International Journal of Legal Medicine

Published by Springer Nature
Online ISSN: 1437-1596
Print ISSN: 0937-9827
Learn more about this page
Recent publications
Days between incident and examination in case group
Days between incident and examination in control group
Circumstances of the examinations
  • Cleo Walz
    Cleo Walz
  • Clara-Sophie Schwarz
    Clara-Sophie Schwarz
  • Karla Imdahl
    Karla Imdahl
  • [...]
  • Tanja Germerott
    Tanja Germerott
Introduction The Istanbul Convention calls for comprehensive care for victims of violence while maintaining forensic standards. After violent crimes, court usable documentation of injuries and securing of evidence is essential to avoid disadvantages for those affected in criminal prosecution. Material and methods This retrospective study compares forensic relevant aspects in clinical forensic examination of victims of physical and sexual violence conducted by clinicians and forensic examiners. Forensic medical reports based on clinical documentation of individuals of all ages in the period from 2015 to 2018 (n = 132) were evaluated in comparison to a control group of examinations conducted by forensic specialists. A comparative statistical evaluation was performed. Results The study revealed statistically significant differences in forensically relevant aspects. In the clinical examinations, full-body examination was performed in only 37.9%, and concealed body sites were examined in 9.8%. Photo documentation was often incomplete (62.4%), without scale (59.1%), blurred (39.7%), or poorly exposed (31.2%). Information on size, color, shape, and texture of injuries was often missing. In about every third examination, the findings were not described purely objective. A body scheme was used only in 8.3% of the clinical cases. Discussion In order to establish nationwide care structures and the forensic standard required in criminal proceedings, intensive involvement of forensic medicine is essential. Standardized examination materials, regular training of medical staff, and telemedical approaches can improve the care for victims of violence regarding criminal prosecution.
Body template used for calculation of body surface affected by desiccation or moist decomposition. Modified after Lund and Browder [31] for assessment of desiccated skin. The anatomical regions (32 in total) were assessed separately and then added together
Histogram of percentual estimation of body surface in the 102 forensic autopsy cases; a) affected by desiccation, b) affected by moist decomposition, and c) unaffected, grouped by post-mortem interval (PMI). Each of the 48 bars represents one unique PMI value. If multiple cases had the same PMI value, the mean percentual estimation of affected body surface for that PMI was calculated. The x-axis is shown as a categorical scale
Bar chart showing percentage of cases exhibiting each type of desiccation observed within the dataset. Cases with “leathery desiccation” could host another desiccation process at the same time, but cases with “Exclusively leathery and parchment-like desiccation” did not exhibit any soft tissue desiccation. No cases with only soft tissue desiccation were observed in the dataset
Distribution of the different desiccation changes expressed as percentages of total number of cases. Ventral and dorsal side combined and represented in one frontal body chart. Details can be found in Fig. S2 to S4
Scatter plot showing distribution of post-mortem interval (PMI) relative to the type of desiccation the cases exhibited, e.g., “Leathery ± soft tissue desiccation” shows the cases that exhibited leathery desiccation on the body surface, with or without underlying soft tissue desiccation
The objective of this study was to evaluate the presence of mummification in an indoor setting, with an emphasis on the forensic perspective. A dataset of 102 forensic autopsy cases was assessed for distribution of desiccation of skin and soft tissue (i.e., sub-cutaneous fat and musculature) and for moist decompositional (i.e., putrefactive) changes. Further, possible correlation with the post-mortem interval (PMI) was evaluated, as well as the effects of clothing coverage of the body. The results indicated that yellow to orange parchment-like desiccated skin was found at significantly shorter PMIs than reddish brown to black leathery desiccated skin, even when soft tissue desiccation was included in the comparative analysis. Clothing appeared to have a significant decelerating effect on the extent of desiccation on the legs, but findings in regard to whole body or torso/arms were inconclusive. A large variation in PMIs was evident as regards fully desiccated skin (PMI 18-217 days), indicating difficulties in PMI estimation due to a variable repressive effect on the decompositional process per se in an indoor setting. For the specific case in forensic practice, no definite conclusion can be drawn from the observed desiccation changes to the PMI. One way forward might be creating a systematic and standardized method for describing different desiccation types, as well as other cooccurring decompositional changes and how they relate to the PMI, as a foundation for a future quantification model.
Overview of the placenta and the umbilical cord (a furcate insertion): the red bracket highlight the accessory and supernumerary vessel; the blue arrow indicates the placental insertion site of the accessory and supernumerary vessel (near this insertion an area of slight hemorrhagic infiltration of the parenchyma of the annex)
Macro- and microscopic view of the umbilical cord nearby the vessel lesion: (a) shows a macroscopic view of the accessory vessel after the section, at the placental insertion site, with a blood collection outside the vascular wall (surrounded in red); (b) shows the microscopic picture of that area, with the corresponding hemorrhagic picture (surrounded in blue)
The furcate insertion of the umbilical cord is an uncommon abnormality, often asymptomatic, potentially dangerous, or lethal for the fetus and the mother. This report shows the case of a healthy 29-year-old patient, at 37 weeks of gestation, admitted to the hospital two days before the due date because of the appearance of uterine contractions; clinical exams were regular. The following day, no fetal movements were perceived, a cardiotocography was performed, showing the absence of fetal heartbeat. A dead fetus was delivered. Autopsy showed furcate insertion of the umbilical cord and the rupture of the umbilical vessel, which caused fetal hemorrhagic shock. Furcate insertion still remains mostly undiagnosed and rarely it can be identified prenatally (only three cases are reported in literature). Future research, mainly in forensic fields, could improve the knowledge about this condition, helping prenatal diagnosis and providing warnings that can prevent similar deaths in the future.
