Homicide Manner-of-Death Classification in Arrest-Related Death.

ABSTRACT: Multiple studies have documented various factors that influence or determine forensic pathologist classification of manner of death. There do not appear to be any published studies on manner of death classification specifically regarding arrest-related deaths (ARDs). The goal of this study was to consider a large body of cases of nonfirearm ARDs to analyze the homicide classification with regards to numerous decedent and practitioner (medical examiner/coroner [ME/C]) variables. We analyzed 1145 US autopsy reports from the years 2006-2020, inclusive, and considered decedent variables of age, ethnicity, height, weight, body mass index, toxicology, and mention of a conducted electrical weapon and ME/C influence variables of gender, country region, and year. We found that the homicide classification likelihood increased by a factor of 1.04-1.05 per year, 1.34-1.37 for a female medical examiner, and 1.4-1.5 going from Southern states to Western states. There is an increasing trend for ME/C to label nonfirearm ARDs as homicides in the United States. The homicide classification is more common in Western states and less common in Southern states, and it was more common with a female ME/C.

Comparison of massively parallel sequencing to capillary electrophoresis for short tandem repeat genotyping of trace DNA.

In forensic genetics, massively parallel sequencing (MPS) offers several advantages over the current golden standard, capillary electrophoresis (CE): additional sequence information, shorter amplicon lengths, and the simultaneous analysis of many markers. These benefits result in a reduced number of reactions necessary while improving the amount of data obtained, thereby conserving valuable sample extracts. This proves particularly advantageous for the analysis of trace DNA. This study assessed the suitability of MPS for short tandem repeat (STR) typing of low template samples compared with results obtained through CE. The MPS genotypes showed higher concordance to reference genotypes, with donor alleles being more frequently assigned to be the major contributor, meeting the requirements for database entry. However, the MPS workflow is more time-consuming and associated with higher costs.

Emergence of SARS-CoV-2 spike protein at the vaccination site.

BACKGROUND: The anti-coronavirus disease 2019 (COVID-19) vaccines are of paramount importance in the fight against the COVID-19 pandemic. Both viral vector- and nucleic acid-based vaccines are known to effectively induce protection against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus by generating high antibody titers and effective T-cell responses to the spike protein they encode. Although these vaccines are being applied worldwide and have been extensively investigated, the immunomorphological events at the vaccination site with respect to SARS-CoV-2 spike protein expression have not yet been described. METHODS: We had the opportunity to examine the deltoid muscles of three men who died shortly after vaccination for unrelated reasons. We examined the vaccination sites histologically and immunohistochemically with various antibodies. Furthermore we incubated two different cell lines with one vaccine and examined the expression of the spike protein. RESULTS: The vaccination sites show a dense lymphohistiocytic interstitial infiltrate which surrounds the small vessels and extends into the perimysium. The spike protein is expressed by histiocytic cells with a dendritic shape that are CD68-positive and CD207-negative, fibrocytes, and very rare S100-positive cells. Interestingly, the skeletal muscle, being constitutively human leukocyte antigen (HLA)-A,B,C-negative, is induced at different levels in each specimen. In a cell culture experiment, we confirmed the ability of fibroblasts and interdigitating dendritic sarcoma cells to express spike protein in vitro after incubation with the Comirnaty vaccine. CONCLUSIONS: Histiocytic cells and fibrocytes are the heralds of spike protein synthesis at the vaccination site. The underlying cause of this apparent cell specifity is unknown. This needs to be investigated in future experiments, for example in an animal model.

Development of a multiplex assay for detection of autosomal and Y-chromosomal STRs, assessment of the degradation state of mitochondrial DNA and presence of mitochondrial length heteroplasmies.

