Tension responses to increased hydrostatic pressure in glycerinated rabbit psoas muscle fibres

A method developed to study the effect of increased hydrostatic pressure on the isometric tension of a single muscle fibre is described and experiments done at room temperature (18-22 degrees C) on glycerinated rabbit psoas muscle fibres are presented. Increase of pressure (range 1-10 MPa) caused little change in tension transducer response when a muscle fibre was relaxed. However, there was a reversible depression of isometric tension with an increase of pressure when a fibre was maximally calcium-activated or in rigor; the depression was around 15% for active tension and 30% for rigor tension, for an increase of pressure of 10 MPa (ca. 100 atm).     

Response surface methodology to supercritical carbon dioxide extraction of astaxanthin from Haematococcus pluvialis

Experimental design was used to investigate the effect of operating temperature (40-80 degrees C), operating pressure (30-50 MPa), and extraction time (1-4h) of supercritical carbon dioxide (SC-CO2) extraction on astaxanthin yields and the extract antioxidant activity (IC50). The ranges of the factors investigated were 40-80 degrees C for the operating temperature (X1), 30-50 MPa for the operating pressure (X2), and 1-4h for the extraction time (X3). The statistical analysis of the experiment indicated that pressure, extraction time, and the interaction between temperature and pressure (X1X2) had significant effect on astaxanthin yields. The central composite design showed that polynomial regression models were in good agreement with the experimental results with the coefficients of determination of 0.924 and 0.846 for astaxanthin yield and antioxidant activity, respectively. The optimal condition for astaxanthin yield within the experimental range of the variables studied was at 70 degrees C, 50 MPa, and 4h. At this condition, the predicted amount of astaxanthin extracted was 23.04 mg/g (2.3 wt% or 83.78% recovery). For the effect of experimental extraction conditions on antioxidant activity, IC50 was used as an index, which is the concentration that gives a 50% reduction in the absorbance of the ABTS free radical. The analysis of the results showed that the interaction between the operating temperature and operating pressure (X1X2) was the only significant factor affecting the extract antioxidant activity. The statistical model gave the minimum point for antioxidant activity at 67 degrees C, 40.3 MPa, and 1.86 h of extraction, at which the value for 1/IC50 was 0.39 l/mg (or IC50 was 2.57 mg/l).     

Fluorimetric determination of sphingosine and its application to natural mixtures of glycosphingolipids

A sensitive estimation of sphingosine, by measurement of the fluorescence of a complex formed with 1-naphthylamino-4-sulfonic acid, is described. The practical range is 5-35 nmoles sphingosine. The method is used to estimate, in terms of sphingosine, amounts of ceramide and glycosphingolipids. The isolation of microamounts (5-30 micro g) of individual glycosphingolipids from a mixture, and their quantitative estimation is described. The percentage composition of a glycosphingolipid mixture from the kidneys of adult C57/BL male mice is given.     

High-pressure stopped-flow spectrometry at low temperatures

A stopped-flow instrument operating over temperature and pressure ranges of +30 to -20 degrees C and 10(-3) to 2 kbar , respectively, is described. The system has been designed so that it can be easily interfaced with many commercially available spectrophotometers of fast response time, with the aid of quartz fiber optics. The materials used for the construction are inert, metal free and the apparatus has proven to be leak free at temperatures as low as -20 degrees C under a pressure of 2 kbar . The performance of the instrument was tested by measuring the rate of reduction of cytochrome c with sodium dithionite and the 2,6-dichloroindophenol/ascorbate reaction. The dead time of the system has been evaluated to be 20, 50, and congruent to 100 ms in water at 20 degrees C, in 40% ethylene glycol/water, and at 20 degrees C and -15 degrees C, respectively. These values are rather pressure independent up to 2 kbar . Application of the bomb was demonstrated using the cytochrome c peroxidase/ethyl peroxide reaction. This process occurred in two phases and an increase in pressure decreased the rates of reactions indicating two positive volumes of activation (delta V not equal to app (fast) = 9.2 +/- 1.5 ml X mol-1; delta V not equal to app (slow) = 14 +/- 1.5 ml X mol-1, temperature 2 degrees C). The data suggest that the fast reaction could involve a hydrophobic bond, whereas the slow process could be associated with a stereochemical change of the protein. The problem of temperature equilibrium for high-pressure experiments is also discussed.     

