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Publicly Available Published by De Gruyter May 1, 2018

Updated review of postmortem biochemical exploration of hypothermia with a presentation of standard strategy of sampling and analyses

Guillaume Rousseau ORCID logo, Pascal Reynier, Nathalie Jousset, Clotilde Rougé-Maillart and Cristian Palmiere


Hypothermia is defined as a core body temperature below 35°C and can be caused by environmental exposure, drug intoxication, metabolic or nervous system dysfunction. This lethal pathology with medico-legal implications is complex to diagnose because macroscopic and microscopic lesions observed at the autopsy and the histological analysis are suggestive but not pathognomonic. Postmortem biochemical explorations have been progressively developed through the study of several biomarkers to improve the diagnosis decision cluster. Here, we present an updated review with novel biomarkers (such as catecholamines O-methylated metabolites, thrombomodulin and the cardiac oxyhemoglobin ratio) as well as some propositional interpretative postmortem thresholds and, to the best of our knowledge, for the first time, we present the most adapted strategy of sampling and analyses to identify biomarkers of hypothermia. For our consideration, the most relevant identified biomarkers are urinary catecholamines and their O-methylated metabolites, urinary free cortisol, blood cortisol, as well as blood, vitreous humor and pericardial fluid for ketone bodies and blood free fatty acids. These biomarkers are increased in response either to cold-mediated stress or to bioenergetics ketogenesis crisis and significantly contribute to the diagnosis by exclusion of death by hypothermia.


Human beings are homeothermic organisms with a regulating body temperature (Tbody). Decrease in Tbody can be poorly tolerated and ultimately lead to death [1]. Mortality studies show that death rate ranges from 38% to 85% from the moderate stage (<32°C) and up to 100% with associated polytrauma [2], [3], [4], [5]. This variability could be related to the studies themselves (from 1987) and the therapeutic approaches of that time. More recently, (between 2003 and 2013), 13,419 deaths were declared in the United-States as being linked to excessive natural cold exposure, with 10% of alcohol or drug poisoning as a contributing cause of death. The annual rates range from 0.3 to 0.5/100,000 persons in the United-States, up to 3.9/100,000 in South Australia and 3.3/100,000 in Sweden [6], [7].

Generalized hypothermia occurs when the Tbody drops below 35°C, in relation to exhausting heat production and failure of heat loss prevention [8]. Five degrees of severity are described: mild (32–35°C), moderate (28–32°C), severe (24–28°C), deep (13–24°C) and irreversible fatal outcome (<13°C) [1]. As Tbody drops, the hypothalamus (thermoregulation center) generates a series of reactions aiming to produce heat and prevent its loss. Heat production involves the secretion of stress response hormones (adrenaline [Ad], noradrenaline [Nad] and cortisol), triggering an overall increase in lipidic metabolism, specifically ketogenesis synthesis [8]. The large inter-individual variability in the establishment of these regulatory mechanisms impacts the diversity of hypothermic presentation.

There are two categories of hypothermia, depending on the quality of thermoregulation mechanisms: primary, when thermoregulatory mechanisms are intact but exceeded by the intensity or duration of cold exposure (mountaineers, those shipwrecked, drowned, etc.); secondary, when these mechanisms are altered, and hypothermia occurs despite temperate environmental temperatures (22–24°C). This may be the case when drug consumption/poisoning (acetaminophen, barbiturate, opioid, tricyclic antidepressant, benzodiazepine, phenothiazine, prazosin, valproic acid,…), ethanol consumption, or in some pathological conditions (thyroid or pituitary insufficiency, neurological disorders, sepsis and extensive burns) [2], [9], [10], [11].

The non-specific macroscopic and microscopic findings described in the clinical literature (Table 1) emphasize the major role that postmortem biochemical investigations might have in identifying hypothermia-related fatalities and helping pathologists to produce more accurate diagnoses [8], [9], [12], [13], [14].

Table 1:

List of main typical autopsy elements of hypothermia.

Autopsy findings
Macroscopic elementsBody and external temperature
Humidity, wind speed
± thermal insulation of housing
Hide-and-die syndrome
Paradoxical undressing
Wischnewski lesions
Frost erythema
Inner knee sign
Hemorrhage infiltrate in core muscle
Microscopic findingsLipid vacuolization of renal tubule epithelial cells, pancreatic cells, hepatocytes, cardiac myocytes, adrenal cells and anterior pituitary gland cells
Expression of adipose differentiation-regulated protein in renal tubule epithelial cells
Degenerative foci in myocardium

On the other hand, findings obtained from postmortem biochemical analyses cannot be considered in isolation, as they may depend on preexisting diseases (which may be responsible for increased or decreased concentrations of specific biochemical markers), and therefore necessitate accurate evaluation of patient conditions, in order to avoid misleading interpretations and mistaking preexisting pathological findings for hypothermia-related results.

The study herein aims to present the different biomarkers studied over the years and discuss their significance. Location and sampling methods are also discussed in some cases.

Forensic biomarkers of hypothermia

Generally speaking, postmortem biochemical investigation results may be different in different studies based on the clinical standard used. This principle applies also to postmortem biochemical investigations pertaining to hypothermia. Indeed, results obtained from biochemical investigations pertaining to catecholamines (and their O-methylated metabolites), cortisol and ketone bodies in suspected hypothermia-related fatalities should always consider and evaluate preexisting conditions (such as catecholamine-secreting tumors, diabetes mellitus, chronic alcohol abuse and stress conditions), in order to avoid erroneous interpretations of the measured valued.

