Lethal Hyperthermia Research Articles

Heat tolerance and evaporative cooling capacity in an arid-zone elephant shrew

Endotherms vary widely in their capacity to defend sublethal body temperature (Tb) during acute heat exposure. Interspecific variation in the upper thermal limits of small mammals remains poorly studied, particularly in taxa other than bats. We hypothesised that rupicolous elephant shrews (Macroscelidae), on account of their occupancy of above-ground rock crevices in hot habitats and rapid cursorial locomotion, have evolved pronounced heat tolerance capacities. To test this hypothesis, we quantified relationships between Tb, resting metabolic rate (RMR) and evaporative water loss (EWL) in western rock elephant shrews (Elephantulus rupestris) exposed to air temperature (Tair) approaching or exceeding Tb under conditions of low humidity representative of the study population’s arid, winter-rainfall habitat in western South Africa. Our data revealed a pronounced capacity to tolerate Tair > Tb, with E. rupestris tolerating Tair up to 48.0 ± 0.1 °C while defending Tb > 7 °C below Tair (maximum Tb = 41.64 ± 0.16 °C). Three behavioural pathways were employed to dissipate heat: open mouth panting, flattening their body posture, and nose-licking. At the highest experimental Tair values achieved, EWL increased 15.05-fold relative to normothermic levels and heat was dissipated evaporatively at rates equivalent to 174%—240% of metabolic heat production. The heat tolerance limit of Tair = 48 °C in E. rupestris is one of the highest yet recorded in a non-volant small mammal, and our data support the notion that elephant shrews possess a pronounced capacity to avoid lethal hyperthermia during acute heat exposure.

Keeping cool with poop: Urohidrosis lowers leg surface temperature by up to 6°C in breeding White storks

AbstractStorks (Ciconiidae) are renowned for defecating on their legs when exposed to high temperatures, a phenomenon known as “urohidrosis.” Previous work suggested that this behavior can cool down the blood supply to the legs and thus prevent hyperthermia in captive storks when overheated. However, no study has quantified the magnitude or duration of its evaporative cooling effect in free‐ranging birds. Here, we combine urohidrosis data with thermal imaging and microclimate data to investigate the thermoregulatory significance of urohidrosis in White storks Ciconia ciconia during the breeding season. We show that urohidrosis can reduce leg surface temperature by up to 6.7°C (4.40 ± 1.04°C). Yet its cooling effect was of short duration (lasting no more than 2.5 min) and decreased with time since defecation. Thus, for urohidrosis to significantly contribute to heat dissipation, storks must perform it repeatedly when overheated. Indeed, an individual can perform up to 11 urohidrosis events per hour; our estimates indicate that repeated urohidrosis could represent a significant amount of heat loss during short time span, equivalent to 4% of daily field metabolic rate for an adult stork. Our results point to an absence of differences in the cooling efficiency of urohidrosis between adults and nestlings, probably because all nestlings were recorded during the last phase of the ontogeny of thermoregulation. Besides, during the hottest days, adult storks delivered water to their nestlings, which might allow them to replenish body water reserves to sustain urohidrosis and other heat dissipation behaviors such as panting or gular fluttering. Our results indicate that urohidrosis might buffer the impact of high temperatures in breeding storks, preventing overheating and lethal hyperthermia. Gaining knowledge about behavioral thermoregulation in the heat is therefore crucial to better predict the future persistence and vulnerability of species under different climate warming scenarios.

Challenges of quantifying direct heat stress effects of climate change on seabirds

The importance of heat stress as a consequence of climate climate change is often overlooked for seabirds. As endotherms, seabirds must actively thermoregulate at temperatures above their thermoneutral zone, or risk lethal hyperthermia. Although essential activities (e.g. foraging, breeding) may be traded off for thermoregulatory behaviors during periods of heat stress, a recent report by Olin et al. (2023; Mar Ecol Prog Ser https://doi.org/10.3354/meps14244, this Theme Section) is one of very few that directly link this to demography. We argue that heat stress effects, which have strong theoretical support, are underreported directly because large-scale mortality events are rare, and small-scale events are hard to identify and easily obscured by indirect trophic effects. Quantifying heat stress effects on seabirds is necessary to understand fully the threats from climate change but requires prioritizing research in the following areas: developing methods to attribute heat mortality, determining baseline levels of heat mortality, elucidating ecological and organismal differences that underlie heat stress sensitivity, investigating the importance of possible sublethal mechanisms, and separating heat stress trade-offs from indirect effects of climate.

