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PubMed2001

Thermal sterilization of heat-sensitive products using high-temperature short-time sterilization.

High-temperature short-time (HTST) sterilization with a continuous-flow sterilizer, developed for this study, was evaluated. The evaluation was performed with respect to (a) the chemical degradation of two heat-sensitive drugs in HTST range (140-160 degrees C) and (b) the microbiological effect of HTST sterilization. Degradation kinetics of two heat-sensitive drugs showed that a high peak temperature sterilization process resulted in less chemical degradation for the same microbiological effect than a low peak temperature process. Both drugs investigated could be sterilized with acceptable degradation at HTST conditions. For the evaluation of the microbiological effect, Bacillus stearothermophilus ATCC 7953 spores were used as indicator bacteria. Indicator spore kinetics (D(T), z value, k, and E(a)), were determined in the HTST range. A comparison between the Bigelow model (z value concept) and the Arrhenius model, used to describe the temperature coefficient of the microbial inactivation, demonstrated that the Bigelow model is more accurate in prediction of D(T) values in the HTST range. The temperature coefficient decreased with increasing temperature. The influence of Ca(2+) ions and pH value on the heat resistance of the indicator spores, which is known under typical sterilization conditions, did not change under HTST conditions.

Geobacillus stearothermophilus↗

PubMed2005

Effect of high temperature and water stress on in vitro germination and growth in isolates of the entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin.

In non-irrigated agricultural fields in tropical zones, high temperature and water stress prevail during the main cropping season. Natural epizootics of Beauveria bassiana on lepidopteran pests occur during winter. Application of B. bassiana during hot months when pest populations are at their climax may prove an effective management strategy. Therefore, 29 isolates of B. bassiana were tested for their ability to germinate and grow in temperature and water availability conditions prevailing during the pest season in these fields. The effect of temperature cycles with 8 h duration of high temperature fluctuating with 16 h duration of lower temperature (similar to field conditions); low water availability; and a combination of these two stress conditions was studied. Germination and growth assays were done at fluctuating temperature cycles of 32, 35, 38, and 42+/-1 degrees C (8 h)/25+/-1 degrees C (16 h) and in media with water stress created by 10, 20, 30, and 40% polyethylene glycol (PEG 6000). Assays set at a continuous temperature of 25+/-1 degrees C with no PEG in the medium served as controls. Stress was assessed as percentage germination or as growth relative to control. Isolates showing 90% growth relative to the control at temperature cycles including high temperatures of 35 and 38+/-1 degrees C were identified. One isolate (ARSEF 2860) had a thermal threshold above 43 degrees C. At 25 degrees C, all but one isolate of B. bassiana showed >90% growth relative to the control in 10% PEG (-0.45 MPa). Some isolates were found with >90% growth relative to control in medium having 30% PEG with water availability (1.33 MPa), nearly equivalent to that in soils which induce permanent wilting point of plants. When isolates that showed >90% growth relative to the control at both stress conditions, were stressed simultaneously, a decrease in growth was observed. Growth was reduced by approximately 20% at 35+/-1 degrees C (8 h)/25+/-1 degrees C (16 h) and 10% PEG and was affected to a greater degree in combinations of harsher stress conditions. The isolate ARSEF 2860 with a thermal threshold of >43 degrees C showed approximately 80% relative growth at a combined stress of 38+/-1 degrees C (8 h)/25+/-1 degrees C (16 h) and 10% PEG. These findings will aid the selection of isolates for use in field trials in hot or dry agricultural climates.

Animals↗

PubMed1989

[Effects of high temperatures on chromosomes of normal and transformed human cells].

The effects of the high temperature on the chromosome of normal and transformed human cells were examined using a temperature gradient incubator (T.G.I., Model TN-212) in culture. The cell nucleus were damaged, specifically fragmentation occurred under the hyperthermic treatments. The diploid cells were passaged at three different temperature conditions ranging from 39.0-41.5 degrees C. It was found that the polyploid cells (predominantly tetraploid) were increased in these conditions compared with the cells cultured at 37 degrees C. We observed the chromosomal aberrations (break, stickiness, fragmentation, etc.) in the cells treated with high temperature for the various periods of time. The results indicated that the transformed cells were more high-temperature sensitive than the normal cells with respect to the chromosomal aberrations. The trend was seen in this experiment, in which the chromatid breaks of HAIN-55 and MKN-1 cell strains occurred more on the large chromosomes such as the ones in group A, B and C under the hyperthermic treatments.

Cell Line, Transformed↗

PubMed1994

Correlation of high-temperature stability of alpha-chymotrypsin with 'salting-in' properties of solution.