Catecholamines and their O-methylated metabolites

The three main catecholamines are Ad, Nad and dopamine (DA), synthesized by chromaffin cells of the adrenal medulla and postganglionic neurons from the sympathetic nervous system. These act as neurotransmitters and regulating hormones. In the context of cold mediated stress, their role is to increase heat production and prevent its loss [15], [16], [17], [18], [19], [20].

Currently, it is known that cold mediated stress is responsible for increasing urinary catecholamines [9], [12], [14], [17], [21], [22], [23], [24], [25]. A sum increase of Ad+Nad in urine higher than 0.1 μg/mL was first described by Hirvonen in 1976, though the difficulty of interpreting urinary catecholamines in relation to cold induced diuresis had already been previously discussed [12]. This latter is thought to be related to an initial increase in renal blood flow resulting from vasoconstriction followed by decreased Tbody, loss of fluid reabsorption capacity by distal tubules and vasopressin action resistance [14]. It is recommended that urinary catecholamine concentrations be adjusted to creatininuria in order to avoid possible misinterpretations and prevent potential underestimation of the result. This should be disclosed in nmol of catecholamines per mmol of creatinine, and is obtained by adapting the concentration of catecholamines to an estimated renal filtration rate using creatininuria [24].

Recent studies have been conducted on the action of catechol-O-methyl transferase on catecholamines, resulting in O-methylated metabolites synthesis of which three exist: metanephrine from Ad, normetanephrine from Nad and methoxytyramine from DA [19], [26]. The study of these markers provides a benefit in terms of in vitro stability [27], [28].

Urine samples must be taken with a sterile needle. Urine acidification is recommended to prevent in vitro degradation, but is not compulsory in the case of immediate freezing. Catecholamines in vitro stability is estimated at 3 weeks and up to several months for their O-methylated metabolites [19], [27], [28], [29], [30], [31]. Postmortem stability, is estimated at 2 to “virtually” 10 days for catecholamines [19], [32]. Thus, the requirement imposed by the modalities of catecholamines urinary sampling, at the risk of an in vitro decay, does not allow a diagnosis of hypothermia to be excluded when normal or slightly increased results are observed.

Historically, urinary catecholamines were interpreted through Ad/Nad ratio analysis [15], [16], [33]. However, the interest in this ratio is put into question as it seems less sensitive and specific than an isolated study of urinary Ad concentrations, for example (Table 1) [18], [19].

According to some authors, Ad injection related to resuscitation does not modify concentrations of urinary catecholamines and their O-methylated metabolites, or at least does not increase their concentrations beyond the thresholds usually described for hypothermia. Age and sex would similarly have no influence on the concentrations of these markers [14], [19], [21].

In vivo, urinary catecholamines are explored in 24 h diuresis because of fluctuation in the excretions during a day, i.e. all activation of the sympaticoadrenal system will stimulate the secretion of catecholamines and thus the excretion. This is impossible in postmortem where only a sample of urine can be taken. Thus, clinical reference intervals are not applicable and new thresholds have been described adapting the conditions of this postmortem sampling and putrefactive phenomena. Palmiere et al. published a multicenter systematic study of urinary catecholamines and their O-methylated metabolites in 2014. Eighty-three fatal hypothermia and 144 control deaths were investigated and ROC curves analyzed, putting forward relevant interpretive thresholds for each parameter (Table 2) [19].

Table 2:

The sensitivity, specificity, positive predictive values (PPV) and negative predictive values (NPV) of urinary catecholamines and their O-methylated metabolites determined according to Palmiere et al. [19].

ParameterCut-off valueSensitivity, %Specificity, %PPV, %NPV, %
Adrenaline (Ad)4072878869
Noradrenaline (Nad)4934810010058
Ratio Ad/Nad0.19253737353

  1. Cut-off values are expressed in nmol/mmol creatinine, except for the Ad/Nad ratio (no units).

Exploring catecholamines and their O-methylated metabolites in serum or vitreous humor are not of interest, their postmortem concentrations are, respectively, discordant (increased or decreased) from blood, or related to cell autolysis [26].

Two urinary samples must be taken, one in a preservative free tube immediately frozen and/or with acidification and another one in a preservative free tube. The first will allow the study of catecholamines and/or their O-methylated metabolites to be carried out and the second creatinine.

Exploring urinary catecholamines and their O-methylated metabolites will allow the diagnosis of a perimortem generalized stress, in this situation a cold mediated stress.


The increase of counter-regulating hormones, such as cortisol, stimulates thermal and energy production. Besides contrary results on adrenal cortex hormone evolution in hypothermia, its perimortem elevation might reflect less of an increase in adrenocorticotrophic hormone (ACTH) than a decrease in metabolism and hepatic clearance [14], [34], [35], [36], [37], [38], [39], [40], [41]. A study concerning the agonal process and blood cortisol concentrations showed that there are no significant differences between instant death and death with prolonged agony, e.g. subdural hematoma [41]. Some teams recommend the exploration of free urinary cortisol and blood cortisol in suspected hypothermia [14], [21], [35], [38], [42].

