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Routine coronal paraffin-sections through the dorsal frontal and parieto-occipital cortex of a total of sixty cases with divergent causes of death were immunohistochemically (IHC) stained with an antibody against TMEM119. Samples of cerebrospinal fluid (CSF) of the same cases were collected by suboccipital needle-puncture, subjected to centrifugation and processed as cytospin preparations stained with TMEM119. Both, cytospin preparations and sections were subjected to computer-assisted density measurements. The density of microglial TMEM119-positive cortical profiles correlated with that of cytospin results and with the density of TMEM119-positive microglial profiles in the medullary layer. There was no statistically significant correlation between the density of medullary TMEM119-positive profiles and the cytospin data. Cortical microglial cells were primarily encountered in supragranular layers I, II, and IIIa and in infragranular layers V and VI, the region of U-fibers and in circumscribed foci or spread in a diffuse manner and high density over the white matter. We have evidence that cortical microglia directly migrate into CSF without using the glympathic pathway. Microglia in the medullary layer shows a strong affinity to the adventitia of deep vessels in the myelin layer. Selected rapidly fatal cases including myocardial infarcts and drowning let us conclude that microglia in cortex and myelin layer can react rapidly and its reaction and migration is subject to pre-existing external and internal factors. Cytospin preparations proved to be a simple tool to analyze and assess complex changes in the CNS after rapid fatal damage. There is no statistically significant correlation between cytospin and postmortem interval. Therefore, the quantitative analyses of postmortem cytospins obviously reflect the neuropathology of the complete central nervous system. Cytospins provide forensic pathologists a rather simple and easy to perform method for the global assessment of CNS affliction.

The aim of this study was to investigate if the biomarkers myelin basic protein (MBP) and neurofilament-H (NF-H) yielded informative value in forensic diagnostics when examining cadaveric cerebrospinal fluid (CSF) biochemically via an enzyme-linked immunosorbent assay (ELISA) and comparing the corresponding brain tissue in fatal traumatic brain injury (TBI) autopsy cases by immunocytochemistry versus immunohistochemistry. In 21 trauma and 19 control cases, CSF was collected semi-sterile after suboccipital puncture and brain specimens after preparation. The CSF MBP (p = 0.006) and NF-H (p = 0.0002) levels after TBI were significantly higher than those in cardiovascular controls. Immunohistochemical staining against MBP and against NF-H was performed on cortical and subcortical samples from also biochemically investigated cases (5 TBI cases/5 controls). Compared to the controls, the TBI cases showed a visually reduced staining reaction against MBP or repeatedly ruptured neurofilaments against NF-H. Immunocytochemical tests showed MBP-positive phagocytizing macrophages in CSF with a survival time of > 24 h. In addition, numerous TMEM119-positive microglia could be detected with different degrees of staining intensity in the CSF of trauma cases. As a result, we were able to document that elevated levels of MBP and NF-H in the CSF should be considered as usefulneuroinjurybiomarkers of traumatic brain injury.

The aim of the present study was a refined analysis of neuroinflammation including TMEM119 as a useful microglia-specific marker in forensic assessments of traumatic causes of death, e.g., traumatic brain injury (TBI). Human brain tissue samples were obtained from autopsies and divided into cases with lethal TBI (n = 25) and subdivided into three groups according to their trauma survival time and compared with an age-, gender-, and postmortem interval-matched cohort of sudden cardiovascular fatalities as controls (n = 23). Brain tissue samples next to cortex contusions and surrounding white matter as well as samples of the ipsilateral uninjured brain stem and cerebellum were collected and stained immunohistochemically with antibodies against TMEM119, CD206, and CCR2. We could document the highest number of TMEM119-positive cells in acute TBI death with highly significant differences to the control numbers. CCR2-positive monocytes showed a significantly higher cell count in the cortex samples of TBI cases than in the controls with an increasing number of immunopositive cells over time. The number of CD206-positive M2 microglial cells increased survival time-dependent. After 3 days of survival, the cell number increased significantly in all four regions investigated compared with controls. In sum, we validate a specific and robustly expressed as well as fast reacting microglia marker, TMEM119, which distinguishes microglia from resident and infiltrating macrophages and thus offers a great potential for the estimation of the minimum survival time after TBI. Electronic supplementary material The online version of this article (10.1007/s00414-020-02384-z) contains supplementary material, which is available to authorized users.

The aim of this pilot study was to investigate the diagnostic potential of TMEM119 as a useful microglia-specific marker in combination with immunostainings for phagocytic function and infiltrating capacity of monocytes in cases of lethal monosubstance intoxications by morphine (MOR), methamphetamine (METH), and of ethanol-associated death (ETH) respectively. Human brain tissue samples were obtained from forensic autopsies of cases with single substance abuse (MOR, n = 8; ETH, n = 10; METH, n = 9) and then compared to a cohort of cardiovascular fatalities as controls (n = 9). Brain tissue samples of cortex, white matter, and hippocampus were collected and stained immunohistochemically with antibodies against TMEM119, CD68KiM1P, and CCR2. We could document the lowest density of TMEM119-positive cells in MOR deaths with highly significant differences to the control densities in all three regions investigated. In ETH and METH deaths, the expression of TMEM119 was comparable to cell densities in controls. The results indicate that the immunoreaction in brain tissue is different in these groups depending on the drug type used for abuse.

Due to its protected anatomical location, cerebrospinal fluid (CSF) is a very stable fluid which undergoes comparatively little change in the early post-mortem phase. While many immunohistochemical markers already established for clinical diagnostic issues in tissue samples obtained by biopsy could meanwhile be translated also to post-mortem tissue, no systematic immunocytochemical investigations have generally been conducted on post-mortem body fluids and for CSF specifically, have not been established at all. CSF as the fluid directly surrounding the brain should also be examined to allow a more detailed characterization of processes in the central nervous system. Comparing traumatized tissue and CSF can complete forensic assessment and complement neuropathological evaluation.