Effects of aortic nerve on hemodynamic response to obstructive apnea in sedated pigs.

In this study we test the hypothesis that aortic nerve traffic is responsible for the pressor response to periodic apneas. In nine intubated, sedated chronically instrumented pigs, periodic obstructive apneas were caused by occlusion of the endotracheal tube for 30 s, followed by spontaneous breathing for 30 s. This was done under control (C) conditions, after section of the aortic nerve (ANS), and after bilateral cervical vagotomy (Vagot). Blood-gas tensions and airway pressure changed similarly under all conditions: PO(2) decreased to 50-60 Torr, PCO(2) increased to approximately 55 Torr, and airway pressure decreased by 40-50 mmHg during apnea. With C, mean arterial pressure (MAP) increased from 111 +/- 4 mmHg at baseline to 120 +/- 5 mmHg at late apnea (P < 0.01). After ANS and Vagot, there was no change in MAP with apneas compared with baseline. Relative to baseline, cardiac output and stroke volume decreased with C but not with ANS or Vagot during apneas. Increased MAP was due to increased systemic vascular resistance. Heart rate behaved similarly with C and ANS, being greater at early interapnea than late apnea. With Vagot, heart rate increased throughout the apnea-interapnea cycle relative to baseline. We conclude that, in sedated pigs, aortic nerve traffic mediates the increase in MAP and systemic vascular resistance observed during periodic apneas. Increase in MAP is responsible for decreased cardiac output and stroke volume. Additional vagal reflexes, most likely parasympathetic efferents, are responsible for interacting with sympathetic excitatory influences in modulating heart rate.     

Effect of ambient temperature on cardiovascular parameters in rats and mice: a comparative approach

Ambient air temperatures (T(a)) of <6 degrees C or >29 degrees C have been shown to induce large changes in arterial blood pressure and heart rate in homeotherms. The present study was designed to investigate whether small incremental changes in T(a), such as those found in typical laboratory settings, would have an impact on blood pressure and other cardiovascular parameters in mice and rats. We predicted that small decreases in T(a) would impact the cardiovascular parameters of mice more than rats due to the increased thermogenic demands resulting from a greater surface area-to-volume ratio in mice relative to rats. Cardiovascular parameters were measured with radiotelemetry in mice and rats that were housed in temperature-controlled environments. The animals were exposed to different T(a) every 72 h, beginning at 30 degrees C and incrementally decreasing by 4 degrees C at each time interval to 18 degrees C and then incrementally increasing back up to 30 degrees C. As T(a) decreased, mean blood pressure, heart rate, and pulse pressure increased significantly for both mice (1.6 mmHg/ degrees C, 14.4 beats.min(-1). degrees C(-1), and 0.8 mmHg/ degrees C, respectively) and rats (1.2 mmHg/ degrees C, 8.1 beats.min(-1). degrees C(-1), and 0.8 mmHg/ degrees C, respectively). Thus small changes in T(a) significantly impact the cardiovascular parameters of both rats and mice, with mice demonstrating a greater sensitivity to these T(a) changes.     

Constitutive penicillinase formation in Staphylococcus aureus owing to a mutation unlinked to the penicillinase plasmid

Previously described penicillinase-constitutive mutations in Staphylococcus aureus are caused by genetic lesions in a regulator gene (or genes) on the penicillinase plasmid in close linkage to the structural gene. This report describes a new class (R2(-)) of penicillinase-constitutive mutants of S. aureus unlinked to the plasmid. By transductional analysis, the penicillinase plasmids in these mutants were wild type. Wild-type plasmids transduced into penicillinase-negative (plasmid loss) derivatives of R2(-) mutants produced penicillinase constitutively in amounts comparable to a fully induced culture of the wild-type strain. Penicillinase production in R2(-) mutants was maximal at 30 to 32 C and was much reduced at 40 C.     

