Assassination of Elisabeth, Empress of Austria, Queen of Hungary, on 10 September 1898: further additions and minor annotations.

Very recently, a comprehensive re-evaluation of the medical facts and observations surrounding the death of Empress Elisabeth "Sisi" of Austria (1837-1898) was published. The Empress was assassinated in Geneva by the anarchist Luigi Luccheni or Lucheni (1873-1910). In parallel to this recent publication, our study group came across an almost unknown letter from Dr. Golay, who was one of the examining doctors of Empress Elisabeth. In this publication we add relevant additions based on this letter and provide further insights regarding the attempts at resuscitation.

Traumatic fractures in an early 19th century museum skeleton suggest the homicide of an old Munich character: the history of "Finessensepperl" (Finesse Joseph).

The well preserved skeleton of Joseph Huber, a very well-known historical character of the 19th century Munich, also nicknamed "Finessen-Sepperl", is the starting point of the reconstruction of life and death of this historical individual. He was known as a postilion d´amour (love's messenger) of the Royal Bavarian capital with numerous comments and anecdotes and a few biographical sketches that indicate he remained well until the last few years of his life where requests for his duties lessened. The skeleton shows a small-sized male individual with almost complete loss of teeth, but otherwise very well-mineralized bone, having suffered from three episodes of trauma - an old-healed incomplete femoral neck fracture leading to severe osteoarthrosis, a clavicle fracture of the medial third with a few weeks old callus formation, and fresh serial rib fractures along with severe skull trauma with fractures of the os temporale and petrosum, presumably leading to intracranial bleeding and finally death. The type and distribution of these latter two injuries are in agreement with a murderous attack - which was retrospectively reported several years after his death, while the old-healed femoral neck fracture may have caused reduction in Joseph´s walking activities but not reduced requests for his services. Paleopathology not only identifies the terminal decline, but also previous diseases of this Old Bavarian character and thereby completes his story.

Human metabolism and excretion kinetics of the surfactant 2,4,7,9-tetramethyl-5-decyne-4,7-diol (TMDD) after oral and dermal administration.

2,4,7,9-Tetramethyl-5-decyne-4,7-diol (TMDD) is a non-ionic surfactant with a wide range of applications. TMDD is considered a high-production chemical and, due to its low biodegradation rate, possesses a potentially high prevalence in the environment. However, despite its widespread use, toxicokinetic data and data on internal exposure to TMDD in the general population are completely lacking. Hence, we developed a human biomonitoring (HBM) method for TMDD. Our approach included a metabolism study with four subjects, who were administered an oral dose of 75 µg TMDD/kg body weight and a dermal dose of 750 µg/kg body weight. Terminal methyl-hydroxylated TMDD (1-OH-TMDD) was previously identified as the main urinary metabolite in our lab. The results of the oral and dermal applications were used to determine the toxicokinetic parameters of 1-OH-TMDD as a biomarker of exposure. Finally, the method was applied to 50 urine samples from non-occupationally exposed volunteers. Results show that TMDD was rapidly metabolized, with an average tmax of 1.7 h and a rapid and almost complete (96%) excretion of 1-OH-TMDD until 12 h after oral dosage. Elimination was bi-phasic, with half-lives of 0.75-1.6 h and 3.4-3.6 h for phases 1 and 2, respectively. The dermal application resulted in a delayed urinary excretion of this metabolite with a tmax of 12 h and complete excretion after about 48 h. The excreted amounts of 1-OH-TMDD represented 18% of the orally administered TMDD dose. The data of the metabolism study demonstrated a fast oral as well as substantial dermal resorption of TMDD. Moreover, the results indicated an effective metabolism of 1-OH-TMDD, which is excreted rapidly and completely via urine. Application of the method to 50 urine samples revealed a quantification rate of 90%, with an average concentration of 0.19 ng/mL (0.97 nmol/g creatinine). With the urinary excretion factor (Fue) derived from the metabolism study, we estimated an average daily intake of 1.65 µg TMDD from environmental and dietary sources. In conclusion, 1-OH-TMDD in urine is a suitable biomarker of exposure to TMDD and can be applied for biomonitoring of the general population.