Blood-brain barrier (BBB) integrity is one of the important elements of central nervous system (CNS) homeostasis. MicroRNAs (miRs) have been demonstrated to play a role in many CNS disorders such as stroke and traumatic brain injury. MiR-212/132 are highly expressed in the CNS but their role at the BBB has not been characterized yet. Thus, we analyzed the expression of miR-212/132 in hypoxic mouse and human brain microvascular endothelial cells (BMEC) as well as in posttraumatic mouse and human brain tissue and serum exosomes. MiR-212/132 expression was detected in brain capillaries by in situ hybridization and was increased up to ten times in hypoxic BMEC. Over-expression of pre-miR-212/132 in BMEC decreased barrier properties and reduced migration of BMEC in the wound healing assay. We identified and validated tight junction proteins claudin-1 (Cldn1), junctional adhesion molecule 3 (Jam3), and tight junction-associated protein 1 (Tjap1) as potential miR-212/132 targets. Over-expression of miRs led to a decrease in mRNA and protein expression of Cldn1, Jam3, and Tjap1, which could be rescued by a respective anti-miR. In conclusion, our study identifies miR-212/132 as critical players at the hypoxic BBB. In addition, we propose three new direct miR-212/132 targets to be involved in miR-212/132-mediated effects on BBB properties. Electronic supplementary material The online version of this article (10.1007/s12975-018-0683-2) contains supplementary material, which is available to authorized users.

In the last few years, quantitative analysis of metabolites in body fluids using LC/MS has become an established method in laboratory medicine and toxicology. By preparing metabolite profiles in biological specimens, we are able to understand pathophysiological mechanisms at the biochemical and thus the functional level. An innovative investigative method, which has not yet been used widely in the forensic context, is to use the clinical application of metabolomics. In a metabolomic analysis of 41 samples of postmortem cerebrospinal fluid (CSF) samples divided into cohorts of four different causes of death, namely, cardiovascular fatalities, isoIated torso trauma, traumatic brain injury, and multi-organ failure, we were able to identify relevant differences in the metabolite profile between these individual groups. According to this preliminary assessment, we assume that information on biochemical processes is not gained by differences in the concentration of individual metabolites in CSF, but by a combination of differently distributed metabolites forming the perspective of a new generation of biomarkers for diagnosing (fatal) TBI and associated neuropathological changes in the CNS using CSF samples.

In this casuistry, two accidents from Germany and Switzerland are presented that happened during the shot of recoilless anti-tank weapons. In both cases, the injuries led to the death of two soldiers: A 22-year-old soldier in Germany was struck by the counter mass of a so-called Davis gun which had been fired by a comrade during a firing exercise; he died from his severe injuries, especially in the abdominal part of the body. As a peculiarity of the wound morphology, it was found to be a thick-layered, metallic, gray material in the wound cavity, which corresponded to the material of the counter mass that was ejected opposite to the shooting direction. The other case took place in Switzerland, where a 24-year-old soldier was seriously injured during an exercise with portable anti-tank rockets. At the time the shot was fired, he stood behind the launcher and was hit by the propulsion jet of the rocket motor. He died as well from his severe injuries, which were located at the chest done by the gas jet and by the very high pressure. In both cases, two different causes of death were present: massive blunt violence in the first case versus a jet of hot gases of very high speed and temperature in the second case.

During the investigative process that typically follows a criminal act, it may prove necessary to work with and analyze evidence that is not recent but old. This could become necessary, for example, when a crime is discovered some time after it was committed or when a cold case is reopened. Due to this need, the present study focused on the detection and visualization of 2-year-old biological traces. To do so, an alternative light source and different filters were used. The optical behavior of 2-year-old samples of blood, semen, urine, saliva, and sweat located on 19 different materials was documented, analyzed, and compared with the optical behavior of the same samples when they were recent [1].

Background Human hippocampal area Cornu Ammonis (CA) 1 is one of the first fields in the human telencephalon showing Alzheimer disease (AD)-specific neuropathological changes. In contrast, CA2 and CA3 are far later affected pointing to functional differences, which may be accompanied by differences in proteome endowment and changes. Methods Human pyramidal cell layers of hippocampal areas CA1, CA2, and CA3 from neurologically unaffected individuals were excised using laser microdissection. The proteome of each individual sample was analyzed and differentially abundant proteins were validated by immuno-histochemistry. Results Comparison of CA1 to CA2 revealed 223, CA1 to CA3 197 proteins with differential abundance, among them we found motor proteins MYO5A and DYNC1H1. Extension of the study to human hippocampus slices from AD patients revealed extensive depletion of these proteins in CA1 area compared to unaffected controls. Conclusion High abundance of motor proteins in pyramidal cell layers CA1 compared to CA2 and CA3 points the specific vulnerability of this hippocampal area to transport-associated changes based on microtubule dysfunction and destabilization in AD.

The concept of spontaneous combustion has huge appeal as an historical fiction but it has no scientific background. However, in some deaths involving fire, a body can burn away virtually completely with minimal thermal destruction to the neighboring environment. We report an extraordinary case of the self-immolation of an elderly woman who set herself on fire with suicidal intentions. The unusual appearance and location of the body closely resembled the phenomenon of so-called spontaneous human combustion because the upper parts of the body were almost totally destroyed by fire, while the legs and surrounding structures remained almost untouched by flames. The results of all investigations proved that the woman set fire to her body using a box of matches and accelerants (concentrated ethanol and a solid firelighter). Near-total combustion of her body subsequently occurred during the postmortem period. The development of alleged spontaneous human combustion requires the following: ignition (external heat source), fuel (molten human fat), a wick (e.g. charred and porous clothing, bedding, or ground), time, and an optimal microclimate for gradual burning.