Distribution of the sample according to age and sex
a–b Photograph and 3D image of a male aged 38 years. c–d Photograph and 3D image of a male aged 80 years (1 cm scale)
95% confidence intervals for age estimation in each phase in dry bones and 3D images
When investigating a death, post-mortem identification provides with results of great legal and humanitarian significance. The effectiveness of the methods used to estimate age depends on the reference population, considering variables such as sex and ancestry. The aim of this study was to validate the Iscan method to estimate age in a Spanish forensic population, comparing the estimates obtained in dry bones and 3D reconstructions created with a surface scanner. We carried out a cross-sectional study on 109 autopsied corpses (67% male), scanning the sternal end of the right fourth rib in a 3D mesh, using an EinScan-Pro® surface scanner (precision: 0.05 mm). Two observers estimated the phases in dry bones and 3D images according to the Iscan method and to the sex of the subject. The mean age was 57.73 years (SD = 19.12 years;18–93 years). The intra-observer agreement was almost perfect in bones (κ = 0.877–0.960) and 3D images (κ = 0.954), while the inter-observer agreement was almost perfect in bones (κ = 0.813) and substantial in 3D images (κ = 0.727). The correlation with the Iscan phases was very strong in bones (Rho = 0.794–0.820; p < 0.001) and strong in 3D images (Rho = 0.690–0.691; p < 0.001). Both sex-adjusted linear regression models were significant (dry bones: R² = 0.65; SEE = ± 11.264 years; 3D images: R² = 0.50; SEE = ± 13.537 years) from phase 4 onwards. An overestimation of age was observed in the first phases, and an underestimation in the later ones. Virtual analysis using a surface scanner in the fourth rib is a valid means of estimating age. However, the error values and confidence intervals were considerable, so the joint use of different methods and anatomical sites is recommended.
Boxplots showing the sex-specific age distribution by category for the five age indicators of rib 1
Descriptions of age-related features on Rib 1 adapted from DiGangi et al. [1] (photos by TMR Houlton). CF = costal face, RH = rib head, TF = tubercle facet
An age-at-death estimation method using the first rib may be particularly advantageous as this rib is relatively easy to identify, not easily damaged postmortem, and associated with less mechanical stresses compared to other age indicators. Previously, mixed results have been achieved using the first rib to estimate age-at-death. This study aimed to develop and test an age-at-death estimation method using the first rib. An identified modern black South African sample of 260 skeletons were used to collect age-related data from the first rib. Multiple linear regression analysis equations were created from this data for male, female, and combined samples. When tested on a hold-out sample, equations generated mean inaccuracies of 7–13 years for point estimates. The 95% confidence intervals contained the true age in 11–33% of individuals depending on the equation used, but wider intervals generated using 95% prediction intervals contained true ages for 100% of individuals. Point estimate inaccuracies are comparable to other age-at-death estimation methods and may be useful if single indicator estimation is unavoidable in the case of missing or damaged bones. However, combined methods that use indicators from many areas of the skeleton are preferable and may reduce interval widths.
Staging according to Olze et al. (2012). Examples from the present cohort
Evaluation of third molar eruption represents an established method for age assessment of living individuals. Different classification systems are available for the radiological assessment of third molar eruption. The aim of this study was to identify the most accurate and reliable classification system for the mandibular third molar eruption on orthopantomograms (OPG). We compared the method of Olze et al. (2012) with the method of Willmot et al. (2018) and a newly derived classification system using OPGs from 211 individuals aged 15-25 years. The assessments were performed by three experienced examiners. One examiner evaluated all radiographs twice. The correlation between age and stage was investigated and the inter-and intra-rater reliability was estimated for all three methods. Correlation between stage and age was similar between classification systems, although higher in the data from males (Spearman's rho ranging from 0.568 to 0.583) than from females (0.440 to 0.446). Inter-and intra-rater reliability measures were similar across methods and invariant on sex, with overlapping confidence intervals, although the highest point estimates for both intra-and inter-rater reliability were for the method by Olze et al. with Krippendorf's alpha values of 0.904 (95% confidence interval 0.854, 0.954) and 0.797 (95% confidence interval 0.744, 0.850). It was concluded that the method of Olze et al. from 2012 is a reliable method for practical application and future studies.
If a dead body is discovered in water, it nearly always raises the question about the cause of death, often associated with the persistent problem to differentiate between a drowning incident and post-mortem immersion. In numerous cases, a reliable confirmation of death by drowning is often only possible by a combination of diagnoses obtained from autopsy and additional investigations. As to the latter, the use of diatoms has been suggested (and debated) since decades. Based on the consideration that diatoms are present in almost every natural waterbody and are unavoidably incorporated when water is inhaled, their presence in the lung and other tissues can provide evidence of drowning. However, the traditional diatom test methods are still subject of controversial discussion and suspected of erroneous outcome, predominantly through contamination. A promising alternative to minimize the risk of erroneous outcome seems to be disclosed by the recently suggested MD-VF-Auto SEM technique. Especially the establishment of a new diagnostic marker (L/D ratio), which represents the factorial proportion between the diatom concentration in lung tissue and the drowning medium, allows for clearer distinction of drowning and post-mortal immersion and is largely robust to contamination. However, this highly elaborated technique requires specific devices which are frequently unavailable. We therefore developed a modified method of SEM-based diatom testing to enable the use on more routinely available equipment. Process steps such as digestion, filtration, and image acquisition were thoroughly broken down, optimized, and ultimately validated in five confirmed drowning cases. Taking certain limitations into consideration, L/D ratio analysis provided promising results, even in cases of advanced decomposition. We conclude that our modified protocol indeed opens a way for a broader use of the method in forensic drowning investigation.
Line plot of number of publications per year
Proportion of unidentified bodies at admission stratified for developing and developed countries
Bar chart indicating key themes used to describe recommendations for improving forensic human identification procedures (numbers represent the number of articles making the recommendation)
While human identification is a crucial aspect of medico-legal investigations, many individuals remain unidentified each year across the world. The burden of unidentified bodies is often referred to when motivating for improved methods of identification, and anatomical teaching, yet the actual burden is somewhat unclear. A systematic literature review was undertaken to identify articles that empirically investigate the number of unidentified bodies experienced. Despite the large number of articles returned, an alarmingly low number (24 articles) provided specific and empirical details on the number of unidentified bodies, demographics and trends thereof. It is possible that this lack of data is due to the variable definition of ‘unidentified’ bodies and the use of alternative terminology such as ‘homelessness’ or ‘unclaimed’ bodies. Nevertheless, the 24 articles provided data for 15 forensic facilities across ten countries of both developed and developing statuses. On average, developing countries experienced more than double (9.56%) the number of unidentified bodies when compared to developed nations (4.40%). While facilities were mandated under different legislations and infrastructures available varied greatly, the most common issue faced is the lack of standardised procedures for forensic human identification. Further to this, the need for investigative databases was highlighted. Through addressing the standardisation of identification procedures and terminology, alongside the appropriate utilisation of existing infrastructure and database creation, the number of unidentified bodies could be significantly reduced globally.
