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Induction of heat, freezing and salt tolerance by heat and salt shock in Saccharomyces cerevisiae.

Stress tolerance of Saccharomyces cerevisiae was examined after exposure to heat and salt shock in the presence or absence of the protein synthesis inhibitor cycloheximide. Cells heat-shocked (37 degrees C for 45 min) in the absence of cycloheximide demonstrated increased tolerance of heat, freezing and salt stress. For cells heat-shocked in the presence of cycloheximide, heat and salt tolerance could still be induced, although at lower levels, while induction of freezing tolerance was completely inhibited. These results indicated that while heat shock proteins (hsps) may contribute to induced heat and salt tolerance they are not essential, although induction of freezing tolerance appears to require protein synthesis. Exposure of cells to salt shock (300 mM NaCl for 45 min) induced stress protein synthesis and the accumulation of glycerol, responses analogous to induction of hsp synthesis and trehalose accumulation in cells exposed to heat shock. Cells salt-shocked in the absence of cycloheximide showed a similar pattern of induced stress tolerance as with heat, with increased tolerance of heat, salt and freezing. Cells salt-shocked in the presence of cycloheximide continued to show induced heat and salt tolerance, but freezing tolerance could not be induced. These results lend support to the hypothesis that hsp synthesis is not essential for induced tolerance of some forms of stress and that accumulated solutes such as trehalose or glycerol may contribute to induced stress tolerance.

Cycloheximide↗

Effect of pre- and post-heat shock temperature on the persistence of thermotolerance and heat shock-induced proteins in Listeria monocytogenes.

The effect of incubation temperature, before and after a heat shock, on thermotolerance of Listeria monocytogenes at 58 degrees C was investigated. Exposing cells grown at 10 degrees C and 30 degrees C to a heat shock resulted in similar rises in thermotolerance while the increase was significantly higher when cells were grown at 4 degrees C prior to the heat shock. Cells held at 4 degrees C and 10 degrees C after heat shock maintained heat shock-induced thermotolerance for longer than cells held at 30 degrees C. The growth temperature prior to inactivation had negligible effect on the persistence of heat shock-induced thermotolerance. Concurrent with measurements of thermotolerance were measurements of the levels of heat shock-induced proteins. Major proteins showing increased synthesis upon the heat shock had approximate molecular weights of 84, 74, 63, 25 and 19 kDa. There was little correlation between the loss of thermotolerance after the heat shock and the levels of these proteins. Thermotolerance of heat shocked and non-heat shocked cells was described by traditional log-linear kinetics and a model describing a sigmoidal death curve (logistic model). Employing log-linear kinetics resulted in a poor fit to a major part of the data whereas a good fit was achieved by the use of a logistic model.

Bacterial Proteins↗

DNA binding of heat shock factor to the heat shock element is insufficient for transcriptional activation in murine erythroleukemia cells.

The heat shock response is among the most highly conserved examples of regulated gene expression, being present in all cellular organisms. Transcriptional activation of heat shock genes by increased temperature or other cellular stresses is mediated by the binding of a heat shock factor (HSF) to a conserved nucleotide sequence (the heat shock element) present in the promoter of heat-inducible genes. Despite the high degree of conservation of this response, embryonic stages of development are characterized by the absence of a heat shock response. Murine erythroleukemia (MEL) cells also lack this response, and we report here a detailed characterization of this defect for one of the most highly conserved of these genes, hsp70. Surprisingly, heat-induced transcriptional activation of this gene does not occur, despite the induction of a protein with the binding specificity of murine HSF. However, the MEL HSF differs slightly in apparent size from the HSF in 3T3 cells, which exhibit a normal heat shock response. These data suggest that activation of mammalian HSF by heat requires at least two separate steps: an alteration of binding activity followed by further modification that activates transcription. MEL cells do not respond to heat shock because they lack the ability to perform this secondary modification. These cells provide a useful system for characterizing heat shock activation in mammals.

Animals↗

Pre-existing inflammatory state compromises heat tolerance in rats exposed to heat stress.