Crania are a reliable source for sex estimation in Euro-Americans, Europeans, and most other populations. Besides morphological assessments, the application of Fordisc® has become a useful tool within the last two decades, creating discriminant functions from morphometric data. Unfortunately, until now, white populations are mostly represented by measurements of American individuals. Therefore, classification rates are lower for European skulls than for Euro-Americans. The aim of this study was to show differences in sexual dimorphism between German and Euro-American crania. Furthermore, their secular change from the nineteenth to the twentieth century has been investigated. Analyses have been performed on glabella subtense (GLS), mastoid height (MDH), and bizygomatic breadth (ZYB). Fordisc® 3.1 was used to study sexual dimorphism and secular change, whereas SAS® was used to perform a two-level ANOVA to test for variation in sex dimorphism. Euro-Americans show greater dimorphism than Germans in all three measurements tested. This larger difference is even increasing from the late nineteenth through the late twentieth century in terms of GLS and MDH, while it stays almost the same in the present Europeans. These results explain the unsatisfying classification rates of German and other European crania on Fordisc®. Data collection for European Fordisc® samples is in progress and should improve the current situation.

In medico-legal literature, only a small number of publications deal with lethal injuries caused by shots with modified guns. This might lead to the conclusion that such cases are extremely rare. However, there are cases again and yet again. During the investigation process, the modified gun is of particular importance since it can show an unusual ballistic behaviour. The present paper reports on a suicide of a 60-year-old man, committed with a modified revolver and a lead bullet. The man had a single gunshot wound with entrance at the right temporal bone. Autopsy revealed that the bullet had fragmented into two major parts. The smaller one stood outside the cranial cavity and pushed its way alongside between the cranial bone and scalp to its end position in the left temporal area. The bigger part entered the cranial cavity and ended in the left parietal lobe. In shots on ballistic soap and on a head-model, the ballistics of the weapon and lead bullet were characterized. The angle necessary for bullet fragmentation was determined by shots on ballistic soap and turned out to be 55°-60° at a velocity of around 200 m/s. This knowledge was transferred to contact shots on a head-model consisting of a layered polyurethane sphere filled with 10 % ballistic gelatine and covered with a skin-like cap almost all around. The resulting injury pattern corresponded to the one of the suicide person. The bigger bullet part entered the skull while the smaller part pushed its way alongside between skin and skull causing an outer contour shot. Furthermore, the revolver was documented firing off two bullets by one trigger pull-a phenomenon of importance for forensic casework the authors have not found reported in forensic literature.

The aim of this study was to figure out a new practically applicable method to distinguish between historical and recent human skeletal remains. Therefore, the optical behavior of bone cross sections was investigated using the combination of two methods: a modification of an already established test (UV-induced fluorescence) and a new method (490 nm-induced fluorescence). We evaluated the areal extent of fluorescence of 30 bone cross sections with known postmortem interval (PMI) using ultraviolet light and 490 nm light. For analysis, the areal extend of fluorescent surface was determined using photos of the samples and an image editing software. The results prove that there is a correlation between PMI and the areal extent of fluorescent surface in both tests. Furthermore, the combination of both methods is a good indicator to distinguish within the forensic relevant post mortem interval between PMI 30 years and PMI 30 years.

Whenever blunt or sharp forces are used in a crime, analysis of bloodstain pattern distribution may provide important information for the reconstruction of happenings. Thereby, attention should be paid to both the crime scene and the clothes of everyone involved in the crime. On dark textiles, though, it is difficult or even impossible for the human eye to detect bloodstains because of the low contrast to the background. However, in the near infrared wavelength range, contrast is considerably higher. Many textiles reflect light beyond a wavelength of 830 nm and thus appear light-colored, whereas blood absorbs the light and appears dark. In our studies, a D7000 NIKON reflex camera modified for infrared photography produced high-resolution photographs visualizing even very small spatter stains on dark textiles. The equipment can be used at any crime scene or lab and provides immediately available and interpretable images. Thus, important findings can be obtained at an early stage of police investigations, as two examples (homicide and attempted homicide) illustrate.

To find out the time since death (PMI) of a human individual, from which only skeletal remains are left is a difficult question to answer, since methods currently available don’t bring concrete answers. Subsequent to our validation study on porcine bones, we focused on human bones analysing if collagen degradation would occur at a predictive rate so that the method may be used for PMI-estimation. We tried to reproduce a method for determining the Co/NCo (Collagen to Non-Collagen) ratio of porcine bones Boaks et al. demonstrated in 2014. Our study sample consisted of 37 human bones from a forensic PMI context and 11 archaeological samples; we prepared thin bone sections of 250 µm; after staining the sections we used spectrophotometry for a portion of the samples and stereomicroscopy and digital imaging to analyze the Co/NCo ratio. Detecting the Co/NCo ratio with spectrophotometry produced results much lower than those published by Boaks et al. (2014) on porcine bone samples and much lower than physiologically expected; a similar result we also got in our validation study on porcine bone samples. We, therefore, applied our new method of stereomicroscopy and digital imaging, which we previously tested on porcine bones. The samples of male individuals showed a significant reduction of the Co/NCo ratio correlating to the PMI. Nevertheless, we still consider analyzing the Co/NCo ration of human bone samples not sufficient for forensic issues. Therefore, greater reference data as well as more experience in practice are needed.

A 31-year-old female asthmatic patient received an infusion of metamizole and tramadol for chronic pain at a GP surgery. After a few minutes, the patient developed breaing difficulties and died in spite of resuscitation measures. The general practitioner was suspected of medical malpractice. Medico-legal investigations confirmed the assumption that death was caused by anaphylacitic shock. In spite of temporary intubation into the oesophagus no evidence of medical malpractice was found, however.