Editorial: Mammalian responses to climate change: From organisms to communities

A particularly powerful component of the Anthropocene extinction is global climate change, which, in concert with other elements of anthropogenic forcing, is driving the reorganization of biotic assemblages everywhere on the planet. Rising ambient temperature challenges the adaptive capacity of organisms and for terrestrial animals it can directly cause lethal hyperthermia or dehydration. More generally however, the indirect effects of climate change permeate all facets of the environment including habitat quality (food, water, refuge availability) and risks from predators, competitors, and pathogens (Fig. 1). Chronic exposure to environmental conditions that exceed critical thresholds at critical times of year ultimately result in a convergence of sublethal fitness costs (Pattinson et al. 2022). Such costs could arise from disturbances to physiological processes and behavioral patterns, which become expressed at the population level as changes in vital rates and habitat occupancy. These in turn drive shifts in species ranges or even extinctions, with consequences at community and ecosystem levels. The papers in this Research Topic explore these interactions and their implications.

Hyperthermia and Heat Stress as Risk Factors for Sudden Infant Death Syndrome: A Narrative Review.

Background and ObjectivesHeat stress and hyperthermia are common findings in sudden infant death syndrome (SIDS) victims. It has been suggested that thermal stress can increase the risk of SIDS directly via lethal hyperthermia or indirectly by altering autonomic functions. Major changes in sleep, thermoregulation, cardiovascular function, and the emergence of circadian functions occur at the age at which the risk of SIDS peaks—explaining the greater vulnerability at this stage of development. Here, we review the literature data on (i) heat stress and hyperthermia as direct risk factors for SIDS, and (ii) the indirect effects of thermal loads on vital physiological functions.ResultsVarious situations leading to thermal stress (i.e., outdoors temperatures, thermal insulation from clothing and bedding, the prone position, bed-sharing, and head covering) have been analyzed. Hyperthermia mainly results from excessive clothing and bedding insulation with regard to the ambient thermal conditions. The appropriate amount of clothing and bedding thermal insulation for homeothermia requires further research. The prone position and bed-sharing do not have major thermal impacts; the elevated risk of SIDS in these situations cannot be explained solely by thermal factors. Special attention should be given to brain overheating because of the head's major role in body heat losses, heat production, and autonomic functions. Thermal stress can alter cardiovascular and respiratory functions, which in turn can lead to life-threatening events (e.g., bradycardia, apnea with blood desaturation, and glottal closure). Unfortunately, thermal load impairs the responses to these challenges by reducing chemosensitivity, arousability, and autoresuscitation. As a result, thermal load (even when not lethal directly) can interact detrimentally with vital physiological functions.ConclusionsWith the exception of excessive thermal insulation (which can lead to lethal hyperthermia), the major risk factors for SIDS appears to be associated with impairments of vital physiological functions when the infant is exposed to thermal stress.

How to Stay Cool: Early Acoustic and Thermal Experience Alters Individual Behavioural Thermoregulation in the Heat