Average DNA yield comparing lysis/purification methods: PrepFiler™ BTA and PrepFiler™ BTA/InnoXtract™ had similar average large and small target yields. Intermountain Forensics/EZ1 had significantly lower yields compared to Intermountain Forensics/InnoXtract™. PrepFiler™ BTA/InnoXtract™ and Intermountain Forensics/InnoXtract™ had similar DNA yields. (Data reported as µ ± σ; * indicates a p value < 0.05; ** indicates a p value < 0.005)
Lysis and digestion method effects on DNA yield: a two-part digestion method resulted in a significantly larger DNA yield for small targets. (Data reported as µ ± σ; * indicates a p value < 0.005)
Challenging skeletal samples (DNA yield): no statistically significant difference was seen when comparing the small target DNA yields for the InnoXtract™ and PrepFiler™ BTA methods
Challenging skeletal samples (STR typing): no significant differences between InnoXtract™ and PrepFiler™ BTA methods. (* indicates samples that failed to amplify)
The InnoXtract™ extraction and purification system is a purification method designed for DNA extraction from low-template samples, specifically rootless hair shafts. Its ability to successfully capture highly fragmented DNA suggests its suitability for use with other challenging sample types, including skeletal remains. However, the lysis and digestion parameters required modifications to successfully optimize the method for this sample type. A two-part digestion was developed utilizing a homebrew digestion buffer (0.5 M EDTA, 0.05% Tween 20, and 100 mM NaCl) and a supplemental lysis with the Hair Digestion Buffer included in the InnoXtract™ kit. Additionally, the magnetic bead volume was modified to improve DNA recovery from these challenging samples. With the altered protocol, the quality and quantity of DNA recovered from InnoXtract™ extracts were comparable to another commercial skeletal extraction method (PrepFiler™ BTA). This modified extraction method successfully purified sufficient amounts of quality DNA from a variety of skeletal samples to produce complete STR profiles. Successful STR typing from surface decomposition, burned, cremated, buried, and embalmed remains indicates the potential of this new method for challenging human identification and missing-person cases.
Sex estimation of skeletal remains is one of the most important tasks in forensic anthropology. The radius bone is useful to develop standard guidelines for sex estimation across various populations and is an alternative when coxal or femoral bones are not available. The aim of the present study was to assess the sexual dimorphism from radius measurements in a French sample and compare the predictive accuracy of several modelling techniques, using both classical statistical methods and machine learning algorithms. A total of 78 left radii (36 males and 42 females) were used in this study. Sixteen measurements were made. The modelling techniques included a linear discriminant analysis (LDA), flexible discriminant analysis (FDA), regularised discriminant analysis (RDA), penalised logistic regression (PLR), random forests (RF) and support vector machines (SVM). The different statistical models showed an accuracy of classification that is greater than 94%. After selection of variables, the accuracies increased to 97%. The measurements made at the proximal part of the radius (sagittal and transversal diameters of the head, and sagittal diameter of the neck), at distal part (maximum width of the distal epiphysis) and of the entire bone (maximum length) stand out among the various models. The present study suggests that the radius bone constitutes a valid alternative for sex estimation of skeletal remains with comparable classification accuracies to the pelvis or femur and that the non-classical statistical models may provide a novel approach to sex estimation from the radius bone. However, the extrapolation of the current results cannot be made without caution because our sample was composed of very aged individuals.
Schematic diagram of the experimental design regarding the process and timing of DNA fingerprint deposition
Comparison of T0, T1, and T2 for the dominant hand (left side of the figure) and non-dominant hand (right side of the figure). T0, before alcohol-based hand sanitizer; T1, immediately after the use of the alcohol-based hand sanitizer; T2, 1 h after the use of alcohol-based hand sanitizer. *, p = 0.0006; **, p = 0.0011
Genotyping results obtained from the dominant and non-dominant hand, shown as percentages on the total number of samples amplified. T0, before alcohol-based hand sanitizer; T1, immediately after the use of the alcohol-based hand sanitizer; T2, 1 h after the use of alcohol-based hand sanitizer
In the last years, forensic research has been focused on touch DNA in order to improve its evidential value in criminal activity investigations as well as to understand the variables impacting touch DNA. One of the emerging variables is represented by the use of alcohol-based sanitizers, which was suggested for hand hygiene during the COVID-19 pandemic. The aims of the present study were to assess the effect of a hand sanitizer on touch DNA deposition, transfer, and recovery and also to evaluate STR typing success, quality of DNA profiles, and personal identification. Before and after the use of an alcohol-based hand sanitizer, 20 volunteers deposited on glass surfaces 120 fingerprints, containing skin-derived or salivary DNA. Samples were quantified by real-time quantitative PCR (q-PCR), and 76 samples yielding > 15 pg/μl were typed for 21 autosomal STRs by GlobalFiler® PCR Amplification Kit. DNA profiles were classified into single source, mixed, and inconclusive profiles, and a LR assessment was performed by comparison to the reference samples using LRmix Studio software. After the use of hand sanitizer, samples yielded lower quantities of recovered transferred DNA, especially considering samples containing salivary DNA (p < 0.05 by Friedman test). All the 76 amplified samples (63.3% of the total) showed at least 10 typed loci, and 83–100% of profiles were consistent with the reference ones on the basis of a LR value ≥ 10⁶. Results showed that, although the hand sanitizer reduces the DNA recovering, touch DNA samples might still be useful for forensic personal identification even when hand sanitizers are used.
