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Kaliszan1
Wiczling2
Penkowski41 Department of Forensic Medicine2 Department of Biopharmaceutics and Pharmacodynamics4 Department of Physics and Biophysics, Medical University of Gdañsk, Gdañsk, Poland 3 Department of Pig Breeding and Production, Agricultural University of Poznañ, Poznañ, Poland
| Abstract |
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(Received 24 March 2005;
accepted after revision 2 June 2005; first published online 8 June 2005)
Corresponding author R. Hauser: Department of Forensic Medicine, Medical University of Gda
sk, ul. Debowa 23, 80-204, Gda
sk, Poland. Email: rohauser{at}amg.gda.pl
| Introduction |
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According to Henssge & Madea (2004), the theoretical basis for using dead body temperature for estimation of the time of death had already been formulated in the mid-19th century by Rainy (1868). Rainy transferred the Newton rule of cooling to forensic medicine and calculated the time of death from the exponential formula relating the difference between the body and ambient temperatures to time.
The single-exponential model was applied by Saram & Webster (1955). That model comprised a fixed term of 45 min added arbitrarily to account for a post-mortem temperature plateau. The model was devised after an analysis of rectal temperatures of 41 judicially hanged prisoners.
The exponential model was a subject of numerous studies leading to either its simplification or its complication. The simplifications, like the so-called rule of thumb, imply a linear drop of rectal temperature with a typical value of 0.8°C h1 (Green & Wright, 1985). Linear regressions, including multivariable regressions, combining physiological with biochemical parameters, were reported by Baccino et al. (1996). These procedures provide only a rough estimate of the time of death and are an unnecessary simplification, especially now that the measured temperature data can be processed according to even the most sophisticated algorithms.
A question arises whether the proposed extensions and complications of the single-exponential model are of real value for the reliability estimation of the time of death in individual cases. Henssge and coworkers (Henssge, 1988; Althaus & Henssge (1999); Henssge et al. 2000a) have developed a rectal temperature-based nomogram method and a relevant software package which take into account the subject's body weight as well as a number of empirical environmental correction factors. However, there are reports in the literature questioning significant correlations between the rates of cooling and the body parameters (Green & Wright, 1985).
Another question is whether the complexity of the necessary temperature data collection actually does help to improve the accuracy of estimation of the time of death. Specifically, it concerns the propositions by Green & Wright (1985) to use two rectal temperatures measured about 1 h apart and by Mall et al. (2004) to employ a continuous temperature recording during a longer time interval post mortem (4.25 h).
Another question is whether and when the rectal temperature measurements are to be preferred over measurements at the other sites of temperature recording. Technical problems of accessibility aside, there may be legal restrictions to invasive methods in some countries. Of particular importance is the dependence of the precision of estimation of time of death on the site of temperature measurement. It should also be tested whether using temperature data from several body sites, like in the triple-exponential approach of al-Alousi et al. (2002), actually improves prediction of th time of death. Perhaps such a complex model is valid in specific cases or for the average case only.
In order to address the questions above, a controlled experiment needs to be carried out under conditions close to real life. Such an experiment would, for many reasons, be difficult to perform in humans but may be done in pigs, which are animals with similar physiology. Here, such an experiment has been designed and performed.
| Methods |
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The pigs were not killed solely for the sake of the experiment. They had been randomly selected from the animals being killed as part of the regular manufacturing process. A surplus supply of pigs is normally used for the manufacture of feeding stuff and such was the final purpose of the experimental animals used here.
The study was conducted in 19 pigs: 13 sows and 6 boars of the Great White Polish species (Wielka Biala Polska) weighing between 81 and 124 kg. Five two-channel thermometers P655 were used for the measurements, connected with pin probes Pt100, class B 1/3 DIN, 100 x 1.4 mm, ending in a a 20 mm temperature sensor, and with probes Pt100, class B, 150 x 3 mm, ending with a 40 mm temperature sensor. The measuring equipment was manufactured by Dostmann electronic GmbH (Wertheim-Reicholzheim, Germany) with the following catalogue numbers: thermometers: 5000-0655; pin probes Pt100, class B 1/3 DIN: 6000-9999; and pin probes Pt100 class B: 6000-1001. Each of the five measuring kits (1 thermometer and 2 probes) was calibrated according to the manufacturer's instructions.
