Peripheral nerve at extreme low temperatures 1: Effects of temperature on the action potential

Hypothermia is an important means of neuroprotection. Understanding the effects of temperature on a physiologic measurement such as the nerve action potential (NAP) is important in monitoring its effects. The effects of hypothermia on the NAP amplitude, conduction velocity, and response to paired pulse stimulation were quantified in a rat sciatic nerve preparation from 37 to 10 °C. The time course of temperature related changes and the effect of repeated cycles of cooling and rewarming are explored using the following measures of the NAP: peak-to-peak amplitude, conduction velocity, duration, area under the curve and response to paired pulse stimuli. The NAP amplitude initially increases as temperature is reduced to 27 °C and then drops to roughly 50% of its baseline value by 16 °C while the area under the curve increases gradually until it begins to decline at 16 °C. Permanent loss of the NAP appears only after cooling below 10 °C for extended periods. Although the dependence of amplitude on temperature is approximately sigmoidal, the conduction velocity declines linearly at a rate of 2.8 m/s/°C. The response to paired pulse stimulation is strongly dependent on both temperature and the interstimulus interval with the responses at shorter interstimulus intervals being more temperature sensitive. With repetitive cycles of cooling and rewarming, the NAP amplitude declines by roughly 4% with every cycle without changes in the temperature at which the NAP amplitude reaches 50% of baseline. Only minor differences in conduction velocity are seen during cooling and rewarming.     

Cryopreservation of carotid artery segments via vitrification subject to marginal thermal conditions: correlation of freezing visualization with functional recovery

Cryopreservation is a well-established technique for long-term storage of viable cells and tissues. However, in recent years, application of established cryobiological principles to the preservation of multicellular tissues and organs has demanded considerable attention to ways of circumventing the deleterious effects of ice and thermal stresses in bulky tissues. As part of a multidisciplinary research program designed to study the interactions of thermo-physical events with tissue preservation, we report here on the implementation of a slow cooling (3 °C/min) and slow warming (62 °C/min) regimen towards scale-up of vitreous preservation of large tissue samples. Specifically, the correlation of thermo-physical events during vitrification of carotid artery segments with function recovery is reported using marginal thermal conditions for achieving vitrification in bulky samples. Moreover, the outcome is compared with a similar study reported previously using a 3-fold higher rate of rewarming (186 ± 13 °C/min). Tissue vitrification using an 8.4 M cryoprotectant cocktail solution (VS55) was achieved in 1 ml samples by imposing a low (2.6 ± 0.1 °C/min) cooling rate, between −40 and −100 °C, and a low rewarming rate (62 ± 4 °C/min) between −100 and −40 °C. Following cryoprotectant removal, the artery segments were cut into 3-4 mm rings for function testing on a contractility apparatus by measuring isometric responses to four agonist and antagonists (norepinephrine, phenylepinephrine, calcium ionophore and sodium nitroprusside). In addition, non-specific metabolic function of the vessel rings was determined using the REDOX indicator alamarBlue. Contractile function, normalized to untreated control samples, in response to the agonists norepinephrine and phenylepinephrine was significantly better in the slowly rewarmed group of carotid segments (74 ± 9% and 62 ± 11%, respectively) than for the more rapidly warmed group 31 ± 7% and 45 ± 15%, respectively). However, EC₅₀ sensitivities were not significantly different between the groups. Thermo-physical events such as ice formation and fractures were monitored throughout the cooling and warming phases using cryomacroscopy with the aid of a purpose-built borescope device. This technique allowed a direct observation of the visual impact of ice formation on specific zones along the blood vessel segment where, in most cases, no ice formation or fractures were observed in the vicinity of the artery segments. However, in specific instances when some ice crystallization was observed to impact the artery segment, the subsequent testing of function revealed a total loss of contractility. The successful vitrification of blood vessel segments using marginal conditions of slow cooling and rewarming, provide essential information for the development of scale-up protocols that is necessary when clinically relevant size samples need to be cryopreserved in an essentially ice-free state. This information can further be integrated into computer simulations of heat transfer and thermo-mechanical stress, where the slowest cooling rate anywhere in the simulated domain must exceed the critical values identified in the current study.     