Post-mortem fundus photographs with quality sufficient to assert presence of absence of retinal hemorrhages suggestive of AHT. a Right eye of a three-month-old-girl, 4.5 h of post-mortem interval (PMI), undetermined cause of death; b Left eye of a 2-month-old boy, 9 h of PMI, cardiac cause of death
Post-mortem fundus photographs of retinal hemorrhages. a, b Right and left eyes of a 4-month-old boy, few bilateral superficial retinal hemorrhages confined to the posterior pole (1Bi and 1Ai) and nonspecific of abusive head trauma (AHT), 3 h of post-mortem interval (PMI), undetermined cause of death; (C) Left eye of a 1-week-old girl, numerous unilateral retinal hemorrhages (2Bi) nonspecific of AHT and explained by birth, 11 h of PMI, deceased by asphyxia; d Left eye of a 1-week-old girl, bilateral retinal hemorrhages (1Ai and 2Bi) nonspecific of AHT and explained by birth, 4 h of PMI, undetermined cause of death
Pathological eye examination with retinal hemorrhages. Pathological examination of the right eye of the 4-month-old boy (undetermined cause of death) with few superficial retinal hemorrhages presented in Fig. 2a and b, nonspecific of abusive head trauma. Microscopic examination of the retina stained with hemalun-eosine (a, magnification × 20; b, magnification × 4) confirmed the presence of superficial retinal hemorrhages under the inner limiting membrane (black arrows). Dotted arrow shows the optic nerve
Post-mortem fundus photographs of macular retinal folds, peripheral retinal folds and papillary vessels enlargement. a Right eye of a three-month-old boy, unique, long, and thin macular retinal fold, peripheral retinal folds, 27 h of post-mortem interval (PMI), deceased by asphyxia; b Left eye of a 3-month-old girl, unique, long and thin macular retinal fold, papillary vessels enlargement, 18 h of PMI, deceased by asphyxia; c Left eye of a 18-month-old boy, multiple, long, and large macular retinal folds, 9 h of PMI, sudden infant death syndrome; d Left eye of a 15-week-old boy, multiple, long and large macular retinal folds, peripheral retinal folds, papillary vessels enlargement, 11 h of PMI, undetermined cause of death
Introduction In the case of sudden unexpected death in infancy (SUDI), eye examination is systematic to detect retinal hemorrhages (RH) that are a crucial hallmark for abusive head trauma (AHT). The aim of this study is to assess the ability of non-invasive post-mortem fundus photographs (PMFP) to detect RH in case of SUDI. Methods Bicentric retrospective analysis of consecutive cases of SUDI under 2 years of age were managed by two French SUDI referral centers with PMFP by RetCam (Clarity Medical Systems USA). PMFP were reviewed randomly, twice, by three independent ophthalmologists blinded for clinical data. Results Thirty cases (60 eyes) were included. Median age was 3.5 months (interquartile [1.6; 6.0]). No child died of AHT. Image quality was sufficient to assert presence or absence of RH in 50 eyes (83%). Sufficient quality rate was significantly higher when the post-mortem interval was inferior to 18 h (91%, 42/46) as opposed to over 18 h (57%, 8/14, p=0.0096). RH were found in six eyes (10%), four children (13%), with excellent inter and intra-raters’ concordance (Cohen’s Kappa from 0.81 [0.56–1.00] to 1.00 [1.00–1.00]). Conclusion PMFP can detect RH in case of SUDI and is a relevant systematic screening test to be carried out as soon as the deceased child arrives in the hospital. It can decrease the need of eye removal for pathological examination, but further studies are needed to define the best decision algorithm.
⁸⁷Sr/⁸⁶Sr (blue circle) and ²⁰⁶Pb/²⁰⁴Pb (black square) per donation (D1–5), and soil ⁸⁷Sr/⁸⁶Sr data. Error bars are smaller than the symbols. Key: –– –– –– = local ²⁰⁶Pb/²⁰⁴Pb signature (black); - - - = local ⁸⁷Sr/⁸⁶Sr signature (blue). Local soil ⁸⁷Sr/.⁸⁶Sr from [26]
A²⁰⁷Pb/²⁰⁴Pb and ²⁰⁸Pb/²⁰⁴Pb ratios. Key: –– –– –– = local ²⁰⁸Pb/²⁰⁴Pb signature (black). B²⁰⁷Pb/²⁰⁶Pb and ²⁰⁸Pb/²⁰⁶Pb ratios of hair, bone, enamel, and dentine samples from donations 1–5. Key: –– –– –– = maximum local ²⁰⁸Pb/²⁰⁶Pb signature (black); - - - = maximum local ²⁰⁷Pb/²⁰⁶Pb signature (blue). Error bars are smaller than the symbols
The isotopic signatures of human tissues can provide valuable information on geographic origin for medicolegal investigations involving unidentified persons. It is important to understand the impact of diagenetic processes on isotopic signatures, as alterations could result in incorrect estimation of geographic origin. This study examines alterations in isotope signatures of different tissues of five human body donors studied throughout decomposition at the Forensic Anthropology Research Facility (FARF), San Marcos, TX. Two body donors were buried, two were placed in open pits, and one was first allowed to naturally mummify and then buried. Remains were recovered after a period of 7–34 months. The preplacement and post-recovery Sr–Pb isotope data of scalp hair, bone (iliac and tibia), and tooth enamel and dentine were compared. The hair samples record significant shifts in Sr-Pb isotope compositions, with hair keratin Pb isotope composition shifting towards the Pb signature of local soil samples. Hair keratin Sr isotope compositions were altered by the burial environment and possibly also by the lab sample cleaning method. The spongy iliac bone samples show inconsistencies in the recoverability of the preplacement Sr–Pb isotope signatures. The post-placement signatures of the buried donors show slight elevation over preplacement signatures. The post-placement signatures of donors placed in open pits are significantly elevated. The tibia and dental samples record the most consistent isotopic data with the least alteration. These more densely mineralised elements show good recoverability of the preplacement isotope signatures in burials and open pits and are thus deemed better targets for forensic investigative purposes.