Cutaneous vascular responses to isometric handgrip exercise

Cutaneous vascular responses to dynamic exercise have been well characterized, but it is not known whether that response pattern applies to isometric handgrip exercise. We examined cutaneous vascular responses to isometric handgrip and dynamic leg exercise in five supine men. Skin blood flow was measured by laser-Doppler velocimetry and expressed as laser-Doppler flow (LDF). Arterial blood pressure was measured noninvasively once each minute. Cutaneous vascular conductance (CVC) was calculated as LDF/mean arterial pressure. LDF and CVC responses were measured at the forearm and chest during two 3-min periods of isometric handgrip at 30% of maximum voluntary contraction and expressed as percent changes from the preexercise levels. The skin was normothermic (32 degrees C) for the first period of handgrip and was locally warmed to 39 degrees C for the second handgrip. Finally, responses were observed during 5 min of dynamic two-leg bicycle exercise (150-175 W) at a local skin temperature of 39 degrees C. Arm LDF increased 24.5 +/- 18.9% during isometric handgrip in normothermia and 64.8 +/- 14.1% during isometric handgrip at 39 degrees C (P less than 0.05). Arm CVC did not significantly change at 32 degrees C but significantly increased 18.1 +/- 6.5% during isometric handgrip at 39 degrees C (P less than 0.05). Arm LDF decreased 12.2 +/- 7.9% during dynamic exercise at 39 degrees C, whereas arm CVC fell by 35.3 +/- 4.6% (in each case P less than 0.05). Chest LDF and CVC showed similar responses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Activation parameters of the blue shift (Shibata shift) subsequent to protochlorophyllide phototransformation

The Shibata shift was analyzed in flash irradiated wheat (Triticum aestivum, L., cult. MV17) leaf homogenates in the pressure range of 0.1 to 500 MPa, at temperatures of 20, 30 and 40 degrees C. The kinetics of the blue shift (called Shibata shift in case of intact leaves) was followed by repeated recording of fluorescence emission spectra after phototransformation. At 20 degrees C, above 100 MPa, the blue shift slowed down remarkably. Two components of the blue shift could be distinguished, one was pressure-dependent and the other was almost pressure-independent. The pressure-independent component can be associated with minor conformational changes of the NADPH:protochlorophyllide oxidoreductase (POR) enzyme, followed by molecular movements of the newly formed chlorophyllide molecules. The calculated activation volume of the pressure-dependent component was 43+/-11 cm(3) mol(-1) at 20 degrees C. This value reflects major molecular reorganizations in the lipid system of the membrane and in the chlorophyllide-protein complexes, and corresponds to changes of the tertiary structure of proteins which can proceed directly or indirectly via structural changes of the membrane lipids. The process was inhibited by 300 and 400 MPa at 30 and 40 degrees C, respectively. The activation volume reduced to 35+/-1.5 cm(3) mol(-1) at 40 degrees C. The decrease of the activation volume with increasing temperature indicates that the blue shift requires loosened lipid structures. The activation energy of the blue shift (measured between 10 and 40 degrees C at atmospheric pressure) was 100+/-20 kJ/mol, indicating that the structural change involves rearrangement of strong molecular interactions.     

High pressure nmr study of dihydrofolate reductase from a deep-sea bacterium Moritella profunda.

