Technical note: Temperature estimation accuracy based on colourimetry of embalmed human and fresh non-human burned bone.

Estimation of the exposure temperature of skeletal remains can be done by means of colourimetry and a previously published decision model for the colourimetric data, resulting in clusters that represent a range of exposure temperature. The method was based on thermally altered freshly burned human skeletal remains. However, in practice the origin or pre-burning condition of (possibly fragmentary) burned remains can be unknown. Further, in order to use the colourimetric analysis, and accompanying decision model, it is important to have a reference or test set. Fresh human material is not available for this purpose in all countries. Hence, the classification accuracy of the decision model was tested for, in some countries more readily available substitutes for fresh bone; embalmed human bone and non-human bone. The model yielded high accuracies for these sample materials, making it possible to create a reference or test set from fresh non-human and embalmed human bone as substitute for human, and also to use the decision model for these deviating samples in practice.

An overview of the heat-induced changes of the chemical composition of bone from fresh to calcined.

When bone is exposed to thermal stress, the chemical composition changes. This affects bone tissue regeneration after surgery, and these changes can also aid in reconstructing ante-, peri-, and post-mortem events in forensic investigations and past activities on cremation practices in archaeology. However, to date, no complete overview exists on the chemical composition of both fresh and thermally altered bone. Therefore, we aimed (i) to present the chemical composition of fresh bone and (ii) to present an overview of heat-induced chemical changes in bone under both reducing and oxidizing conditions. From the overview, it became clear that some chemical changes occur at a consistent temperature, independent of exposure duration, meaning there is a temperature threshold. However, the occurrence of other chemical changes appeared to be more inter-experimentally variable, and therefore, it is recommended to further investigate these changes.

Subaerial Decomposition of Small-Sized Remains in The Netherlands: Important Findings Regarding the PMI of a Four-Year Taphonomic Study.

Studying post-mortem changes based on signs of decomposition (e.g., using scoring models) is one of the methods used in scientific studies to relate observable changes to the post-mortem interval (PMI). The majority of the studies on cadaver decomposition are based on large cadavers. There is limited literature on the decomposition pattern and rate of small cadavers, even though it is at least as important to be able to estimate the PMI for infants and subadults. Therefore, it is crucial to acquire knowledge of the decomposition process of child-sized remains. To fill this knowledge gap, a season-based subaerial outdoor decomposition study was conducted with small pig cadavers at the Forensic Anthropological Outdoor Research Facility located in Den Ham, The Netherlands, over a period of 4 years. Den Ham is located in the eastern part of the Netherlands, close to the German border, and has a temperate maritime climate, with a Cfb classification according to the Köppen-Geiger system. Salient findings were acquired during the decomposition study, specifically regarding a deviating decomposition rate during winter and the subsequent spring, reproducibility, the effect of body weight, post-mortem movement, the effect of heavy rainfall on insect activity, delayed bloating, the interaction of different insect species, and invertebrate activity. This article includes a systemic review of the results obtained during this four-year decomposition study and discusses the impact of the findings on the estimation of the PMI.

Temperature-specific spectral shift of luminescing thermally altered human remains.

Human bone has shown to have luminescent properties that remain throughout the phases of cremation, with the exception of fully carbonized bone, when excited with a narrow band light source. During this research, an alternate light source (420-470nm, peak at 445nm) was used to visualize and investigate latent details relevant for forensic investigations of human remains recovered at fire scenes. As fire is a destructive force, it induces a vast variety of physical and chemical alterations to all components of the bone, making the subsequent analysis and interpretation of burned human remains challenging. A spectral shift in emission bandwidth, from green to red, was previously observed when the exposure temperature increased from 700 to 800 °C. This spectral shift was reproduced on a total of 10 human forearms, divided into 20 segments, by burning at 700 °C and 900 °C in an ashing furnace. The shift of emission bandwidth caused only by an increase in temperature was furthermore investigated by colorimetric analysis, proving the spectral shift to be significant. By easily quantifying the spectral shift, substantiation is provided for the use of this technique in practice to improve the interpretation of heat induced changes of bone.

Mechanical or thermal damage: differentiating between underlying mechanisms as a cause of bone fractures.