A correlation between the stability of alpha-chymotrypsin against irreversible thermal inactivation at high temperatures (long-term stability) and the coefficient of Setchenov equation as a measure of salting-in/out efficiency of solutes in the Hofmeister series has been found. An increase in the concentration of salting-in solutes (KSCN, urea, guanidinium chloride, formamide) leads to a many-fold decrease of the inactivation rate of the enzyme. In contrast, addition of salting-out solutes has a small effect on the long-term stability of alpha-chymotrypsin at high temperatures. The effects of solutes are additive with respect to their salting-in/out capacities; the stabilizing action of the solutes is determined by the calculated Setchenov coefficient of solution. The correlation is explained by a solute-driven shift of the conformational equilibrium between the 'low-temperature' native and the 'high-temperature' denatured forms of the enzyme within the range of the kinetic scheme put forward in the preceding paper in this journal: irreversible inactivation of the high-temperature form proceeds much more slowly compared with the low-temperature form.

Chymotrypsin↗

PubMed2004

Linear carbon dioxide in the high-pressure high-temperature crystalline phase IV.

High-temperature IR absorption spectra of solid CO2 in phases II and IV were measured in a resistive heated diamond anvil cell up to 30 GPa. The spectral structures of the bending mode, observed in high quality thin crystalline samples, and of the IR lattice phonons, measured for the first time between 80 and 640 K, are discussed using group theory arguments. According to this analysis the claimed bent molecular geometry of CO2 in phase IV can be unambiguously ruled out. Furthermore, the structures of both phases II and IV have been identified, among those so far proposed, as orthorhombic.

Journal Article↗

PubMed2025

De novo transcriptome assembly and gene expression analysis of Cnidium officinale under high-temperature conditions.

BACKGROUND: The medicinal plant Cnidium officinale (CO) is widespread in Northeast Asia and vulnerable to heat stress. The naturally occurring composition of pharmacological ingredients of CO results in overall physiological consequences; therefore, it is crucial to have a comprehensive understanding of metabolic response to ambient heat in terms of acclimation to estimate how much CO is exposed to threatening environmental conditions. RESULTS: Transcriptome analysis is critical for understanding the consequences of long-term physiological adaptation of CO to abiotic stress. However, transcriptome analysis on this species, particularly under prolonged stress conditions, has remained limited. We employed a temperature gradient tunnel (TGT) to subject CO to high-temperature exposure for four months, enabling us to observe the cumulative effects of heat and assess its acclimation mechanisms. In the absence of genome sequencing data, we performed de novo transcriptome assembly and compared DEGs from temperature treatment plots of a TGT and a growth chamber (GC). Since interpreting transcriptomic data can be complex, we employed a sequential analytical approach, including DEG clustering, GO enrichment, KEGG pathway mapping, miRNA-target gene analysis, and multiple rounds of RNA sequencing validation. DEGs were classified into two categories: genes exhibiting significant fold changes and genes showing significant count changes rather than fold changes. Then, we analyzed the functional roles of DEGs to determine which pathways respond to ambient and stressful high temperatures and validated the findings through cross-comparison with GC. Additionally, we conducted miRNA analysis to investigate post-transcriptional regulation under high temperatures. CO grown under higher ambient temperatures exhibited slight upregulation of pathways related to protein stability and turnover, ABA biosynthesis, and energy production, such as photosynthesis and oxidative phosphorylation. However, under extreme heat stress, most metabolic pathways were downregulated except for those involved in transcription, translation, oxidative phosphorylation and the biosynthesis of cutin, suberin, and wax. CONCLUSION: This study demonstrated that proper clustering of genes based on expression levels and fold changes in two different experimental conditions, along with pathway mapping, may provide a comprehensive understanding of CO's response to heat stress. These insights could contribute to future research on heat tolerance and crop improvement.

Gene Expression Profiling↗

PubMed2006

On-line coupling of high temperature GPC and 1H NMR for the analysis of polymers.

The on-line coupling of gel permeation chromatography (GPC) and 1H NMR operating at temperatures up to 130 degrees C is presented. A NMR flow probe with a cell volume of 120 microL and a stop-flow valve are developed for on-flow and stop-flow NMR measurements at high temperatures. To maintain high and constant temperatures through the whole probe, the flow probe contains two separate heating circuits. A modified stop-flow valve is developed as a control device for enabling on-flow and stop-flow experiments at high temperature conditions. Heated transfer lines connect the flow probe with the high temperature GPC system. Due to their semicrystalline nature, polyolefins can be studied by liquid chromatography only at temperatures above 100 degrees C. The novel high temperature GPC-NMR system is used for the separation of complex polyolefins regarding their molar mass and for the analysis of different chemical structures. Blends of polyethylene, poly(methyl methacrylate), and ethylene-methyl methacrylate copolymers are separated according to the molar masses of the components. The compositions of the components are directly studied by on-line NMR. Moreover, the chemical composition distribution of an ethylene-methyl methacrylate copolymer sample is analysed. Differences between results of on-flow and stop-flow measurements are discussed.