[The Iceman : Life scenarios and pathological findings from 30 years of research on the glacier mummy "Ötzi"]. / Der Mann aus dem Eis : Lebensszenario und Pathologische Befunde aus 30 Jahren Forschung an der Gletschermumie "Ötzi".

The comprehensive investigation of the excellently preserved mummy of Ötzi, the Iceman, and his equipment over the last 30 years has provided a wealth of information about the life and disease of this late Neolithic individual. This research has indicated that his origin was from a local southern Alpine population, that he grew up in the valleys of the Southern Alps, and that he had considerable local mobility. He had well-balanced nutrition with a mixed vegetable and animal diet. He was very mobile in the alpine terrain and of athletic constitution. The Iceman suffered from mild to moderate degenerative joint disease primarily of the right hip joint, slight spondylosis of the cervical and lumbar spine, a minor focal (premature) arteriosclerosis, lung anthracosis and possibly silicosis, previous pleuritic inflammation (possibly of post-specific origin), intestinal infections of the stomach by Helicobacter pylori and Trichuris trichiura worm infestation in the intestines, a mild osteomalacia of cancellous bone, and diverse pathologies of his teeth with dental caries and periodontitis, as well as hair anomalies. The presence of borreliosis is still under debate. As potential remedies, the Iceman carried some anthelmintic substances with him: a birch polypore and an anthelmintic fern. The numerous tattoos may also have had therapeutic effects. Finally, the last days of Ötzi could be reconstructed quite precisely: his gastrointestinal content indicates that the Iceman moved from Alpine heights to a lower location and then again up to the glacier region where he died. During this journey he encountered two attacks: the first, several days before his death, lead to a stabbing wound in his right hand; the second was an arrow hit that wounded the Iceman lethally at his left axilla by laceration of the subclavian artery.

Development of a human biomonitoring method for assessing the exposure to ethoxyquin in the general population.

Ethoxyquin (EQ) is commonly used as an antioxidant in animal feeds. Although EQ is not permitted for usage in food products for humans within the EU, residues of EQ and its transformation products could be determined in food of animal origin. Despite its widespread use and concerns on its toxicological profile, no information about the systemic exposure to EQ in the general population is available. Hence, we developed a human biomonitoring (HBM) method for EQ. Our approach included a metabolism study with five subjects, who were administered an oral dose of 0.005 mg EQ/kg body weight. Unchanged EQ and the major metabolite 2,2,4-trimethyl-6(2H)-quinolinone (EQI) were identified as urinary excretion products of EQ. While small amounts of EQ could be determined in high concentrated samples from the metabolism study only, 28.5% of the orally applied EQ dose could be recovered as EQI. Toxicokinetic parameters were determined for EQI, the potential biomarker of exposure. In addition, an analytical method for EQI (LOQ = 0.03 µg/L) in urine based on UHPLC-MS/MS comprising enzymatic glucuronide hydrolysis and salt-assisted liquid-liquid extraction was developed, validated and applied to 53 urine samples from the general population. EQI could be quantified in 11 (21%) of the samples in levels up to 1.7 µg/L urine, proving the suitability of the developed method for the intended purpose.

Detectability of bloodstains after machine washing.

Bloodstains on textiles can provide useful information for the forensic reconstruction of a crime. Surprisingly, little is known about the applicability of bloodstain traces after a textile was machine washed. In this study, we investigated the effect of machine washing on bloodstains on both cotton and polyester cloths. The influence of the washing detergent, the type of washing machine, the washing temperature, and the duration of drying of the bloodstain prior to washing as well as the drying temperature was investigated. Additionally, the molecular analyses of a subsample of the experiments were conducted. We found that although the primary morphology of the traces is often blurred, the presence of blood on the textiles can still be detected in many cases. Blood can also be transmitted to previously blood-free textiles during the washing process, leading to a positive Luminol or Combur® reaction of these samples. When traces of blood can be detected via the Luminol reaction, a molecular identification of the blood donor was successful in 28% of the cases.

[Forensic postmortem examination service: challenges and opportunities]. / Forensischer Leichenschaudienst: Herausforderungen und Chancen.

