Forensic pathological study on temporal appearance of dendritic cells in skin wounds.

Dendritic cells (DCs) can essentially contribute to innate and adaptive immune system in various organs. A double-color immunofluorescence analysis was carried out with anti-CD11c and -HLA-DRα antibodies to detect DCs in 53 skin wounds (their postinfliction intervals: group I, 0-3 days; group II, 4-7 days; group III, 9-14 days; and group IV, 17-21 days). CD11c+HLA-DRα+ DCs were first observed in skin wounds with postinfliction intervals of 3 days, and the DC numbers were found to be elevated in skin wounds with the subsequent increase in postinfliction intervals. Semi-quantitative morphometric analyses showed that the DC number was the highest in the 12-day-old wound. More than 50 DCs were present in 8 of 10 samples (80%) in group II and 14 of 16 samples (87.5%) in group III, and there was no difference between the two groups. Thus, the presence of DCs in a skin wound was possibly estimated as postinfliction intervals of at least 3 days. Furthermore, when a skin wound contained > 50 DCs, its age would be judged as 4-14 days. Collectively, the appearance of DCs in human skin wounds may provide useful information in determining the age of a wound.

Immunohistochemical analysis on aquaporin-1 and aquaporin-3 in skin wounds from the aspects of wound age determination.

Immunohistochemical investigation of aquaporin (AQP)1 and AQP3 was performed in human skin wounds obtained from forensic autopsy cases. A total of 55 human skin wounds of different postinfliction intervals were collected as follows: group I, 0-3 days (n = 16); II, 4-7 days (n = 11); III, 9-14 days (n = 16); and IV, 17-21 days (n = 12). In uninjured skin samples, AQP1 and AQP3 could be slightly detected in dermal vessels and keratinocytes, respectively. The percentage of AQP1+ vessels and the number of AQP3+ keratinocytes were apparently elevated in accordance with wound ages. The number of AQP3+ keratinocytes was distinctly evident in groups II and III. Morphometrically, both AQP1+ vessel area and AQP3+ cell number were markedly increased in group II, compared with other three groups. With regard to forensic safety, AQP1+ vessel area of over 5% would imply wound ages of 4-12 days. Moreover, the positive area of > 15% would suggest wound age of 7-10 days. Especially, most samples of skin wounds aged 5-10 days except for only one sample (a 10-day-old wound) showed AQP3+ cell number of > 300, and the remaining other samples had that of < 300. Thus, the AQP3+ cell number of > 300 would indicate wound ages of 5-10 days. Collectively, immunohistochemical analyses of AQP1 and AQP3 in human skin wounds would support the objective accuracy of wound age determination.

Detection of endothelial progenitor cells in human skin wounds and its application for wound age determination.

Endothelial progenitor cells (EPCs), a newly identified cell type, are bone marrow-derived progenitor cells that co-express stem cell markers and vascular endothelial growth factor (VEGF) receptor (Flk-1). In this study, a double-color immunofluorescence analysis was carried out using anti-CD34 and anti-Flk-1 antibodies to examine the time-dependent appearance of EPCs, using 52 human skin wounds with different wound ages (Group I, 0-1 days; Group II, 2-6 days; Group III, 7-14 days; and Group IV, 17-21 days). In wound specimens with an age of less than one day, CD34(+)/Flk-1(+) EPCs were not detected. EPCs were initially observed in wounds aged two days, and their number was increased in lesions with advances in wound age. In morphometrical analysis, the average number of EPCs was the highest in the wounds of Group III. Especially, 20 out of 21 wounds aged 7-12 days had >20 EPCs, and all wound samples with postinfliction intervals of 14-21 days had <15 EPCs. These observations at least showed that >20 EPCs would indicate a wound age of 7-12 days. Taken together, our observations indicate the detection of EPCs would be useful for wound age determination.

Immunohistochemical analysis on MMP-2 and MMP-9 for wound age determination.