In vivo, blood cortisol is explored at a precise time related to cyclic secretion following a nychthemeral rhythm, e.g. the upper secretion is at around 8 a.m. causing awakeness and the lower secretion is at around 12 a.m. to maintain sleep statement. Blood clinical reference intervals are not applicable in postmortem because generally the time of death is only suspected or simply unknown and postmortem fluctuation [14]. Bańka et al. [41] proposed a cut-off value of 300 ng/mL for blood cortisol using ultra high performance liquid chromatography. As catecholamines, free urinary cortisol is explored on 24 h diuresis because of the cyclic nychthemeral secretion. Again, in postmortem only a sample of urine can be taken. So, forensic pathologists and biochemists must consider a single samples, unknown postmortem interval and again inapplicable clinical reference intervals. One study observed a free urinary concentration above 250 nmol/L in all hypothermia fatalities explored and below 240 nmol/L for the control group which included cases with and without injuries as well as sudden and protracted death (analytically performed by radioimmunoassay) [14].

A high urine or blood cortisol is not a typical sign of fatal hypothermia and may reflect the body’s reaction to the stress in general [14]. Interpreting cortisol’s concentration in postmortem can be very difficult and must take into account the circumstances of death, and macroscopic and microscopic findings during and after autopsy exploration.

One peripheral blood sample with one urine sample must be put in preservative free tubes to allow the study of cortisol and free urinary cortisol.

Exploring blood and urine cortisol, as catecholamines and their O-methylated metabolites, will allow the diagnosis of a perimortem generalized stress, in this situation a cold mediated stress.

Ketones bodies

Exposure to cold is characterized by significant stress reactions that enhance catecholamine and counter-regulatory hormone release. Enhanced fat catabolism and increased ketone body production are the metabolic consequences of hypothermia-induced secretion of insulin antagonist hormones. Acetoacetate and β-hydroxybutyrate (HB) are energy-rich compounds that transport energy from the liver to other tissues. They can be interconverted by the enzyme β-HB dehydrogenase. Acetone is generated through the decarboxylation of acetoacetate (either spontaneously or through the enzyme acetoacetate decarboxylase) and is generally considered of little metabolic significance [9], [43], [44], [45], [46], [47].

Acetone production increases due to postmortem decompositional changes. On the other hand, these are not associated with HB production, whose concentration does not increase after death but may at most decrease as a result of spontaneous molecule degradation [48], [49], [50].

Increased HB can be explored in serum, vitreous humor and pericardial fluid (not recommended in urine) [14], [44], [45]. To these different matrices, Palmiere and Werner [51] have recently added the synovial fluid contained in the knee. This is useful when no other matrix is available due to sampling absence or insufficient volume.

Ketones should be explored in blood because the concentration balance with some other matrix (e.g. vitreous humor or pericardial fluid) takes several hours [14], [44], [45], [46]. However, there may be an interest in analyzing all these matrices in order to estimate agonal period duration (e.g. a patient with a long period of agony may have similar HB values in each matrix).

To the different matrices that can be explored in postmortem for their HB concentration, “liver homogenates” can be added. Within a long postmortem delay and thus taphonomic processes, there is a disappearance of blood, vitreous humor and pericardial fluid though the liver persists. In this context, Palmiere et al. [52] published preliminary but promising results in 2013 regarding the exploration of HB in “liver homogenates”. In the future, pathologies with hyperketonemia, in our case hypothermia, could be explored despite long postmortem intervals.

Ketones must always be explored in conjunction with ethanol, as the latter is able to inhibit ketogenesis. Hence, a diagnosis of hypothermia cannot be reversed on a lack of increase of the ketone bodies, especially in the presence of ethyl intoxication [44].

Peripheral blood sampling should therefore be carried out in sodium fluoride tubes whereas vitreous humor and/or pericardial fluid should be sampled in preservative free tube as these will allow ketone bodies to be best explored, especially HB and ethanol.

Exploring ketone bodies will allow the diagnosis of a stimulating ketogenesis which occurs during hypothermia.

Free fatty acids (FFAs)

As for Bańka et al. “Elevated levels of FFAs in the blood result from activation of the sympathetic-adrenal system and enhancement of lipolysis as one of the mechanism maintaining heat homeostasis. FFAs play an essential role in counteracting the effects of body cooling as, together with ketone bodies, they are main energy substrate metabolized by the nervous tissue, even before glucose” [20].

In cases of death from hypothermia, Bańka et al. observed an increase in FFA levels in femoral vein blood by 425%. They also pointed out that the most relevant FFAs are: palmitic, stearic and oleic acid. Using a one-step analytical procedure, gas chromatography coupled to mass spectrometry with a negative chemical ionization, they postulated a cut-off threshold at >0.2 mmol/L for each of these three compounds [20].

Palmiere et al. also observed a significant elevation of FFAs in blood for deaths by hypothermia. For their part, using an enzymatic colorimetric method, they observed concentrations ranging from 1.6 to 1.9 mmol/L (mean 1.8 mmol/L), compared to 1.1–1.7 mmol/L (mean 1.4 mmol/L, control group) which differ largely from clinical reference intervals (0.1–0.6 mmol/L) [14].

Both groups of researchers concluded that elevated blood FFAs cannot be considered a pathognomonic symptom of hypothermia due to a lack of specificity (they may also be increased in starvation, uncontrolled diabetes mellitus, hyperthyroidism and adrenal pheochromocytoma cases) [14], [20].