Climate change is pushing organisms closer to their physiological limits. Animals can reduce heat exposure – and the associated risks of lethal hyperthermia and dehydration – by retreating into thermal refuges. Refuge use nonetheless reduces foraging and reproductive activities, and thereby potentially fitness. Behavioural responses to heat thus define the selection pressures to which individuals are exposed. However, whether and why such behavioural responses vary between individuals remains largely unknown. Here, we tested whether early-life experience generates inter-individual differences in behavioural responses to heat at adulthood. In the arid-adapted zebra finch, parents incubating at high temperatures emit “heat-calls,” which adaptively alter offspring growth. We experimentally manipulated individual early life acoustic and thermal experience. At adulthood, across two summers, we then repeatedly recorded individual panting behaviour, microsite use, activity (N = 2,402 observations for 184 birds), and (for a small subset, N = 23 birds) body temperature, over a gradient of air temperatures (26–38°C), in outdoor aviaries. We found consistent inter-individual variation in behavioural thermoregulation, and show for the first time in endotherms that early-life experience contributes to such variation. Birds exposed prenatally to heat-calls started panting at lower temperatures than controls but panted less at high temperatures. It is possible that this corresponds to a heat-regulation strategy to improve water saving at high temperature extremes, and/or, allow maintaining high activity levels, since heat-call birds were also more active across the temperature gradient. In addition, microsite use varied with the interaction between early acoustic and thermal experiences, control-call birds from cooler nests using the cooler microsite more than their hot-nest counterparts, whereas the opposite pattern was observed in heat-call birds. Overall, our study demonstrates that a prenatal acoustic signal of heat alters how individuals adjust behaviourally to thermal challenges at adulthood. This suggests that there is scope for selection pressures to act differently across individuals, and potentially strengthen the long-term fitness impact of early-life effects.

Mortality among birds and bats during an extreme heat event in eastern South Africa

AbstractHeat‐related mortality events involving birds and bats are projected to occur more frequently as a result of anthropogenic global heating. Reports of mass mortalities associated with extreme heat have, over the last decade, mostly involved Australian birds and pteropodid flying‐foxes. Here, we report a mortality event involving ~110 birds and fruit bats in eastern South Africa in early November 2020 when maximum air temperatures (Tmax) reached 43–45°C and relative humidities were 21–23%. The mortalities included 47 birds of 14 species, all but three of which were passerines, and ~60 Wahlberg’s epauletted fruit bats (Epomophorus wahlbergi). This mortality event occurred on a single very hot day preceded by several cooler days (Tmax = 37–39°C at one location) and involved weather conditions similar to those associated with at least one recent flying‐fox die‐off in Australia. The disproportionately high representation of passerines among the avian mortalities supports recent predictions that songbirds are more vulnerable to lethal hyperthermia on account of the relative inefficiency of panting as an avenue of evaporative heat dissipation. As far as we are aware, this is the first documented heat‐related mortality event involving wild birds and bats in southern Africa.

Regularly drinking desert birds have greater evaporative cooling capacity and higher heat tolerance limits than non‐drinking species

Abstract Surface water is a critical resource for many birds inhabiting arid regions, but the implications of regular drinking and dependence on surface water for the evolution of thermal physiology remain largely unexplored. We hypothesized that avian thermoregulation in the heat has evolved in tandem with the use of surface water and predicted that (a) regularly drinking species have a greater capacity to elevate rates of evaporative water loss (EWL) compared to non‐drinking species, and (b) heat tolerance limits (HTLs) are higher among drinking species. To test these predictions, we quantified thermoregulatory responses to high air temperature (Ta) in 12 species of passerines from the South African arid zone and combined these with values for an additional five species. We categorized each species as either: (a) water‐dependent, regularly drinking, or (b) water‐independent, occasional‐/non‐drinking. In support of our predictions, both conventional and phylogenetically independent analyses revealed that evaporative scope (the ratio of maximum EWL to minimum thermoneutral EWL) during acute heat exposure was significantly higher among drinking species. Moreover, evaporative scope was significantly and positively related to HTL (i.e. maximum air temperature tolerated before the onset of severe hyperthermia). These findings offer new insights into the co‐evolution of water dependence, movement ecology and thermal physiology in the context of trade‐offs between dehydration avoidance and resistance to lethal hyperthermia in hot desert habitats. A free Plain Language Summary can be found within the Supporting Information of this article.

Avian mortality risk during heat waves will increase greatly in arid Australia during the 21st century.