a Axial MRI shows correct bilateral placement of dental cotton rolls filled with water (arrows) between the molars. The cotton rolls delineated the upper and lower molars and stabilized the bite. b, c Axial MRI through the upper jaw shows unsegmented and segmented tooth 28. Green for hard tooth tissue, blue for predentine, and red for pulp. d, e Coronal MRI through the upper jaw shows unsegmented and segmented tooth 28. Green for hard tooth tissue, blue for predentine, and red for pulp. f, g Sagittal MRI through the upper jaw shows unsegmented and segmented tooth 28. Green for hard tooth tissue, blue for predentine, and red for pulp. h 3D rendering of the segmentation, with a wedge of approximately one quarter removed to visualize the pulp. Green for hard tooth tissue, blue for predentine, and red for pulp
Age distribution of 67 healthy volunteers 44 females (F) and 23 males (M)
Tissue volumes for pulp, predentine, and hard tooth tissue for each sex. The box represents the upper and lower quartiles, the vertical thick lines the median, and the vertical thin lines of the whiskers represent maximum and minimum. Outliers are marked as circles
The regression model with the response variable natural logarithm of the ratio (pulp + predentine)/total applied to the upper 3rd molars for both sexes (p = 3.4 × 10⁻⁹) on y-axis against chronological age as explanatory variable on x-axis. The expectations are shown as solid lines and 95% confidence intervals as dashed curves (red for females, black for males). The sexes have the same slope but different intercept. Observed data are shown as circles
a–d The expectation (dashed black oblique line) for males (a) and females (b) and 95% prediction interval (dashed red oblique lines) of the natural logarithm of (pulp + predentine)/total applied to the upper 3rd molars (y-axis) against chronological age (x-axis). The prior age distribution (14.0–23.0 years) is shown as solid vertical black lines. The 18-year threshold is shown as a vertical dashed line. The color coded solid horizontal lines in black, red, green, and blue illustrate four different ratios of (pulp + predentine)/total, from the natural logarithm of hypothetical measurements in individuals. The posterior age distributions for males (c) and females (d), after applying Bayes theorem. The age distribution curves are color-coded (black, red, green, and blue) and correspond to the hypothetical ratios in a and b. The probabilities of being older than 18 years for each ratio are shown in the legends. The area under each curve is 100% of all probabilities (area equal to 1). For the blue curve, the probability for being older than 18 years is 97% for males taken the corresponded hypothetical blue color-coded vertical line in a. The age distribution curves are limited by the prior age distribution (14.0–23.0 years). The 18-year threshold is shown as a vertical dashed line
Purpose Our aim was to investigate tissue volumes measured by MRI segmentation of the entire 3rd molar for prediction of a sub-adult being older than 18 years. Material and method We used a 1.5-T MR scanner with a customized high-resolution single T2 sequence acquisition with 0.37 mm iso-voxels. Two dental cotton rolls drawn with water stabilized the bite and delineated teeth from oral air. Segmentation of the different tooth tissue volumes was performed using SliceOmatic (Tomovision©). Linear regression was used to analyze the association between mathematical transformation outcomes of the tissue volumes, age, and sex. Performance of different transformation outcomes and tooth combinations were assessed based on the p value of the age variable, combined or separated for each sex depending on the selected model. The predictive probability of being older than 18 years was obtained by a Bayesian approach. Results We included 67 volunteers (F/M: 45/22), range 14–24 years, median age 18 years. The transformation outcome (pulp + predentine)/total volume for upper 3rd molars had the strongest association with age (p = 3.4 × 10⁻⁹). Conclusion MRI segmentation of tooth tissue volumes might prove useful in the prediction of age older than 18 years in sub-adults.
Correlation between chronological age and methylation at the eight CpG sites in all 230 buccal swap samples
On the left: RMSE from repeated cross-validation of age prediction models on the training set given different cut-off values for a unisex model (first row), a model for sex 0 = men (second row), and a model for sex 1 =women (third row); vertical line indicates the cut-off value achieving the smallest RMSE; horizontal lines indicate RMSEs for the standard linear model (dotted), the default cut-off value (dashed), and the minimal value (solid). On the right: shape of the transformation function linking chronological and epigenetic age with the standard cut-off (vertical line)
The seven CpG site age estimation model predicting age for both training and validation set
Deviation of estimation errors from chronological age for the training and validation set. Boxplots represent the estimation error deviation for the training and validation set. Circles represent outliers in the corresponding set
Agreement between estimated and chronological age in the validation set. The chronological age was plotted against the difference between estimated age and chronological age. Each data point represents one analyzed sample within the validation set (n = 69). The dotted line represents the mean difference; the dashed lines show the upper and lower limits of agreement (1.96 × SD and −1.96 × SD, respectively). SD standard deviation
DNA methylation patterns change during human lifetime; thus, they can be used to estimate an individual’s age. It is known, however, that correlation between DNA methylation and aging might not be linear and that the sex might influence the methylation status. In this study, we conducted a comparative evaluation of linear and several non-linear regressions, as well as sex-specific versus unisex models. Buccal swab samples from 230 donors aged 1 to 88 years were analyzed using a minisequencing multiplex array. Samples were divided into a training set (n = 161) and a validation set (n = 69). The training set was used for a sequential replacement regression and a simultaneous 10-fold cross-validation. The resulting model was improved by including a cut-off of 20 years, dividing the younger individuals with non-linear from the older individuals with linear dependence between age and methylation status. Sex-specific models were developed and improved prediction accuracy in females but not in males, which might be explained by a small sample set. We finally established a non-linear, unisex model combining the markers EDARADD, KLF14, ELOVL2, FHL2, C1orf132, and TRIM59. While age- and sex-adjustments did not generally improve the performance of our model, we discuss how other models and large cohorts might benefit from such adjustments. Our model showed a cross-validated MAD and RMSE of 4.680 and 6.436 years in the training set and of 4.695 and 6.602 years in the validation set, respectively. We briefly explain how to apply the model for age prediction.