The external examination of a corpse is regulated by federal law and presents physicians and police with a series of challenges. Mostly GPs, but practically every licensed physician, are obliged to complete death certificates, resulting in a very large number of potential physicians, which at the same time means only a small number of cases for each individual. Consequently, this sensitive topic often lacks the experience needed.As already shown in several studies, only very limited possibilities for the correct determination of the cause of death are generally present at the inquest. The legal provisions also represent a certain basic problem, from which further pitfalls can arise.In Munich, the medical association, in cooperation with the Institute of Legal Medicine, organizes a 24­h service with at least one physician on standby, which ensures quality assurance and/or enhancement of this postmortem service through continuous education and further training as well as monthly meetings. The principles and considerations concerning the practicability of such a system will be discussed in this paper.

Low-dose CT in body-packers: delineation of body packs and radiation dose in a porcine model.

PURPOSE: To compare low-dose computed tomography (CT) with standard CT and conventional radiography (CR) regarding delineation of body packs and radiation dose. METHODS: Nine samples of illicit drugs including cocaine, heroin, and hashish were positioned in the rectum of a 121.5 kg pig cadaver. Each sample was scanned on a 64-row MDCT with 120 kV: one standard modulated pelvic protocol (STD), and without modulation at 80 mA (LD80), 30 mA (LD30), and 10 mA (LD10). Additionally, conventional abdominal anterior-posterior radiographs (77 kV and 106 ± 13 mA) were taken. Body pack characteristics (wrapping, content, shape) were rated independently by two radiologists and summarized to a delineation score from 0 to 9 with scores ≥6 representing sufficient delineation. Mean delineation scores were calculated for CR and CT protocols. These were additionally differentiated for readings in soft tissue (S), lung (L), user defined, variable window settings (V), and in cumulative window evaluation including all the other window settings (SLV). Effective doses were calculated (mSv). RESULTS: The CR delineation score was insufficient (3.1 ± 2.5; 2.4 ± 0.3 mSv). For CT, the SLV window setting performed best (p < 0.01). Its score significantly (p < 0.01) declined with decreasing effective radiation doses: STD (8.8 ± 0.5; 10.6 mSv), LD80 (8.2 ± 0.7; 2.6 mSv), LD30 (6.8 ± 1.3; 1.0 mSv), and LD10 (4.6 ± 1.9; 0.3 mSv). Thus, LD30 was the protocol using the lowest but sufficient dose. Moreover, for LD30 further differentiation between the particular window settings resulted in scores of 6.4 ± 1.3 (L), 6.3 ± 1.2 (V), and 3.1 ± 1.0 (S). CONCLUSIONS: With appropriate window settings, low-dose CT at 30 mA allowed for sufficient body-pack delineation below the dose of CR, which itself performed insufficient.

Wound ballistic evaluation of the Taser® XREP ammunition.

The Taser® eXtended Range Electronic Projectile (XREP®) is a wireless conducted electrical weapon (CEW) designed to incapacitate a person from a larger distance. The aim of this study was to analyze the ballistic injury potential of the XREP. Twenty rounds were fired from the Taser®X12 TM shotgun into ballistic soap covered with artificial skin and clothing at different shooting distances (1-25 m). One shot was fired at pig skin at a shooting distance of 10 m. The average projectile velocity was 67.0 m/s. The kinetic energy levels on impact varied from 28-52 J. Depending on the intermediate target, the projectiles penetrated up to 4.2 cm into the ballistic soap. On impact the nose assembly did not separate from the chassis, and no electrical activation was registered. Upon impact, a skin penetration of the XREP cannot be excluded. However, it is very unlikely at shooting distances of 10 m or more. Clothing and a high elasticity limit of the target body area can significantly reduce the penetration risk on impact.

Determination of a specific metabolite for the non-ionic surfactant 2,4,7,9-Tetramethyl-5-decyne-4,7-diol (TMDD) by UPLC-MS/MS.