Peripheral blood sampling should therefore be carried out in preservative free tubes as these will allow FFAs to be best explored, especially palmitic, stearic and oleic acid.

Other biomarkers

Thrombomodulin (TM) is a transmembrane protein expressed in the endothelial cells of most blood vessels. TM functions essentially as anticoagulant and anti-inflammatory regulator by acting as a cofactor for thrombin. Severe cold stress has been shown to cause changes in the expression and secretion of TM [53]. Pakanen et al. have described that lethal hypothermia is associated with low myocardial TM transcript and TM protein levels as well as low urinary TM level compared with other causes of death. They also demonstrate a postmortem stability of TM during at least 24 h. Using urinary TM as a biomarker, with a cut-off at 15.5 ng/mL, they found sensitivity and specificity at 70.8% and 70.3%, respectively [54].

In fatal hypothermia, right cardiac blood is darker than left cardiac blood. This color difference could result from an increased oxyhemoglobin ratio (%O2-Hb) in left cardiac blood. %O2-Hb can be measured using CO-oximetry, spectrophotometry or gas chromatography. Kanto-Nishimaki et al., using co-oximetry, identified that %O2-Hb in left cardiac blood is significantly higher than in right cardiac blood (63.39% average for left cardiac blood versus 18.97% for right). Their study concluded that the difference between left and right cardiac blood could be a significant biological finding in death due to hypothermia [55].

Thyrotropin (TSH) and ACTH increase to produce heat should there be a loss. After this initial increase, there is a secondary decrease due to lipid disorders and/or depletion of reserves and central production capacities. Though concentrations vary according to the postmortem period, they are often lower [56], [57].

Although vitreous magnesium was widely used in the early 1960s during biochemical investigations for hypothermia, neither it nor vitreous calcium are recommended today due to inconsistent result reliability [14], [21], [58], [59].

Chromogranin A is a glycoprotein present in the secretory granules of endocrine and neuroendocrine cells. Immunohistochemical staining of the hypothalamus shows a decrease of its staining in the case of hypothermia while its concentration rises in cerebrospinal fluid. These findings would indicate a terminal stage of dysfunction involving the secretion of chromogranin A for cases of death by hypothermia with prolonged agony [60].

Information regarding biochemical markers that have already been studied but have not demonstrated their significance in the exploration of hypothermic deaths include: urinary histamine, urinary serotonin, blood, vitreous and urinary glucose and blood, vitreous and endocan [12], [14], [61].

Sampling and analyses strategy

This updated review of the literature allows proposing that the most adapted strategy to identify biomarkers of hypothermia would currently retain the following sampling and analyses (Table 3).


Postmortem exploration of biochemical markers, also called thanatobiochemistry refer to the study of biomarkers that contribute to the understanding of the origin of a death. This discipline, which appeared in the middle of the 20th century, has been gaining interest over the last several years due to the diversifications of its implications [62], [63], [64], [65]. The postmortem biochemical exploration of hypothermia, presented here, is an example of these implications.

The increase of catecholamines and their O-methylated metabolites is related to cold mediated stress but can also be identified in pheochromocytoma. Catecholamines and their O-methylated metabolites must be explored in the urine because of discordant conclusions in blood and insufficient exchange in vitreous humor. Related to a cold induced diuresis with dilution-effect some authors recommend expressing the results by a ratio Adr/Nad, which is increased, with different thresholds between the studies (1 or 0.19) [22], [33]. As with Sadler and Pounder [24], we recommend adjusting the catecholamines’ results to creatininuria as is done in vivo, in order to correlate levels to an estimated renal filtration rate thus, preventing the dilution effect and because of less specificity and sensitivity of this ratio [19].

Cortisol increases during hypothermia related to cold mediated stress. The specificity of this biomarker seems lower than for catecholamines as it increases in all stress situations. Anyway, its exploration is important to obtain an additional argument for a stress-related death, such as hypothermia. Cortisol can be explored in blood or urine with a higher statistical significance in urine [14]. Notably, no study focused on the correlation between urinary free cortisol and creatininuria.

Ketogenesis is stimulated during hypothermia to allow energy and thermal production as ketone bodies are easily metabolized. This finding is not a specificity of hypothermia and is observed in all situations of ketoacidosis and some metabolic dysfunctions [43]. Nevertheless, ketone bodies must be explored to identify stimulating ketogenesis as occurs during hypothermia.

FFA are increased in death from hypothermia because of the activation of the sympathetic-adrenal system and the enhancement of lipolysis as one of the mechanism maintaining heat homeostasis. FFA play an essential role in counteracting the effects of body cooling as, together with ketone bodies, they are the main energy substrate metabolized by the nervous tissue, even before glucose [20].

The parameters described in the section “Other biomarkers” have not been yet confirmed by different research teams or found in consistent result reliability or simply have no statistical significance. These are the reasons why they did not figure in our strategy of sampling and analyses.

Sample collection is crucial in thanatobiochemistry as postmortem redistribution can quantitatively affect some parameters, e.g. catecholamines’ levels are much higher in cardiac than peripheral blood [66]. That is why we recommend the exploration of peripheral blood as many studies have focused on this matrix instead of cardiac blood. Blood, urine and vitreous humor must be punctured gently with a sterile syringe to avoid contamination and the dilution effect.