The current focus in most routine forensic casework is detection of autosomal or gonosomal Short Tandem Repeats (STRs). With increasing degradation, STR analysis tends to be less successful up to complete failure. For challenging samples such as telogen hair roots and shafts, touch DNA samples or skeletal remains, mitochondrial DNA (mtDNA) analysis provides a powerful tool. Determination of DNA quantity is an important part in the casework workflow. Several ready-to-use kits are commercially available for nuclear DNA targets. However, quantification of mtDNA targets requires the establishment of an in-house method. Some assays even contain assessment of degradation, which alleviates the choice of target enrichment for sequencing through medium or small amplicons. As Sanger-type Sequencing (STS) still remains the golden standard in many laboratories, identification of heteroplasmies in C-tract regions prior to the sequencing reaction is advantageous. Firstly, primer selection can be expanded with primers binding near the C-tract and secondly, determination of the dominant variant is straightforward. All those quantity (nuclear and mtDNA) and quality (degradation and length heteroplasmies) evaluations usually require at least two separate reactions. Therefore, the aim of this project was the combination of all these targets in one multiplex assay using capillary electrophoresis to spare valuable sample extract. Amplification of representative autosomal and Y-chromosomal STRs allows estimate of success of (Y-)STR analysis. Simultaneously, five length heteroplasmies in the mitochondrial control region are targeted as well as three conservative regions of differing fragment lengths for assessment of the mitochondrial degradation state. Based on the outcome of this assay, forensic examiners can decide if STR analysis may be suitable. In case of absent STR peaks, appropriate proceeding of mtDNA sequencing can be determined.

Best of both: A simultaneous analysis of mRNA and miRNA markers for body fluid identification.

The body fluid identification of traces found at crime scenes is crucial in relation of the circumstance of crime. For this reason, the body fluid identification (BFI) by molecular biological methods has been increasingly investigated in recent decades. Especially the use of messenger RNA (mRNA) has been established and validated by various studies. mRNAs can resist degradation for several decades under dry and dark environmental conditions, but degradation increases greatly e.g., in humid environments and UV radiation. In contrast, the shorter and protein-protected micro RNAs (miRNAs) are less susceptible to degradation, but not all potential markers are tissue-specific. The aim of this study was to develop a simultaneous mRNA/miRNA multiplex assay to take advantage of both types of RNA. The final assay was tested for various body fluids, dilutions, and mixtures. To demonstrate the advantage of a combined mRNA/miRNA assay, older and mostly degraded samples were examined and compared to an established mRNA assay. Initial results from degraded samples show that tissue-specific miRNAs expected could be detected for 93% of the degraded samples compared to mRNA markers with 25% of the mRNA assay. The result is a simultaneous mRNA/miRNA multiplex assay on capillary electrophoresis (CE) for the first time.

[Suicidal gas embolism in hospital]. / Suizidale Gasembolie im Krankenhaus.

Air embolisms are a common entity seen in clinical practice after traumatic or iatrogenic events. Cases of a suicidally induced air embolism are rare. The connectivity of air-carrying and fluid-carrying tubing systems allow large gas volumes to be infused in a short period of time, usually with fatal outcome. In such cases, the use of computed tomography prior to autopsy is mandatory and provides a comprehensive visualization of introduced gas volumes. We present an unusual case of a suicidal gas embolism, in which a man in hospital connected a tube supplying oxygen to a venous catheter.

Casework-related DNA transfer on footwear in consideration of the shedder status.

DNA evidence on shoes can play an important role in solving a variety of crimes. We investigated the transfer, persistence, prevalence and recovery of DNA (DNAtppr) on shoes (sneakers) and their soles in realistic handling scenarios taking into account the shedder status. This study aims to increase the understanding of the expected composition of DNA profiles and their probative value, providing a basis for activity level assessments. Samples were analyzed using a direct lysis method, suggesting its versatility and increasing the DNA typing success compared to previous studies on footwear. The data showed surface-dependent background DNA (bDNA) levels on shoe soles and prevalence of bDNA on the upper parts of the shoe. The owner of the shoe was allocatable to the mixture for almost every shoe and sampling location. Alternating scenarios of shoe handling were simulated through different pairs of shedders to distinguish shoe owner and subsequent user. Secondary users were attributable to DNA mixtures regardless of shedder status after wearing shoes a single time. The influence of the shedder status follows specific trends in this context. However, particularly intermediate shedders show inconsistent results. The prevalence of bDNA appears to have a greater effect on the impact of the shedder status on DNA profile composition than previously reported. The data help researchers to better resolve suspect statements and determine if a person of interest wore the shoes relevant to the investigation.

Persistence of blood (DNA/RNA) on shoe soles under varying casework related conditions.