Straight after killing (by electrical current), the animals were placed in a specially assigned room of the slaughterhouse with the abdominal surface facing 150 mm-high wooden gratings, in an isolated room approximately 200 m3 in volume.
After applying an eyelid retractor in order to obtain a wide lid slit, the eyeballs were stabilized with stabilization pincets. Two 100 mm pin probes were inserted into the sclera, into the nasal quadrants of the left eyeballs, 3 mm away from the corneal limbus, passing through the pars plana of the ciliary body into the vitreous chamber, and further on, posteriorly and laterally from the optic nerve head, until a depth of 22 mm was reached.
The next two 100 mm pin probes were inserted into the soft tissues of the right orbits at the medial canthus, passing along the medial rectus muscle towards the superior orbital fissure, until a depth of 25 mm was reached. After the probes had been inserted, the pincets and the eyelid retractor were removed and the eyelids were closed.
The entire lengths of two 150 mm probes were inserted into the muscles of the left rumps from the insertion site at the central portion of the rump.
Two other 150 mm probes were inserted into the rectum up to their handles.
Ambient temperature was measured using a probe located 500 mm above the ground.
The nine probes used in each set of measurements were located in parallel to the ground, and their handles were stabilized in the grips of the stands. It was also ensured that the probes, after their settlement, did not cause the eyelids to open, which were naturally closed in all the animals tested.
The thermometers were connected to a computer preset to record the transmitted temperature values at a frequency of one sample every 30 s (for construction of graphs every tenth time point was used).
The recording device was switched on 75 min following a necessary unified preparation procedure. The registration of the measurements was completed 2325 h after the animals were killed.
The results were processed with Microsoft Excel 2000 (Microsoft Corporation, USA) using appropriate statistical procedures: Matlab® Software version 7.0 (The Math Works, Inc., Natick, MA, USA).
| Results |
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Figure 2 represents the records at 5 min intervals of the temperature difference between the individual pig eyeball (T) and the environment (TE) for 21 eyeballs tested. It must be noted here that for both pigs tested on days 1 and 7 we measured the temperature in both eyeballs, whereas on day 2 we did not use the eyeballs for temperature measurements (see Table 1).
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In Fig. 2, a mathematical model is given along with the plots of the mean exponential relationships between the eyeball and ambient temperature difference and time for 21 eyeballs. The statistical quality of the model derived is very high, as quantified by the values of S.S.E., r2 and R.M.S.E. The meaning of these parameters is as follows. The S.S.E. denotes the sum of squares due to error. This statistics measures the total deviation of the actual value from the fit to the calculated one. The r2 measures how successful the fit is in explaining the variation of the data. The R.M.S.E. is the root mean squared error.
The physical meaning of the coefficients of the exponential model is as follows. Coefficient a is a starting temperature difference, i.e. the extrapolated difference between the temperature of the eyeball and the environment at the time of death (t = 0). Coefficient b denotes the rate constant of the temperature decrease.
Analysing the data from Table 1, it may be noted that, for example, pig 1 studied on day 4 (pig 4_1) would have an extrapolated to the time of death temperature of eyeball, TD, of 36.6°C, because a = 15.1°C when TE = 21.5°C. Hence, TD TE = 15.1°C means that TD = 36.6°C. That value appears to be reasonable. Unfortunately, we were unable to find any literature report on normal eyeball temperature in pigs. We refrained from performing an experiment in vivo, assuming that the mean values of a = 15.2°C and TE = 21.0°C provide a reliable TD or normal physiological temperature in pig eyeball of 36.2°C. That temperature might be a bit higher in pigs than in humans since the body temperature in pigs is reported (Prost, 1985; Klont & Lambooy, 1995; Hanneman et al. 2004) to be within 3840°C, whereas in humans it is commonly recognized as 36.637.5°C.