Development models of necrophagous insects are applied in forensic entomology for post-mortem interval estimation. Such estimates may be used as scientific evidence in legal investigations. For this reason, it is important that the models are valid and that the expert witness is aware of their limitations. Necrodes littoralis L. (Staphylinidae: Silphinae) is a necrophagous beetle species that frequently colonizes human cadavers. Temperature models of development for the Central European population of these beetles were recently published. In this article, we present results of the laboratory validation study for these models. Errors of beetle age estimation differed significantly between the models. Thermal summation models yielded the most accurate estimates, and the isomegalen diagram least accurate estimates. Errors of the beetle age estimation varied across beetle developmental stages and rearing temperatures. In general, most development models of N. littoralis were satisfactorily accurate in estimating beetle age under laboratory conditions; therefore, the study provides initial evidence to support their validity in forensic cases.
Schematic representation of age of the enrolled patients
Schematic comparison between the claimed typology of injuries by patients and the diagnoses made by clinicians
Schematic representation of the injuries observed by clinicians on the patients
Schematic representation of the frequency of sampling clothes (on the left) and taking photos (on the right)
Schematic flowchart of the essential medicolegal measures
Most victims of physical violence sooner or later will access a hospital or medical cabinet because of that violence, and in particular emergency departments (EDs). This paper aims to analyze the performance of emergency ward clinicians in the forensic management of such victims by examining the activities carried out and the data reported. A total of 991 medical records were extrapolated from the database of the ED of the Policlinico of Milan in an average pre-pandemic 1-year activity. For each medical record, 16 parameters were analyzed in-depth including epidemiological data, information on the type of violent actions, injuries, and time between the infliction of the lesion and access to the ED. In the vast majority of cases, all the actions with medicolegal implications had been neglected by health professionals causing loss of data not only for the justice system but especially for correctly interpreting what happened and taking appropriate measures to protect the patient/victim. Hence, given that clinicians in EDs are busy with non-forensic clinical tasks (and rightly so), it should be ensured that there be specific forensic clinical personnel. However, it is crucial that when unfortunately there can be no forensic staff, at least the clinicians who work in the ED are properly trained to correctly apply essential medicolegal measures. Overall, timely and informed medical and forensic intervention is possible and necessary for the improvement and maintenance of the mental and physical health of victims of violence.
a, b Skin on chest and lower back (left image) and skin on left breast (right image), with partially missing mammilla, scarred areola, and burns, most likely from cigarettes
Discolorations, in part like tram-line bruises on thighs and buttocks
We report the case of a woman in her thirties who suffered an esophageal rupture while participating in extreme sadomasochistic practices. After herself seeking help in a hospital for complaints alleged to be from a fall, she was initially diagnosed with several broken ribs and a pneumothorax. The cause of the pneumothorax was later discovered to be an esophageal rupture. When confronted with this atypical injury for a fall, the woman admitted to have accidentally swallowed an inflatable gag, which her partner had afterwards inflated. In addition to the esophageal rupture, the patient also had numerous other externally visible injuries of various ages, reportedly also from sadomasochistic acts. Although an in-depth police investigation was conducted and a “slave contract” was found, the woman’s consent to the extreme sexual practices performed by her life partner could not be substantiated conclusively. The man was convicted for intentional infliction of serious as well as dangerous bodily injury and sentenced to a long term in prison.
In the context of the coronavirus disease (COVID-19) pandemic, measures were taken to protect the population from infection. These were almost completely lifted in several countries in the spring of 2022. To obtain an overview of the spectrum of respiratory viruses encountered in autoptical routine case work, and their infectivity, all autopsy cases at the Institute of Legal Medicine in Frankfurt/M. with flu-like symptoms (among others) were examined for at least 16 different viruses via multiplex PCR and cell culture. Out of 24 cases, 10 were virus-positive in PCR: specifically, 8 cases with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), 1 with respiratory syncytial virus (RSV), and 1 with SARS-CoV-2 and the human coronavirus OC43 (HCoV-OC43), as a double infection. The RSV infection and one of the SARS-CoV-2 infections were only detected due to the autopsy. Two SARS-CoV-2 cases (postmortem interval of 8 and 10 days, respectively) showed infectious virus in cell culture; the 6 other cases did not show infectious virus. In the RSV case, virus isolation by cell culture was unsuccessful (Ct value of 23.15 for PCR on cryoconserved lung tissue). HCoV-OC43 was measured as non-infectious in cell culture, with a Ct value of 29.57. The detection of RSV and HCoV-OC43 infections may shed light on the relevance of respiratory viruses other than SARS-CoV-2 in postmortem settings; however, further, more extensive studies are needed for a robust assessment of the hazard potential due to infectious postmortem fluids and tissues in medicolegal autopsy settings.