Blunt force traumas by footwear can result in severe and even fatal head and upper body injuries. Oftentimes, footwear impressions are only partially available and evidential value is limited. DNA evidence on shoe soles could provide crucial evidence helping to solve crimes by linking target DNA to the activity of interest. Little is known about the persistence and detectability of biological material post such offenses and the interplay of factors affecting the analytical success. In this study, we assessed the persistence of blood on shoe soles under varying parameters such as blood location, different sneakers, weather condition, gait, amount of blood, underground and step count. We applied an optimized DNA/RNA workflow adapted to micro-traces without constraints for the primary DNA pipeline. There is a high probability to link donor DNA to the shoe sole for up to 300-400 steps, regardless of the underground, blood location, and amount of blood. Depending on the sole material and the degree of abrasion of the sole, a longer blood persistence can be observed. Considering blood, 98.2% of the initial DNA amount (1 µl initial blood volume) was lost after 100 steps walked on sole areas that are in constant contact with the ground. Proportion of foreign DNA was marginal (avg. 4.4 alleles), minimizing the probability of unintentional DNA transfer in this context. RNA typing showed high specificity but lower sensitivity than presumptive tests used for body fluid identification (BFI). Luminol is essential for targeted sampling on shoe soles, as latent blood traces (>100-200 steps) provided sufficient biological material for DNA/RNA typing. The generated data help to address the activity of interest and evaluate probabilities about prevalence of target DNA important for casework implications and assessments on activity level.

Development and validation of an mRNA-based multiplex body fluid identification workflow and a rectal mucosa marker pilot study.

Molecular identification of body fluids and tissues is crucial in order to understand the circumstances of crimes. For that reason, molecular investigations used to identify body fluids/tissues have increasingly been examined recently. Various studies have proved that messenger RNA (mRNA) profiling is a sensitive and robust method for body fluid/tissue identification. The forensically relevant body fluids/tissues blood, semen, saliva, vaginal secretion, menstrual blood and skin have all been detected successfully by applying suitable mRNA assay. However, rectal mucosa, which can be found as evidence in sexual assault cases, has been neglected in forensic investigations. So far there is no mRNA marker to detect rectal mucosa, although anal penetration occurs in a large number of sexual assaults (23.2% of female victims and 50% of male victims). In this study, specific and sensitive mRNA markers for forensically relevant body fluids were adapted and validated in an mRNA multiplex assay for routine casework. This included the implementation of a DNA/RNA re-extraction method for automated extraction that can be integrated into casework without loss of DNA. This re-extraction method and the mRNA multiplex assay were tested using casework samples. PCR-primers were designed for the identification of rectal mucosa and the more effective marker MUC12 was integrated into an extended multiplex assay. The result of our study is a highly specific and sensitive mRNA multiplex assay plus an automated DNA/RNA re-extraction method, that can be integrated into casework and identify rectal mucosa for the first time.

The diversity of shedder tests and a novel factor that affects DNA transfer.

Since the first shedder test was formulated almost 20 years ago, a plethora of different test strategies has emerged. The amount of data generated so far is considerable. However, because of the limited reproducibility of its results, the reliability of the shedder concept is frequently questioned. This study provides a literature overview of applied shedder tests that capture the diversity of the concept. It is pointed out to what extent different classification criteria, workflows, and trace evaluation can impair the classification outcome. The robustness of shedder status was assessed by applying a promising approach established by Fonneløp et al. (Forensic Sci Int Genet 29:48-60, 21). Data provide similar results to those in recent studies but also ambiguous shedder classifications. The applied shedder test was adapted based on our own as well as the reviewed data. With novel classification parameters, promising results were achieved. This study reveals uncertainties and inconsistencies of the shedder concept. Recommendations for harmonization and transparency are proposed. Implementation of the recommendations may result in an increased impact on casework and transfer studies, including activity-level assessments. Furthermore, this study shows that moisturizers affect participants' shedder status as well as DNA transfer. The impact appears to remain relevant even 60 min post ointment application but depends greatly on the type of moisturizer applied.

Electrical weapons and rhabdomyolysis.