Intense heat waves are occurring more frequently, with concomitant increases in the risk of catastrophic avian mortality events via lethal dehydration or hyperthermia. We quantified the risks of lethal hyperthermia and dehydration for 10 Australian arid-zone avifauna species during the 21st century, by synthesizing thermal physiology data on evaporative water losses and heat tolerance limits. We evaluated risks of lethal hyperthermia or exceedance of dehydration tolerance limits in the absence of drinking during the hottest part of the day under recent climatic conditions, compared to those predicted for the end of this century across Australia. Increases in mortality risk via lethal dehydration and hyperthermia vary among the species modelled here but will generally increase greatly, particularly in smaller species (~10–42 g) and those inhabiting the far western parts of the continent. By 2100 CE, zebra finches’ potential exposure to acute lethal dehydration risk will reach ~ 100 d y−1 in the far northwest of Australia and will exceed 20 d y−1 over > 50% of this species’ current range. Risks of dehydration and hyperthermia will remain much lower for large non-passerines such as crested pigeons. Risks of lethal hyperthermia will also increase substantially for smaller species, particularly if they are forced to visit exposed water sources at very high air temperatures to avoid dehydration. An analysis of atlas data for zebra finches suggests that population declines associated with very hot conditions are already occurring in the hottest areas. Our findings suggest that the likelihood of persistence within current species ranges, and the potential for range shifts, will become increasingly constrained by temperature and access to drinking water. Our model adds to an increasing body of literature suggesting that arid environments globally will experience considerable losses of avifauna and biodiversity under unmitigated climate change scenarios.

High air temperatures induce temporal, spatial and social changes in the foraging behaviour of wild zebra finches

Understanding how heatwaves affect organisms is becoming an important issue in animal behaviour, given the changing climate. Exposure to high air temperatures can lead to lethal hyperthermia, when individuals are no longer able to maintain body temperature within their optimal physiological range. Animals will rapidly adjust their behaviour, prioritizing heat dissipation through activities such as drinking and sitting in shade to maintain their body temperature over other activities, such as foraging. Here, we used an automated logging system to consider both the spatial and temporal foraging patterns under a range of different air temperatures at an individual level, in a strictly granivorous species in the wild. We continuously monitored individual foraging activity of wild zebra finches, Taeniopygia guttata, a species well adapted to arid conditions, in an Australian arid area across two heatwave events. High air temperatures significantly reduced foraging activity, with the extent of this effect depending on the time of day. They also led to a significant decrease in the number of birds foraging together and to birds spending a higher proportion of their foraging activity close to a water supply. As temperatures exceeded 35 °C we saw a significant escalation of heat dissipation behaviour. Our results indicate that extreme air temperatures significantly affected temporal, social and spatial characteristics of zebra finch foraging behaviour and these are likely to adversely reduce an individual's capacity to forage efficiently, and consequently its food intake in the short term, while also potentially having implications for both reproduction and survival in the long term.

118 - Induction of peroxiredoxins by heat shock is mediated by reactive oxygen species and Nrf2

Hyperthermia is a promising anticancer treatment used in combination with radiotherapy and/or chemotherapy. It uses heat (>42°C) to kill cancer cells. However, lower temperatures (39-41°C) induce thermotolerance, an adaptive survival response (hormesis) that could interfere with toxicity of cancer treatments. Exposure to low doses of stress (e.g. heat shock, reactive oxygen species (ROS)) can upregulate cellular defenses such as heat shock proteins, antioxidants and anti-apoptosis proteins, which protect cells against subsequent exposure to toxic stressors. Although hyperthermia has proven effective in clinical trials, there are still many things to learn about its cellular and molecular mechanisms. This study examines the role of ROS and antioxidants in cellular stress responses to mild (40°C) and lethal (42-43°C) hyperthermia. Hyperthermia (42-43°C) caused apoptosis and activation of caspases 9 and 3, which were decreased in thermotolerant (40°C) cancer cells. Mild and lethal hyperthermia both increased ROS levels in cells, relative to 37°C. Exposure to mild and lethal hyperthermia increased protein expression of the antioxidants peroxiredoxin-2 (Prx2) and Prx3. Expression of the hyperoxidized form Prx-SO3 increased only at higher temperatures (>42°C). The increases in Prx3 and Prx-SO3 levels at 42°C were inhibited by the antioxidants PEG-catalase and MnTBAP. In thermotolerant (40°C) cells, transcription factor Nrf2 was upregulated. The heat–induced increase in Prx3 expression was diminished in shRNA Nrf2 cells. Furthermore, heat (40-43°C) increased levels of reduced glutathione (GSH), which were inhibited by PEG-catalase and MnTBAP. These results suggest that heat-induced ROS were involved in the induction of antioxidants and shed some light on the role of antioxidants in cellular responses induced by heat shock at both mild and lethal temperatures. Improved knowledge about induction of adaptive survival responses by low-dose heat shock could have potential to protect against toxicity of multiple xenobiotics.

