spectra showed a gradual increase with time in absorbance in the shorter wavelength region (400-600 nm). The 430 nm absorbance (ligand field transition of Fe3+ increased more with time at higher temperatures. Assuming diffusional transport in plane sheets, apparent diffusion coefficients were determined at temperatures of 600-800 degrees C. The activation energy for this diffusion in olivine is 208 +/- 17 kJ/mol. This activation energy value is similar to those for the metal vacancy diffusion in olivine. This newly developed in situ high-temperature visible microspectroscopy can provide kinetic measurements of visible spectral change of materials at high temperatures such as volcanic materials.

Algorithms↗

[Effects of daytime sub-high temperature on greenhouse tomato growth, development, yield and quality].

The study on the effects of daytime sub-high temperature (30 and 35 degrees C) in spring and autumn on the growth, development, yield and quality of greenhouse tomato in northeast China showed that compared with the plant growing under feasible temperature 25 degrees C, three days' daytime sub-high temperature increased the relative growth significantly, and made the plant spindling. Seven days or more sub-high temperature made the plant grow fast, with thinner leaves and presenility. Under sub-high temperature, the harvest time was earlier, the fruit weight and the 2nd and 3rd inflorescence lessened, the quality of fruit decreased, and the yield decreased significantly. Sub-high temperature had a strong influence on the normal growth and development of greenhouse tomato. The longer the plant suffered sub-high temperature from flower bud differentiation to fruit maturing, the worse the fruit quality, and the lower the yield would be.

Biomass↗

Growth performance and concentrations of thyroid hormones and growth hormone in plasma of broilers at high temperatures.

Responses of broiler chickens to a high ambient temperature (35 degrees C) were measured in two experiments. In one experiment temperatures were increased abruptly from 21 degrees C to a daily range of 21-35 degrees C whereas, in the other, temperatures were increased more gradually over 6 days. The high temperatures were maintained for 5 h/day. In both experiments, birds exposed to the high temperatures ate less food and gained less liveweight than birds maintained at 21 degrees C. Efficiency of food conversion to liveweight gain and body composition were not affected by high temperature but there was a tendency for thyroid weight to decrease. Overall, the plasma concentration of triiodothyronine (T3) decreased and the plasma concentration of thyroxine (T4) increased, resulting in a decreased T3/T4 molar ratio, during exposure to high temperature. The concentration of plasma growth hormone, but not plasma reverse T3, was increased by high temperature. The initial responses to increased temperature were variable, with birds exposed more gradually adjusting relatively well until the maximum temperature was increased to 35 degrees C. All heated birds readjusted quickly to the daily reduction in temperature to 21 degrees C.

Animals↗

[Relationship between daily high temperature and mortality in Kyushu, Japan].

We analyzed the relationship between daily high temperature and mortality in Kyushu, Japan, 1972-1990. Mortality rate was obtained by dividing the number of deaths in a certain daily high temperature category by person-days in the same category. The results were as follows: (1) With daily high temperature on the x-axis and mortality rate on the y-axis, the relationship between the two variables appeared to be V-shaped. The minimum mortality rate was observed at the 28 degrees C-33 degrees C level. (2) Among all deaths at 33+ degrees C level, the proportion of deaths from "excessive heat" never exceeded 0.4%. This suggested that heat stroke is not a major contributor to the V-shape formation. (3) The V-shape was observed even in 1990, suggesting that the rapid spread of air conditioners did not alter the relationship between daily high temperature and mortality rate. (4) For the 0-4 years age group, mortality was not temperature dependent. However, the 5-14 years age group showed high mortality rates at 33+ degrees C. This may be attributed to outdoor activities during summer. After 30 years of age, the overall mortality rates rose with age, with greater increases at lower ambient temperature levels. As a result, the 65+ years age group had the greatest temperature dependence. (5) Deaths from respiratory diseases, circulatory diseases, and senility were highly temperature dependent, whereas death from cancer was not temperature dependent. Deaths from digestive diseases, infectious diseases, and accidents showed moderate temperature dependence.

Adolescent↗