In many forensic cases, the existence of a traumatic brain injury (TBI) is an essential factor, and the determination of the survival time is nearly as important as the determination of whether or not a trauma exists. Since it is known that glucose uptake increases in injured brain cells in order to perpetuate the neuronal integrity, this study focuses on the pathomechanism of brain glucose supply via sodium/glucose cotransporters 1 and 2 (SGLT1, SGLT2) following traumatization. Human cerebrum tissue of male and female individuals who died following TBI was taken from the contusional and contralateral regions, as well as from individuals deceased due to cardiac arrest (control group). Total SGLT1 and SGLT2 protein expression was analyzed by immunoblotting comparing injured and non-injured tissue. The immunoreactivity in contusional cerebral cortex region began to increase 3 to 7 h following traumatization. We found that both SGLT1 and SGLT2 protein expression increased significantly 37 h post-injury compared to the control group. SGLT1 rose significantly at 52 h post-injury and peaked significantly at 72 h, while SGLT2 rose significantly at 52 and 72 h after injury. By compiling these data, we predict a standard operator via SGLT expression as a comparative expression assertion to determine post-injury survival time for unknown cases. Our result suggests that SGLT1 and SGLT2 protein expression may be useful in forensic practice as an effective target to analyze the existence of a TBI and to determine the time of the traumatization.

Because biological traces often play an important role in the investigation process of criminal acts, their detection is essential. As they are not always visible to the human eye, tools like a forensic light source or infrared photography can be used. The intention of the study presented was to give advice how to visualize biological traces best. Which wavelengths and/or filters give the best results for different traces on different fabrics of different colors? Therefore, blood (undiluted and diluted), semen, urine, saliva, and perspiration have been examined on 29 different materials.

Protrusion of the tongue is a common, though often neglected finding in fire fatalities. According to a study recently published by Bernitz et al., it is an indicator of vital burning. This statement has been doubted repeatedly. Retrospective analysis of 61 fire fatalities from our own autopsy material did not show any statistically significant increased incidence of tongue protrusion in deaths with vital exposure to heat. Similarly, there was no correlation with the degree of destruction by the fire in general or the extent of cervical burning. Further prospective studies seem to be necessary also with regard to the pathophysiological processes.

Studying secular changes on human skulls is a central issue in anthropological research, which is however insufficiently investigated for modern German populations. With our study, we focus on morphological cranial variations within Germans during the nineteenth and twentieth centuries. To study this, we recorded different facial landmarks from a cohort study of about 540 German individuals of different age and sex by calculating their cranial size, shape dimensions, and cranial module and cranial capacity to get information about variations occurring during the decades. According to this, measured variables for Germans and Americans, to which we compared our results, were maximum cranial length (glabello-occipital length), basion-bregma height (BBH), basion-nasion length (BNL), maximum cranial breadth (XCB), and cranial base breadth (AUB). Cranial size was calculated as the geometric mean of GOL, BBH, and XCB. Samples were organized into quarter century birth cohorts, with birth years ranging from 1800 to 1950. One-way ANOVA was used to test for variation among cohorts. Over the past 150 years, Americans and Germans showed significant parallel changes, but the American cranium remained relatively higher, with a longer cranial base, as well as narrower than the German cranium. Our results should also lead to the extension of the range of populations listed and investigated for Fordisc®, a forensic software to identify unknown individuals as from their skeletal remains or just parts of them. Fordisc cannot provide a satisfying identification of European individuals yet because the database is missing enough European reference samples.

In the present study, a series of reflectance spectra of postmortem lividity, pallor, and putrefaction-affected skin for 195 investigated cases in the course of cooling down the corpse has been collected. The reflectance spectrometric measurements were stored together with their respective metadata in a MySQL database. The latter has been managed via a scientific information repository. We propose similarity measures and a criterion of similarity that capture similar spectra recorded at corpse skin. We systematically clustered reflectance spectra from the database as well as their metadata, such as case number, age, sex, skin temperature, duration of cooling, and postmortem time, with respect to the given criterion of similarity. Altogether, more than 500 reflectance spectra have been pairwisely compared. The measures that have been used to compare a pair of reflectance curve samples include the Euclidean distance between curves and the Euclidean distance between derivatives of the functions represented by the reflectance curves at the same wavelengths in the spectral range of visible light between 380 and 750 nm. For each case, using the recorded reflectance curves and the similarity criterion, the postmortem time interval during which a characteristic change in the shape of reflectance spectrum takes place is estimated. The latter is carried out via a software package composed of Java, Python, and MatLab scripts that query the MySQL database. We show that in legal medicine, matching and clustering of reflectance curves obtained by means of reflectance spectrometry with respect to a given criterion of similarity can be used to estimate the postmortem interval.

A 23-year-old woman was mortally afraid of dental interventions and decided to have her four wisdom teeth removed by outpatient surgery under endotracheal anaesthesia. According to the files, the patient was categorized as ASA I and Mallampati II, and surgery was considered an elective routine intervention. Soon after initiation of anaesthesia, O2 saturation and blood pressure dropped, and the young woman died shortly afterwards in spite of immediate resuscitation measures. At first, an allergic reaction to succinylcholine, which had been administered as a muscle relaxant, was suspected. Autopsy and histological examination showed haemorrhagic pulmonary oedema and a defined lesion in the midportion of the oesophageal mucosa in spite of correct placement of the endotracheal breathing tube. Ultimately, misintubation into the oesophagus, which had not been noticed at first, was determined as cause of death.