PCA model: PMI increases from the upper right-hand corner to the bottom left-hand corner (panel A). Investigating the loading plot (panel B), the decreasing of glucose, pyruvate, and 3-OH-butyrate with the increasing of PMI is observed, whilst late PMI samples (48–84 h) are characterised by higher levels of taurine, choline, creatine, hypoxanthine, ethanolamine, and succinate. Samples of the training set are indicated as “training” whereas samples of the test set as “test”
Regression model for ordinal data: confusion matrices obtained calculating the training set (A), performing cross-validation (B) and predicting the test set (C)
The SR obtained for the ordinal regression model (SRordinal) and that of the regression model (SR) are reported in the same plot; both the ordinal and the regression models discovered 3-hydroxybutyrate, negatively correlated to PMI, and alanine, glutamate, and glycine, positively correlated to PMI, as significantly relevant in predicting PMI (the profiles of these metabolites are reported in Fig. S2 of Supplementary Materials). The SR values have been multiplied by the sign of the Pearson correlation coefficient calculated between PMI and metabolite concentration; dashed red lines indicate the thresholds of SR at level α = 0.05
Potassium concentration ([K⁺]) vs. PMI; the dashed line indicates the regression line estimated by linear regression
Potassium concentration vs. quantified metabolites: SR plot; threonine, choline, alanine, hypoxanthine, taurine, creatine, glutamate, and glycine, positively correlated to [K⁺], and glucose and 3-hydroxybutyrate, negatively correlated to [K⁺], were discovered as relevant. The SR values have been multiplied by the sign of the Pearson correlation coefficient calculated between [K⁺] and metabolite concentration; dashed red lines indicate the thresholds of SR at level α = 0.05
Introduction The estimation of post-mortem interval (PMI) remains a major challenge in forensic science. Most of the proposed approaches lack the reliability required to meet the rigorous forensic standards. Objectives We applied ¹H NMR metabolomics to estimate PMI on ovine vitreous humour comparing the results with the actual scientific gold standard, namely vitreous potassium concentrations. Methods Vitreous humour samples were collected in a time frame ranging from 6 to 86 h after death. Experiments were performed by using ¹H NMR metabolomics and ion capillary analysis. Data were submitted to multivariate statistical data analysis. Results A multivariate calibration model was built to estimate PMI based on 47 vitreous humour samples. The model was validated with an independent test set of 24 samples, obtaining a prediction error on the entire range of 6.9 h for PMI < 24 h, 7.4 h for PMI between 24 and 48 h, and 10.3 h for PMI > 48 h. Time-related modifications of the ¹H NMR vitreous metabolomic profile could predict PMI better than potassium up to 48 h after death, whilst a combination of the two is better than the single approach for higher PMI estimation. Conclusion The present study, although in a proof-of-concept animal model, shows that vitreous metabolomics can be a powerful tool to predict PMI providing a more accurate estimation compared to the widely studied approach based on vitreous potassium concentrations.
H&E staining in rat skeletal muscle samples. a Normal skeletal muscles as a control. b PMNs are detected at 1 day after contusion (arrows). c Round-shaped MNCs (arrows) and long spindle-shaped cells (arrowheads) are present in the injured tissue at 3 days after contusion. d, e FBCs (arrowheads) concomitant with regenerated multinucleated myotubes (arrows) are observed in the areas of contusion at 5 and 9 days after contusion. f Some FBCs (arrowheads), fibrotic tissue, and regenerated multinucleated myotubes (arrows) were detectable in the wound zones at 14 days after injury. Scale bar, 50 µm
IHC staining results in rat skeletal muscle samples. a Representative IHC staining results of Pax7 at different posttraumatic intervals, and C represents the result obtained from the normal skeletal muscle as a control. b Representative IHC staining results of MyoD at different posttraumatic intervals. c Representative IHC staining results of myogenin at different posttraumatic intervals. Pax7, MyoD and myogenin immunoreactivities were all detected in the nucleus. Scale bar, 50 µm
Statistical analysis results of the positive-cell numbers and ratios in immunohistochemistry. All data were presented as mean ± SD and analyzed using one-way ANOVA. a, b, c *p < 0.05 (vs control group), #p < 0.05 (vs preceding posttraumatic group). d *p < 0.05 (vs 1 day group post-injury), #p < 0.05 (vs preceding posttraumatic group). e *p < 0.05 (vs 9 days group post-injury), #p < 0.05 (vs 1 day group post-injury). f *p < 0.05 (vs 14 days group post-injury), #p < 0.05 (vs preceding posttraumatic group)
Western blotting assays. a Representative immunoblotting results of Pax7, MyoD, myogenin, IGF-1, and GAPDH at different posttraumatic intervals in rat skeletal muscle specimens, the molecular weight of each band is indicated. Lane C represents the result of the control skeletal muscle sample. b Relative intensity of Pax7 to GAPDH. c Relative intensity of MyoD to GAPDH. d Relative intensity of myogenin to GAPDH. e Relative intensity of IGF-1 to GAPDH. All values are expressed as the mean ± SD (n = 5). *p < 0.05 (vs control group), #p < 0.05 (vs preceding posttraumatic group)
qPCR assays from rat skeletal muscle specimens. a Relative quantity of Pax7 mRNA expression. b Relative quantity of Myod1 mRNA expression. c Relative quantity of Myog mRNA expression. d Relative quantity of Igf1 mRNA expression. All values are expressed as the mean ± SD (n = 5). *p < 0.05(vs control group), #p < 0.05 (vs preceding posttraumatic group)
From the perspective of forensic wound age estimation, experiments related to skeletal muscle regeneration after injury have rarely been reported. Here, we examined the time-dependent expression patterns of multiple biomarkers associated with satellite cell fate, including the transcription factor paired box 7 (Pax7), myoblast determination protein (MyoD), myogenin, and insulin-like growth factor (IGF-1), using immunohistochemistry, western blotting, and quantitative real-time PCR in contused skeletal muscle. An animal model of skeletal muscle contusion was established in 30 Sprague–Dawley male rats, and another five rats were employed as non-contused controls. Morphometrically, the data obtained from the numbers of Pax7 + , MyoD + , and myogenin + cells were highly correlated with the wound age. Pax7, MyoD, myogenin, and IGF-1 expression patterns were upregulated after injury at both the mRNA and protein levels. Pax7, MyoD, and myogenin protein expression levels confirmed the results of the morphometrical analysis. Additionally, the relative quantity of IGF-1 protein > 0.92 suggested a wound age of 3 to 7 days. The relative quantity of Pax7 mRNA > 2.44 also suggested a wound age of 3 to 7 days. Relative quantities of Myod1, Myog, and Igf1 mRNA expression > 2.78, > 7.80, or > 3.13, respectively, indicated a wound age of approximately 3 days. In conclusion, the expression levels of Pax7, MyoD, myogenin, and IGF-1 were upregulated in a time-dependent manner during skeletal muscle wound healing, suggesting the potential for using them as candidate biomarkers for wound age estimation in skeletal muscle.
In the context of further impact tests with various striking weapons against the skull, it turned out that the manufacturer had incorrectly calibrated the force measuring plate, which was used in our earlier experiments. When the tests were carried out again under the same conditions, the measurement results were significantly higher.