This study investigated the roles of endotoxemia and heat-induced tissue damage in the pathology of heat stroke. In groups of eight, male Wistar rats were treated with heat exposure only (HE), or heat exposure with turpentine (T+HE), dexamethasone (D+HE), and turpentine and dexamethasone combined (TD+HE). The rats remained sedated for 2 h after receiving the respective treatments, followed by heat exposure until the core temperature (T(c)) was 42 degrees C for 15 min; control rats received turpentine (T), dexamethasone (D), and turpentine and dexamethasone (TD) without heat stress. Blood samples were collected before treatment (baseline I), after 2 h of passive rest (baseline II), at T(c) 40 degrees C (T40), and 15 min after achieving T(c) 42 degrees C (T42). No rats died in the nonheat-stressed groups. Survival rate was lowest in the TD+HE rats (37.5%), followed by the HE (62.5%), T+HE (75%), and D+HE (100%) rats (P < 0.05). The duration of survival at T42 degrees C was shortest in the TD+HE rats (9.9 +/- 6.2 min) (P < 0.01), followed by the T+HE (11.3 +/- 6.1 min) and the HE (12.2 +/- 4 min) (P < 0.05) rats. The increase in plasma IL-6 concentrations was highest in the T+HE (352%) and HE (178%) rats (P < 0.05). D+HE treatment suppressed the increases in plasma aspartate transaminase, alanine aminotransferase, and IL-6 and LPS concentrations during severe heat stress. Heat stroke can be triggered by endotoxemia or heat-induced tissue damage, and preexisting inflammation compromises heat tolerance, whereas blocking endotoxemia increases heat tolerance.

Alanine Transaminase↗

Heat preconditioning attenuates renal injury in ischemic ARF in rats: role of heat-shock protein 70 on NF-kappaB-mediated inflammation and on tubular cell injury.

Although heat preconditioning has been known to be protective in various types of injury, the precise molecular mechanism for this is unclear. Recent observations that indicate that previous heat shock has an anti-inflammatory, antiapoptotic effect led to this investigation of the in vivo effect of heat preconditioning on NF-kappaB activation and inflammation and also on tubular cell injury in ischemic acute renal failure (ARF). Heat preconditioning provided marked functional protection and also reduced histologic evidence of tubular necrosis. Ischemia/reperfusion-induced NF-kappaB activation was suppressed by heat preconditioning with a subsequent decrease in monocyte chemoattractant protein-1 expression and inflammatory cell infiltration. Heat preconditioning also suppressed the accumulation of phosphorylated inhibitory kappaBalpha (IkappaBalpha) with a resultant depletion of cytoplasmic IkappaBalpha, indicating that heat preconditioning blocked the activation of the IkappaB kinase complex. Tubular cell apoptosis, determined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining, also was decreased by heat preconditioning, and this was accompanied by decreased caspase 3 activation. Among several heat-shock proteins (HSP), HSP-70 was induced primarily by heat preconditioning. Inhibition of HSP-70 by quercetin almost completely reversed the functional protection that was provided by heat preconditioning. These data provide evidence that HSP-70 affords protection via inhibition of NF-kappaB-mediated inflammation and also inhibition of the cell death pathway in ischemic ARF. Further elucidation of the cytoprotective mechanism of stress proteins could facilitate new target or drug development in the treatment of ARF.

Acute Kidney Injury↗

Alterations in specific and general protein synthesis after heat shock in heat-sensitive mutants of CHO cells and their wild-type counterparts.