We performed immunohistochemical study combined with morphometrical analyses in order to examine the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 using 55 human skin wounds of different ages: group I, 0-3 days (n = 16); II, 4-7 days (n = 11); III, 9-14 days (n = 16); and IV, 17-21 days (n = 12). Immunopositive reactions for MMP-2 were observed in all human skin specimens including uninjured skin as control. The number of MMP-2(+) macrophages was significantly increased in accordance with wound ages. In contrast to MMP-2, no MMP-9(+) signals were detected in uninjured and wound specimens aged less than 1 day. However, the number of MMP-9(+) macrophages profoundly appeared in groups II and III. Morphometrically, in all of wound samples aged 9-12 days, MMP-2(+) cell number was more than 20. On the contrary, most of the remaining samples had <20 positive cells. However, only one sample (a 7-day-old wound) showed 21 positive cells. Thus, with regard to practical applicability with forensic safety, MMP-2(+) macrophages of >20 would indicate a wound age of 7-12 days. Additionally, 10 out of 12 wound specimens aged 9-12 days showed the MMP-2(+) cell number of >25, implying that MMP-2(+) cell number of >25 would indicate the wound age of 9-12 days. On the contrary, all wound samples aged 3-14 days except for only one sample had MMP-9(+) cell number of >30, indicating that MMP-9(+) cell number of >30 would indicate the wound age of 3-14 days. Collectively, MMP-2 seemed to be more distinct marker, compared with MMP-9.

Immunohistochemical analysis on cyclooxygenase-2 for wound age determination.

Immunohistochemical study combined with morphometry was carried out to examine the expression of cyclooxygenase-2 (COX-2) using 60 human skin wounds of different ages: group I, 0-4 h (n = 11); II, 8 h-2 days (n = 21); III, 3-9 days (n = 14); and IV, 12-21 days (n = 14). In wound specimens aged 2 h to 2 days, anti-myeloperoxidase-positive neutrophils observed at the wound site expressed immunopositive reaction to COX-2. In wound specimens of more than 3 days, CD68-positive macrophages as well as neutrophils were positively immunostained with anti-COX-2. In group II, all 21 wound samples had COX-2-positive ratios of >40 %, and 15 out of them showed >50 %. In group III, only three wound samples with the postinfliction intervals of 3 days showed positive ratios of 40-50 % and the remaining 11 cases less than 40 %. In groups I and IV, all 25 wound specimens had COX-2-positive ratio of <40 %. With regard to the practical applicability with forensic safety, these observations suggested that a COX-2-positive ratio of >40 % indicated a wound age of 8 h to 3 days. Moreover, COX-2-positive ratios, considerably exceeding a ratio of 50 %, indicate a wound age of 8 h to 2 days. Collectively, COX-2 would be a useful marker for the determination of early wound age.

Detection of fibrocytes in human skin wounds and its application for wound age determination.

Fibrocytes, a newly identified cell type, are bone marrow-derived mesenchymal progenitors that coexpress hematopoietic cell antigens and fibroblast products. In this study, a double-color immunofluorescence analysis was carried out using anti-CD45 and anti-collagen type I antibodies to examine the time-dependent appearance of fibrocytes, using 53 human skin wounds with different wound ages (group I, 0-3 days; group II, 4-7 days; group III, 9-14 days; and group IV, 17-21 days). In wound specimens with an age of less than 3 days, CD45+/collagen type I+ fibrocytes were not detected. The fibrocytes were initially observed in wounds aged 4 days, and their number increased in lesions with advances in wound age. In a semiquantitative morphometrical analysis, the average number of fibrocytes was highest in the wounds of group III. These findings imply that human skin wounds containing fibrocytes are at least 4 days old. Moreover, a fibrocyte number of over 10 indicates a wound age between 9 and 14 days (i.e., group III). Based on the average number of fibrocytes in each group, a fibrocyte number of over 15 more strongly suggests a wound age of 9-14 days. Together, our observations indicate the participation of fibrocytes in wound healing of human skin inducing the accumulation of extracellular matrix components, and therefore, detection of fibrocytes could be a useful marker for wound age determination.

Aberrant expression of histo-blood group A type 3 antigens in vascular endothelial cells in inflammatory sites.

Histo-blood group ABH antigens are widely distributed in human tissues. The epitopes of ABH antigens are carried by at least four different peripheral core isotypes of internal carbohydrate backbones (type 1-4). Each type of ABH antigen is expressed tissue specifically, and aberrant expression of ABH antigens is often observed during oncogenesis. We immunohistochemically examined the expression of A type 3 antigens in wounded and diseased skin tissues (A and AB blood groups). In uninjured skin, the expression of A type 3 antigens was restricted to the eccrine sweat gland. In addition to the sweat glands, A type 3 antigens were found in vascular endothelial cells of the wound sites. The extent of A type 3 antigens expression related to postinfliction intervals. A significantly higher expression rate of A type 3 antigens in endothelial cells was also observed in diseased skin, suggesting that inflammation might induce A type 3 antigen expression in endothelial cells. Double-color immunofluorescence staining of the specimens showed that von Willebrand factor (vWF) was a core-protein of A type 3 determinants aberrantly expressed in endothelial cells in inflamed tissues, suggesting that aberrant expression of A type 3 antigens is involved in stabilization of vWF in inflammation.