As to their optical properties, the components of human skin can be divided into two different categories: the light-scattering components shown as peaks and those absorbing light appearing as dips in the reflectance spectrum. As the post-mortem interval progresses, the concentration of scatterers and absorbers and thus the reflectance spectra change due to post-mortem tissue breakdown and degradation. Based on a total number of 532 reflectance spectrometric measurements in 195 deceased, a characteristic change in the reflectance spectra could be documented in the post-mortem course. Subsequently, an algorithm to calculate the post-mortem interval was developed by analysing the reflectance spectrometric extrema.

Immunohistochemical studies about the presence of fibronectin in the lungs were performed in a group of 73 fire victims (63 cases of intravital and 10 cases of postmortem burn) as well as in an unselected control group of 55 individuals not exposed to fire before death. The cases of intravital burn showed a significantly stronger fibronectin expression than the control cases and the cases of postmortem burn. Fibronectin was mainly present in macrophages of the peribronchial lung parenchyma and, not associated with cells, in the matrix of peribronchial tissue. Our findings suggest that higher levels of fibronectin expression in the lung tissue of burn victims compared to fire-unrelated deaths may serve as an indicator of an early intravital inflammatory response to fire damage.

A 23-year-old woman traveling in the front passenger seat of a car was killed in a traffic accident involving two rear-end collisions. After the second rear-end collision, the car caught fire. While the driver could be rescued, the pregnant passenger remained inside the car. After the fire had been extinguished, a charred body was found in a pugilistic attitude with an exposed abdominal cavity and a ruptured anterior wall of the gravid uterus. Between the woman's legs, an almost unburned fetus was found with body measurements corresponding to a gestational age of 28 weeks. The placenta had remained inside the uterine cavity and was still connected with the fetus via the umbilical cord. In conclusion, the rupture of the pregnant uterus with expulsion of the fetus had to be considered a secondary event due to the initial heating and subsequent shrinking of the uterine wall.

During the autopsy of fire victims, one often sees intensive reddening of the inner vascular layers resembling changes due to putrefaction. To determine the frequency of this phenomenon and the circumstances under which it occurs, in particular the presence of putrescence, signs of vitality and the extent of destruction by the fire, the fire fatalities autopsied at the Institutes of Legal Medicine in Freiburg, Halle and Würzburg were analyzed. In the concrete study, 259 fire deaths were investigated retrospectively and 21 prospectively. The study material included 191 males and 68 females aged 0 to 93 years. Vital exposure to the fire was found in 70%, perimortal exposure in 23 % and postmortem exposure in 7% of the cases. Red discoloration of the intima of the central vessels was seen in 106 victims (41%). This finding is most likely caused by postmortem heat-induced hemolysis, which occurs when the temperature in the body rises above 52 °C due to the effect of external heat. Therefore, a reddish discoloration of the intima cannot be regarded as sign of vitality.

In a case of aggravated extortion under threat of force, the medicolegal expert was asked to give an opinion on the drying time of human saliva. Fresh traces of saliva were applied to composite stones similar to those at the scene of the crime and their drying behavior was examined in a climate test chamber. Air temperature and air humidity during the experiment corresponded to the values of the measuring station located nearest to the crime scene. After one hour and 40 minutes no salivary spots were discernible on the stones any more. Depending on various influence factors, the traces could have originated both at the time of the offence and within a time window of up to 3 hours before.

In terms of physics, the skin can be regarded as an optically turbid medium in which the light is mainly scattered by the collagen fibers, mitochondria and cell nuclei, whereas the absorption is determined by the content of reduced hemoglobin, oxyhemoglobin, bilirubin, and melanin. When the measuring geometry and the illumination spectrum are known, the optical characteristics of the skin can be approximately described by the diffusion and absorption coefficients. These values define the diffusion and absorption probability per unit distance traveled for each wavelength. Based on these parameters, a mathematical skin model was developed with the help of Monte Carlo simulations. By implementing the absorption coefficient of carbon monoxide hemoglobin (CO-Hb) into the skin model, the authors wanted to investigate whether this method is suitable to determine the CO-Hb concentration from spectral reflectance curves of livores. The investigations performed on 28 deaths from CO poisoning so far showed that this is generally possible. In almost all cases, the actual CO-Hb values could be estimated correctly by using the Monte Carlo simulations.

The most important signs of vitality in burned bodies are soot deposits in the respiratory tract, the esophagus and the stomach as well as elevated CO-Hb values in the blood. But these findings show only that a person was exposed to fire fumes while alive; they are no indicator for vital heat exposure. As the external findings in burned bodies are often not very conclusive due to postmortem consumption by the fire, the internal findings are all the more important. Macroscopic signs that hot fumes were inhaled may be edematous swelling and vesicular detachment of the mucosa in the pharynx, the larynx and/or the upper section of the esophagus. As histological changes in the respiratory tract soot deposits, vesicular detachment of the epithelium, hyperemia and edema of the tracheal and bronchial mucosa as well as increased secretion of mucus have been described. These findings may partly be absent although the victim was alive during the fire, which can make diagnosis more difficult. Especially in peracute deaths vitality findings are usually sparse. The significance of the macroscopic and histological parameters of vitality was systematically investigated on the basis of our own autopsy material from the years 1996-2002 (88 cases) and compared with the statements found in the literature. In our study material, the vitality parameters were incomplete in 23% of the burned corpses. In 3% of the cases the question whether there was an antemortem heat exposure could not be answered.

On a day in November, the body of a 31-year-old man was found near a swimming lake with two open and partly emptied fish tins lying next to him. Further investigations showed that the man had been allergic to fish protein and suffered from severe depression and drug psychosis. Already some days before the suicide, he had repeatedly asked for fish to kill himself. Although the results of the chemical and toxicological examinations were negative, the autopsy findings and histological tests suggest that death was caused by an anaphylactic reaction.