We have investigated the effect of pressure and temperature on the structural and thermodynamic stability of a protein dihydrofolate reductase from a deep-sea bacterium Moritella profunda in its folate-bound form in the pressure range between 3 and 375 MPa and the temperature range between -5 and 30 degrees C. The on-line cell variable pressure 1H NMR spectroscopy has been used to analyze the chemical shift and signal intensity in one-dimensional 1H NMR spectra. Thermodynamic analysis based on signal intensities from protons in the core part indicates that the thermodynamic stability of Moritella profunda DHFR is relatively low over the temperature range between -5 and 30 degrees C (deltaG0=15.8 +/- 4.1 kJ/mol at 15 degrees C), but is well adapted to the living environment of the bacterium (2 degrees C and 28 MPa), with the maximum stability around 5 degrees C (at 0.1 MPa) and a relatively small volume change upon unfolding (deltaV= 66 +/- 19 ml/mol). Despite the relatively low overall stability, the conformation in the core part of the folded protein remains intact up to approximately 200 MPa, showing marked stability of the core of this protein.     

Effects of environmental temperature on the venodilatory response to nitroglycerin.

The venodilatory response to nitroglycerin (0.8 mg sublingually) was measured in 10 healthy young male volunteers in a cool [24.3 +/- 0.6 degrees C skin temperature (Tsk)] and a warm environment (34.7 +/- 0.2 degrees C Tsk). Nitroglycerin caused mean arterial pressure to fall and heart rate to rise in both the cool (105 +/- 2 to 96 +/- 3 mmHg; 55 +/- 3 to 62 +/- 3 beats/min) and the warm environment (87 +/- 3 to 81 +/- 3 mmHg; 66 +/- 4 to 75 +/- 3 beats/min), but the fall in pressure was greater in the cool than in the warm environment. Forearm blood flow was reduced and forearm vascular resistance elevated in the cool (117 +/- 19 units; 1.15 +/- 0.08 ml.100 cc arm-1.min-1) compared with the warm environment (15 +/- 3 units; 8.60 +/- 1.89 ml.100 cc arm-1.min-1). Nitroglycerin caused forearm vascular resistance to fall in the cool but had no effect in the warm environment. Venous distensibility (increase in venous volume per 30-mmHg increase in venous pressure) was twice as great in the warm as in the cool environment (3.90 +/- 0.27 vs. 1.88 +/- 0.23 ml/100 cc arm). However, the venodilatory effect of nitroglycerin was similar in the cool and warm environments (0.79 +/- 0.10 vs. 0.67 +/- 0.13 ml/100 cc arm, respectively). Arterioles are not dilated by nitroglycerin in the warmer environment, but the venodilatory effect of nitroglycerin is quantitatively similar in the two environments.     

Gastroesophageal dynamics during immersion in water to the neck.

This investigation was undertaken to study the effect of hydrostatic pressure on gastroesophageal dynamics during immersion in thermoneutral water to the neck. In 5 healthy male subjects (normal end-expiratory), gastric pressure (PG), esophageal pressure (PE), location and pressure of distal esophageal sphincter (des), location of respiratory inversion point (RIP), and gastroesophageal pH gradient were measured standing in air (A), standing in water to the neck (B), and standing in air with abdominal compression (C). The pressure was measured with a Honeywell esophageal catheter (model 31) with built-in pressure transducer. A Beckman stomach pH electrode (no. 39042) was positioned adjacent to the pressure transducer. PG increased from 4.6 +/- 0.6 (SE) mmHg in A to nearly 20 mmHg in B and C, while PE increased from -6.0 +/- 0.8 mmHg in A to -0.8 +/- 1.0 and -3.4 +/- 0.9 mmHg in B and C, respectively. However, PDES was always 11-15 mmHg higher than PG. The superior limit of DES was displaced cephalad by indicating a stretching of DES and a shortening of the esophagus. Qualitatively similar findings were obtained in C. In all experiments, the esophageal pH remained above 6, and no alteration in the amplitude of primary peristaltic waves was seen. It is concluded that a head-out immersion with increased gastroesophageal pressure gradient predisposes to gastric reflux in the absence of a competent DES mechanism.     