The rates of general and specific protein synthesis were studied in two heat-sensitive strains of CHO cells (Harvey and Bedford, Radiat. Res. 113, 526-542, 1988), both of which show a reduced ability to develop thermotolerance following an initial 45 degrees C heat shock. After various labeling periods with [35S]methionine, wild-type and mutant labeled proteins were separated by one- and two-dimensional polyacrylamide gel electrophoresis. Autoradiograms showed differences in levels of synthesis of several proteins after a 45 degrees C heat shock. In particular, these were in the hsp-70 group referred to as hsp-70a, b, and c, having molecular weights of 76, 73, and 72 kDa and isoelectric focusing pH values of 5.7, 5.5, and 5.7, respectively. Of particular note were changes in the hsp-70c region of the autoradiograms. We found that there was perhaps a low level of synthesis of hsp-70c in unheated wild-type cells but none was detectable in the mutant lines. After an isosurvival (approximately 10%) pulse of 45 degrees C heat there was a gradual increase in the synthesis of hsp-70c for wild-type but a smaller increase for the heat-sensitive strain 36 (HS-36) cells. In contrast, for HS-23 cells there was a very large initial increase by 5 to 7 h after the heat pulse and then a rapid decrease to undetectable levels by 11 to 13 h. The inhibition and recovery of general protein synthesis for both mutant and wild-type cells was also measured following various heat treatments at 45 degrees C. We observed that inhibition and resumption to a "normal" rate of protein synthesis for HS-23 cells paralleled the same response observed for the wild-type 10B2 cells. In sharp contrast, the time for recovery from the inhibition of protein synthesis for HS-36 cells was severely reduced for all heating times tested. Our results show that the period of delay before resumption of protein synthesis after heating does not always correlate with heat sensitivity or the degree of thermotolerance development. Several explanations for these observations are possible. One is that while synthesis of certain heat-shock proteins may indeed be responsible for the development of thermotolerance, the timing of the synthesis of these proteins in relation to the period of inhibition of general protein synthesis is crucial to such development.(ABSTRACT TRUNCATED AT 400 WORDS)

Acclimatization↗

Heat sensitivity, thermotolerance, and profile of heat shock protein synthesis of human myelogenous leukemias.

In anticipation of using single or fractionated hyperthermia treatment in ex vivo purging of leukemic bone marrow in the clinic, we have compared the hyperthermic sensitivity, kinetics of thermotolerance, and heat-shock protein synthesis in three human myelogenous leukemic cell lines. In terms of heat sensitivity, the chronic myelogenous leukemic cell line K562 was found to be the most resistant. The Dos of the 43, 44, and 45 degrees C heat survival curves were 22, 13, and 6 min, respectively. HL-60 and KG-1, both acute myelogenous leukemic lines, however, were found to be several fold more sensitive to the cytotoxic effects of heat. The Dos of the 43, 44, and 45 degrees C heat survival curves for HL-60 were 7.6, 5.6, and 2 min and for KG-1 were 5.7, 4.5, and 1.7 min, respectively. All cell lines developed thermotolerance. However, K562 developed more tolerance which lasted for longer times. For K562 cells at priming heat doses of 45 degrees C/10 min, 42 degrees C/2 h, or 41 degrees C/2 h thermotolerance was maximum at 4 to 6 h and began to decay at 24 h. HL-60 and KG-1 cells showed some thermotolerance at the priming doses of 45 degrees C/5 min or 42 degrees C/30 min and had fully decayed by 24 h. K562 cells synthesized Mr 70,000 heat shock protein for over 24 h following the 45 degrees C/10 min heat shock, while HL-60 and KG-1 synthesized Mr 70,000 heat shock protein for 2-4 h for the same amount of cell kill. These studies suggest that most human leukemias may be extremely sensitive to the cytotoxic effects of heat, and in vitro purging of leukemias from bone marrow specimens by heat needs to be further studied both by in vitro and in vivo model systems.

Cell Survival↗

Thiol reducing reagents inhibit the heat shock response. Involvement of a redox mechanism in the heat shock signal transduction pathway.