2,4,7,9-Tetramethyldec-5-yne-4,7-diol (TMDD) is a non-ionic surfactant commonly used as defoaming agent and numerous other applications. Effluents of wastewater treatment plants have been identified as one of the main sources of TMDD emissions into the environment. Due to its broad application in various fields, TMDD was selected for the development of a biomonitoring method for assessing human exposure within the frame of the cooperation project of the German Federal Ministry for the Environment, Nature Conservation, Building and Nuclear Safety (BMUB) and the German Chemical Industry Association (VCI) in 2020. This study aimed to identify a urinary metabolite for TMDD by UPLC-Q-Orbitrap-MS which can be used as a biomarker of TMDD exposure. Monohydroxylated TMDD (1-OH-TMDD) was deciphered as the most prominent metabolite of TMDD in humans in a series of in vitro and in vivo experiments. In a next step, a quantitative method for the determination of 1-OH-TMDD was developed and validated. Quantification was achieved by isotope dilution using D3-1-OH-TMDD as internal standard. The method is characterized by a simple sample clean-up procedure and an enzymatic hydrolysis of possible metabolite conjugates with ß-glucuronidase. Method validation was performed according to international guidelines for bioanalytical method validation. The method proved its robustness, precision, accuracy and sensitivity for the intended purpose, i.e. the assessment of TMDD exposure in the general population by means of human biomonitoring.

Adipositas and metabolic bone disorder in a 16th century Upper Austrian infant crypt mummy-An interdisciplinary palaeopathological insight into historical aristocratic life.

We describe here the results of a multidisciplinary study on an infant mummy from 16th century Upper Austria buried in the crypt of the family of the Counts of Starhemberg. The macroscopic-anthropological, radiological (whole-body CT scan), histological (skin tissue), and radiocarbon isotope investigations suggested a male infant of 10-18 months' age, most likely dying between 1550 and 1635 CE (probably Reichard Wilhelm, 1625-1626 CE), that presented with evidence of metabolic bone disease with significant bilateral flaring of costochondral joints resembling "rachitic rosary" of the ribs, along with straight long bones and lack of fractures or subperiosteal bleeding residues. Although incompletely developed, the osteopathology points toward rickets, without upper or lower extremities long bone deformation. The differential diagnosis is vitamin C deficiency (scurvy) (also with an incomplete presentation, although overlap between both disorders may be present). As additional pathology, there was significantly enlarged subcutaneous fat tissue (thickness more than 1 cm at the navel and thighs and longitudinal creases of the skin) along with a histologically enlarged subcutaneous fat layer consistent with infantile adipositas as a coincident disorder. Finally, remnants of lung tissue with pleural adhesion of the right lung indicate possibly lethal pneumonia, a disease with an increased prevalence in vitamin D deficient infants. Ultimately, the skull presented with extensive destruction of the bones of the base and dislocation of the bones of the skull squama. These changes, however, are most likely post-mortal pseudopathology, the result of a burial in a flat, narrow coffin because there were no bone fractures or residues of bleeding/tissue reaction that would have occurred whilst the patient was alive.

Trauma of bone and soft tissues in South American mummies-New cases provide further insight into violence and lethal outcome.

There exist numerous reports on violence in South American populations which shed a particular light on life and living conditions in those historic communities. Most studies have been performed on collections of isolated skulls. Whole-body investigations especially on well-preserved mummified human remains are rare. In the present study we investigated three South American mummies predating the Colonial Spanish period. The "Marburg" man lived between 996 and 1147 CE and was buried in typical burial bundle. The analysis of the textiles, ceramics and fishing tools associated with his naturally mummified body suggests that he most likely originated from the Arica region in Northern Chile and was possibly part of a fishing community. The "Delémont" natural mummies belong to an adult male and an adult female, respectively. The mummies, the textiles and grave goods were investigated. The ceramics suggest a provenance from the Arequipa region, supposing that all the artifacts were originally associated with the two mummies. The Delémont male mummy is 14C dated between 902 and 994 CE and the "Delémont" female mummy 14C dated between 1224 and 1282 CE. All mummies underwent Multidetector Computed Tomography which showed evidence of trauma, some of which were interpreted as evidence of interpersonal violence. An interdisciplinary approach was applied with the particular intention to identify trauma sequels and to evaluate their paleo-forensic potential. Evidence of violence was identified in the two male individuals. Our study provides evidence that the interdisciplinary investigation of well-preserved human remains may detect much more frequent traces of intentional trauma than previously thought. Particularly, trauma against the body may not be identified in studies on skulls alone, and trauma residues of internal organs/soft tissues will only be seen in mummies. We therefore add further evidence of two cases of (lethal) trauma in pre-colonial South-American male individuals.