Photothermal and photodynamic activity of polymeric nanoparticles based on α-tocopheryl succinate-RAFT block copolymers conjugated to IR-780.

Multifunctional polymeric nanoparticles (NPs) that combine imaging and therapeutic properties are highly valuable in cancer treatment. In this paper we describe the development of NPs that are fluorescent in the near-infrared (NIR). This is important for their visualization in living tissues that present low absorption and low autofluorescence in this wavelength region (between 700 and 1000nm). Moreover, NPs present photothermal and photodynamic properties when NIR irradiated: the NPs produce an efficient increment of temperature and increase the intracellular reactive oxygen species (ROS) when laser irradiated at 808nm. These tuneable photoinduced properties make the NPs highly cytotoxic after NIR irradiation and provide a new tool for highly precise cancer treatment.

Potentiation of 5-methoxy-N,N-dimethyltryptamine-induced hyperthermia by harmaline and the involvement of activation of 5-HT1A and 5-HT2A receptors

5-Methoxy-N,N-dimethyltryptamine (5-MeO-DMT) and harmaline are serotonin (5-HT) analogs often abused together, which alters thermoregulation that may indicate the severity of serotonin toxicity. Our recent studies have revealed that co-administration of monoamine oxidase inhibitor harmaline leads to greater and prolonged exposure to 5-HT agonist 5-MeO-DMT that might be influenced by cytochrome P450 2D6 (CYP2D6) status. This study was to define the effects of harmaline and 5-MeO-DMT on thermoregulation in wild-type and CYP2D6-humanized (Tg-CYP2D6) mice, as well as the involvement of 5-HT receptors. Animal core body temperatures were monitored noninvasively in the home cages after implantation of telemetry transmitters and administration of drugs. Harmaline (5 and 15 mg/kg, i.p.) alone was shown to induce hypothermia that was significantly affected by CYP2D6 status. In contrast, higher doses of 5-MeO-DMT (10 and 20 mg/kg) alone caused hyperthermia. Co-administration of harmaline (2, 5 or 15 mg/kg) remarkably potentiated the hyperthermia elicited by 5-MeO-DMT (2 or 10 mg/kg), which might be influenced by CYP2D6 status at certain dose combination. Interestingly, harmaline-induced hypothermia was only attenuated by 5-HT1A receptor antagonist WAY-100635, whereas 5-MeO-DMT- and harmaline-5-MeO-DMT-induced hyperthermia could be suppressed by either WAY-100635 or 5-HT2A receptor antagonists (MDL-100907 and ketanserin). Moreover, stress-induced hyperthermia under home cage conditions was not affected by WAY-100635 but surprisingly attenuated by MDL-100907 and ketanserin. Our results indicate that co-administration of monoamine oxidase inhibitor largely potentiates 5-MeO-DMT-induced hyperthermia that involves the activation of both 5-HT1A and 5-HT2A receptors. These findings shall provide insights into development of anxiolytic drugs and new strategies to relieve the lethal hyperthermia in serotonin toxicity.