We evaluated the effects of thiol-reducing agents on the heat shock response in human and rodent cells in culture. Using HeLa cells as an example, we demonstrated that dithiothreitol (DTT,2mM) inhibited the heat (42 degrees C) induced increase in the synthesis of heat shock proteins (HSPs), abundance of mRNA of hsp 70, hsp 70 gene promoter activity, and the heat shock factor (HSF) DNA binding activity. This effect of DTT was specific and attributable to its reducing activity; oxidized DTT was ineffective, and other thiol reducing compounds had the same effect as DTT. Time course and dose-response studies showed that DTT significantly inhibited the heat shock induction of heat shock element binding activity with no preincubation and that 0.6 and 1-2 mM DTT gave half-maximal and maximal inhibition, respectively. The effect of DTT was reversible; removal of the DTT-containing medium prior to heat shock rendered the cells fully responsive. Analysis of the effects of DTT on the regulation and function of HSF suggests that DTT blocked an early and important step in the activation process without having a direct effect on the HSF protein. Thus, DTT inhibited the heat-induced trimerization, phosphorylation, and nuclear translocation of HSF and was also effective against a number of other reagents that are known to activate HSF. On the other hand, DTT did not block the response induced by heat shock at 45 degrees C, and in vitro addition of DTT failed to modulate the DNA binding activity of activated HSF present in cell extracts, suggesting that the HSF protein itself is unlikely to be a direct target of action of DTT. These results, together with the observation that activation of HSF DNA binding activity was attenuated under an anoxic condition and that hydrogen peroxide mimicked the effects of heat shock, suggest the involvement of a redox mechanism as an early and important step in the heat shock signal transduction pathway.

Cell Nucleus↗

Effect of heat stress on development in vitro and in vivo and on synthesis of heat shock proteins in porcine embryos.

The present study was conducted (1) to examine the effect of an acute increase in ambient temperature on the development of porcine day 6 embryos in culture and after transfer to recipient gilts, and (2) to analyze intracellular production of heat shock proteins (hsps). The viability of porcine day 6 embryos following a temporary acute elevation in ambient temperature (at 42 degrees-45.5 degrees C and for 10-180 min) was examined. Synthesis of 70 kDa hsp (hsp70) and 90 kDa hsp (hsp90) was determined by SDS-PAGE and Western blot analysis in porcine day 6 embryos subjected to heat stresses. Nonheat-stressed embryos were considered as control. Significantly higher numbers of viable nuclei were observed in treatment groups of 42 degrees C-10 min (236.6 +/- 71.4; P < 0.05) and 43 degrees C-30 min (276.8 +/- 89.4; P < 0.005) compared to control (173.9 +/- 53.9). The 42 degrees C-180 min group (158.0 +/- 27.1 microns) had a greater increase in diameter after 24 hr in culture following heat stress compared to control (82.5 +/- 47.3 microns), while heat stress with 43 degrees C for > or = 60 min, 44 degrees-44.5 degrees C for > or = 30 min, or 45 degrees-45.5 degrees C for > or = 10 min impaired their survival, as assessed by differences in number of viable nuclei. The embryos subjected to heat stresses under the conditions of 42 degrees C-180 min, 43 degrees C-10 min, 43 degrees C-30 min, 44 degrees C-10 min, or 45 degrees C-10 min developed to normal piglets after transfer to recipient gilts. Overall pregnancy rate was 75% (6/8), and farrowing rate 62.5% (5/8). Of heat-stressed embryos transferred, 59% (36/61) developed to normal piglets. Heat-stress conditions of 42 degrees C for 180 min, 43 degrees C for 30 min, 44 degrees C for 10 min, and 45 degrees C for 10 min were determined as critical with respect to the in vitro and in vivo survival of porcine embryos. Porcine day 6 embryos constitutively synthesized hsp70 even without heat stress, while hsp90 was detected only at trace level. Neither hsp70 nor hsp90 levels increased in the embryos subjected to heat stresses. In conclusion, porcine day 6 embryos could continue to develop in vivo or during in vitro culture after exposure to acute and temporary rise in temperature. However, no increase of hsp70 and hsp90 was observed in the heat-stressed porcine embryos, while hsp70 was detected in the nonheat-stressed porcine embryos. The precise mechanism of the thermotolerance was unclear.

Animals↗

Heat-induced proteolysis of HSF causes premature deactivation of the heat shock response in Nb2 lymphoma cells.