Exploring ethanol consumption is also crucial when a diagnosis of lethal hypothermia is suspected. First, chronic or acute alcohol consumption are risk factors of hypothermia for many reasons: acceleration of heat loss, general vasodilatation, sedation stimulation, inhibition of shuddering abilities, denutrition and hypothalamic dysfunction. Secondly, ethanol has a antiketonemic action which mainly influences acetone levels. Thus, some authors postulated that a normal or subnormal result of ketone bodies cannot exclude hypothermia, particularly in cases of alcohol consumption [9]. However, it seems that ethanol intake does not influence the levels of urinary catecholamines, blood FFAs, blood cortisol or urinary free cortisol [14], [20], [41].

The last point is the most important: these different biological markers (just like the macroscopic and microscopic elements) make it possible to carry out a diagnosis of hypothermia but not a specific death by hypothermia. Only the absence of arguments for another etiology, after a thorough per- and post-autopsy exploration (histological, toxicological and imaging), leads to the conclusion of death by hypothermia. Several diagnostic findings (macroscopic, microscopic and biological) and an absence of arguments for any other lethal origin are required to carry out this diagnosis [67].


A death by hypothermia remains a diagnosis by exclusion according to a cluster of arguments. Biochemical markers take an increasingly important place in such decision clusters. The studies reviewed here have considerably increased the knowledge of such biomarkers, by demonstrating the relevance or not of each of them. For the most pertinent biomarkers, their postmortem stability, the better biological fluid in which to dose them, as well as their specificity and limits, have been progressively affirmed by these studies. Despite the complexity to develop clinical biochemistry adapted to forensic medicine, the potential of such approach remains considerable.

Table 3:

Strategy of sampling and analyses to identify biomarkers of hypothermia.

MatricesSamplingAnalysesInterfering conditions/disturbing factors
Peripheral bloodSodium fluoride tubeHydroxybutyrate and/or acetoneKetoacidosis (diabetic, alcoholic, starvation and septic)
EthanolInhibit ketogenesis
Preservative free tubeCortisolAll generalized stress reaction
Free fatty acidsStarvation, uncontrolled diabetes mellitus, hyperthyroidism, adrenal pheochromocytoma
UrinesPreservative free tube immediately frozen and/or with acidificationCatecholamines and/or their O-methylated metabolitesAdrenal pheochromocytoma
Preservative free tubeFree cortisolAll generalized stress reaction
Creatinine(for proper interpretation of urinary catecholamines)
Vitreous humorSodium fluoride tubeEthanolInhibit ketogenesis
Hydroxybutyrate and/or acetoneRetinal cells and ketoacidosis (diabetic, alcoholic, starvation and septic)
Pericardial fluidPreservative free tubeHydroxybutyrate and/or acetoneKetoacidosis (diabetic, alcoholic, starvation and septic)

Corresponding author: Guillaume Rousseau, MS, Département de Biochimie et Génétique, Centre Hospitalier Universitaire d’Angers, 4 rue Larrey, 49933 Angers, France, Phone: +33 2 41 35 36 37, Fax: +33 2 41 35 40 17

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


1. Briot R, Menthonnex E, Brun J, Anglade D, Girardet P, Jacquot C. Hypothermies accidentelles. EMC (Elsevier Masson SAS, Paris), Médecine d’urgence, 25-030-D-20, 2007.10.1016/S1959-5182(07)73287-7Search in Google Scholar

2. Peek GJ, Davis PR, Ellerton JA. Management of severe accidental hypothermia. In: Vincent JL, editor. Yearbook of intensive care and emergency medicine. Springer, New York, NY, 2008:147–59.10.1007/978-3-540-77290-3_14Search in Google Scholar

3. Danzl DF, Pozos RS, Auerbach PS, Glazer S, Goetz W, Johnson E, et al. Multicenter hypothermia survey. Ann Emerg Med 1987;16:1042–55.10.1016/S0196-0644(87)80757-6Search in Google Scholar

4. Jurkovich GJ, Greiser WB, Luterman A, Curreri PW. Hypothermia in trauma victims: an ominous predictor of survival. J Trauma 1987;27:1019–24.10.1097/00005373-198709000-00011Search in Google Scholar

5. Vassal T, Benoit-Gonin B, Carrat F, Guidet B, Maury E, Offenstadt G. Severe accidental hypothermia treated in an ICU: prognosis and outcome. Chest 2001;120:1998–2003.10.1378/chest.120.6.1998Search in Google Scholar PubMed

6. Bright FM, Winskog C, Walker M, Byard RW. A comparison of hypothermic deaths in South Australia and Sweden. J Forensic Sci 2014;59:983–5.10.1111/1556-4029.12451Search in Google Scholar PubMed

7. Meiman J, Anderson H, Tomasallo C, Centers for Disease Control and Prevention (CDC). Hypothermia-related deaths – Wisconsin, 2014, and United States, 2003–2013. Morb Mortal Wkly Rep 2015;64:141–3.Search in Google Scholar

8. Turk EE. Hypothermia. Forensic Sci Med Pathol 2010;6:106–15.10.1007/s12024-010-9142-4Search in Google Scholar PubMed

9. Palmiere C, Teresiński G, Hejna P. Postmortem diagnosis of hypothermia. Int J Legal Med 2014;128:607–14.10.1007/s00414-014-0977-1Search in Google Scholar PubMed