[Suicidal gunshot to the nape from a small-bore rifle]. / Suizid durch Gewehrschuss in den Nacken.

A 55-year-old man was found dead on the bed lying on his side in a pool of blood with a bullet entrance hole in the nape. Behind his body, a semi-automatic rifle Remington Nylon, cal. .22 LR, was detected. As the gunshot entrance site was rather unusual for suicide, a forensic autopsy was performed, which showed a contact shot to the nape with the bullet path running upward to the left frontal area. The man had been treated with citalopram for delusional depression, so that a suicidal act seemed plausible, and the autopsy and criminalistic findings were also compatible with this assumption. A remarkable fact of the present case is that a long firearm had been used. Reports on suicidal shots to the nape are comparatively rare in the medicolegal literature and usually refer to pistols or revolvers.

Analysis of tryptophan hydroxylase I and II mRNA expression in the human brain: a post-mortem study.

Dysregulation of brain serotonin transmission is an important contributing factor in many psychiatric disorders. Tryptophan hydroxylase (TPH), the rate limiting enzyme in the biosynthesis of serotonin plays a major role as candidate gene in several psychiatric disorders. Recently, a second TPH isoform (TPH2) was identified in mice which was exclusively expressed in the brain. Due to the lack of data about its anatomic expression in humans we performed a mRNA expression analysis comparing TPH1 and TPH2 mRNA in several regions of the human brain (cortex, thalamus, hypothalamus, hippocampus, amygdala, cerebellum and raphe nuclei). The study was performed with post-mortem specimens obtained from eight individuals with sudden deaths, not directly involving CNS diseases. Our results demonstrate that the mRNA of both genes is expressed in each investigated brain region with variations between the brain areas, as well as between the particular genes. The major finding of this study was the high expression level of TPH2 mRNA in the raphe nuclei ( approximately 4-fold more abundant than that of TPH1). The raphe nuclei showed the highest TPH2 mRNA levels at all, compared to the other regions (7-fold higher levels on average). To our knowledge, this is the fist study which demonstrates the localization of TPH1 and TPH2 mRNA in different regions of the human brain. Our findings provide further support for a duality of the serotonergic system and may open up new research strategies for the analysis of the repeatedly observed disturbances in the serotonergic system in patients suffering from several psychiatric disorders.

Comparative proteomic analysis with postmortem prefrontal cortex tissues of suicide victims versus controls.

BACKGROUND: The origin of suicidal behaviour is multifactorial including genetic, neurobiological and psychosocial correlates. Although there is no doubt that serotonin has a central role, the overall genetic findings with candidate genes of the serotonergic pathway are relatively inconsistent and suggests that other, yet unidentified, genes and gene products are also contributing to the vulnerability of suicidality. Proteomics is a powerful method to investigate modifications in protein expression. METHODS: We performed comparative proteomic analysis with prefrontal cortex tissues of 17 suicide victims and 9 controls. RESULTS: Applying two dimensional gel electrophoresis and image analysis we detected five protein spots to differ significantly in intensities between both groups. Three of them appeared only in suicide victims and could be identified by means of MALDI-TOF-MS analysis and protein database search as alpha crystallin chain B (CRYAB), glial fibrillary acidic protein (GFAP) and manganese superoxide dismutase (SOD2). CRYAB belongs to the low molecular heat shock proteins and GFAP is known as a marker of astrocytic activation in gliosis. SOD2 is a major antioxidant enzyme protecting cells against oxidative injury. Two further spots revealed higher intensities in the control group but had no unambiguous protein to match. CONCLUSIONS: Our findings suggest that proteins, being involved in glial function, neurodegeneration and oxidative stress neuronal injury, might also have an impact upon the neurobiological cascade leading to suicidality. As animal data provide evidence for an up-regulation of GFAP synthesis in astrocytes due to alterations in 5-HT levels, similar mechanisms of interaction might also be relevant in humans.