It has been suggested that an application of a conducted electrical weapon (CEW) might cause muscle injury such as rhabdomyolysis and an acute inflammatory response. We explored this hypothesis by testing the effects of electrical weapons on circulating markers of inflammation and muscle damage. In a prospective study, 29 volunteers received a full-trunk 5-s TASER® X26(E) CEW exposure. Venous blood samples were taken before, 5 min after, and at 24 h following the discharge. We tested for changes in serum levels of C-reactive protein (CRP), alkaline phosphatase (ALP), myoglobin, albumin, globulin, albumin/globulin ratio, aspartate and alanine aminotransferase, creatine kinase, total protein, bilirubin, and lactic acid dehydrogenase. Uncorrected CRP and myoglobin levels were lower in the immediate post exposure period (CRP levels 1.44 ± 1.39 v 1.43 ± 1.32 mg/L; p = 0.046 and myoglobin 36.8 ± 11.9 v 36.1 ± 13.9 µg/L; p = 0.0019) but these changes were not significant after correction for multiple comparisons. There were no changes in other biomarkers. At 24 h, CRP levels had decreased by 30% to 1.01 ± 0.80 mg/L (p = 0.001 from baseline). ALP was unchanged immediately after the CEW application but was reduced by 5% from baseline (66.2 ± 16.1 to 62.7 ± 16.1 IU/L; p = 0.0003) at 24 h. No other biomarkers were different from baseline at 24 h. A full-trunk electrical weapon exposure did not lead to clinically significant changes in the acute phase protein levels or changes in measures of muscle cellular injury. We found no biomarker evidence of rhabdomyolysis.

A comparative study of conducted electrical weapon incapacitation during a goal-directed task.

Conducted electrical weapons (CEW) are ubiquitous in law enforcement given their unique ability to physically incapacitate violently resisting subjects. Early use of animal models to study CEW incapacitation effectiveness (e.g. porcine model with 4-limb strain gauges) proved to be poorly predictive of human incapacitation effectiveness. In a previously published human study, we developed a methodology for the prospective assessment of the incapacitation effectiveness of CEWs in highly motivated human subjects. Here we use this methodology in Part 1 to compare the incapacitation effectiveness of the newly released Axon® (formerly TASER® International) T(ASER) 7 to the TASER X26E, the "gold standard", and the TASER X2, a current model. The T7 has a new "adaptive cross-connect" technology that may improve incapacitation effectiveness in the scenario of small spreads between probe pairs, a common cause of weapon "failure" in the field. In Part 2, we use our methodology to test the functionality of the T7 cross-connect technology by comparing different bay and probe configurations. This is the first published study in the literature comparing different CEW models using this human model. For Part 1, 29 subjects completed the study and had data available for analysis. For Part 2, 21 subjects completed the study and had data available for analysis. The subjects were motivated to complete the task of reaching a suspended martial arts dummy 3.4 m (11 ft) away while being exposed "under power" to the CEW. In Part 1, subjects were assigned to 1 of 6 groups with probe spreads of 10, 20, and 30 cm (4, 8, 12 in). Subjects were exposed to a "control" CEW (either the X2 or X26E) and the T7 on alternating sides. Exposures with the X2 and T7 included 2 bay-exposures. In Part 2, 21 subjects were assigned to 1 of 5 groups of different T7 bay and probe configurations all with a theoretical effective spread of 30 cm (12 in). Subjects were rated on their progress towards successfully reaching the dummy and on the extent of limb incapacitation using a quasi-blinded expert-observer scoring panel based on high-speed video review. In Part 1, all CEW models achieved maximal or near-maximal subject control with the 30 cm probe spread. With probe spreads of 10 and 20 cm the pooled data showed the T7 to be superior to the X2 for goal achievement (p < 0.001) and limb incapacitation (p = 0.002) mostly driven by differences seen with the 10 cm spread (2-bay exposures). The T7 was non-inferior to the X26E. In Part 2, there was no statistical significance between the limb capture scores, but there was a statistically significant difference in goal scores. The results overall validate that the T7 CEW cross-connect feature performed as expected. The T7 adaptive cross-connect feature with two simultaneous deployed probe pairs demonstrated a significant improvement in incapacitation effectiveness compared to the current X2 CEW with two simultaneously deployed probe pairs. Small probe spreads are a common reason for limited incapacitation effectiveness in the field and this study suggests the T7 may offer an improvement in this scenario. The T7, with single-bay exposures, was non-inferior to the single-bay X26E.

Electrical weapons, hematocytes, and ischemic cardiovascular accidents.