Nb2-11 cells, a prolactin (PRL)-dependent T-lymphoma cell line, display an unusual response to heat stress characterized by the lack of expression of inducible hsp70 mRNA transcripts and a reduction in the levels of constitutively expressed heat shock protein (HSP) genes. This aberrant heat shock response appears to result from heat-induced proteolytic fragmentation of heat shock factor (HSF). In this report, we have investigated processes that promote HSF fragmentation and identified characteristics of a protease that may be responsible for this effect. Cycloheximide did not affect HSF fragmentation of heat-shocked Nb2-11 cells suggesting that proteases responsible for this proteolysis are constitutively expressed and become activated by the heat shock conditions. PRL protected Nb2-11 cells from heat-induced fragmentation whereas sodium butyrate (NaBT) rendered a fragmentation-resistant cell line (Nb2-SFJCD1 cells) sensitive to HSF proteolysis. Heat-induced HSF fragmentation in Nb2-11 cells was not affected by pretreating cultures with several serine protease inhibitors. However, a dose-dependent decrease in HSF fragmentation was achieved by pretreating cultures with iodoacetamide, a cysteine protease inhibitor that is active in apoptosis. Apparently, the heat shock response in Nb2 cells is attenuated by a mechanism that involves the premature deactivation of HSF by its selective proteolysis. Attenuation of this critical cellular stress response may be an important contributor to the progression of hormone-dependent tumors possibly by influencing apoptotic processes known to regulate the activity of these cells.

Animals↗

Activation of heat-shock transcription factor 1 in heated Chinese hamster ovary cells is dependent on the cell cycle and is inhibited by sodium vanadate.

Inducible heat-shock protein 70 (HSP72) is expressed in a cell cycle-specific manner in Chinese hamster ovary (CHO) cells after heating for 15 min at 45.0 degrees C, with the highest level in S-phase cells. Since heat shock induces the transcription of heat-shock proteins through the transactivation of heat-shock elements (HSEs) by heat-shock factor HSF1, we wished to determine whether the cell cycle-specific expression of HSP72 was regulated at the level of transcription. The levels of HSF1 did not vary through the cell cycle, as measured by polyclonal antibodies and flow cytometry. The binding of HSF1 to the heat-shock element was measured with the gel mobility shift assay using cell extracts from Hoechst 33342-labeled heated cells sorted from G1, S and G2/M phases. The HSF1-HSE binding activity was twofold higher in S phase than in G1 or G2/M phase. When CHO cells were exposed to 10 microM sodium vanadate, an inhibitor of tyrosine phosphatase, for 24 h before heat shock, the binding of HSF1 to HSE was reduced by a factor of 2 and the level of HSP72 was greatly reduced. The HSF1 binding to HSE was completely eliminated by using anti-HSF1 antibody in the gel mobility shift assays. Antibodies against HSP73 did not reduce the HSF1-HSE binding activity, but antibodies against HSP40 actually increased the binding activity. These results support the hypothesis that cell cycle-dependent binding of HSF1 to HSE is the cause of the cell cycle-specific expression of HSP72 in heated CHO cells and is regulated by phosphorylation.

Animals↗

Sensitization to x-rays by sodium arsenite or heat in normal cells and in cells with an induced tolerance for heat and arsenite.