Estimation of time since death by heat-flow Finite-Element model. Part I: method, model, calibration and validation.

The determination of the time since death which often represents the presumed time of an offence plays an important role in medico-legal practice. In the early postmortem phase analyses of postmortem cooling provide the most accurate estimates. Empirical models of postmortem cooling are methodically restricted to standard conditions while heat flow models can in principle be applied to any complex cooling situations. The main problem having so far prevented heat flow models from being used in practice was the difficulty of solving the heat transfer equation for complex geometrical, initial and boundary conditions. This problem is now overcome by using the Finite-Element-Method as a numerical procedure. The study presents a three-dimensional Finite-Element-Model of the human body containing various tissue compartments with different thermal tissue properties. The initial temperature field is modelled inhomogeneously with a temperature gradient between body core and shell. Heat loss by conduction, convection and radiation as well as heat gain by supravital activity or irradiation from external sources can be simulated. One model parameter, the decrease rate of the supravital energy production, was calibrated and the model successfully validated using the experimentally verified empirical model by Marshall and Hoare.

Estimation of time since death by heat-flow Finite-Element model part II: application to non-standard cooling conditions and preliminary results in practical casework.

The present paper is part of a study investigating the application of the Finite-Element-Method to temperature-based death time determination. Part I introduced a three-dimensional Finite-Element model of the human body containing different tissue compartments with different thermal tissue properties. The initial temperature distribution is modelled inhomogeneously with a gradient between core and shell. Boundary conditions such as heat loss by convection or radiation as well as heat gain by supravital energy production or irradiation can be modelled. One model parameter, the decrease rate of the supravital energy production, was calibrated using the empirical model by Marshall and Hoare. Validation was successful using the Marshall and Hoare model as well for standard cooling situations. Part II now concentrates on the application of the model to non-standard cooling conditions and in practical casework. The parameters that have to be recorded at the crime scene are discussed. Special attention has to be paid to the insertion depth of the rectal temperature probe and to the thickness of the clothing material. Five real cases are presented, where the time of death is meanwhile known. The different steps of adjusting the standard Finite-Element model to the actual body and environmental conditions are described. In all cases, the victims were completely clothed. In one case, the victim was turned from a ventral to a dorsal position. In another case, the victim was killed and transported at room temperature in still air and then deposited outside in a much lower temperature and in slight wind. The model produced plausible results in all cases. The Finite-Element cooling curves are compared to the curves produced by the empirical model of Henssge. In two cases with strong body constitutions, the time since death was considerably overestimated by the Henssge model, in the other cases slightly. The results indicate that the heat transfer approach by Finite Elements is suited to considerably improve death time estimation.

Application of the BioRobot EZ1 in a forensic laboratory.

In a forensic laboratory, the routine application of an automated DNA extraction and purification robot has to fulfil several conditions, like producing reproducible DNA's of sufficient quantity and quality from all the different forensic biological stains relevant to various carrier materials. In this study, the suitability of the BioRobot EZ1 system from QIAGEN (Hilden, Germany), which offers fully automated extraction and purification of nucleic acids using magnetic bead technology, was tested. In summary, the DNA's obtained from the BioRobot EZ1 for different forensic relevant biological materials showed a quantity and quality comparable to those of the forensic standard protocols normally used in our laboratory. The system saves time, because there is no need of any further purification or concentration step after the automated DNA extraction. It can also be used as a replacement for time consuming organic extractions. A disadvantage of the system was the unsteady quality of the chemical regencies used by the robot. Nineteen different lots were tested with a self designed test system.

mtDNA investigations after differential lysis.

In order to gain information regarding the ethnic origin of an unknown offender in a murder case it was necessary to sequence the HV1 and 2 regions of the mitochondrial DNA (mtDNA). The only evidentiary material that could be linked to the suspect was DNA, extracted from spermatozoa after differential lysis. The observed mtDNA sequences were identical to the sequences of the victim. Therefore, we had to check if this was a coincidence or the result of a technical limitation of the testing procedure. Subsequently, we performed a systematic study. In cases with complete separation of sperm and female cells it wasn't possible to obtain a mtDNA sequence for the sperm fraction. This phenomena is based on the loss of the sperm's flagellum and mid-piece during the first lysis step and a concomitant loss of the sperm's mitochondria. In our murder case, a minor carry-over of female cells to the sperm fraction was responsible for the sequencing result.