10. Nixdorf-Miller A, Hunsaker DM, Hunsaker JC. Hypothermia and hyperthermia medicolegal investigation of morbidity and mortality from exposure to environmental temperature extremes. Arch Pathol Lab Med 2006;130:1297–304.10.5858/2006-130-1297-HAHMIOSearch in Google Scholar PubMed

11. Lim C, Duflou J. Hypothermia fatalities in a temperate climate: Sydney, Australia. Pathology (Phila) 2008;40:46–51.10.1080/00313020701716466Search in Google Scholar PubMed

12. Hirvonen J. Necropsy findings in fatal hypothermia cases. Forensic Sci 1976;8:155–64.10.1016/0300-9432(76)90059-5Search in Google Scholar

13. Nara A, Nagai H, Yamaguchi R, Makino Y, Chiba F, Yoshida K, et al. An unusual autopsy case of lethal hypothermia exacerbated by body lice-induced severe anemia. Int J Legal Med 2016;130:765–9.10.1007/s00414-015-1266-3Search in Google Scholar

14. Palmiere C, Bardy D, Letovanec I, Mangin P, Augsburger M, Ventura F, et al. Biochemical markers of fatal hypothermia. Forensic Sci Int 2013;226:54–61.10.1016/j.forsciint.2012.12.007Search in Google Scholar

15. Lapinlampi TO, Hirvonen JI. Catecholamines in the vitreous fluid and urine of guinea pigs dying of cold and the effect of postmortem freezing and autolysis. J Forensic Sci 1986;31:1357–65.10.1520/JFS11913JSearch in Google Scholar

16. Hanhela R, Hollmen A, Huttunen P, Hirvonen J. Plasma catecholamines, corticosterone, glucose and fatty acids concentrations and mean arterial pressure and body temperature in haemorrhagic hypovolaemia, hypothermia and a combination of these in the rabbit. Acta Physiol Scand 1990;139:441–9.10.1111/j.1748-1716.1990.tb08945.xSearch in Google Scholar

17. Hirvonen J, Huttunen P. Hypothermia markers: serum, urine and adrenal gland catecholamines in hypothermic rats given ethanol. Forensic Sci Int 1995;72:125–33.10.1016/0379-0738(95)01694-ESearch in Google Scholar

18. Ishikawa T, Quan L, Michiue T, Kawamoto O, Wang Q, Chen J-H, et al. Postmortem catecholamine levels in pericardial and cerebrospinal fluids with regard to the cause of death in medicolegal autopsy. Forensic Sci Int 2013;228:52–60.10.1016/j.forsciint.2013.02.008Search in Google Scholar PubMed

19. Palmiere C, Teresiński G, Hejna P, Mangin P, Grouzmann E. Diagnostic performance of urinary metanephrines for the postmortem diagnosis of hypothermia. Forensic Sci Med Pathol 2014;10:518–25.10.1007/s12024-014-9585-0Search in Google Scholar PubMed

20. Bańka K, Teresiński G, Buszewicz G. Free fatty acids as markers of death from hypothermia. Forensic Sci Int 2014;234:79–85.10.1016/j.forsciint.2013.10.028Search in Google Scholar PubMed

21. Palmiere C, Mangin P. Postmortem biochemical investigations in hypothermia fatalities. Int J Legal Med 2013;127:267–76.10.1007/s00414-012-0738-ySearch in Google Scholar PubMed

22. Hirvonen J, Huttunen P. Increased urinary concentration of catecholamines in hypothermia deaths. J Forensic Sci 1982;27:264–71.10.1520/JFS11481JSearch in Google Scholar

23. Hirvonen J, Huttunen P, Nuutinen L, Pekkarinen A. Catecholamines and free fatty acids in plasma of patients undergoing cardiac operations with hypothermia and bypass. J Clin Pathol 1978;31:949–55.10.1136/jcp.31.10.949Search in Google Scholar

24. Sadler DW, Pounder DJ. Urinary catecholamines as markers of hypothermia. Forensic Sci Int 1995;76:227–30.10.1016/0379-0738(95)01814-XSearch in Google Scholar

25. Kernbach-Wighton G, Saternus K. Postmortem biochemical estimations in cases of fatal hypothermia (catecholamines and volatiles). Rom J Leg Med 2007;15:32–8.Search in Google Scholar

26. Hervet T, Teresiński G, Hejna P, Descloux E, Grouzmann E, Palmiere C. Catecholamines and their O-methylated metabolites in vitreous humor in hypothermia cases. Forensic Sci Med Pathol 2016;12:163–9.10.1007/s12024-016-9764-2Search in Google Scholar

27. Willemsen JJ, Ross HA, Lenders JW, Sweep FC. Stability of urinary fractionated metanephrines and catecholamines during collection, shipment, and storage of samples. Clin Chem 2007;53:268–72.10.1373/clinchem.2006.075218Search in Google Scholar

28. Roberts NB, Higgins G, Sargazi M. A study on the stability of urinary free catecholamines and free methyl-derivatives at different pH, temperature and time of storage. Clin Chem Lab Med 2010;48:81–7.10.1515/CCLM.2010.017Search in Google Scholar