In this study we compared sensitization to x-rays by heat or sodium arsenite and the effect of an induced heat or arsenite resistance on radiosensitization. Treatment of Reuber H35 hepatoma cells with either heat or arsenite causes a dose-dependent radiosensitization. Based on a comparison of isosurvival doses for arsenite and heat, arsenite causes a stronger enhancement of the radiosensitivity. Radiosensitization increases exponentially with increasing sensitizer dose. It is gradually lost when the time interval between irradiation and treatment with heat or arsenite increases, depending on the treatment sequence. For x-rays prior to heat, radiosensitization disappears approximately twice as fast as in the reverse case. Arsenite radiosensitization shows approximately the same kinetics for an isoeffective combination, but slightly longer times are needed for the complete clearance of the interaction. As with heat, an exposure to arsenite induces a stress response in cultured cells which results in the development of an increased tolerance towards a second exposure. Heat and arsenite induce self- as well as cross-tolerance. The reduction in arsenite or heat toxicity in tolerant cells is correlated with a reduction in radiosensitization. The mechanisms for heat and arsenite cytotoxicity appear to be different. A combination of non-toxic doses of heat and arsenite has a synergistic effect on the cytotoxicity. One hour incubation with 0.02 mM arsenite at 41 degrees C has the same cytotoxicity as 0.2 mM after 3 h incubation at 37 degrees C, and the amount of radiosensitization induced by these treatments is approximately the same.

Animals↗

Quantifying the combined effects of the heating time, the temperature and the recovery medium pH on the regrowth lag time of Bacillus cereus spores after a heat treatment.

The purpose of this study was to quantify the lag time of re-growth of heated spores of Bacillus cereus as a function of the conditions of the heat treatment: temperature, duration and pH of the recovery medium. For a given heating temperature, curves plotting lag times versus time of heating show more or less complex patterns. However, under a heating time corresponding to a decrease of 2 decimal logarithms of the surviving populations of spores, a linear relationship between the lag time of growth and the time of the previous heat treatment can be observed. The slope of this linear relationship followed itself a Bigelow type linear relationship, the slope of which yielded a zeta-value very close to the observed conventional z-value. It was then concluded that the slope of the regrowth lag time versus the heating time followed a linear relationship with the sterilisation value reached in the course of the previous heat treatment. A sharp effect of the pH of the medium which could be described by a simple "secondary" model was observed. As expected, the observed intercept of the linear relationship between lag time and heating time (lag without previous heating) was dependent on only the pH of the medium and not on the heating temperature.

Adaptation, Physiological↗

Heat capacity of proteins. II. Partial molar heat capacity of the unfolded polypeptide chain of proteins: protein unfolding effects.

Using the heat capacity values for amino acid side-chains and the peptide unit determined in the accompanying paper, we calculated the partial heat capacities of the unfolded state for four proteins (apomyoglobin, apocytochrome c, ribonuclease A, lysozyme) in aqueous solution in the temperature range from 5 to 125 degrees C, with an assumption that the constituent amino acid residues contribute additively to the integral heat capacity of a polypeptide chain. These ideal heat capacity functions of the extended polypeptide chains were compared with the calorimetrically determined heat capacity functions of the heat and acid-denatured proteins. The average deviation of the experimental functions from the calculated ideal ones in the whole studied temperature range does not exceed the experimental error (5%). Therefore, the heat-denatured state of a protein, in solutions with acidic pH preventing aggregation, approximates well the completely unfolded state of this macromolecule. The heat capacity change caused by hydration of amino acid residues upon protein unfolding was also determined and it was shown that this is the major contributor to the observed heat capacity effect of unfolding. Its value is different for different proteins and correlates well with the surface area of non-polar groups exposed upon unfolding. The heat capacity effect due to the configurational freedom gain by the polypeptide chain was found to contribute only a small part of the overall heat capacity change on unfolding.

Apoproteins↗

In vivo efficacy of heated and non-heated humidifiers during nasal continuous positive airway pressure (nCPAP)-therapy for obstructive sleep apnoea.