When inspecting the scene the circumstances under which a body is found and the findings on the body surface may give first clues to premortal drug abuse. Besides fresh and/or old injection marks tattoos, underweight and signs of physical neglect, especially after long-term abuse, are mentioned in literature. The incidence of such externally visible physical signs of drug abuse was systematically investigated in 100 consecutive drug deaths occurring from 1995 to 1997. Sixty-eight percent of the bodies were found in the apartments of the deceased or those of friends, 11% in public restrooms or the washrooms of restaurants. The so-called body dumping was seen in three cases. In 61% externally visible, fresh injection marks were found; on dissection of the subcutaneous veins, residues of previous injections were found in 95% of the cases. Tattoos were present in 63%. In 8% there was clear evidence of physical neglect. Eighty-six percent of the drug victims had a normal nutritional status.

The highly putrefied corpse of an 80-year-old man was found in the apartment which he had rented to a prostitute. A package of Viagra 25 was found beside the corpse and three tablets were missing. Autopsy revealed severe coronary artery sclerosis as well as signs of previous myocardial infarctions. For the detection and identification of sildenafil and three metabolites in urine and tissue samples, solid-phase extraction, LC/MS and MS/MS methods were developed. Blood was not available for toxicological analysis due to the putrefaction. For method development, urine from a volunteer who had ingested 25 mg sildenafil was collected over 8 h, and three metabolites were identified by MS/MS. These metabolites were also found in the victim's urine. These findings prove that sildenafil was taken some time prior to death, but the causality of sildenafil intake and fatal cardiac failure could not be proven, since no blood was available for analysis. However, the administration of sildenafil was contraindicated due to several previous myocardial infarctions.

In hanging and ligature strangulation, the noose mostly causes a mark or groove which is formed partly by compression of the skin and partly by abrasion with loss of the upper epidermal layers. The horny scales abraded from the neck may be transferred to the strangulation device or to the interposed textiles where they are sometimes visible at stereomicroscopic examination or even to the naked eye as silver-grey particles. The morphologic features of the epidermal transfer due to hanging and ligature strangulation is demonstrated by 14 case examples. The biological traces may be sufficient for comparative DNA typing by means of PCR-based methods. In 9 out of the 14 cases, genomic DNA typing was successful. Analysis of mtDNA succeeded in another two cases, although genomic DNA could not be detected. Beside the accumulation of solid epidermic particles the paper describes deposition of serous and fatty tissue fluid at the ligature (mainly adjacent to skin ridges).

The present case deals with the unusual suicide method of a 36-year-old man who fastened one end of a nylon rope to a tree, guided the other end into a van through the open tailgate and placed the loop round his neck. Then he stepped on the accelerator. Before, he had marked the point on the ground where the rope would tighten. As the rope tightened complete decapitation occurred at a speed of about 35 km/h. Autopsy showed a nearly circular abrasion zone around the site of transection slightly ascending towards the nape, a fracture of the cervical spine between the 3rd and 4th vertebra and a fracture of the thoracic spine between the 7th and 8th vertebra. The test for air embolism of the heart was positive. Macroscopically, no evidence of blood aspiration was found. Histological investigation showed general anaemia and minor blood aspiration in the lungs. Wound morphology was largely in line with the injury patterns described after decapitation in the literature. However, our results differed in that blood aspiration was discernible only under the microscope and there was a second fracture of the spine. Decapitation as a suicide method is an expression of enormous autoaggression and is categorized as a "hard" suicide method. It is used predominantly by men and its occurrence in the spectrum of suicidal actions is rare. Police investigations revealed that the man had led a sort of double life with a sexually motivated background and had suffered from depressive episodes.

Spectrophotometric measurements were performed on intra- and/or subcutaneous bruises occurring in direct temporal connection with peracute fatal trauma. The purpose of these measurements was to determine whether the visual colour impression of a fresh traumatic extravasation can give information on the localisation of the haemorrhage in a certain tissue layer. After visual assessment of the colour of the bruise, the spectral reflectance curves and the CIE-L*a*b* colour values were determined with the help of a diode array spectrophotometer. The localisation and size of the haemorrhages in the cutis and/or subcutis were evaluated morphologically after incision of the skin. It was confirmed that there is a relationship between the colour impression and the localisation of the bruise. Bruises localised near the surface have a more reddish appearance while bruises in deeper layers give a more bluish colour impression. An explanation may be found in the optical characteristics of skin. Blood localised in the subcutis appears blue on the surface due to scattering processes in the dermis (Rayleigh scattering), as the blue wavelengths of the light are scattered (and thus reflected) to a greater extent than the red wavelengths.

The time-dependent inflammatory cell reaction in human cortical contusions has been investigated during the first 30 weeks after blunt head injury. Immunohistochemical staining was carried out using CD 15 for granulocytes and LCA, CD 3 and UCHL-1 for mononuclear leucocytes. In order to provide reliable data for a forensic wound age estimation, the intensity of the cellular reaction was evaluated with a quantitative image analysis system. CD 15-labelled granulocytes were detectable earliest 10 min after brain injury, whereas significantly increased numbers of mononuclear leucocytes occurred in cortical contusions after a postinfliction interval of at least 1.1 days (LCA), 2 days (CD 3) or 3.7 days (UCHL-1), respectively.

A 28-year-old man sustained two shotgun injuries of the left inguinal region from a distance of about 1.5 m by simultaneous discharge of both shells from a sawn-off double-barrelled 16-bore shotgun (diameter of the lead pellets, 4 mm). The first X-ray examination carried out soon after hospital admission showed a single embolized pellet near the right margin of the cardiac silhouette. Eight months later, the man committed suicide by drug intoxication. At autopsy, the embolized pellet was found embedded between the pectinate muscles of the right atrium. On the basis of the reported case and with reference to the pertinent literature, the paper points out the medico-legal aspects of venous bullet/pellet embolism and the risk of lead poisoning after shotgun injury.