Inducible and constitutive HSP70s confer synergistic resistance against metabolic challenges

Chaperonic proteins, including inducible HSP70 (HSP70i) and constitutive HSP70 (HSC70), have been implicated as essential players in the cellular adaptive protection. Ensuing studies demonstrated that overexpression of either protein individually protects against thermal and oxidative challenges. The present study aimed to determine whether a concurrent overexpression of both HSC70 and HSP70i confers a better metabolic protection than the expression of each protein alone. Using a rat heart-derived H9c2 cardiac myoblast cell line, we found that HSP70i was rapidly induced within 2–8h following a mild thermal preconditioning (43°C for 20min) in both parental cells and an established H9/70c clonal sub-line overexpressing HSC70. The level of HSP70i protein in heat pretreated H9/70c clonal cells reached only 50% of that in heat pretreated H9c2 parental cells. Nevertheless, protection against lethal hyperthermia, menadione (an oxidant) and hydrogen peroxide (H2O2) exposure in the pretreated H9/70c clonal cells was significantly higher than the sum of protection afforded by the early induction of HSP70i in the pretreated parental cells and protection afforded by the pre-existing HSC70 in the H9/70c cells without preconditioning. Using dosimetric analysis, we also found that menadione resistance in the pretreated parental cells increased linearly with cellular HSP70i level (10–300ng/mg total protein). However, the resistance in the pretreated H9/70c cells showed a biphasic relationship with cellular HSP70i level; when HSP70i concentration reached >250ng/mg protein, survivability after menadione exposure was markedly enhanced. Similar results were observed in H9c2 cells genetically manipulated to overexpress both HSC70 and HSP70i. The survival benefit against lethal hyperthermia, oxidant treatment, and hypoxia/reoxygenation conferred by a concerted HSC70 and HSP70i overexpression was greater than the sum of benefits contributed by individual protein overexpression. Together, these findings suggest that HSC70 and HSP70i may complement each other in a synergistic manner to preserve cellular integrity during metabolic challenges.

Climate of a South African fur seal (Arctocephalus pusillus pusillus) breeding island off the south-east coast of South Africa

The Bird Island in Algoa Bay supports a breeding colony of South African (Cape) fur seals, Arctocephalus pusillus pusillus (n = 1000–3000) on Black Rocks which is the easternmost extreme of their breeding range and so it is likely to be the most environmentally marginal of the existing fur seal colonies. Conditions on Bird Island for seal pups are not as benign as they might first appear, particularly with regard to air temperatures. Climatic conditions on Bird Island were recorded in 1993–1995 and are compared to the conditions at Port Elizabeth Airport 63 km W-SW from the Bird Islands. Daily maximum and minimum temperatures, sea surface temperatures (SSTs) and rainfall were compared to concurrent Port Elizabeth data. Wind speeds and directions and sea swell heights at Bird Island were also recorded. Air temperatures ranged from 9°C to 33°C, SSTs ranged from 12°C to 23°C, total annual rainfall was ≈ 600 mm and there were typically relatively strong winds (13–15 knots/7–8 m s− 1) predominantly W-SW and E-NE. Intermittent periods of calm hot weather during summer pupping season would lead to potentially lethal hyperthermia (heat stroke). In 1993–1995, about 50% of summer days were potentially lethal to seal pups in the open by hyperthermia if the wind speed is near zero. Eddying of the Agulhas Current and strong winds leads to irregular favourable small-scale upwelling events, but incursions of warm water in hot still weather occurred in January–March 1993 and November–December 1993 and likely lead to starvation of some pups as well as endangering seal pups by hyperthermia. Detailed climate data are usually not available for seal colony sites, but we found good correlations between the offshore Bird Islands and climate data collected from the nearest permanent weather station (Port Elizabeth Airport) making it possible to infer long-term changes in conditions on Bird Island based upon Port Elizabeth data. We show that modest (2°C) increases in average summer temperatures would greatly increase the number of potentially lethal hot days.

A comparison of methylphenidate-, amphetamine-, and methamphetamine-induced hyperthermia and neurotoxicity in male Sprague–Dawley rats during the waking (lights off) cycle