Upper airway dryness is a frequent side-effect of nasal continuous positive airway pressure therapy (nCPAP) in obstructive sleep apnoea (OSA). In this situation, heated or non-heated passover humidifiers are often added to the nCPAP-therapy. The efficacy of these two modes in terms of increasing the absolute humidity of the inspired air in vivo has so far not been established. The present investigation was therefore designed to compare various heated and non-heated passover humidifiers in terms of the their ability to increase the absolute humidity in the inspired air during nCPAP. In six healthy test individuals, nCPAP-therapy at pressures of 5 mbar and 10 mbar was simulated, and the relative humidity and temperature of the air within the tube at the junction between CPAP tube and mask were measured. In each test person, measurements were carried out both with and without the two heated (HC 100, Fischer&Paykel Inc., New Zealand and HumidAire, ResMed Ltd., Australia) and two non-heated (Oasis and Humidifier, both from Respironics Inc., U.S.A.) passover humidifiers under steady-state conditions. The absolute humidity was calculated from the relative humidity and temperature measurements. The mean (SD) absolute humidity (gm(-3)) in the steady-state was significantly (P<0.05 higher with each of the humidifiers than that calculated when no humidifier was used. The relevant figures were as follows: no humidifier: 10(-2) (1.8) gm(-3) (at 5 mbar)/9.8 (1.8) gm(-3) (at 10 mbar); Humidifier: 16.4 (0.97)/15.6 (1.26); Oasis: 17.3 (0.97)/ 16.7 (0.93); HC100: 26.5 (1.40)/26.2 (1.23); HumidAire: 31.8 (2.50)/30.9 (2.64). The mean increase in absolute humidity (in gm(-3)) with the aid of the heated humidifiers was 16.3 (5 mbar) gm(-3)/16.4 (10 mbar) gm(-3) with HC100 and 21.6/21.1 with HumidAire, and in both cases was clearly and significantly (P=0.028) higher in comparison with the non-heated humidifiers--6.2/5.8 with Humidifier and 7.2/6.9 with Oasis. In terms of the absolute humidity achieved within the CPAP tube system, the heated humidifiers were clearly superior to the non-heated humidifiers. These results were, however, obtained under laboratory conditions, and therefore cannot be translated unreservedly to the situation represented by long-term CPAP-treatment. Furthermore, it is possible that the smaller humidification capacity of the non-heated humidifiers may still suffice to meet the requirements of clinical use in terms of effectively preventing dry airways under CPAP treatment. This point, however, needs further investigation on the basis of long-term clinical studies.

Adult↗

The effect of heat on amino acids for growing pigs. 1. A comparison of ileal and faecal digestibilities of amino acids in raw and heat-treated field peas (Pisum sativum cultivar Dundale).

Three experiments were conducted to examine the effect of heating field peas (Pisum sativum cultivar Dundale) on (1) proximate analysis and total amino acid composition, (2) ileal and faecal digestibilities of amino acids, and (3) digestible energy content. Alternative techniques for assessing ileal and faecal digestibilities and digestible energy respectively, were also investigated. Forced-air dehydrators were used to heat field peas at temperatures of 110 degrees, 135 degrees, 150 degrees or 165 degrees. In the first experiment the apparent ileal and faecal digestibilities of amino acids and the faecal digestibility of energy in the raw and heated field peas were determined using pigs fitted with 'T'-shaped cannulas. In the second, apparent ileal digestibility of amino acids and the faecal digestibility of energy were determined using the direct ileal and rectal sampling technique. This involved a single collection of digesta and faeces from the digestive tract of the pig while it was anaesthetized. The faecal digestibilities of amino acids and energy were determined using total faeces collection in the third experiment. In all experiments the respective field-pea treatments comprised 400 g/kg sugar-based diets and were the only source of amino acids. Heat significantly decreased the lysine (14.6-8.7 g/kg; P < 0.001), cystine (3.2-2.6 g/kg; P < 0.01) and arginine (16.7-14.5 g/kg; P < 0.05) contents of the heated peas. The 'reactive' lysine content of the field peas, as measured using the Silcock technique, was decreased by 0.11 and 0.30 with the application of heat at 150 degrees and 165 degrees respectively. Heat treatments did not alter the ileal digestibility of most amino acids. Only aspartic acid (0.72-0.58), glutamic acid (0.80-0.65) and the basic amino acids, lysine (0.79-0.56) and arginine (0.85-0.75), showed a significant linear decrease (P < 0.05) in ileal digestibility over the heat treatments, determined using the ileal cannulation procedure. Heating significantly (P < 0.05) decreased faecal digestibility for all amino acids. Faecal digestibility was consistently greater than ileal digestibility for the raw field peas; however, this difference decreased with heat application until faecal digestibility was equal or less than ileal digestibility at the 165 degrees treatment. Heat linearly depressed digestible energy, diet dry-matter digestibility and diet energy digestibility. Losses in lysine, cystine and arginine are likely to be due to early and advanced Maillard reactions. Considerable binding of the remaining lysine also occurred as indicated by a decline in Silcock-reactive lysine.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗

The effect of heat on amino acids for growing pigs. 2. Utilization of ileal-digestible lysine from heat-treated field peas (Pisum sativum cultivar Dundale).

Two growth experiments were conducted to determine the effect of heat on the utilization of ileal-digestible lysine from field peas (Pisum sativum cultivar Dundale) fed to growing pigs. Five lysine-deficient diets (0.36 g ileal-digestible lysine/MJ digestible energy (DE)) were formulated using raw field peas, and field peas heated to either 110 degrees, 135 degrees, 150 degrees, or 165 degrees for 15 min respectively in a forced-air dehydrator. Additional diets were formulated with supplements of free lysine to verify that lysine was limiting in the diets containing the raw peas, and peas heated to 150 degrees or 165 degrees. The growth performance and retention of ileal-digestible lysine by pigs given the diets was determined over the 20-45 kg growth phase. Heat had a significant quadratic effect (P < 0.01) on growth rate, with responses declining from 543 g/d with pigs given the raw peas, to 407 g/d for those given the peas heated to 165 degrees. Similarly, crude protein deposition declined in a quadratic manner (P < 0.001) from 76 to 36 g/d for pigs fed on raw peas and peas heated to 165 degrees respectively. Retention of ileal-digestible lysine was 0.85 in the pigs given the raw field peas and declined in a quadratic manner (P < 0.001) with the application of heat to 0.48 in those pigs given the peas heated to 165 degrees. Pigs fed on field peas heated to 165 degrees had increased (P < 0.05) liver weights. The results indicate that heat applied to protein concentrates, even at mild temperatures, renders lysine in a form that is apparently absorbed but inefficiently utilized by the growing pig. Consequently, ileal digestibility values for lysine in heat-processed meals are unsuitable for diet formulations.

Amino Acids↗

Heating of indoor dust causes reduction in its ability to stimulate release of IL-8 and TNFalpha in vitro compared to non-heated dust.

UNLABELLED: Dust is a major contaminant of the indoor air environment and may affect human health. Indoor dust accumulates on surfaces including heaters and light fixtures, and will be heated when these devices are used. Heat treatment of the dust may change its biologic properties and in this study we simulated the heat treatment with a dust-heating model (50-250 degrees C). The residual and the non-heated dust from seven samples were tested in cultures of fresh peripheral blood mononuclear cells and in A549 cell culture using the release of TNFalpha and IL-8, respectively, as effect indicators. The endotoxin-content and the particle size distribution of the residual and the non-heated dust suspensions were determined for some of the samples. We found that the residual dust had less ability to induce the release of TNFalpha and IL-8. The cytokine decline pattern was similar for all the dust tested and could partly be explained by the reduction in endotoxin content or possibly by inhibitory decomposition products. No correlation was found between the measured particle size distribution and the decreased cytokine levels. The results in this study suggest that the residual dust promotes reduced cytokine response and thereby a possibly lower inflammation reaction in the airways if suspended and inhaled compared with the non-heated dust. PRACTICAL IMPLICATIONS: Accumulation of indoor dust on electric heaters and light fixtures may produce a bad odor when switched on in the cold season and some people claim respiratory distress during such events. To investigate to what extent the residuals of heated indoor dust represent a health hazard, we measured the effect in cell cultures before and after heat treatment of the dust. The in vitro results imply that the residual dust will cause a lower proinflammatory response in the airways if suspended and inhaled compared with non-heated dust. This is partly explained by heat destruction of inflammatory components in the dust.

Air Pollutants↗