Response of recombinant Chinese hamster ovary cells to hyperosmotic pressure: effect of Bcl-2 overexpression

In an attempt to use the hyperosmotic pressure for improved foreign protein production in recombinant Chinese hamster ovary (rCHO) cells, the response of rCHO cells producing a humanized antibody (SH2-0.32-(Delta)bcl-2 cells) to hyperosmotic pressure was determined in regard to cell growth and death, and antibody production. Further, the feasibility of Bcl-2 overexpression in improving rCHO cell viability under hyperosmotic pressure was also determined by comparing control cells (SH2-0.32-(Delta)bcl-2) with Bcl-2 overexpressing cells (14C6-bcl-2). After 3 days of cultivation in the standard medium (294 mOsm x kg(-1)), the spent medium was exchanged with the fresh media with various osmolalities (294-640 mOsm x kg(-1)). The results obtained show that hyperosmotic pressure inhibited cell growth in a dose-dependent manner, though 14C6-bcl-2 cells were less susceptible to hyperosmotic pressure than SH2-0.32-(Delta)bcl-2 cells. At 522 mOsm x kg(-1), SH2-0.32-(Delta)bcl-2 cells underwent a gradual cell death mainly through apoptosis due to the cytotoxic effect of hyperosmotic pressure. In contrast, Bcl-2 overexpression in 14C6-bcl-2 cells could delay the apoptosis induced by 522 mOsm x kg(-1) by inhibiting caspase-3 activation. Bcl-2 overexpression could also improve the cellular membrane integrity of 14C6-bcl-2 cells. When subjected to hyperosmotic pressure, the specific antibody productivity of SH2-0.32-(Delta)bcl-2 cells and 14C6-bcl-2 cells was increased in a similar extent. As a result, the final antibody concentration achieved in 14C6-bcl-2 cells at 522 mOsm x kg(-1) was 2.5-fold higher than that at 294 mOsm x kg(-1). At 580 mOsm x kg(-1), acute hyperosmotic pressure induced the rapid loss of viability in both SH2-0.32-(Delta)bcl-2 and 14C6-bcl-2 cells through necrosis rather than through apoptosis. Taken together, Bcl-2 overexpression and optimized hyperosmotic pressure could improve the antibody production of rCHO cells.     

Partial phase diagram of aqueous bovine carbonic anhydrase: analyses of the pressure-dependent temperatures of the low- to physiological-temperature nondenaturational conformational change and of unfolding to the molten globule state

At 1.0 atm pressure and in 150 mM sodium phosphate (pH = 7.0), bovine carbonic anhydrase undergoes a nondenaturational conformational change at 30.3 degrees C and an unfolding transition from the physiological conformer to the molten globule state at 67.4 degrees C. The pressure dependences of the temperatures of these transitions have been studied under reversible conditions for the purpose of understanding DeltaH degrees , DeltaS degrees , and DeltaV for each conformational change. Temperatures for the low-temperature to physiological-temperature conformational change T(L-->P) are obtained from physiologically relevant conditions using slow-scan-rate differential scanning calorimetry. Temperatures for the physiological-temperature conformation to molten globule state conversion T(P-->MG) are obtained from differential scanning calorimetry measurements of the apparent transition temperature in the presence of guanidine hydrochloride extrapolated to zero molar denaturant. The use of slow-scan-rate differential scanning calorimetry permits the calculation of the activation volume for the conversion of the low-temperature conformer to the physiological-temperature conformer DeltaV(double dagger)(L-->P). At 1.0 atm pressure, the transition from the low-temperature conformer to the physiological-temperature conformer involves a volume change DeltaV(L-->P) = 15 +/- 2 L/mole, which contrasts with the partial unfolding of the physiological-temperature conformer to the molten globule state (DeltaV(P-->MG) = 26 +/- 9 L/mole). The activation volume for this process DeltaV(double dagger)(L-->P) = 51 +/- 9 L/mole and is consistent with a prior thermodynamic analysis that suggests the conformational transition from the low-temperature conformation to the physiological-temperature conformation possesses a substantial unfolding quality. These results provide further evidence the structure of the enzyme obtained from crystals grown below 30 degrees C should not be regarded as the physiological structure (the normal bovine body temperature is 38.3 degrees C). These results should therefore have implications in any area that seeks to correlate the crystal structure of bovine carbonic anhydrase to physiological function.     