Parallel DNA amplification by convective polymerase chain reaction with various annealing temperatures on a thermal gradient device

We present a thermal gradient convective polymerase chain reaction (PCR) for parallel DNA amplification with different annealing temperatures. The thermal gradient for microfluidic gradient PCR is produced by an innovative fin design whose formation principle is given. Without the need for a pump, the buoyancy forces continuously circulate reagents in a closed loop through different thermal zones, which brings self-actuated convective-flow PCR. In our prototype, we measured a temperature difference of about 45 °C along the gradient direction on the copper flake (45 × 40 × 4 mm). When the temperature of the hot zone is 90-97 °C and the temperature of the cold zone is 60-70 °C, the convection triggered two-temperature amplification of 112-bp fragment of Escherichia coli DNA. The time for amplification is less than 45 min. Interestingly, parallel DNA amplification with different annealing temperatures ranging from 60 to 70 °C was performed by this method. The PCR thermocycler demonstrated herein can be further scaled down and the loop length can be further reduced, and therefore the PCR times can be further reduced. These devices are suited as a platform for a new generation of low-power, portable DNA analysis systems.     

Effects of acclimation and egg-incubation temperature on selected temperature by hatchling western painted turtles (Chrysemys picta bellii)

The ability of hatchling turtles to detect environmental temperature differences and to effectively select preferred temperature is a function that critically impacts survival. In some turtle species, temperature preference may be influenced by embryonic and post-hatching conditions, such as egg-incubation and acclimation temperature. We tested for effects of embryonic incubation temperature (27.5 °C, 30 °C) and acclimation temperature (20 °C, 25 °C) on the selected temperature and movement patterns of 32 Chrysemys picta bellii (Reptilia: Emydidae) hatchlings in an aquatic thermal gradient of 14-34 °C and in single-temperature (20 °C, 25 °C) control tests. Among 10-11 month old hatchlings, acclimation temperature and egg-incubation temperature influenced temperature selection and movement patterns. Acclimation temperature affected activity and movement: in thermal gradient and single-temperature control tests, 25 °C-acclimated turtles relocated between chambers significantly more frequently than individuals acclimated to 20 °C. Acclimation temperature also affected temperature selection: 20 °C-acclimated turtles selected a specific temperature during gradient tests, but 25 °C-acclimated turtles did not. Among 20 °C-acclimated turtles, egg-incubation temperature was inversely related to selected temperature: hatchling turtles incubated at 27.5 °C selected the warmest temperature available (34 °C); individuals incubated at 30 °C selected the coldest temperature (14 °C). These results suggest that interactions of environmental conditions may influence post-hatching thermoregulatory behavior in C. picta bellii, a factor that ultimately affects fitness.     

Response of hatchling wood turtles (Glyptemys insculpta) to an aquatic thermal gradient

Effective thermoregulation and the ability to select preferred temperature is an important factor influencing fitness in hatchling and juvenile turtles. Six-month-old Glyptemys insculpta acclimated to 20 °C selected the warmest temperature available and avoided the coldest temperature available in a gradient of 12–27 °C. Turtles visited fewer chambers and switched chambers in the gradient tank less frequently when the gradient was present than during control tests. Mean selection of chambers differed between control and gradient tests across all temperatures except at 21 °C, the temperature closest to the acclimation and control temperature (20 °C).     