Huntington's Disease (HD): Neurodegeneration of Brodmann's Primary Visual Area 17 (BA17).

Huntington's disease (HD), an autosomal dominantly inherited polyglutamine or CAG repeat disease along with somatomotor, oculomotor, psychiatric and cognitive symptoms, presents clinically with impairments of elementary and complex visual functions as well as altered visual-evoked potentials (VEPs). Previous volumetric and pathoanatomical post-mortem investigations pointed to an involvement of Brodmann's primary visual area 17 (BA17) in HD. Because the involvement of BA17 could be interpreted as an early onset brain neurodegeneration, we further characterized this potential primary cortical site of HD-related neurodegeneration neuropathologically and performed an unbiased estimation of the absolute nerve cell number in thick gallocyanin-stained frontoparallel tissue sections through the striate area of seven control individuals and seven HD patients using Cavalieri's principle for volume and the optical disector for nerve and glial cell density estimations. This investigation showed a reduction of the estimated absolute nerve cell number of BA17 in the HD patients (71,044,037 ± 12,740,515 nerve cells) of 32% in comparison with the control individuals (104,075,067 ± 9,424,491 nerve cells) (Mann-Whitney U-test; P < 0.001). Additional pathoanatomical studies showed that nerve cell loss was most prominent in the outer pyramidal layer III, the inner granular layers IVa and IVc as well as in the multiform layer VI of BA17 of the HD patients. Our neuropathological results in BA17 confirm and extend previous post-mortem, biochemical and in vivo neuroradiological HD findings and offer suitable explanations for the elementary and complex visual dysfunctions, as well as for the altered VEP observed in HD patients.

Single nucleotide polymorphism and haplotype analysis of a novel tryptophan hydroxylase isoform (TPH2) gene in suicide victims.

BACKGROUND: Tryptophan hydroxylase, the rate-limiting enzyme in the biosynthesis of serotonin, represents a major candidate in numerous genetic association analyses of suicidal behavior; however, the results are so far inconclusive. Recently, a second tryptophan hydroxylase isoform (TPH2) was identified in mice, which was exclusively present in the brain. In a previous postmortem study of our own group, we could demonstrate that TPH2 is also expressed in the human brain but not in peripheral tissues. METHODS: We performed single nucleotide polymorphisms, haplotypes, and linkage disequilibrium studies on 263 suicide victims and 266 healthy control subjects with 10 single nucleotide polymorphisms in the TPH2 gene. RESULTS: Significant association was detected between one single nucleotide polymorphism (p = .004, global p = .01) and suicide. Additionally, haplotype analysis also produced support for association (p < .0001, global p = .0001). CONCLUSIONS: This is the first report about an association between TPH2 gene polymorphisms and completed suicide. Our findings provide evidence for an involvement of genetic variants in the TPH2 gene in suicidal behavior. These results might open up new research strategies for the analysis of the observed disturbances in the serotonergic system in several other psychiatric disorders.

Regional mRNA expression of a second tryptophan hydroxylase isoform in postmortem tissue samples of two human brains.

Tryptophan hydroxylase (TPH) as rate limiting enzyme in the biosynthesis of serotonin plays a major role as candidate gene in several psychiatric disorders. Recently a second TPH isoform (TPH2) was identified in mice, which was exclusively expressed in the brain. We investigated whether the mRNA of the human homologue of this new TPH2 isoform is expressed in the human brain but not in peripheral tissues. The study was performed with postmortem specimen obtained from two subjects who died on cardiovascular failure. TPH2 mRNA levels were determined by quantitative real time RT-PCR. TPH2 mRNA was exclusively present in the human brains but not in the investigated peripheral tissues. Our finding may open up new research strategies for the analysis of the repeatedly observed disturbances in the serotonergic system in patients suffering from several psychiatric disorders.

Simulating irradiation power density on body surface in postmortem cooling.