The changes occurring during cremation were watched and documented in 15 undissected bodies to be cremated. It was found that at temperatures between 670 and 810 degrees C the body showed the "pugilistic attitude" after about 10 minutes. After 20 minutes the calvaria was free from any soft tissue and fissures of the tabula externa could be noticed. The body cavities became visible after approximately 30 minutes, so that the organs were exposed. Forty minutes after cremation had started, the internal organs were severely shrunken and showed a net-like or sponge-like structure. After about 50 minutes the extremities were destroyed to an extent leaving only the torso which broke apart after 1-1.5 hours. The complete incineration of a human body took about 2-3 hours.

Based on a recent case, in which an expert opinion had to be prepared, the question was investigated if fractures of the base of the skull can result from the influence of heat on the human skull. Neither the retrospective analysis of autopsy records nor the prospective examination of charred bodies revealed any cases with heat-induced fractures of the base of the skull. Observation of cremations showed that the changes caused by the fire followed certain rules: fractures of the calvaria were seen after approximately 20 min; the base of the skull became exposed after about 45 to 60 min. In none of the 20 cremations watched could any fractures of the base of the skull be detected.

Purpose The presented case deals with an unusual suicide by carbon monoxide poisoning. In a car parked in a highway rest area, the body of a middle-aged man was found. In the open trunk of the car there was a gas-powered generator which was switched on, but no longer running. The tank was three quarters full. At autopsy, bright-red livores, cherry-pink fingernails, cherry-red blood and salmon-red skeletal musculature were found. According to the toxicological analysis performed during autopsy, the COHb content in the corpse blood was 68%. Methods To reconstruct the event, the emergency generator was started again in the man's car. By means of measuring probes placed in the interior of the car, carbon monoxide, carbon dioxide and oxygen were measured and recorded in a concentration–time curve; the concentration of cyanide was measured at the end of the experiment. Results The lower explosion limit of 500 ppm CO was reached after 30 s already. For technical reasons, no further values could be recorded. After about 14 min the engine started stuttering with approximately 14 vol.% of oxygen in the air, but continued to run at a lower speed until the experiment was stopped after 25 min. The final concentration of cyanide was 7.5 ppm. Conclusion In view of the rapid CO increase in the interior of the vehicle it is to be assumed that the victim lost consciousness very fast.

Bernitz et al. published a paper on the protrusion of the tongue in burned bodies and were of the opinion that this was an indicator of vital burning [1]. The authors reported that in 63 of 107 fire fatalities, protrusion of the tongue was observed, whereas in 27 cases, this phenomenon was not seen. The authors compared this parameter with the presence of soot deposits in the respiratory tract and an elevated CO-Hb content in the corpse blood—the characteristic parameters of vitality. They found that in the vast majority of the cases with protrusion of the tongue, soot aspiration and CO-Hb values >10 % were also present. According to the authors, subsequently performed statistical tests (Fisher’s exact test, Mann–Whitney test, Kolgorov–Smirnov test) confirmed that protrusion of the tongue was a sign of vitality. This is a very bold conclusion. The authors ignore in their argumentation that most fire-related deaths show signs of vitality [1–6]. This has not only been demonstrated in various studies, but is also true in the 107 cases presented by Bernitz et al. [1]. Statistical comparison of two groups of such different sizes (vital versus avital burning) is questionable. However, in the discussion, this methodological problem is not even mentioned. Moreover, protrusion of the tongue and soot aspiration are findings very often seen in fire fatalities. For that reason alone, the probability that both parameters occur simultaneously is very high, although the mere coincidence of two parameters does not mean that they are of equal validity— in this case, that they are signs of vital exposure to fire. Apart from looking for a correlation between tongue protrusion and internal signs of vitality, one could then also look for a correlation between charring of the skin, pugilistic attitude or fire-related amputations and internal signs of vitality. In all probability, this approach would also produce positive correlations. By the way, the application of statistical methods would show the same significant results if the hypothesis were “Protrusion of the tongue is no vital sign and thus simultaneous soot aspiration is not either.” This obviously absurd statement could also be proved by the data material and tests used by the authors. In addition, there is no pathophysiological explanation whatsoever why protrusion of the tongue should be a sign of vitality. The statement that, after death, no physiological processes are going on which might explain protrusion of the tongue, does not really help, as it goes without saying that physiological processes always occur before death. Obviously, the authors were also aware (as the last sentence of the discussion chapter shows) that the pathophysiological processes should also be recorded. On the other hand, the assumption mentioned in the introduction that this may be a postmortem artefact due to the heat-related shrinking of the cervical soft tissue [7] is not addressed either in the methodology part or in the discussion. The essential parameter in this context would be to what extent the cervical soft tissue was burned, but this was not even recorded in the present study.

Our current research in the framework of an interdisciplinary project focuses on modelling the dynamics of the hemoglobin reoxygenation process in post-mortem human skin by reflectance spectrometry. The observations of reoxygenation of hemoglobin in livores after postmortem exposure to a cold environment relate the reoxygenation to the commonly known phenomenon that the color impression of livores changes from livid to pink under low ambient temperatures. We analyze the spectra with respect to a physical model describing the optical properties of human skin, discuss the dynamics of the reoxygenation, and propose a phenomenological model for reoxygenation. For additional characterization of the reflectance spectra, the curvature of the local minimum and maximum in the investigated spectral range is considered. There is a strong correlation between the curvature of specra at a wavelength of 560 nm and the concentration of O2-Hb. The analysis is carried out via C programs, as well as MySQL database queries in Java EE, JDBC, Matlab, and Python.