Comparative mortality of diapausing and nondiapausing larvae of Plodia interpunctella (Lepidoptera: Pyralidae) exposed to monoterpenoids and low pressure

Monoterpenoids and low pressure have each been demonstrated to cause mortality of stored-product insect pests. The current report investigated the prospects of integrating the two methods in the management of diapausing and nondiapausing larvae of Plodia interpunctella (Hübner). In a separate experiment, the larvae were exposed to 35.5 mmHg in Erlenmeyer flasks at 19 and 28 degrees C for times ranging from 30 min to 96 h. Another set of experiments was conducted to investigate the toxicity of exposing P. interpunctella larvae to monoterpenoids including E-anethole, estragole, S-carvone, linalool, L-fenchone, geraniol, gamma-terpinene, and DL-camphor alone or in combination with low pressure (50 mmHg). Lethal times (LT) determined by subjecting time-mortality data to probit analyses were shortened to half when both diapausing and nondiapausing larvae were exposed to low pressure at 28 degrees C compared with 19 degrees C. Exposure of diapausing larvae to a monoterpenoid alone, with the exception of DL-camphor and estragole, at a concentration of 66.7 microl/1L of volume required > 30 h to generate 99% mortality at 19.0 +/- 0.8 degrees C. However, the LT99 values for diapausing and nondiapausing larvae exposed to combinations of DL-camphor or estragole and low pressure were considerably shortened. Combinations involving the rest of the monoterpenoids investigated and low pressure did not generate LT99 that were shorter than those of the control, which was low pressure only. These results suggest that integrating low pressure with DL-camphor or estragole could be a new method for the control of diapausing larvae of P. interpunctella at cooler temperatures.     

Effect of tracheal smooth muscle tone on collapsibility of immature airways

To test the influence of smooth muscle tone on extremely immature airways, tracheal segments (n = 19) were excised from premature lambs at 114-121 days gestation and mounted in a chamber filled with Krebs solution. Inflation (Si) and collapsing (Sc) compliance were determined by altering transmural pressure from 30 to 0 Torr and -30 to 0 Torr, respectively, both during control (C) and after acetylcholine (ACh) administration (experimental, E). Flow (V = 2-15 l/min) was then introduced through the tracheal lumen while chamber pressure (Pc) was increased from 0 to 30 Torr and driving pressure (Pd) was recorded for both C and E conditions. Tracheae were found to be extremely compliant; both Si and Sc were significantly (P less than 0.005) lower after ACh administration. Resistance to airflow (R = Pd/V) was also significantly (P less than 0.05) lower after ACh administration at each compressive pressure and each flow value. These results suggest that the highly compliant preterm trachea exhibits pressure-flow characteristics similar to a Starling resistor, and the effects of compressive pressures may be attenuated by ACh-induced smooth muscle contraction. Comparison of these results with data from adult and newborn animals suggests a developmental difference in tracheal mechanics and pressure-flow relationships, as well as in the way airway function is altered by smooth muscle stimulation.     