29. Chan EC, Wee PY, Ho PC. Evaluation of degradation of urinary catecholamines and metanephrines and deconjugation of their sulfoconjugates using stability-indicating reversed-phase ion-pair HPLC with electrochemical detection. J Pharm Biomed Anal 2000;22:515–26.10.1016/S0731-7085(99)00308-8Search in Google Scholar

30. Miki K, Sudo A. Effect of urine pH, storage time, and temperature on stability of catecholamines, cortisol, and creatinine. Clin Chem 1998;44:1759–62.10.1093/clinchem/44.8.1759Search in Google Scholar

31. Hervet T, Grouzmann E, Grabherr S, Mangin P, Palmiere C. Determination of urinary catecholamines and metanephrines in cardiac deaths. Int J Legal Med 2016;130:995–1001.10.1007/s00414-015-1303-2Search in Google Scholar PubMed

32. Ishikawa T, Inamori-Kawamoto O, Quan L, Michiue T, Chen J-H, Wang Q, et al. Postmortem urinary catecholamine levels with regard to the cause of death. Leg Med Tokyo Jpn 2014;16:344–9.10.1016/j.legalmed.2014.07.006Search in Google Scholar PubMed

33. Pakanen L, Kortelainen M-L, Särkioja T, Porvari K. Increased adrenaline to noradrenaline ratio is a superior indicator of antemortem hypothermia compared with separate catecholamine concentrations. J Forensic Sci 2011;56:1213–8.10.1111/j.1556-4029.2011.01805.xSearch in Google Scholar

34. Okada Y, Miyai K, Iwatsubo H, Kumahara Y. Human growth hormone secretion in normal adult subjects during and after exposure to cold. J Clin Endocrinol Metab 1970;30:393–5.10.1210/jcem-30-3-393Search in Google Scholar

35. Wilkerson JE, Raven PB, Bolduan NW, Horvath SM. Adaptations in man’s adrenal function in response to acute cold stress. J Appl Physiol 1974;36:183–9.10.1152/jappl.1974.36.2.183Search in Google Scholar

36. Stoner HB, Frayn KN, Little RA, Threlfall CJ, Yates DW, Barton RN, et al. Metabolic aspects of hypothermia in the elderly. Clin Sci Lond Engl 1980;59:19–27.10.1042/cs0590019Search in Google Scholar

37. Leppäluoto J, Korhonen I, Huttunen P, Hassi J. Serum levels of thyroid and adrenal hormones, testosterone, TSH, LH, GH and prolactin in men after a 2-h stay in a cold room. Acta Physiol Scand 1988;132:543–8.10.1111/j.1748-1716.1988.tb08363.xSearch in Google Scholar

38. Wittert GA, Or HK, Livesey JH, Richards AM, Donald RA, Espiner EA. Vasopressin, corticotrophin-releasing factor, and pituitary adrenal responses to acute cold stress in normal humans. J Clin Endocrinol Metab 1992;75:750–5.Search in Google Scholar

39. Mallet ML. Pathophysiology of accidental hypothermia. QJM Mon J Assoc Physicians 2002;95:775–85.10.1093/qjmed/95.12.775Search in Google Scholar

40. Woolf PD, Hollander CS, Mitsuma T, Lee LA, Schalch DS. Accidental hypothermia: endocrine function during recovery. J Clin Endocrinol Metab 1972;34:460–6.10.1210/jcem-34-3-460Search in Google Scholar

41. Bańka K, Teresiński G, Buszewicz G, Mądro R. Glucocorticosteroids as markers of death from hypothermia. Forensic Sci Int 2013;229:60–5.10.1016/j.forsciint.2013.03.003Search in Google Scholar

42. Finlayson NB. Blood cortisol in infants and adults: a postmortem study. J Pediatr 1965;67:248–52.10.1016/S0022-3476(65)80247-5Search in Google Scholar

43. Rousseau G, Simard G, Homedan C, Reynier P, Rougé-Maillart C. Exploration biologique des décès par acidocétose diabétique: revue de la littérature. Rev Med Leg 2017;8:116–22.10.1016/j.medleg.2017.05.002Search in Google Scholar

44. Teresiński G, Buszewicz G, Madro R. The influence of ethanol on the level of ketone bodies in hypothermia. Forensic Sci Int 2002;127:88–96.10.1016/S0379-0738(02)00106-8Search in Google Scholar

45. Teresiński G, Buszewicz G, Madro R. Biochemical background of ethanol-induced cold susceptibility. Leg Med Tokyo Jpn 2005;7:15–23.10.1016/j.legalmed.2004.07.002Search in Google Scholar

46. Teresiński G, Buszewicz G, Madro R. Acetonaemia as an initial criterion of evaluation of a probable cause of sudden death. Leg Med Tokyo Jpn 2009;11:18–24.10.1016/j.legalmed.2008.08.002Search in Google Scholar

47. Teresiński G, Buszewicz G, Dabrowski W, Madro R. Ketone bodies in controlled intraoperative hypothermia- preliminary study. Arch Med Sadowej Kryminol 2007;57:389–93.Search in Google Scholar

48. Iten PX, Meier M. Beta-hydroxybutyric acid – an indicator for an alcoholic ketoacidosis as cause of death in deceased alcohol abusers. J Forensic Sci 2000;45:624–32.10.1520/JFS14739JSearch in Google Scholar