To investigate the differences between pre- and post-fire fractures, 30 human forearm bones were subjected to either blunt-force impact, burning, or both. Bones, covered in soft tissue and wrapped in clothing, were burned in a reconstructed house fire. The burning context and dynamics led to differential burning, that was equal amongst the three groups. To evaluate the damage to the bones, a data collection sheet was developed based on the current literature on fracture features. To analyze the relation between exposure temperature and fracture class and occurrence, color was measured to estimate the exposure temperature. Observable and measurable changes on bone, and fracture surfaces, were macro- and microscopically analyzed. Many features overlapped between the three groups, and thus are not usable for differentiation. Fractures caused by blunt force impact (post-mortem, pre-fire) showed a rough fracture surface with smoothness in curved/slanted regions near the margin after burning, while heat-induced bone fractures showed a smooth fracture surface. The margins and surface of bone fractures that originated after the fire (indirect heat-induced) were evenly discolored, whereas heat-induced bone fractures showed uneven discoloration of the fracture margin and surface. Although there were generally more heat-induced fractures in the 450-700 °C range, no other distinctive trend was observed between exposure temperature and class of fractures.

The time of death in Dutch court; using the Daubert criteria to evaluate methods to estimate the PMI used in court.

When a capital crime is committed the post-mortem interval (PMI) is of particular importance in investigating a suspect's alibi in court. A forensic expert can use different methods to estimate the PMI. This research focuses on who is considered an expert in court and whether the methods used to estimate the PMI are reliable. In this study, the methods used to estimate the PMI and the experts consulted, available in Dutch jurisprudence, in the period 2010-2019 were investigated. Ninety-four judicial cases were included and multiple experts and methods of estimating the PMI were found. As part of this study, the methods that were used to estimate the PMI in court were subjected to the Daubert criteria. Of these methods, only the Henssge nomogram and entomological methods met the Daubert criteria. However, the methods are only useful when applied by the right forensic expert and in the right manner. Unfortunately, this was not always the case.

Phosphorescence of thermally altered human bone.

Bone has photoluminescent characteristics that can aid the analysis of thermally altered human skeletal remains as part of the forensic anthropological investigation. Photoluminescence stands collectively for fluorescence and phosphorescence. Because the difference in lifetime between fluorescence and phosphorescence is usually in the range of nano- to microseconds, it is only possible to visually determine whether bone phosphoresces when the lifetime is long enough to be observed. For this study, a distinction was made between long-decay and short-decay phosphorescence. So far, it was unknown whether (thermally altered) human bone emits long-decay phosphorescence after being illuminated and, thus, whether phosphorescence contributes to the observed photoluminescence. If so, whether the observable phosphorescence is dependent on temperature, exposure duration, surrounding medium, bone type, skeletal element, and excitation light and could aid the temperature estimation of heated bone fragments. In this study, bone samples were subjected to heat in the range of from room temperature to 900 °C for various durations in either air or adipose as surrounding medium. In addition, different skeletal elements of a human cadaver were recollected after cremation in a crematorium. Both sample collections were illuminated with light of different bandwidths and visually inspected for phosphorescence and photoluminescence. The samples were scored by means of a scoring index for the intensity of long-decay phosphorescence and photographically documented. The results show that thermally altered human bone fragments do phosphoresce. The observed phosphorescence is more dependent on temperature than on exposure duration, surrounding medium or skeletal element. Of the used wavelength bands, ultraviolet light provided the most temperature-related information, showing changes in both phosphorescence intensity and emission spectrum. Long-decay phosphorescence and fluorescence with short-decay phosphorescence coincide; however, there are also temperature-dependent differences. It is therefore concluded that phosphorescence contributes to the observable photoluminescence and that the visibly observable phosphorescent characteristics can aid the temperature estimation of cremated human skeletal fragments.

Amsterdam Research Initiative for Sub-surface Taphonomy and Anthropology (ARISTA) - A taphonomic research facility in the Netherlands for the study of human remains.

A taphonomic research facility for the study of human remains was recently realized in Amsterdam, the Netherlands, to systematically investigate the decomposition of the human body under known conditions. Governmental authorization was obtained to make use of the body donation program of the Amsterdam University Medical Centers, location Academic Medical Center, for this specific purpose. In contrast to the small number of comparable initiatives elsewhere, this facility specifically allows for the study of buried bodies e.g. with the use of telemetry and remote sensing. Here, we discuss the concept of body donation in the Netherlands, its role in taphonomic research, and the sequence of events that preceded the realization of this facility, which is the first of its kind in Europe. In addition to offering novel research options to the scientific community, we hope that it will also pave the way for the successful realization of similar initiatives in other locations.

Colourimetric analysis of thermally altered human bone samples.