The dispersion of the eyeball temperature time courses among the pigs tested is illustrated in Fig. 2. The data for only one pig cross the 95% confidence limit provided by the model given in Fig. 2. The single-exponential model appears to be reliable and can be used to estimate the time elapsing from death based on the eyeball temperature measured. For the purpose of such estimation, a simple algebraic transformation of the model into the following form seems convenient:
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Figure 3, illustrating data for the orbit soft tissues, shows a close similarity to the corresponding figure for the eyeball (Fig. 2). Coefficients a and b of the exponential equation for individual pigs (Table 1), as well as those of the general model (Fig. 2), are closely similar to those derived for the eyeballs. This would mean that both the extrapolated temperature at death, a, and the rate constant of temperature decrease, b, are similar, as expected because of the close vicinity of temperature measurement sites. However, the dispersion of temperature decrease time courses is evidently larger for the orbit soft tissues (Fig. 3) than for the eyeballs (Fig. 2). This is quantitatively evidenced by a larger R.M.S.E. value for the orbit soft tissues than for the eyeballs (0.705 versus 0.589). Also, the intervals between the lower and the upper death time 95% confidence limits, corresponding to the same temperature differences, T TE, are a bit larger in the case of the orbit soft tissues compared to the eyeballs (Table 3).
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Table 4 shows that errors in estimation of the time of death from the single-exponential model employing rectal temperature are larger than in the case of the eyeballs and the orbit soft tissues at earlier periods after death. However, rectal temperature data allow estimations of the time of death after longer periods of time (up to 30 h), whereas no time estimations beyond 20 h since death are possible from temperature measurements in the eyeballs and in the orbit soft tissues.
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| Discussion |
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The deviations from the single-exponential model, as illustrated in Figs 4 and 5, are small. However, the intersubject diversity is large enough to exclude a more complex two-exponential model claimed by Marshall & Hoare (1962) to improve the reliability of predictions of the time of death. Of course, mathematically a two-exponential model cannot produce worse predictions than a simpler single-exponential model. However, at least in the case of the data considered, the second exponential term provides no statistically significant improvement of description of the mean temperature decrease time course. In such a situation, there is no practical advantage to employing a more complex model.
It must be noted here that considering a single curve for a given animal in Figs 25, one would expect a better fit of the measured data points to a two-exponential model than to a single-exponential one. From our analysis (Table 1) it turns out that in fact that is not the case. Values of r2 close to 1 and low values of R.M.S.E. are evidence that the monoexponential model is good enough for individual cases. The scatter of the individual cooling curves plotted 75 min after death for 19 pigs makes the increase of correlation obtained with a second exponential term insignificant due to the lack of statistical significance of the second exponential term.
A question arises whether the missing temperature data for the first 1.25 h after death would form a plateau on the mean cooling curves derived. This question is, in fact, academic in nature, because either answer, yes or no, would not support the practical use of the two-exponential model, leaving aside technical difficulties regarding the collection of the necessary reproducible experimental data directly after death. If, indeed, the initial plateau did exist, no estimation of the time of death could be done within that period other than the plateau length. Here, this would be the case between 0 and 1.25 h. If no plateau existed, the differences between the lower and the upper 95% confidence limits for time since death estimated from the single-exponential model would be 1.5 h at best (Tables 2-5).
From the practical point of view, for a forensic medicine specialist, the two-exponential model describing the body temperature fall after death is of no use. This model may of course provide a better description of a given individual body site temperature time course, especially with regard to such measurement sites as the rectum or the muscles. However, its predictive value with respect to a biological object not used to derive the specific two-exponential model is not higher than that of a simpler, thermodynamically based monoexponential model. In our opinion, the often cited advantages of the two-exponential model might be statistical artefacts.
In view of the marked scatter of the cooling curves for the body sites illustrated in Figs 25, the statistical significance of various correction factors to the exponential models recommended for use in forensic medicine also seems disputable (Henssge, 1988). While these correction factors could work for a given artificial phantom subjected to various controlled environmental conditions, it is highly unlikely that those corrections could significantly increase the reliability of estimation of the time of death of individually diversified mammals.