Optimisation of a microwave pretreatment of wheat straw for methane production

This study aims at the optimisation of a microwave pretreatment for wheat straw solubilisation and anaerobic biodegradability. The maximum yield of methane production was obtained at 150 °C with an improvement of 28% compared to an untreated sample. In addition, at this temperature, the time to reach 80% of the methane volume obtained from untreated straw was about 35%. The study of ramp time and holding time at targeted temperature showed that they had no improvement effect. Thus, the best conditions are the highest heating rate for a final temperature 150 °C without any holding time. The reading of energy consumed by pretreatment and energy overproduced by pretreated samples showed that increasing tVS amount and heating rate led to a saving of energy consumption. Nevertheless, to obtain a positive energy balance, a microwave device should consume less than 2.65 kJ/g_tVS.     

Akt and phosphoinositide regulation and wortmannin-dependent induction of phospho-Akt in 3T3-L1 adipocytes on cold exposure followed by rewarming

We exposed 3T3-L1 adipocytes to cooling followed by rewarming and examined phospho-molecule induction, including that of phospho-Akt, and the mechanism underlying the induction in these exposed cells; the viability did not deteriorate. Cooling at 4 °C increased the phospho-Thr-308 Akt level. On subsequent rewarming at 37 °C, phospho-Ser-473 Akt expression was induced in a wortmannin (PI3K inhibitor)-dependent and anisomycin (ribotoxic agent)-independent manner without decrease in the phospho-Thr-308 Akt level, and Akt-mediated phosphorylation and PI 3,4,5-trisphosphate accumulation were induced. Our results suggest that cooling followed by rewarming induces phospho-molecules through different responses of regulatory molecules, including the target molecule, to changes in temperature.     

The effect of thermopreference on circadian thermoregulation in sprague-dawley and fisher 344 rats

Interstrain differences in thermoregulation of rats are important in biomedical research because subtleties in thermoregulatory sensitivities may greatly affect data collected. Little is known regarding how individual rodent strains differentially utilize behavioral thermal preference to regulate core temperature (Tc). Sprague-Dawley (SD) and Fischer 344 (F344) rats are known to have differences in thermoregulation including heat tolerance and are useful models to study interstrain differences in thermoregulation. Adult male SD and F344 rats of similar body size were implanted with radiotelemetry thermoprobes (DSI) to measure Tc and MA and housed in either a longitudinal temperature gradient with an ambient temperature (Ta) range of ∼15–40 °C to measure selected Ta (STa) or control environment maintained at a Ta of 23 °C. When continuously monitored for 48 h, Tc and MA increased at night, while STa decreased, according to their normal circadian cycle in both strains. SD rats were more active than F344 rats throughout the circadian cycle (SD gradient: day=12.9±1.2 m/h, night=32.1±2.4 m/h; F344 gradient: day=4.1±0.6 m/h, night=16.8±1.8 m/h; p<0.05 interstrain and circadian effects). The STa of each strain was greater during the daytime (SD: 26.4±0.2 °C; F344: 27.8±0.3 °C) than at night (SD: 24.7±0.3 °C; F344: 25.7±0.3 °C) confirming past studies that thermopreference during the day and night is greater than standard room temperature (∼23 °C). Correlations between MA and Tc suggest that MA has a greater effect on Tc in the F344 but not the SD strain when housed in a temperature gradient. There were significant strain differences in Tc depending on whether rats were housed in a temperature gradient. That is, the control F344 rats had a lower Tc during the transition from dark to light compared to rats housed in a gradient. Tc of the SD strain was unaffected by housing in the gradient. Rats are typically housed at a standard room temperature of 23 °C. However, the results demonstrate that when given the opportunity to behaviorally thermoregulate in a temperature gradient, the F344 strain selects a warmer environment that affects the regulation of Tc. This may be important in the experimenters' choice of ambient temperatures to house and study rats and other rodents.     