Reflectance spectrometry is a fast and reliable method for the characterization of human skin if the spectra are analyzed with respect to a physical model describing the optical properties of human skin. For a field study performed at the Institute of Legal Medicine and the Freiburg Materials Research Center of the University of Freiburg, a scientific information repository has been developed, which is a variant of an electronic laboratory notebook and assists in the acquisition, management, and high-throughput analysis of reflectance spectra in heterogeneous research environments. At the core of the repository is a database management system hosting the master data. It is filled with primary data via a graphical user interface (GUI) programmed in Java, which also enables the user to browse the database and access the results of data analysis. The latter is carried out via Matlab, Python, and C programs, which retrieve the primary data from the scientific information repository, perform the analysis, and store the results in the database for further usage.

Complete or incomplete decapitation as a consequence of suicidal hanging is very rare, few cases having been reported in the worldwide literature. Posthanging decapitation is typically related to a drop of several meters. Three cases of complete decapitation and one case of incomplete decapitation by suicidal hanging are reported with particular emphasis on internal findings and vital reaction patterns. Personal, circumstantial, autopsy, and toxicological data were analyzed to define basic characteristics of such extreme injuries. The crucial factor for the state of decapitation itself is the kinetic energy of the falling body, the strength of the human neck tissue, and the diameter and elasticity of the used ligature. Results of our case study suggest Simon's hemorrhage and air embolism as useful autopsy findings in posthanging beheading cases. Simon's hemorrhage was demonstrated in three cases of four. The test for air embolism was positive in all four cases.

An 88-year-old woman committed suicide by drinking a toxic amount of highly concentrated alcohol and setting two rooms of her flat on fire. As there was not enough oxygen, the fire went out, however. At autopsy, no thermal lesions were found on the body, but soot depositions in the airways and a COHb value of 14% pointed to the inhalation of fire fumes. The ethanol concentration in femoral blood was 6.62 per mille. The gastric mucosa was fixed by the ingested alcohol and showed hardly any autolytic changes despite a post-mortem interval of five days. Congener analysis of the gastric contents and the femoral blood indicated the uptake of a fruit distillate or its foreshot.

Pesticides are used to protect plants all over the world. Their increasing specificity has been due to utilization of differences in biochemical processes, and has been accompanied by lower human toxicity. Nevertheless cases of poisoning are still observed. While certain toxic substances are provided with characteristic dyes or pigments to facilitate easy identification, no overview of pesticide colors exists. The lack of available product information prompted us to explore the colors and dyes of pesticides registered in Germany, most of which are commercially available worldwide. A compilation of the colors and odors of 207 pesticide products is presented. While some of the substances can be identified by their physical characteristics, in other cases, the range of possibilities can be narrowed by their nature and color.

Aims To investigate incipient inflammatory changes as first steps in the development of a systematic inflammatory response syndrome in the lungs of burn victims. Methods At the Institute of Legal Medicine of the University Hospital of Freiburg a collection of 40 forensic autopsy cases of burn victims was established that had died within 1 h after fire exposure. This group was compared with a total of 48 autopsy cases in three control groups (postmortem burns vs deaths from haemorrhagic shock vs railway suicide deaths). In all cases, immunohistochemical studies of lung tissue probes were performed to detect the presence of pro-inflammatory mediators using antibodies against tumour necrosis factor α (TNF-α), interleukin-8 (IL-8) and inter-cellular adhesion molecule 1 (ICAM-1). Results The lungs of burn victims showed a significantly higher extent of intra-alveolar oedema than the other groups. Immunohistochemically, macrophages in all groups mostly showed a distinct expression of TNF-α, but not of IL-8 or ICAM-1. Interestingly, intravascular erythrocytes often showed a positivity of TNF-α that was strongest in the group of burn victims and differed significantly from all the control groups. Conclusions In burn victims with short survival times of ≤1 h after fire exposure, the immunohistochemical expression profiles of TNF-α, IL-8 and ICAM-1 in the lungs were not altered enough to prove an instant inflammatory reaction in these cases. Nevertheless, the positive reaction of TNF-α in erythrocytes of burn victims may indicate the beginning of a non-specific immune response to fire-induced inhalation trauma.

Sudden unexpected death from a natural (pathological) cause often occurs under suspicious circumstances suggesting foreign intervention, so that a forensic autopsy is deemed necessary. In adults, a significant share of these deaths is due to spontaneous ruptures of the aorta. In figures, dissections of the ascending aorta and the aortic arch (mostly associated with secondary perforation into the pericardium) account for a large percentage followed by arteriosclerotic aneurysms of the abdominal aorta (with retroperitoneal and sometimes intraperitoneal hemorrhage). The authors report on two fatal aortic ruptures in which the hemorrhage showed an unusual route to spread. In the first case an 80-year-old man with an arteriosclerotic aneurysm of the abdominal aorta died from acute intestinal bleeding via an aortojejunal fistula. In a 42-year-old man, a dissecting hemorrhage in the wall of the aortic arch resulted in the compression of the branching sites of the brachiocephalic trunk, the left common carotid artery and the left subclavian artery, while the volume of blood in the pericardium was rather small (60 ml). On the basis of these examples, the medicolegal aspects of spontaneous aortic ruptures are discussed.

Complete relaxation can be achieved by floating in a darkened, sound-proof relaxation tank filled with salinated water kept at body temperature. Under these conditions, meditation exercises up to self-hypnosis may lead to deep relaxation with physical and mental revitalization. A user manipulated his tank, presumably to completely cut off all optical and acoustic stimuli and accidentally also covered the ventilation hole. The man was found dead in his relaxation tank. The findings suggested lack of oxygen as the cause of death.