Whodunit? A reconstruction study in a case of one-handed strangulation.

A 27-year old woman reported an attack by her ex-partner. According to her, he suddenly started to strangle her with his left hand, using a claw-like grip against her throat. After 30-60 seconds the victim reacted by kicking the attacker in the groin, thereby disrupting the strangulation. During the court hearing, pictures of the strangulation marks taken by the police were shown as evidence. From a forensic viewpoint, the pictures and the victim's statements did present several inconsistencies, suggesting the possibility of self-inflicted injuries. The ex-partner was found guilty. The defense appealed against the sentence and demanded a thorough forensic expertise on the origin of the strangulation marks. To identify the possible origin of the strangulation marks considering the victim's statements and the presented strangulation marks, a reconstructive study with 26 participants (25 attackers, 1 victim) was carried out. In the study, the expected strangulation marks did show a vertical, C-shaped pattern on the study subject's neck and throat, while the wounds on the victim's neck were aligned horizontally on the right side of the neck. These results show that the strangulation marks on the neck of the victim did not correspond to the claw-like grip at the throat as described by the victim. In this light, the possibility of self-inflicted injuries is discussed.

HPO-driven virtual gene panel: a new efficient approach in molecular autopsy of sudden unexplained death.

BACKGROUND: Molecular autopsy represents an efficient tool to save the diagnosis in up to one-third of sudden unexplained death (SUD). A defined gene panel is usually used for the examination. Alternatively, it is possible to carry out a comprehensive genetic assessment (whole exome sequencing, WES), which also identifies rare, previously unknown variants. The disadvantage is that a dramatic number of variants must be assessed to identify the causal variant. To improve the evaluation of WES, the human phenotype ontology (HPO) annotation is used internationally for deep phenotyping in the field of rare disease. However, a HPO-based evaluation of WES in SUD has not been described before. METHODS: We performed WES in tissue samples from 16 people after SUD. Instead of a fixed gene panel, we defined a set of HPO terms and thus created a flexible "virtual gene panel", with the advantage, that recently identified genes are automatically associated by HPO terms in the HPO database. RESULTS: We obtained a mean value of 68,947 variants per sample. Stringent filtering ended up in a mean value of 276 variants per sample. Using the HPO-driven virtual gene panel we developed an algorithm that prioritized 1.4% of the variants. Variant interpretation resulted in eleven potentially causative variants in 16 individuals. CONCLUSION: Our data introduce an effective diagnostic procedure in molecular autopsy of SUD with a non-specific clinical phenotype.

Chronic active non-lethal human-type tuberculosis in a high royal Bavarian officer of Napoleonic times-a mummy study.

In paleopathology, morphological and molecular evidence for infection by mycobacteria of the M. tuberculosis complex (MTC) is frequently associated with early death. In the present report, we describe a multidisciplinary study of a well-preserved mummy from Napoleonic times with a long-standing tuberculous infection by M. tuberculosis senso stricto who died at the age of 88 years of focal and non-MTB related bronchopneumonia. The well-preserved natural mummy of the Royal Bavarian General, Count Heinrich LII Reuss-Köstritz (1763-1851 CE), was extensively investigated by macro- and histomorphology, whole body CT scans and organ radiography, various molecular tissue analyses, including stable isotope analysis and molecular genetic tests. We identified signs for a long-standing, but terminally inactive pulmonary tuberculosis, tuberculous destruction of the second lumbar vertebral body, and a large tuberculous abscess in the right (retroperitoneal) psoas region (a cold abscess). This cold abscess harboured an active tuberculous infection as evidenced by histological and molecular tests. Radiological and histological analysis further revealed extensive arteriosclerosis with (non-obliterating) coronary and significant carotid arteriosclerosis, healthy bone tissue without evidence of age-related osteopenia, evidence for diffuse idiopathic skeletal hyperostosis and mild osteoarthrosis of few joints. This suggests excellent living conditions correlating well with his diet indicated by stable isotope results and literary evidence. Despite the clear evidence of a tuberculous cold abscess with bacterioscopic and molecular proof for a persisting MTC infection of a human-type M. tuberculosis strain, we can exclude the chronic MTC infection as cause of death. The detection of MTC in historic individuals should therefore be interpreted with great caution and include further data, such as their nutritional status.