49. Kadis P, Balazic J, Ferlan-Marolt V. Alcoholic ketoacidosis: a cause of sudden death of chronic alcoholics. Forensic Sci Int 1999;103:S53–9.10.1016/S0379-0738(99)00074-2Search in Google Scholar

50. Palmiere C, Mangin P, Werner D. Postmortem distribution of 3-beta-hydroxybutyrate. J Forensic Sci 2014;59:161–6.10.1111/1556-4029.12265Search in Google Scholar PubMed

51. Palmiere C, Werner D. Post-mortem β-hydroxybutyrate determination in synovial fluid. Forensic Sci Int 2014;241:e28–30.10.1016/j.forsciint.2014.04.034Search in Google Scholar PubMed

52. Palmiere C, Mangin P, Werner D. Preliminary results on the postmortem measurement of 3-beta-hydroxybutyrate in liver homogenates. Int J Legal Med 2013;127:943–9.10.1007/s00414-013-0870-3Search in Google Scholar PubMed

53. Kaija H, Pakanen L, Uusitalo J, Nikkilä S, Kortelainen M-L, Porvari KS. Changes in cardiac thrombomodulin and heat shock transcription factor 1 expression and peripheral thrombomodulin and catecholamines during hypothermia in rats. Stress Amst Neth 2014;17:504–11.Search in Google Scholar

54. Pakanen L, Kaija H, Kortelainen M-L, Särkioja T, Porvari K. Victims of lethal hypothermia have decreased levels of thrombomodulin in myocardium and urine. Int J Legal Med 2015;129:289–96.10.1007/s00414-014-1138-2Search in Google Scholar PubMed

55. Kanto-Nishimaki Y, Saito H, Watanabe-Aoyagi M, Toda R, Iwadate K. Investigation of oxyhemoglobin and carboxyhemoglobin ratios in right and left cardiac blood for diagnosis of fatal hypothermia and death by fire. Leg Med Tokyo Jpn 2014;16:321–5.10.1016/j.legalmed.2014.06.010Search in Google Scholar PubMed

56. Ishikawa T, Quan L, Li D-R, Zhao D, Michiue T, Hamel M, et al. Postmortem biochemistry and immunohistochemistry of adrenocorticotropic hormone with special regard to fatal hypothermia. Forensic Sci Int 2008;179:147–51.10.1016/j.forsciint.2008.04.023Search in Google Scholar PubMed

57. Ishikawa T, Michiue T, Zhao D, Komatsu A, Azuma Y, Quan L, et al. Evaluation of postmortem serum and cerebrospinal fluid levels of thyroid-stimulating hormone with special regard to fatal hypothermia. Leg Med Tokyo Jpn 2009;11:S228–30.10.1016/j.legalmed.2009.02.049Search in Google Scholar PubMed

58. Mant AK. Autopsy diagnosis of accidental hypothermia. J Forensic Med 1969;16:126–9.Search in Google Scholar

59. Mant AK. Some post-mortem observations in accidental hypothermia. Med Sci Law 1964;4:44–6.10.1177/002580246400400111Search in Google Scholar PubMed

60. Yoshida C, Ishikawa T, Michiue T, Quan L, Maeda H. Postmortem biochemistry and immunohistochemistry of chromogranin A as a stress marker with special regard to fatal hypothermia and hyperthermia. Int J Legal Med 2011;125:11–20.10.1007/s00414-009-0374-3Search in Google Scholar PubMed

61. Descloux E, Augsburger M, Teresiński G, Hejna P, Grouzmann E, Scarpelli MP, et al. Endocan concentrations in postmortem serum, vitreous humor and urine in victims of lethal hypothermia. J Forensic Leg Med 2017;50:39–43.10.1016/j.jflm.2017.07.002Search in Google Scholar PubMed

62. Coe JI. Postmortem chemistries on human vitreous humor. Am J Clin Pathol 1969;51:741–50.10.1093/ajcp/51.6.741Search in Google Scholar PubMed

63. Coe JI. Postmortem chemistry update. Emphasis on forensic application. Am J Forensic Med Pathol 1993;14:91–117.10.1097/00000433-199306000-00001Search in Google Scholar PubMed

64. Palmiere C, Mangin P. Postmortem chemistry update part I. Int J Legal Med 2012;126:187–98.10.1007/s00414-011-0625-ySearch in Google Scholar PubMed

65. Palmiere C, Mangin P. Postmortem chemistry update part II. Int J Legal Med 2012;126:199–215.10.1007/s00414-011-0614-1Search in Google Scholar PubMed

66. Zhu B-L, Ishikawa T, Michiue T, Li D-R, Zhao D, Quan L, et al. Postmortem serum catecholamine levels in relation to the cause of death. Forensic Sci Int 2007;173:122–9.10.1016/j.forsciint.2007.02.013Search in Google Scholar PubMed

67. Byard RW, Bright FM. Lethal hypothermia – a sometimes elusive diagnosis. [Editorial] Forensic Sci Med Pathol 2017, [Epub ahead of print], DOI: 10.1007/s12024-017-9916-z.10.1007/s12024-017-9916-zSearch in Google Scholar PubMed

Received: 2018-02-09
Accepted: 2018-04-03
Published Online: 2018-05-01
Published in Print: 2018-10-25

©2018 Walter de Gruyter GmbH, Berlin/Boston

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