Previous studies focusing on amphetamine (AMPH), methamphetamine (METH) and methylphenidate (MPH) neurotoxicity have almost exclusively been conducted in rodents during the light cycle, which is when most rodents sleep. There are virtually no studies that have simultaneously compared the effects of these three stimulants on body temperature and also determined serum stimulant levels during exposure. The present study compared the effects of MPH, AMPH and METH treatment on body temperature and neurotoxicity during the waking (dark) cycle of the rat. This was done to more effectively replicate stimulant exposure in waking humans and to evaluate the relative risks of the three stimulants when taken inappropriately or non-therapeutically (e.g., abuse). Four subcutaneous injections (4×), at 2h intervals, were used to administer each dose of the stimulants tested. Several equimolar doses for the three stimulants were chosen to produce plasma levels ranging from 3 times the highest therapeutic levels (no effect on body temperature) to those only attained by accidental overdose or intentional abuse in humans. Either 4×2.0mg/kg AMPH or 4×2.2mg/kg METH administered during the waking cycle resulted in peak serum levels of between 1.5 and 2.5μM (4 to 5 times over maximum therapeutic levels of METH and AMPH) and produced lethal hyperthermia, 70% striatal dopamine depletions, and neurodegeneration in the cortex and thalamus. These results show that METH and AMPH are equipotent at producing lethal hyperthermia and neurotoxicity in laboratory animals during the wake cycle. Administration of either 4×2.2 or 4×3.3mg/kg METH during the sleep cycle produced lower peak body temperatures, minimal dopamine depletions and little neurodegeneration. These findings indicate that administration of the stimulant during the waking cycle compared to sleep cycle may significantly increase the potency of amphetamines to produce hyperthermia, neurotoxicity and lethality. In contrast, body temperature during the waking cycle was only significantly elevated by MPH at 4×22mg/kg, and the serum levels producing this effect were 2-fold (approximately 4.5μM) greater on a molar basis than hyperthermic doses of AMPH and METH. Thus, AMPH and METH were equipotent on a mg/kg body weight basis at producing hyperthermia and neurotoxicity while MPH on a mg/kg body weight basis was approximately 10-fold less potent than AMPH and METH. However, the 10-fold lower potency was in large part due to lower plasma levels produced by MPH compared to either AMPH or METH.

Gene expression analysis characterizes antemortem stress and has implications for establishing cause of death

Within the field of forensic pathology, determination of the cause of death depends upon identifying physical changes in the corpse or finding diagnostic laboratory abnormalities. When such perturbations are absent, definitive assignment of a cause of death may be difficult or impossible. An example of such a problem is sudden infant death syndrome (SIDS), a common cause of neonatal mortality that does not produce physical findings or laboratory abnormalities. Although respiratory failure as a cause of SIDS represents the most widely held hypothesis, sudden cardiac death and hyperthermia have also been advanced as possible causes. We hypothesize that each of these physiological stresses would produce a different pattern of premortem gene expression and that these patterns of gene expression would remain evident in tissues collected postmortem. If these patterns were sufficiently distinctive, they could be used to identify the cause of death. Using an infant mouse model, we compared gene expression patterns in liver tissue after sudden death, lethal hyperthermia, and lethal hypoxia. Each of these conditions produced readily distinguishable differences in gene expression patterns. With the K-nearest neighbor classification algorithm, only 10 genes are necessary to correctly classify samples. If the liver tissue was not harvested immediately after death, additional alteration in gene expression patterns resulted; however, these alterations did not affect the group of genes used to classify the samples. Our findings suggest that gene expression analysis from tissues collected postmortem may provide useful clues about certain physiologic stresses that may precede death.

Thermotolerance induced at a fever temperature of 40 °C protects cells against hyperthermia-induced apoptosis mediated by death receptor signalling

Mild temperatures such as 40 degrees C are physiological and occur during fevers. This study determines whether mild thermotolerance induced at 40 degrees C can protect HeLa cells against activation of the death receptor pathway of apoptosis by lethal hyperthermia (42-45 degrees C). Protein expression of heat shock proteins (Hsps) 27, 32, 60, 72, 90, and 110 was increased in thermotolerant cells (3 h, 40 degrees C). Lethal hyperthermia (42-43 degrees C) caused cell death by apoptosis, but at 45 degrees C there was a switch to necrosis. Mild thermotolerance protected cells against heat-induced apoptosis (Annexin V labelling). Hyperthermia induced apoptosis through generation of reactive oxygen species (ROS) and death receptor signalling. The antioxidant polyethylene glycol-catalase abrogated increased expression of Fas death ligand and caspase-8 activation in response to lethal hyperthermia (42-43 degrees C). Mild thermotolerance attenuated the heat induction of ROS and FasL, which were initiating events in death receptor activation and signalling. Mild thermotolerance inhibited early events in hyperthermia-induced death receptor apoptosis such as Fas-associated death domain (FADD) translocation to membranes, caspase-8 activation, and tBid translocation to mitochondria. Downstream events in apoptosis such as caspase-3 activation, cleavage of PARP and ICAD, and chromatin condensation were also diminished in thermotolerant cells. It is important to improve knowledge about adaptive responses induced by exposure to mild stresses, such as fever temperatures, which can protect cells against subsequent exposure to lethal stress.