Chromatography, Gel↗

PubMed2003

Evolutionary changes in heat-inducible gene expression in lines of Escherichia coli adapted to high temperature.

The involvement of heat-inducible genes, including the heat-shock genes, in the acute response to temperature stress is well established. However, their importance in genetic adaptation to long-term temperature stress is less clear. Here we use high-density arrays to examine changes in expression for 35 heat-inducible genes in three independent lines of Escherichia coli that evolved at high temperature (41.5 degrees C) for 2,000 generations. These lines exhibited significant changes in heat-inducible gene expression relative to their ancestor, including parallel changes in fkpA, gapA, and hslT. As a group, the heat-inducible genes were significantly more likely than noncandidate genes to have evolved changes in expression. Genes encoding molecular chaperones and ATP-dependent proteases, key components of the cytoplasmic stress response, exhibit relatively little expression change; whereas genes with periplasmic functions exhibit significant expression changes suggesting a key role for the extracytoplasmic stress response in the adaptation to high temperature. Following acclimation at 41.5 degrees C, two of the three lines exhibited significantly improved survival at 50 degrees C, indicating changes in inducible thermotolerance. Thus evolution at high temperature led to significant changes at the molecular level in heat-inducible gene expression and at the organismal level in inducible thermotolerance and fitness.

Adaptation, Physiological↗

PubMed1998

Acquisition, storage, and review of safety data from a commercial system for high temperature, short time pasteurization.

A high temperature, short time (HTST) pasteurization system was equipped with electronic sensors to determine the temperature, pressure, flow rate, and position of the flow diversion valve. A computer for data acquisition was wired to the sensors to monitor and to record processing conditions related to public health. The processing conditions were stored in safety files on the hard drive of the computer, transferred weekly to a tape drive, and stored. The processing conditions of the HTST system were monitored for 270 d to determine the accuracy and reliability of the data acquisition system. The size of the HTST safety files ranged from 6.2 to 9.1 MB when the sensors were monitored every second. The file size was reduced to < 1.8 MB when the monitoring frequency was increased to every 5 s. To determine accuracy, the temperatures recorded by the data acquisition system were compared with the temperatures recorded by an electronic recorder controller. To determine reliability, changes in the position of the flow diversion valve were examined to identify process deviations and were compared with the event marker on circular charts. The review of the data file by the actual time method was an effective alternative to the electronic recorder controller for monitoring the completeness of data in the safety files. Off-line review to determine reliability required approximately 10 min/d of records.

Animals↗

PubMed2004

Protein folding failure sets high-temperature limit on growth of phage P22 in Salmonella enterica serovar Typhimurium.

The high-temperature limit for growth of microorganisms differs greatly depending on their species and habitat. The importance of an organism's ability to manage thermal stress is reflected in the ubiquitous distribution of the heat shock chaperones. Although many chaperones function to reduce protein folding defects, it has been difficult to identify the specific protein folding pathways that set the high-temperature limit of growth for a given microorganism. We have investigated this for a simple system, phage P22 infection of Salmonella enterica serovar Typhimurium. Production of infectious particles exhibited a broad maximum of 150 phage per cell when host cells were grown at between 30 and 39 degrees C in minimal medium. Production of infectious phage declined sharply in the range of 40 to 41 degrees C, and at 42 degrees C, production had fallen to less than 1% of the maximum rate. The host cells maintained optimal division rates at these temperatures. The decrease in phage infectivity was steeper than the loss of physical particles, suggesting that noninfectious particles were formed at higher temperatures. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a decrease in the tailspike adhesins assembled on phage particles purified from cultures incubated at higher temperatures. The infectivity of these particles was restored by in vitro incubation with soluble tailspike trimers. Examination of tailspike folding and assembly in lysates of phage-infected cells confirmed that the fraction of polypeptide chains able to reach the native state in vivo decreased with increasing temperature, indicating a thermal folding defect rather than a particle assembly defect. Thus, we believe that the folding pathway of the tailspike adhesin sets the high-temperature limit for P22 formation in Salmonella serovar Typhimurium.