At this moment, no method is available to objectively estimate the temperature to which skeletal remains have been exposed during a fire. Estimating this temperature can provide crucial information in a legal investigation. Exposure of bone to heat results in observable and measurable changes, including a change in colour. To determine the exposure temperature of experimental bone samples, heat related changes in colour were systemically studied by means of image analysis. In total 1138 samples of fresh human long bone diaphysis and epiphysis, varying in size, were subjected to heat ranging from room temperature to 900 °C for various durations and in different media. The samples were scanned with a calibrated flatbed scanner and photographed with a Digital Single Lens Reflex camera. Red, Green, Blue values and Lightness, A-, and B-coordinates were collected for statistical analysis. Cluster analysis showed that discriminating thresholds for Lightness and B-coordinate could be defined and used to construct a model of decision rules. This model enables the user to differentiate between seven different temperature clusters with relatively high precision and accuracy. The proposed decision model provides an objective, robust and non-destructive method for estimating the exposure temperature of heated bone samples.

Evaluation of postmortem biochemical markers: Completeness of data and assessment of implication in the field.

Throughout the years an increase has been observed in research output on biochemical markers for determining the postmortem interval (PMI). However, to date, a complete overview is missing on the results of postmortem biochemical markers (PBM's) for PMI estimation. In this paper, literature was reviewed in order to identify the knowledge lacunae of PBM research from a practical point of view. A three-step approach was undertaken in order to achieve the set goal. Literature was collected, the PBM's were evaluated for completeness by means of a scorings index based on set criteria, and PBM's were subsequently evaluated in light of the Daubert &Frye criteria for scientific evidence in court. Seven PBM's were found to be well investigated, from which potassium had the highest completion score. However, none of these PBM's could be qualified as suitable for court evidence. Further, this study revealed that the majority of PBM's (94%) is not well investigated. Consequently, these PBM's did not meet Daubert &Frye criteria. In order to improve the assessment for use of PBM's as evidence in court regarding PMI estimation, PBM's should be investigated more thoroughly and data should be made readily available.

Temperature estimations of heated bone: A questionnaire-based study of accuracy and precision of interpretation of bone colour by forensic and physical anthropologists.

The colour of thermally altered bone, recovered from archaeological and forensic contexts, is related to the temperature(s) to which it was exposed. As it is heated bone changes in colour from ivory white, to brown and black, to different shades of grey and chalky white. It should be possible to estimate exposure temperature based on visually observable changes in colour. In forensic casework the temperature that human remains have been subjected to can reveal information about the existence and nature of foul play. Therefore, it is important to understand the accuracy and precision of visual methods of temperature estimation. Twenty-eight forensic and/or physical anthropologists estimated the temperature that fourteen bone samples had been subjected to based only on their colour via an online questionnaire. Bone samples shown in the questionnaire ranged from unheated to having been heated at 1200°C. Respondents were given two options to base their estimates on, resulting in a multiple response analysis. The results suggest it is difficult to identify the correct temperature range based solely on colour. Most respondents felt confident enough to opt for a single option, which may have contributed to a relatively high number of incorrect estimates. Low accuracy and precision were found for most of the temperature ranges, especially in the lower and middle categories. This study demonstrates that caution should be taken in the reliance upon temperature estimates of thermally induced colour changes in bone and the need for further research and improved methods.

Luminescence of thermally altered human skeletal remains.

Literature on luminescent properties of thermally altered human remains is scarce and contradictory. Therefore, the luminescence of heated bone was systemically reinvestigated. A heating experiment was conducted on fresh human bone, in two different media, and cremated human remains were recovered from a modern crematory. Luminescence was excited with light sources within the range of 350 to 560 nm. The excitation light was filtered out by using different long pass filters, and the luminescence was analysed by means of a scoring method. The results show that temperature, duration and surrounding medium determine the observed emission intensity and bandwidth. It is concluded that the luminescent characteristic of bone can be useful for identifying thermally altered human remains in a difficult context as well as yield information on the perimortem and postmortem events.

Technical note: unsafe rectal temperature measurements due to delayed warming of the thermocouple by using a condom. An issue concerning the estimation of the postmortem interval by using Henßge's nomogram.

In some cases, in the Netherlands, an additional layer is being added to the thermocouple, used to measure the rectal temperature in medicolegal death investigations. Because of this deviation from the standard method, questions arose regarding the accuracy and precision of the measured temperature. Therefore, a cooling experiment was carried out on a round body made of agar with an average thermal conductivity of 0.454 W/(m °C) while measuring the temperature with and without an additional layer around the thermocouple for three different starting temperatures: 36, 30, and 27 °C. The results show a significant difference between the measured values for the first 5 min when comparing with and without the additional layer. Further, a decrease in precision is present within the first minutes when using an additional layer. Therefore, it is concluded that it is best to measure the rectal temperature without an additional layer around the thermocouple and caution should be taken when measuring with an additional layer.