Certain authors believe that taking into account two temperature measurements about 1 h apart (Green & Wright, 1985) or a continuous recording of temperature for a certain period (4.25 h) after death (Mall et al. 2004) will improve estimation of the time of death. This might indeed work, but complicates the procedure for estimation of the time of death. The single-exponential equation describing post-mortem temperature decrease at individual body sites is readily interpretable in physical terms. Coefficient a denotes the temperature difference between a given measurement site and the environment, T TE, extrapolated to t = 0, i.e. the moment of death. Hence, the temperature at death, TD, can be calculated from a = TD TE. Coefficient a, once derived, may be adjusted to any ambient temperature if TD is known. Assuming the mean temperature of the environment during the experiments to be TE = 21°C (Fig. 1), one may calculate the mean pig eyeball temperature at death from the equation characterized in Fig. 2. Hence, a = TD TE becomes 15.2 = TD 21 or TD = 36.2°C. Similarly, the extrapolated temperature at death would be 36.6°C in the orbit soft tissues (Fig. 3), 41.5°C in the rectum (Fig. 4) and 41.3°C in the muscles (Fig. 5).
We did not manage to find any data in the literature literature on the physiological eyeball temperature of pigs. In humans, the mean eyeball temperature is reported to be about 3435°C (Charman & Jennings, 2000; Jurowski et al. 2004). The value of the pig eyeball temperature of 36.2°C, as estimated from our models, appears to be reliable. Also, the value 36.6°C for the orbit soft tissues is reasonable. These values seem to agree with the value of 36.6°C for tympanic temperature, which may be estimated from the graph included in the report on temperature measurement in swine by Hanneman et al. (2004). Hence, the single-exponential model might fully account for the changes in temperature of the eyeball and the orbit soft tissues from the moment of death. That is because of the lack of any plateau in the cooling plots for these body sites.
The extrapolated temperature at death in the rectum (41.5°C) and in the muscles (41.3°C) appears to be higher than physiological temperature. Veterinary reports (Prost, 1985; Klont & Lambooy, 1995; Hanneman et al. 2004) quote pig temperature as ranging from 38.0 to 40.0°C in the rectum and from 38.5 to 40.2°C in the muscles. These values are slightly lower than those obtained by extrapolation from our single-exponential model. Thus, a plateau phase on the temperature decrease time course for the rectum and muscles may exist during the early phase after death. This deviation from the single-exponential model is minor and of no practical consequence for the reliability of estimation of the time of death. Introducing a second exponential term to the cooling equation to account for the probable initial plateau would neither increase the range of the actual estimation period nor the precision of estimations.
Coefficient b of the relationships presented in Figs 25 denotes the rate constant of the cooling process in individual body sites. Its mean value is 0.113 h1 for the eyeballs, 0.111 h1 for the orbit soft tissues, 0.058 h1 for the rectum and 0.064 h1 for the muscles. Thus, the rate constant of temperature decrease in the pig's eyeball is nearly twice that in the rectum. This is due to the anatomical and localization differences between the temperature measuring sites and, consequently, their different thermal conductivities.
While coefficient a depends on the actual ambient temperature only, coefficient b may be affected by various body and environmental parameters such as the body mass, body surface area, obesity, environmental humidity, airflow, clothing, etc. The question is how strongly b depends on individual factors and whether the corrections due to individual factors significantly affect the estimation of the time of death where a large intersubject diversity is present.
In our opinion, the practical usefulness of the otherwise rational corrections is highly disputable. The most commonly accepted factor to affect the cooling speed is the object's body mass (Henssge, 1988). Our correlation analysis does not support the assumed practical value of that factor for improving predictions of the time of death. Table 6 shows that correlations between the pig's body mass (ranging from 81 to 124 kg) and the cooling rate constant b are generally low. In the case of the eyeball cooling equation, the correlation between b and the body mass is at the level of r2 = 0.037, which is practically non-existent. There is a higher, but still very weak, corresponding correlation in the case of the rectum, r2 = 0.365. Still, taking an individual case into consideration, one has little chance to improve the estimation of the time of death by using an exponential equation corrected for the individual's body mass.