BACKGROUND: There have been case reports following the use of a conducted electrical weapon (CEW) suggesting that these devices might affect coagulation or thrombosis in at-risk individuals. The aim of this manuscript therefore is firstly to explore this hypothesis by reviewing each of these cases and secondly to report the results of a prospective study exploring a priori the effects of electrical weapons on hematocytes in a group of human volunteers. METHODS: First, we systematically reviewed all cases of adverse outcomes following CEW discharge that could be due to an effect on coagulation or thrombosis, with particular focus on the clinical scenario and its relationship with the weapon discharge. Second, we assessed hematocyte levels in venous blood from 29 volunteers before, 5 min after, and 24 h after receiving a full-trunk 5-s TASER® X26(E) CEW exposure. RESULTS: Following extensive review of the literature, we found 3 relevant case reports of possible vascular thromboembolic clinical events after CEW exposure, specifically a case of ischemic stroke, and 2 cases of ST-segment elevation myocardial infarctions. Review of these published cases failed to establish a plausible linkage to the CEW beyond a temporal association with significant emotional and physiological stress from a violent struggle. Our prospective study of biomarker change following CEW discharge revealed acutely increased values for WBC (white blood cells), specifically lymphocytes and monocytes, and a raised platelet count. Neutrophil levels decreased as a percentage of WBC. While these changes were statistically significant at 5 min, all results remained within established reference ranges. At 24 h, all values had returned to baseline except total WBC which decreased to slightly below baseline but was still within the normal reference range. CONCLUSIONS: A review of clinical cases, of ischemic or thrombotic events revealed no direct association with the CEW discharge. A full-trunk electrical weapon exposure did not lead to hematocyte changes beyond normal clinically expected variations in similar acute response scenarios. The case report and biomarker data do not support the hypothesis that a CEW discharge is associated with changes likely to promote coagulation or thrombus formation.

The physiologic effects of a new generation conducted electrical weapon on human volunteers at rest.

Axon Enterprise, Inc. (Axon) released its newest generation conducted electrical weapon (CEW), the T7, in October 2018. In order to compare the effects of this new CEW to prior generations, we used our previously described methodology to study the physiologic effects of CEWs on human volunteers at rest. This was a prospective, observational study of human subjects consisting of two parts. Part 1 was testing a single cartridge (2-probe) exposure. Subjects received a 10-s exposure from the T7 to the back with a 30 cm (12 in.) spread between the two probes. Part 2 was testing a simultaneous two-cartridge (4-probe) exposure. Subjects received a 10-s exposure from the T7 to the back with two cartridges with a 10 cm (4 in.) spread between each probe pair. The probe pairs were arranged cephalad to caudal such that the distance between the top probe of the first cartridge and the bottom probe of the second cartridge was 30 cm (12 in.). Vital signs were measured immediately before and after the exposure. Continuous spirometry was performed. ECG monitoring was performed immediately before and after the exposure. Venous pH, lactate, potassium, CK, catecholamines, and troponin were measured before and immediately after the exposure, at 1-h post-exposure, and again at 24 h. 11 subjects completed part 1 of the study. 9 subjects completed part 2 of the study. No subjects had a dysrhythmia or morphology change in the surface ECG. There were no statistical changes in vital signs pre- and post-exposure. While subjects did not have a statistical change in spirometry parameters pre-exposure to exposure except for a small drop in PETCO2, there was an increase in minute ventilation after the exposure that could have several explanations. A similar pattern was seen with prior generation weapons. No subject had elevated troponin levels. Other blood parameters including venous pH, lactate, potassium, CK, and catecholamines had changes similar to prior generation weapons. Comparison of the data for the single-cartridge exposures against the simultaneous two-cartridge exposures yielded no difference in vital signs, but the minute ventilation was higher for the two-cartridge exposures. The blood data, where there was a difference, was mixed. In our study, the physiologic effects of the Axon T7 are modest, consistent with the electrically-induced motor nerve-driven muscle contraction, and were similar to prior generation weapons.

A comparative brief on conducted electrical weapon safety.

The variety and high number of published research articles on conducted electrical weapons (CEW) provides a detailed, yet in some parts inconclusive overview of medical aspects of CEW. Due to different research approaches and the use of dissimilar test subjects, an assessment of possible health risks of CEW is limited. The present work provides a brief on CEW safety based on currently available animal, computer and human research data. Using the medical database PubMed, articles published on this topic are critically evaluated and compared with each other. Special focuses are the differences and similarities of human and animal research as well as computer simulation programs. The authors explain why some studies are more reliable than others and give their expert opinion on the safety of CEW. The body of data that have been reviewed provides reasonable support for the safety of CEW.