Preparation of crystalline starch nanoparticles using cold acid hydrolysis and ultrasonication

Waxy maize starch in an aqueous sulfuric acid solution (3.16 M, 14.7% solids) was hydrolyzed for 2–6 days, either isothermally at 40 °C or 4 °C, or at cycled temperatures of 4 and 40 °C (1 day each). The starch hydrolyzates were recovered as precipitates after centrifuging the dispersion (10,000 rpm, 10 min). The yield of starch hydrolyzates depended on the hydrolysis temperature and time, which varied from 6.8% to 78%. The starch hydrolyzed at 40 °C or 4/40 °C exhibited increased crystallinity determined by X-ray diffraction analysis, but melted in broader temperature range (from 60 °C to 110 °C). However, the starch hydrolyzed at 4 °C displayed the crystallinity and melting endotherm similar to those of native starch. The starch hydrolyzates recovered by centrifugation were re-dispersed in water (15% solids), and the dispersion was treated by an ultrasonic treatment (60% amplitude, 3 min). The ultrasonication effectively fragmented the starch hydrolyzates to nanoparticles. The hydrolyzates obtained after 6 days of hydrolysis were more resistant to the ultrasonication than those after 2 or 4 days, regardless of hydrolysis temperatures. The starch nanoparticles could be prepared with high yield (78%) and crystallinity by 4 °C hydrolysis for 6 days followed by ultrasonication. Scanning electron microscopy revealed that the starch nanoparticles had globular shapes with diameters ranging from 50 to 90 nm.     

Myocardial mechanical dysfunction and calcium overload following rewarming from experimental hypothermia in vivo

Rewarming patients from accidental hypothermia are regularly complicated with cardiovascular instability ranging from minor depression of cardiac output to fatal circulatory collapse also termed “rewarming shock”. Since altered Ca²⁺ handling may play a role in hypothermia-induced heart failure, we studied changes in Ca²⁺ homeostasis in in situ hearts following hypothermia and rewarming. A rat model designed for studies of the intact heart in a non-arrested state during hypothermia and rewarming was used. Rats were core cooled to 15 °C, maintained at 15 °C for 4 h and thereafter rewarmed. As time-matched controls, one group of animals was kept at 37 °C for 5 h. Total intracellular myocardial Ca²⁺ content ([Ca²⁺]_i) was measured using ⁴⁵Ca²⁺. Following rewarming we found a significant reduction of stroke volume and cardiac output compared to prehypothermic control values as well as to time-matched controls. Likewise, we found that hypothermia and rewarming resulted in a more than six-fold increase in [Ca²⁺]_i to 3.01 ± 0.43 μmol/g dry weight compared to 0.44 ± 0.05 μmol/g dry weight in normothemia control. These findings indicate that hypothermia-induced alterations in the Ca²⁺-handling result in Ca²⁺ overload during hypothermia, which may contribute to myocardial failure during and after rewarming.     

Determination of the γ-secretase inhibitor MK-0752 in human plasma by online extraction and electrospray tandem mass spectrometry (HTLC–ESI-MS/MS)

A sensitive and rapid HTLC–ESI-MS/MS method with an advanced online sample preparation was developed for determination of the γ-secretase inhibitor MK-0752 in human plasma using an internal standard. Plasma samples (100 μL) were diluted and injected directly onto an online extraction column (Cohesive Cyclone MAX 0.5 mm × 50 mm, >30 μm), the sample matrix was washed out with an aqueous solution, and retained analytes were eluted out and transferred directly to the analytical column (Phenomenex Gemini 3μ C18 110A, 50 mm × 2.0 mm at 50 °C) for separation using a gradient mobile phase. The eluted analytes were then detected on an API-3000 LC–MS/MS System with ESI and a negative multiple reaction monitoring mode. The monitored ion transitions were m/z 441 → 175 for MK-0752 and 496 → 175 for the internal standard. Online extraction recoveries were 81%. The method was validated and was linear in the range of 0.05–50 μg/mL. Within-day and between-day precisions were < 8.6%, and accuracies were 0.7 and 7.1%. This method was applied to the measurement of plasma MK-0752 levels in a Phase I study of pediatric patients with recurrent or refractory brain tumors.     