Some authors propose a multiexponential equation with individual exponential terms accounting for the temperature in specific body sites (al-Alousi et al. 2002). Such equations are claimed to improve the estimation of the time of death. The approach appears problematic from the thermodynamic point of view, however. If single-exponential equations best describe the cooling of individual sites, as some authors have correctly noticed (al-Alousi et al. 2002), then perhaps the mean of times since death calculated from these simple equations could reduce the error of the estimation. However, the multiexponential equations make no clear physical sense. We managed to formally derive and test multivariable equations for estimation of the time of death taking into account the ratio of temperature differences in individual body sites and the environment. Hence, for example, denoting the actual temperature at time t in the eyeball by T1 and the corresponding temperature in the rectum by T2, we have:
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The fit of our experimental data to the above model was very poor, R.M.S.E. = 1.600.
For the eyeball temperature data sets we also derived a two-exponential equation of the form proposed by al-Alousi et al. (2002):
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The R.M.S.E. for that equation was 1.412. The corresponding values were 0.589 in the case of the eyeball, 0.705 for the orbit soft tissues, 0.936 for the rectum and 0.868 for the muscles. Again, the most simple and thermodynamically valid model appeared to be the best one.
The results of the present study may have direct practical consequences for forensic medicine. The analysis of the data contained in Figs 25 and Tables 1-5 indicates that the precision of the estimation of time of death decreases functionally with time. The single-exponential model correctly predicts the increasingly larger differences between the actual and the calculated time of death, especially in case of the upper 95% confidence limit. Therefore, with time, a larger overestimation than underestimation of the time of death is likely. Our study demonstrates that over the first 13 h after death, the highest accuracy estimations of time of death are provided by measurements of eyeball temperature. This period is characterized by the difference between its temperature (T) and the ambient temperature (TE) = 3.5 (± 1.2)°C. From 13 h post mortem onwards, this accuracy starts to be superseded by the accuracy of muscle temperature measurements, for which this period is defined by the values T TE = 8.9 (± 1.7)°C. From 14 h post mortem, the accuracy of eyeball temperature measurements is superseded by the accuracy of rectal temperature measurements, for which this period is defined by the values T TE = 9.2 (± 1.8)°C. Between 13 and 23.5 h post mortem, the accuracy of the estimation of time of death is highest for muscle temperature measurements, defined by the values T TE = 8.9 (± 1.7)°C and T TE = 4.6 (± 1.7)°C, while from approximately 24 h post mortem onwards, until the cooling of the body is complete, it becomes highest for rectal temperature measurements, for which this period is defined by the values T TE = 5.2 (± 1.8)°C. The accuracy of the measurements of orbit soft tissue temperatures throughout the entire recording period was worse than that for the eyeball measurements. Starting approximately 10.5 h post mortem, defined by T TE = 4.8 (± 1.4)°C, this started to be superseded by the accuracy of muscle temperature measurements.
In summary, the tests have shown that eyeball and orbital tissue temperature measurements may play a key role for estimation of the time of death over the several-hour-long dynamic period occurring directly after death during which the dead body cools off, and that thereafter these measurements should be replaced by muscle or rectal temperature measurements. The results prove the usefulness of the eyeball and the orbit soft tissues as sites for temperature measurement in dead bodies in order to assess the time of death.
The anatomy and physiology of the swine and the human eyeball is similar. This similarity must also apply to the time course of the post-mortem temperature decrease. Arguments to support selection of the eyeballs include: absence of any influence of clothing on their temperature; the practical lack of intersubject variability with respect to thermal capacity of the eyeballs resulting from the homogenicity of the structure and the anatomical location, which leads to identical thermal properties; and the influence of ambient temperature in different individuals depends on eyelid thickness, which is related to the degree of obesity only to a minor extent. This should enable comparison of results obtained from different corpses. The above reasoning applies for the most part to the orbit soft tissues. With longer times from death, however, temperature measurements in the rectum and in the muscles prove superior to measurements in the eyeball and the orbit soft tissues.
The proposition to use eyeball temperature in forensic medicine can be supported by the minor effect of intersubject variables (body size, clothing, etc.) on the cooling rate. The invasive nature of the test may limit its use in some countries for regulatory reasons. However, this obstacle may be omitted using non-invasive infrared laser thermometers providing that the reading will not be too much affected by external conditions, such as wind.
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