Bacteriophage P22↗

PubMed2004

High-temperature sustainability of strong fiber Bragg gratings written into Sb-Ge-codoped photosensitive fiber: decay mechanisms involved during annealing.

An antimony--germanium- (Sb--Ge-) codoped fiber specially designed for the fabrication of fiber Bragg gratings (FBGs) with high temperature sustainability has been developed. The photosensitivity and the high-temperature sustainability of FBGs that have been written into this fiber were tested. The results obtained showed that the FBG written into this fiber has a very high temperature sustainability of 900 degrees C. A decay mechanism that involves cation hopping is presented to explain the observed high temperature sustainability of the grating written into this fiber.

Journal Article↗

PubMed1996

Reversible Inhibition of Tomato Fruit Gene Expression at High Temperature (Effects on Tomato Fruit Ripening).

The reversible inhibition of three ripening-related processes by high-temperature treatment (38[deg]C) was examined in tomato (Lycopersicon esculentum L. cv Daniella) fruit. Ethylene production, color development, and softening were inhibited during heating and recovered afterward, whether recovery took place at 20[deg]C or fruit were first held at chilling temperature (2[deg]C) after heating and then placed at 20[deg]C. Ethylene production and color development proceeded normally in heated fruit after 14 d of chilling, whereas the unheated fruit had delayed ethylene production and uneven color development. Levels of mRNA for 1-aminocyclopropane-1-carboxylic acid oxidase, phytoene synthase, and polygalacturonase decreased dramatically during the heat treatment but recovered afterward, whereas the mRNA for HSP17 increased during the high-temperature treatment and then decreased when fruit were removed from heat. As monitored by western blots, the HSP17 protein disappeared from fruit tissue after 3 d at 20[deg]C but remained when fruit were held at 2[deg]C. The persistence of heat-shock proteins at low temperature may be relevant to the protection against chilling injury provided by the heat treatment. Protein levels of 1-aminocyclopropane-1-carboxylic acid oxidase and polygalacturonase also did not closely follow the changes in their respective mRNAs. This implied both differences in relative stability and turnover rates of mRNA compared to protein and nontranslation of the message that accumulated in low temperature. The results suggest that high temperature inhibits ripening by inhibiting the accumulation of ripening-related mRNAs. Ripening processes that depend on continuous protein synthesis including ethylene production, lycopene accumulation, and cell-wall dissolution are thereby diminished.

Journal Article↗

PubMed1998

The stability of salt bridges at high temperatures: implications for hyperthermophilic proteins.

Salt bridges have been proposed to play a crucial role in promoting hyperthermostability in proteins, yet they appear to make little contribution to protein stability at room temperature. The latter point has been rationalized previously on the basis that the association of two charged molecules to form a salt bridge incurs a substantial desolvation penalty, which is seldom completely compensated by favourable interactions within the salt bridge and with the rest of the protein. Here a continuum solvation model is used to investigate how this same argument applies at temperatures more appropriate to hyperthermophiles. The solvation model employed was previously parameterised to reproduce the hydration free energies of neutral and charged amino acid side-chains in the temperature range from 5-100 degreesC. A key result of the previous work was that the hydration free energies of charged side-chains are more adversely affected by increasing temperature than are the hydration free energies of hydrophobic side-chains of identical size and shape (isosteres). As is shown here, a direct consequence of the temperature dependence of the hydration free energies is that at high temperatures the desolvation penalty for formation of a salt bridge is markedly reduced in magnitude. As a result, the argument that relative to hydrophobic isosteres, salt bridges destabilise proteins, may no longer be true at high temperatures. We demonstrate this point first in the setting of a small model system, but then also show that the same argument is likely to carry over to real proteins. We compare three hyperthermophilic proteins with their mesophilic homologues and find that hydration effects preferentially stabilise the hyperthermophiles at high temperatures. When the hydration effects are incorporated into a model for the free energy of folding of the proteins, it is found that in each case, the hyperthermophile is predicted to remain stable to a temperature 20-25 deg.C higher than the corresponding mesophile. Higher thermal stability for the hyperthermophile is obtained even if the mesophile is more stable at room temperature. The results obtained therefore suggest one possible way in which the apparently destabilising effects of salt bridges at room temperature can be reconciled with their increased abundance in hyperthermophilic proteins.

Enzyme Stability↗

PubMedDate not supplied

HPLC separations with micro-bore columns using high-temperature water and flame ionization detection.