Cardiac and skeletal muscle effects of electrical weapons : A review of human and animal studies.

Conducted Electrical Weapons (CEWs) are being used as the preferred non-lethal force option for police and special forces worldwide. This new technology challenges an exposed opponent similarly to the way they would be challenged by physical exercise combined with emotional stress. While adrenergic and metabolic effects have been meta-analyzed and reviewed, there has been no systematic review of the effects of CEWs on skeletal and cardiac muscle. A systematic and careful search of the MedLine database was performed to find publications describing pathophysiological cardiac and skeletal muscle effects of CEWs. For skeletal muscle effects, we analyzed all publications providing changes in creatine kinase, myoglobin and potassium. For cardiac effects, we analyzed reported troponin changes and arrhythmias related to short dart-to-heart-distances. Conducted electrical weapons satisfy all relevant electrical safety standards and there are, to date, no proven electrocution incidents caused by CEWs. A potential cardiovascular risk has been recognized by some of the experimental animal data. The effects on the heart appear to be limited to instances when there is a short dart-to-heart-distance. The effect on the skeletal muscle system appears to be negligible. A responsible use of a CEW on a healthy adult, within the guidelines proposed by the manufacturer, does not imply a significant health risk for that healthy adult.

[Fatal skiing accidents: a forensic analysis taking the example of Salzburg]. / Tödliche Skiunfälle--eine gerichtsärztliche Übersicht am Beispiel des Landes Salzburg.

The rising popularity of Alpine skiing in recent years has led to an increase of skiing accidents, some with fatal outcome. In this paper, all fatal skiing accidents from the autopsy material of the Institute of Forensic Medicine of the Paris Lodron University Salzburg were evaluated and compared with statistical data of the Alpine Police. In the wintertime of 2005/2006 until 2013/2014, 22 deadly skiing accidents were autopsied. The age of the male and female victims ranged between 12 and 71 years. The main cause of death was craniocerebral and chest trauma. A relevant blood alcohol concentration was detected in only one case. Together with trauma-biomechanical and technical experts, forensic medicine serves as a necessary clarification interface between the investigating authorities and the judiciary. Determining the cause and manner of death as well as reconstructing the accident is the main task of the forensic pathologist. The present study shows that in the county of Salzburg, only a small percentage of fatal skiing accidents is evaluated from a forensic and trauma-biomechanical point of view. Thus the possibilities of an interdisciplinary accident analysis are not always fully utilized.

Homicide by hanging: A case report and its forensic-medical aspects.

We report a rare case of homicide by hanging. The postmortem examination resulted in a verdict of death by suicidal hanging and the Public Prosecutor's Office released the body for burial. After intervention by the relatives police investigations were resumed. Based on evidence impossible to reconcile with the results of the postmortem examination and requiring further clarification, an autopsy was ordered. The results of the postmortem could not be brought in line with a suicidal hanging and were further substantiated by DNA analysis. The scenario put forward by the defense claiming a secondary transfer of trace evidence onto the ligature and the victim's clothes was excluded because of the distribution pattern and the trace evidence ratio. The defendant was sentenced to 20 years of prison for homicide. The verdict was confirmed by the Supreme Court and commuted to 18 years.

Characteristics of backspatter on the firearm and shooting hand--an experimental analysis of close-range gunshots.

Contact and close-range gunshot injuries can produce bloodstains on the shooting hand and weapon. In this study, backspatter was analyzed in experimental horizontal pistol shots into blood-filled sponges. The shooting distances were set between 0 and 45 cm. With increasing shooting distance, there was a significant decrease in the degree of backspatter. Microspatter could be detected on the weapon and shooting hand up to a shooting distance of 40 cm. They were predominantly located on thumb and index finger as well as the radial dorsal aspects of the hand. Fine spray of microspatter combined with elongated droplets in the shape of exclamation marks was found at contact and 2-cm distance shots. An angular calculation based on the shape of the bloodstains was possible up to a shooting distance of 2 cm. No bloodstains were detected beyond 45 cm.