Effect of positive-pressure breathing on cardiovascular and thermoregulatory responses to exercise

Five healthy male volunteers performed 20 min of both seated and supine cycle-ergometer exercise (intensity, 50% maximal O2 uptake) in a warm environment (Tdb = 30 degrees C, relative humidity = 40-50%) with and without breathing 10 cmH2O of continuous positive airway pressure (CPAP). The final esophageal temperature (Tes) at the end of 20 min of seated exercise was significantly higher during CPAP (mean difference = 0.18 +/- 0.04 degree C, P less than 0.05) compared with control breathing (C). The Tes threshold for forearm vasodilation was significantly higher (P less than 0.05) during seated CPAP exercise than C (C = 37.16 +/- 0.13 degrees C, CPAP = 37.38 + 0.12 degree C). The highest forearm blood flow (FBF) at the end of exercise was significantly lower (P less than 0.05) during seated exercise with CPAP (mean +/- SE % difference from C = -30.8 +/- 5.8%). During supine exercise, there were no significant differences in the Tes threshold, highest FBF, or final Tes with CPAP compared with C. The added strain on the cardiovascular system produced by CPAP during seated exercise in the heat interacts with body thermoregulation as evidenced by elevated vasodilation thresholds, reduced peak FBF, and slightly higher final esophageal temperatures.     

Trolox C, a lipid-soluble membrane protective agent, attenuates myocardial injury from ischemia and reperfusion

The lipophilic antioxidant Trolox C, a vitamin E analog, was administered to isolated, buffer-perfused rabbit hearts subjected to 25 min of global stop-flow ischemia and 30 min of reperfusion. In six hearts, Trolox C (200 microM) was infused for 15 min immediately prior to ischemia and for the first 15 min of reperfusion. Six control hearts received only vehicle. Gas chromatography analysis confirmed that effective myocardial levels of Trolox were attained. At 30 min reperfusion, the recovery of left ventricular developed pressure was 56 +/- 3% of baseline in control hearts versus 70 +/- 4% in Trolox-treated hearts (p < .01). There was also significant improvement in recovery of Trolox-treated hearts in diastolic pressure and both maximum and minimum values of the first derivative of left ventricular pressure (dP/dt). Creatine phosphokinase release into the coronary effluent at 30 min of reperfusion was 16.5 +/- 8.4 IU/min in untreated and 6.3 +/- 1.0 IU/min (p < .05) in Trolox-treated hearts. Thus Trolox C, a lipophilic antioxidant, attenuated myocardial injury during stop-flow ischemia and reperfusion.     

The effects of skin pressure by clothing on circadian rhythms of core temperature and salivary melatonin

The present experiment investigated the effects of skin pressure by foundation garments (girdle and brassiere) on the circadian rhythms of core temperature and salivary melatonin. Ten healthy females (18-23 years) maintained regular sleep-wake cycles for a week prior to participation in the experiment. The experiments were performed from June to August 1999 using a bioclimatic chamber controlled at 26.5 degrees C +/- 0.2 degrees C and 62% +/- 3% RH. Ambient light intensity was controlled at 500 lux from 07:30 to 17:30, 100 lux from 17:30 to 19:30, 20 lux from 19:30 to 23:30; there was total darkness from 23:30 to 07:30. The experiment lasted for 58h over three nights. The participants arose at 07:30 on the first full day and retired at 23:30, adhering to a set schedule for 24h, but without wearing foundation garments. For the final 24h of the second full day, the subjects wore foundation garments. Rectal and leg skin temperatures were measured continuously throughout the experiment. Saliva and urine were collected every 4h for the analysis of melatonin and catecholamines, respectively. Skin pressure applied by the foundation garments was in the range 11-17 gf/cm2 at the regions of the abdomen, hip, chest, and back. The main results were as follows: (1) Rectal temperatures were significantly higher throughout the day and night when wearing foundation garments. (2) The nocturnal level of salivary melatonin measured at 03:30 was 115.2 +/- 40.4 pg/mL (mean +/- SEM, N = 10) without and 51.3 +/- 18.4 pg/mL (mean +/- SEM, N = 10) with foundation garments. (3) Mean urinary noradrenaline excretion was significantly lower throughout the day and night when wearing foundation garments (p < .05), but mean urinary adrenaline excretion was not different. The results suggest that skin pressure by clothing could markedly suppress the nocturnal elevation of salivary melatonin, resulting in an increase of rectal temperature.