Previous work demonstrates that a flame ionization detector (FID) may be used as a detector for high-temperature water separation. However, the relatively high flow rate of the eluent required by standard high-performance liquid chromatography columns causes instability of the FID. In this work, micro-bore columns are packed with poly(styrene-divinylbenzene) (PRP-1) or Daiso gel octadecylsilane-bonded phase particles. Because micro columns require low volume flow rates, the eluent used in high-temperature water chromatography does not cause instability of the FID. Separation of carbohydrates, amino acids, and other organic acids and bases is performed on two micro-bore columns using a homemade high-temperature water chromatograph with FID. Both isothermal and programmed temperatures are used in this work. The limit of detection and the linear range are also determined for amino acids tested.

Journal Article↗

PubMed2006

[Effects of short-term high temperature stress on flag leaf photosystem II functions of high-yielding wheat at late grain-filling stage].

By using fast OJIP chlorophyll a ( Chl a) fluorescence transients, this paper studied the effects of short-term high temperature stress on the flag leaf photosystem II (PS II ) functions of high-yielding winter wheat cultivar 01-35 at its late grain filling stage. The results showed that with the temperature raised from 37 degrees C to 43 degrees C , the extent and velocity of QA reduction enhanced gradually, being 23. 89% and 24. 09% higher at 43 degrees C than at room temperature, respectively, which suggested that the electron transport from QA to QB was inhibited. The electron acceptor pool of PS 1I including QA Q, and PQ at 43 degreesC decreased to 47. 4% of that at room temperature, indicating its being damaged. The electron donor side of PS II was not damaged from 37 degrees C to 43 degrees C. When temperature raised to 46 degrees C , the extent and velocity of QA reduction were 13. 95% and 20. 48% higher than those at room temperature, respectively, but decreased obviously, compared with those at 43 degrees C. In contrast, the electron acceptor pool of PS II at 46 degrees C had no obvious change, compared with that at 43 degrees C, indicating that the electron donor side of PS II at 46 TC was damaged. Comparing with cultivar Lumai 14, the light harvesting complex ( LHC U) of high-yielding wheat cultivar 01-35 could harvest more sun energy, and distributed more absorbed energy into electron transport, suggesting that the LHC II of cultivar 01-35 could endure a certain higher temperature, and maintain higher electron transport capacity.

Edible Grain↗

PubMed1993

The helical repeat of DNA at high temperature.

The increasing number of studies on thermophilic organisms addressed the question of DNA double helix parameters at high temperature. The present study shows that the helix rotation angle per base pair omega of an unconstrained DNA decreases linearly upon temperature increase, up to the premelting range. In the ionic conditions tested, this rule extends to temperatures up to 85 degrees C, which is a common growth temperature for many hyperthermophilic organisms. In addition, the torsional constant K of DNA decreases with temperature, indicating that the energy required to modify the DNA twist is lower at high temperature. These findings have several implications for people working on the structure and enzymology of DNA at high temperature.

Animals↗

PubMed1988

Yeast cell viability under conditions of high temperature and ethanol concentrations depends on the mitochondrial genome.

Wine yeasts manifest simultaneously a high tolerance to ethanol, thermotolerance, and a high resistance to the mutagenic effects of ethanol on the mitochondrial genome. The transfer of mitochondria from these strains to laboratory yeasts demonstrate that this genome influences the above parameters, since thermotolerance, ethanol-growth tolerance, and the frequency of rho- mutants were either totally or partially modified in the laboratory recipient strain. When the death rate and the rate of formation of rho- mutants were measured under extreme conditions of inhibitory ethanol concentrations and high temperature, a perfect correlation was found between these parameters, and both of them were dependent on the strain of mitochondrial genome. Thus, the transfer of wine yeast mitochondria leads to a lower death rate, and a simultaneous increase in thermotolerance and ethanol tolerance in the recipient strain. These results demonstrate the role that viability plays under conditions of high temperatures and high ethanol concentrations. The greater stability of the rho+ phenotype shown by the wine yeast mitochondrial genome may be responsible for the increased viability conferred by these mitochondria.

Cell Division↗

PubMed2002

Polymorphism of the CNS active drug Org 13011: the application of high temperature analysis to detect new polymorphs.

The polymorphism of the CNS active compound Org 13011 was studied using different crystallisation methods (i.e. different solvents and cooling rates). The samples were analysed by Raman, solid state NMR, X-ray powder diffraction (XRPD) and thermal analysis. This led to the characterisation of two crystalline forms A and B. Further high temperature analysis using Raman, XRPD, solid state NMR and DSC revealed another two (high temperature) crystalline forms C and D. The transitions to the high temperature crystalline forms occur at temperatures of about 60 degrees C. This study shows that the application of high temperature experiments is useful and can lead to the discovery of new crystalline forms.

Central Nervous System Agents↗