Response of juvenile diamond-backed terrapins (Malaclemys terrapin) to an aquatic thermal gradient

In many ectotherms, selection of environmental thermal niches may positively affect growth, nutrient assimilation rates, immune system function, and ultimately survival. Temperature preference in some turtle species may be influenced by environmental conditions, including acclimation temperature. We tested for effects of acclimation temperature (22 °C, 27 °C) on the selected temperature and movement patterns of 14 juvenile Malaclemys terrapin (Reptilia: Emydidae) in an aquatic thermal gradient of 14–34 °C and in single-temperature (22 °C, 27 °C) control tests. Among 8–10 month old terrapins, acclimation temperature influenced activity and movement patterns but did not affect temperature selection. In thermal gradient and single-temperature control tests, turtles acclimated to 27 °C used more tank chambers and relocated between chambers significantly more frequently than individuals acclimated to 22 °C. However, acclimation temperature did not affect temperature selection: both 22- and 27 °C-acclimated turtles selected the warmest temperature (34 °C), and avoided the other temperatures available, during thermal gradient tests. These results suggest that young M. terrapin are capable of detecting small temperature increments and prefer warm temperatures that may positively influence growth and metabolism.     

Cryopreservation of periodontal ligament cells with magnetic field for tooth banking

The purpose of this study was to establish a long-term tooth cryopreservation method that can be used for tooth autotransplantation. Human periodontal ligament (PDL) cells were frozen in 10% dimethyl sulfoxide (Me₂SO) using a programmed freezer with a magnetic field. Cells were cryopreserved for 7 days at −150 °C. Immediately after thawing, the number of surviving cells was counted and the cells were cultured; cultured cells were examined after 48 h. Results indicated that a 0.01 mT of a magnetic field, a 15-min hold-time, and a plunging temperature of −30 °C led to the greatest survival rate of PDL cells. Based on these findings, whole teeth were cryopreserved under the same conditions for 1 year. The organ culture revealed that the PDL cells of cryopreserved tooth with a magnetic field could proliferate as much as a fresh tooth, although the cells did not appear in the cryopreserved tooth without a magnetic field. Histological examination and the transmission electron microscopic image of cryopreserved tooth with a magnetic field did not show any destruction of cryopreserved cells. In contrast, severe cell damage was seen in cells frozen without a magnetic field. These results indicated that a magnetic field programmed freezer is available for tooth cryopreservation.     

Rewarming rates of two large hibernators: Comparison of a monotreme and a eutherian

We measured body temperatures in two large hibernating mammals, the eutherian alpine marmot (Marmota marmota) and the egg-laying echidna (Tachyglossus aculeatus) from unrestrained animals in their natural environment. In both species hibernation is broken every 13 days on average by rewarming to euthermic temperatures. We found that the time course of a rewarming could be closely fitted with a sigmoid curve, allowing calculation of peak rewarming rate and corresponding body temperature. Maximum rewarming rates were twice as high in marmots as in echidnas (12.1±1.3 °C h⁻¹, n=10 cf. 6.2±1.2 °C h⁻¹, n=10). Peak rewarming rates were positively correlated with body temperature in echidnas, but negatively correlated in marmots.     

Effect of age and body size on selected temperature by juvenile wood turtles (Glyptemys insculpta)

In an aquatic thermal gradient of 15–30 °C, 3-, 6-, and 12-month-old juvenile wood turtles (Glyptemys insculpta) acclimated to 20 °C selected the warmest temperature available (30 °C) and avoided the coldest temperatures available (15 and 18 °C). Mean selection of chambers differed between control and gradient tests across all temperatures except 27 °C. Turtles of all age classes relocated between chambers less often when the gradient was present than during control tests. Six- and 12-month-old turtles selected 30 °C more frequently, and selected colder temperatures less frequently, than 3-month-old turtles, suggesting that the ability to select preferred temperatures is better developed in older hatchlings.     