Human metabolism and urinary excretion kinetics of the UV filter Uvinul A plus® after a single oral or dermal dosage.

Hexyl 2-[4-(diethylamino)-2-hydroxybenzoyl]benzoate, better known under its trading name Uvinul A plus® is a UV filter mainly used in sunscreens, but also present in other cosmetic products with a maximum concentration of 10% (w/w) according to the EU directive. In this study we investigated the human metabolism after a single oral and a single dermal dose of Uvinul A plus®, respectively. Samples collected within 72 h of administration were analyzed with a newly developed UHPLC-MS/MS method. Results of the study revealed three major urinary metabolites, namely 2-(4-amino-2-hydroxybenzoyl)benzoic acid (AHB), 2-(4-(ethylamino)-2-hydroxybenzoyl)benzoic acid (EHB) and 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid (DHB), representing 52% of the administered oral dose. The three major metabolites are further converted into four minor metabolites with an additional hydroxyl group in the aniline moiety. Toxicokinetic parameters (amount excreted, tmax, elimination constant and half-life t1/2) and conversion factors were determined for the three major metabolites. The conversion factors were used to estimate the mean daily exposure to Uvinul A plus® in spot urine samples from 58 volunteers not intentionally exposed to Uvinul A plus® derived from a pilot study. The three major metabolites were quantifiable in 26% of the samples. In 35% of the samples, at least one major metabolite could be quantified. The daily systemic exposure to Uvinul A plus® was estimated to approximately 8.1-9.3 µg/d by applying the combined conversion factor for all three major metabolites. In conclusion, a very low systemic exposure to DHHB was observed with regard to the no observed adverse effect level (NOAEL) as an established threshold for chronic uptake.

Assignment of unknown persons to their geographical origin by determination of stable isotopes in hair samples.

Analysing the isotope ratio of light elements in human tissue of an unknown person helps to reconstruct the life history with regard to geographical origin and/or typical food supply. In this study the isotope ratios of the bio-elements in hair samples of 111 persons from 13 different countries all over the world were measured with the aim of provenance determination. The results indicated that individuals from Costa Rica and Brazil can be differentiated from typical European individuals by delta(13)C, Australians by delta(34)S and delta(2)H in hair samples. The combination and evaluation of the data by multivariate statistical analysis considerably improved origin assignment. Investigation of hair samples from a number of individuals from one particular region (southern Germany) yielded remarkable variation of isotopic values indicating different nutritional habits. The possibilities and limitations of this method in its current state are demonstrated and discussed.

Intraosseous needles in pediatric cadavers: Rate of malposition.

AIM OF THE STUDY: Intraosseous vascular access is a commonly conducted procedure especially in pediatric resuscitation. Very high success rates for intraosseous (IO) devices are reported. Aim of the study was to describe the rates of malposition of intraosseous needles (ION) in pediatric cadavers via post-mortem computed tomography (PMCT). METHODS: 212 consecutive pediatric cadavers underwent PMCT, of which 38 cadavers had visible ION and were included in the study. They were divided into two subgroups depending on their age (n = 22 infant cadavers (age <1 year) and n = 16 child cadavers (age ≥1 year)). Two independent readers evaluated the number and position of ION. RESULTS: In 22 infant cadavers 34 ION were found. Malposition of at least one ION was visible in 14 subjects (64%), among which 7 cadavers (32%) even had no correctly placed ION, thus being without established vascular access. Overall, 16 of the 34 used ION devices (47%) were in malposition. 23 ION were found in 16 child cadavers. In 8 subjects (50%) at least one ION was malpositioned, among which 3 cadavers (19%) had no correctly placed ION, resulting in a complete absence of vascular access. Overall, 9 of the 23 ION devices (39%) were malpositioned. CONCLUSION: Our study showed relatively high malposition rates for ION devices in pediatric cadavers which was not to be assumed regarding the success rates of 80% and higher in previous literature. This should be clarified by further studies in living patients.