Irradiation poses a major problem to determining the time since death by temperature-based methods. Neither empirical nor heat-flow postmortem cooling models have so far been able to assess irradiation. Heat-flow models seem overall better suited to calculate irradiation because of their direct relation to the physics of heat transfer. An implementation of irradiation boundary conditions in heat-transfer models requires the knowledge of the irradiation power density on the body surface. The present study develops formulae and implements them in a computer program to simulate the radiation power density on a semi-cylindrical body surface coming from irradiation by a rectangular radiant heater nearby or from the sun. The formulae are valid for deliberate geometrical arrangements of either body and radiant heater or body and sun. In case of the radiant heater scenario shading functions for the shading of the semi-cylinder by itself and by the rear panel of the radiant heater are developed. In case of the sun scenario only the shading by the semi-cylinder is relevant. In examplary analyses of typical irradiation scenarios the power density coming from a 2000W radiant heater nearby on the body surface amounted to a maximum of 418W/m2, the radiation power density originating from sunlight on a clear summer afternoon in middle-Europe amounted to a maximum of 422W/m2.

Modelling postmortem surface cooling in continuously changing environmental temperature.

Heat loss depends on the temperature gradient between body surface and environment. Skin cooling data in the forensic literature are scarce and models for skin cooling have not been developed. The dependence on the environmental temperature is a general problem in modelling postmortem cooling processes; most models of rectal cooling are therefore restricted to constant ambient temperatures. Since surface in contrast to core temperatures are highly sensitive to changes of ambient temperature, a model for skin cooling has to take into account such changes. The present study provides an estimator for the time-dependent function of the temperature decrease of the skin and presents a model of the cooling process. The formulae are developed on the basis of skin cooling data of the exposed skin of the forehead in a 40-year-old female (163 cm, 62.1 kg). The single exponential Newtonian model for the surface temperature T(S) valid for constant environmental temperature T(E):T(S)(t)=(T(S)(0)-T(E))e(-lambda(t))+T(E) is localized to small time intervals. By Taylor series expansions a differential equation directly providing an estimator for the temperature decrease rate lambda is derived. The solution of this differential equation represents the extended Newtonian model valid for non-constant environmental temperatures and non-constant temperature decrease rates. The extended model is tested successfully by reinserting the estimated values for the temperature decrease rate: the reconstructed and the measured skin temperature decrease curves completely overlap each other. The temperature decrease rate is a function of the difference between skin and environmental temperature and of the actual change of the skin temperature. A scatter plot of this function shows a structured cloud of points lying in one plane. The temperature decrease rate can thus be parametrized by a simple affine equation with three coefficients determined by linear regression. Inserting the affine equation in the extended Newtonian model leads to an inhomogeneous, non-linear differential equation which is solved by recursion. With knowledge of the initial temperature and the course of the environmental temperature the decrease of the skin temperature can be predicted with very good results. The model is validated with good results in 12 further experimental skin cooling curves of ten different individuals.

Supravital energy production in early post-mortem phase - estimate based on heat loss due to radiation and natural convection.

The temperature-based determination of the time since death in the early post-mortem (pm) period plays an important role in medico-legal practice. In contrast to the common opinion according to which convection and conduction are mainly responsible for post-mortem heat loss, a considerable part of energy is emitted by thermal radiation. The present paper concentrates on the heat loss due to radiation and natural convection. Since both heat transfer mechanisms depend on the temperature gradient between skin and environment, the skin temperature was measured in corpses of different constitution (lean, medium and obese) and its decrease fitted by a single-exponential model. Heat loss due to radiation was calculated according to the non-linearized form of the law of Stefan and Boltzmann, heat loss due to natural convection according to the semi-empirical thermodynamic laws; the shape of the body in supine position was approximated to a semi-cylinder of finite length. The power due to radiation ranged between 386kJ/h (lean) and 550kJ/h (obese), that due to natural convection between 307kJ/h (lean) and 429kJ/h (obese) initially. Cumulative energy loss amounted to 2167kJ (lean) and 4239kJ (obese) by radiation and 1485kJ (lean) and 2922kJ (obese) by natural convection up to 20h pm. The energy loss due to radiation plus natural convection initially exceeded the energy loss due the decrease of the energy content of the body (mass x heat capacity x temperature decrease). This surplus can be explained only by exothermal processes in the phase of intermediary life and directly provides lower bounds for supravital energy production. Cumulative supravital energy ranges between 1139kJ up to 5h pm in the lean and 2516kJ up to 10h pm in the obese corpses. The courses of supravital energies and powers are presented as functions of time. Under standard conditions like still air (no forced convection) and insulating ground (little conductive heat transfer), the lower bounds represent estimates for total supravital energy production.