The use of cryomicroscopy in guppy sperm freezing

The present study employed cryomicroscopy to derive an optimal sperm freezing protocol for guppy (Poecilia reticulata) sperm. Evaluation criteria during the freezing-thawing process were assessed for nucleation temperature (Tn), temperature when more than 50% of sperm display bending mid-piece (Tb), temperature when more than 80% of sperm stop moving (Tm), thawing temperature (Tt), and post-thaw motility. We compared four different cryoprotectants: 5% N-dimethyl formamide (DMF), 6% methanol (MEOH), 10% dimethyl sulfoxide (DMSO), and 14% glycerol, as well as glycerol at different concentrations of 7-50%; cooling and rewarming rates ranged from 5 to 100 °C/min. The protocol that yielded the highest post-thaw motility was samples suspended in 14% glycerol, cooled at 25 °C/min, and thawed at 100 °C/min, which was in complete agreement with our previous findings derived from a controlled-rate freezer. In addition, Tb and Tm were found to be negatively correlated with post-thaw motility, suggesting their possible role in predicting freezing success. The present study for the first time demonstrated the usefulness of cryomicroscopy in deriving an optimal sperm freezing protocol for aquatic species.     

Cooling rate optimization for zebrafish sperm cryopreservation using a cryomicroscope coupled with SYBR14/PI dual staining

The Zebrafish has gained increased popularity as an aquatic model species in various research fields, and its widespread use has led to numerous mutant strains and transgenic lines. This creates the need to store these important genetic materials as frozen gametes. Sperm cryopreservation in zebrafish has been shown to yield very low post-thaw survival and many protocols suffer from great variability and poor reproducibility. The present study was intended to develop a freezing protocol that can be reliably used to cryopreserve zebrafish sperm with high post-thaw survival. In particular, our study focused on cooling protocol optimization with the aid of cryomicroscopy. Specifically, sperm suspended in 8% DMSO or 4% MeOH were first incubated with live/dead fluorescent dyes (SYBR14/PI) and then cooled at various rates from 4 °C to different intermediate stopping temperatures such as −10, −20, −30 and −80 °C before rewarming to 35 °C at the rate of 100 °C/min. %PI-positive (dead) cells were monitored throughout the cooling process and this screening yielded an optimal rate of 25 °C/min for this initial phase of freezing. We then tested the optimal cooling rate for the second phase of freezing from various intermediate stopping temperatures to −80 °C before plunging into liquid nitrogen. Our finding yielded an optimal intermediate stopping temperature of −30 °C and an optimal rate of 5 °C/min for this second phase of freezing. When we further applied this two-step cooling protocol to the conventional controlled-rate freezer, the average post-thaw motility measured by CASA was 46.8 ± 6.40% across 11 males, indicating high post-thaw survival and consistent results among different individuals. Our study indicates that cryomiscroscopy is a powerful tool to devise the optimal cooling conditions for species with sperm that are very sensitive to cryodamage.     

Evaluation of cryoprotectant and cooling rate for sperm cryopreservation in the euryhaline fish medaka Oryzias latipes