Sprague–Dawley rats display metabolism-mediated sex differences in the acute toxicity of 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy)

The use of the amphetamine derivative 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) has been associated with unexplained deaths. Male humans and rodents are more sensitive to acute toxicity than are females, including a potentially lethal hyperthermia. MDMA is highly metabolized to five main metabolites, by the enzymes CYP1A2 and CYP2D. The major metabolite in rats, 3,4-methylenedioxyamphetamine (MDA), also causes hyperthermia. We postulated that the reported sex difference in rats is due to a sexual dimorphism(s). We therefore determined (1) the LD50 of MDMA and MDA, (2) their hyperthermic effects, (3) the activities of liver CYP1A2 and CYP2D, (4) the liver microsomal metabolism of MDMA and MDA, (5) and the plasma concentrations of MDMA and its metabolites 3 h after giving male and female Sprague-Dawley (SD) rats MDMA (5 mg.kg{sup -1} sc). The LD50 of MDMA was 2.4-times lower in males than in females. MDMA induced greater hyperthermia (0.9 deg. C) in males. The plasma MDA concentration was 1.3-fold higher in males, as were CYP1A2 activity (twice) and N-demethylation to MDA (3.3-fold), but the plasma MDMA concentration (1.4-fold) and CYP2D activity (1.3-fold) were higher in females. These results suggest that male SD rats are more sensitive to MDMA acute toxicity than are females, probablymore » because their CYP1A2 is more active, leading to higher N-demethylation and plasma MDA concentration. This metabolic pathway could be responsible for the lethality of MDMA, as the LD50 of MDA is the same in both sexes. These data strongly suggest that the toxicity of amphetamine-related drugs largely depends on metabolic differences.« less

Sprague–Dawley rats display metabolism-mediated sex differences in the acute toxicity of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy)

The use of the amphetamine derivative 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) has been associated with unexplained deaths. Male humans and rodents are more sensitive to acute toxicity than are females, including a potentially lethal hyperthermia. MDMA is highly metabolized to five main metabolites, by the enzymes CYP1A2 and CYP2D. The major metabolite in rats, 3,4-methylenedioxyamphetamine (MDA), also causes hyperthermia. We postulated that the reported sex difference in rats is due to a sexual dimorphism(s). We therefore determined (1) the LD50 of MDMA and MDA, (2) their hyperthermic effects, (3) the activities of liver CYP1A2 and CYP2D, (4) the liver microsomal metabolism of MDMA and MDA, (5) and the plasma concentrations of MDMA and its metabolites 3 h after giving male and female Sprague–Dawley (SD) rats MDMA (5 mg.kg − 1 sc). The LD50 of MDMA was 2.4-times lower in males than in females. MDMA induced greater hyperthermia (0.9 °C) in males. The plasma MDA concentration was 1.3-fold higher in males, as were CYP1A2 activity (twice) and N-demethylation to MDA (3.3-fold), but the plasma MDMA concentration (1.4-fold) and CYP2D activity (1.3-fold) were higher in females. These results suggest that male SD rats are more sensitive to MDMA acute toxicity than are females, probably because their CYP1A2 is more active, leading to higher N-demethylation and plasma MDA concentration. This metabolic pathway could be responsible for the lethality of MDMA, as the LD50 of MDA is the same in both sexes. These data strongly suggest that the toxicity of amphetamine-related drugs largely depends on metabolic differences.