Medaka Oryzias latipes is a well-recognized biomedical fish model because of advantageous features such as small body size, transparency of embryos, and established techniques for gene knockout and modification. The goal of this study was to evaluate two critical factors, cryoprotectant and cooling rate, for sperm cryopreservation in 0.25-ml French straws. The objectives were to: (1) evaluate the acute toxicity of methanol, 2-methoxyethanol (ME), dimethyl sulfoxide (Me₂SO), N,N-dimethylacetamide (DMA), N,N-dimethyl formamide (DMF), and glycerol with concentrations of 5%, 10%, and 15% for 60 min of incubation at 4 °C; (2) evaluate cooling rates from 5 to 25 °C/min for freezing and their interaction with cryoprotectants, and (3) test fertility of thawed sperm cryopreserved with selected cryoprotectants and associated cooling rates. Evaluation of cryoprotectant toxicity showed that methanol and ME (5% and 10%) did not change the sperm motility after 30 min; Me₂SO, DMA, and DMF (10% and 15%) and glycerol (5%, 10% and 15%) significantly decreased the motility of sperm within 1 min after mixing. Based on these results, methanol and ME were selected as cryoprotectants (10%) to evaluate with different cooling rates (from 5 to 25 °C/min) and were compared to Me₂SO and DMF (10%) (based on their use as cryoprotectants in previous publications). Post-thaw motility was affected by cryoprotectant, cooling rate, and their interaction (P ? 0.000). The highest post-thaw motility (50 ± 10%) was observed at a cooling rate of 10 °C/min with methanol as cryoprotectant. Comparable post-thaw motility (37 ± 12%) was obtained at a cooling rate of 15 °C/min with ME as cryoprotectant. With DMF, post-thaw motility at all cooling rates was ?10% which was significantly lower than that of methanol and ME. With Me₂SO, post-thaw motilities were less than 1% at all cooling rates, and significantly lower compared to the other three cryoprotectants (P ? 0.000). When sperm from individual males were cryopreserved with 10% methanol at a cooling rate of 10 °C/min and 10% ME with a rate of 15 °C/min, no difference was found in post-thaw motility. Fertility testing of thawed sperm cryopreserved with 10% methanol at a rate of 10 °C/min showed average hatching of 70 ± 30% which was comparable to that of fresh sperm (86 ± 15%). Overall, this study established a baseline for high-throughput sperm cryopreservation of medaka provides an outline for protocol standardization and use of automated processing equipment in the future.     

The use of a temperature-profiled position transducer for the study of low-temperature growth in Gramineae

A device is described for measuring linear extension of grass leaves during controlled cooling and heating of the growing region. The instrument was employed to investigate the sensitivity to temperature of the expanding third and fourth leaves of Lolium temulentum L. seedlings. Using a stepped temperature profile it was established that there was no lag in the response of growth rate to rapid changes in temperature below 16°C. If cooling was continued to the point where growth ceased (1°C) but no further, then rates of growth on rewarming were enhanced over the chilling range and reverted to the original rate at 20°C. Cooling to successively lower subzero temperatures before rewarming abolished the hysteretic enhancement, progressively raised the temperature at which growth resumed and decreased the rate of extension until, at-5.3°C, no recovery occurred. The temperature sensitivity of growth, measured as Q₁₀, was essentially constant when cooling from 20°C to 5°C, with 5°C-grown leaf tissue exhibiting a higher mean Q₁₀ than tissue developed at 20°C. The possible physiological significance of these data is discussed.Abbreviations LVDT linear variable displacement transformer - Pe, Fx temperatures at which growth ceases during cooling and resumes during rewarming     

A study of human skin and surface temperatures in stable and unstable thermal environments

Skin temperature is a common physiological parameter that reflects human thermal responses. The purpose of this research was to investigate the effects of radiant temperature on human skin temperature and surface temperature in stable and unstable thermal environments. For a clothed human body, the skin temperature is the surface temperature of the skin, while the surface temperature is the outer surface temperature of the clothes. For this aim, the radiant temperature from 26 to 38 °C and then from 38 to 26 °C was controlled in three different ways; in stable condition keeping stable above 40 min, in unstable condition at a rate of 2 °C/5 min, and in another unstable condition at a rate of 2 °C/10 min. Experimental data showed that at the same radiant temperature, the local skin/surface temperatures during the radiant temperature decrease were higher compared to those during the radiant temperature increase. During the radiant temperature increase/decrease, the increments/decrements of the mean skin temperature and the mean surface temperature decreased gradually from the stable condition, 2 °C/10 min to 2 °C/5 min. Compared to surface temperature, the faster the radiant temperature changed, the more obviously the change in skin temperature was delayed. These data demonstrated that the human body has physiological adaptability to unstable thermal environments.