The most common pulmonary findings in opiate-related fatalities are congestion and oedema, as well as acute and/or chronic alveolar haemorrhage, the cause of which is thought to be a damage to the capillary endothelium related to ischemia. Human vascular mesenchymal stromal cells (vMSCs) play a fundamental role in tissue regeneration and repair after endothelial cell injury, and they express opioid receptors. The aim of this study was to assess the effect of in vitro morphine exposure on the physiological activity and maintenance of human vMSCs. vMSCs were obtained from abdominal aorta fragments collected during surgery repair and were exposed to incremental doses (0.1 mM, 0.4 mM, 0.8 mM and 1 mM) of morphine sulphate for 7 days. The effect was investigated through cell viability assessment, proliferation assay, reactive oxygen species (ROS) detection assay, senescence-associated β-galactosidase assay, senescent-related markers (p21WAF1/CIP1 and p16INK4) and the apoptosis-related marker caspase 3. Moreover, an ultrastructural analysis by transmission electron microscopy and in vitro vascular differentiation were evaluated. Results showed a decrease of the cellular metabolic activity, a pro-oxidant and pro-senescence effect, an increase in intracellular ROS and the activation of the apoptosis signalling, as well as ultrastructural modifications and impairment of vascular differentiation after morphine treatment of vMSC. Although confirmation studies are required on real fatal opiate intoxications, the approach based on morphological and immunofluorescence methodologies may have a high potential also as a useful tool or as a complementary method in forensic pathology. The application of these techniques in the future may lead to the identification of new markers and morphological parameters useful as complementary investigations for drug-related deaths.
Overview of variables used for statistical analysis with their encoding and the respective data sources. Dependent variables = DV. Dichotomous variable encoded yes or no = y/n. Continuous variables = numeric value. Interval scaled variables with values 1 to 5 = interval scaled 1–5. Psychosocial Centre for Refugees = PSC. Istanbul Protocol = IP
Overview of study participants. Istanbul Protocol = IP. Psychosocial Centre for Refugees = PSC
Comparison of IP grading between Franceschetti et al. [14], Aarts et al. [13] and the study at hand
Sums of different types of violence mentioned during forensic medical examinations
As part of a third-party funded project, expert opinions according to the Istanbul Protocol (IP) standards were compiled in Germany on a larger scale for the first time. The assessment process was initiated for 130 project participants. Statistical analysis on numerous variables was performed to test the impact of the expert opinions, foremost of the forensic medical expert opinions, on the individuals’ asylum proceedings. The variables were drawn from forensic medical expert opinions and reports of findings, questionnaires for the study participants’ counsellors in the project and a query on the asylum status of the study participants. Regression analysis and bivariate analysis on two dependent variables—subjective impact on the asylum procedure from the counsellors’ point of view and objective change in the asylum status—were performed to test for an influence on asylum procedures. No statistically significant results were obtained for the objective change of the study participants’ asylum status. For the subjective dependent variable, a positive prediction was possible when simultaneously controlling for the independent variables introduction of a forensic medical expert opinion and highest IP grade ; a negative prediction was possible when simultaneously controlling for the independent variables introduction of a forensic medical expert opinion and use of IP grading . Apart from the statistical analysis, a positive effect of the assessment on the psychosocial well-being of the study participants could be demonstrated. The results differed from other European studies which demonstrated a correlation between the objective outcome of an asylum procedure (asylum status) and, for example, specific types of violence or the number of documented injuries. Differences also occurred in the use of the plausibility grades proposed by the IP, which questions their use in cases in which the reported torture happened a relevant time ago. Therefore, compiling individually worded evaluations instead of using the IP grading system—if possible, by an experienced forensic physician—is recommended in this scenario. Still, the assessment of alleged torture experiences should follow the IP guidelines, since psychological assessments are of especially high importance in cases with healed physical injuries and since the results also demonstrated a positive effect on the psychosocial well-being of the study participants.
Birth-related fractures are an important differential diagnosis of child abuse in early infancy. While fractures associated to vaginal deliveries are well known, cesarean section is not necessarily known to cause such injuries. Nevertheless neonatal fractures have been described after cesarean sections. To give an overview over the frequency and typical locations of such fractures, the appearance of symptoms and the timespan until diagnosis, a literature research was conducted via Google scholar and Pubmed, using the key words “cesarean section” and “fractures”. Birth-related fractures after cesarean sections are rare but can occur, with the long bones being particularly affected. Therefore, birth injuries should always be considered in the forensic medical assessment of fractures in early infancy, even after cesarean section. To enable a differentiation between birth trauma and physical abuse, birth and operation records should be checked for surgical manoeuvres, possible difficulties during the procedure or other risk factors. Birth-related fractures are usually detected early; in rare cases, the diagnosis is made only weeks after birth.
Background The out-of-hospital cardiac arrest (OHCA) in the young may be associated with a genetic predisposition which is relevant even for genetic counseling of relatives. The identification of genetic variants depends on the availability of intact genomic DNA. DNA from autopsy may be not available due to low autopsy frequencies or not suitable for high-throughput DNA sequencing (NGS). The emergency medical service (EMS) plays an important role to save biomaterial for subsequent molecular autopsy. It is not known whether the DNA integrity of samples collected by the EMS is better suited for NGS than autopsy specimens. Material and methods DNA integrity was analyzed by standardized protocols. Fourteen blood samples collected by the EMS and biomaterials from autopsy were compared. We collected 172 autopsy samples from different tissues and blood with postmortem intervals of 14–168 h. For comparison, DNA integrity derived from blood stored under experimental conditions was checked against autopsy blood after different time intervals. Results DNA integrity and extraction yield were higher in EMS blood compared to any autopsy tissue. DNA stability in autopsy specimens was highly variable and had unpredictable quality. In contrast, collecting blood samples by the EMS is feasible and delivered comparably the highest DNA integrity. Conclusions Isolation yield and DNA integrity from blood samples collected by the EMS is superior in comparison to autopsy specimens. DNA from blood samples collected by the EMS on scene is stable at room temperature or even for days at 4 °C. We conclude that the EMS personnel should always save a blood sample of young fatal OHCA cases died on scene to enable subsequent genetic analysis.
Pictures during the autopsy. A Large bilateral retroperitoneal hematoma. B Complete rupture of the infrarenal aorta and the left renal artery (►). Bleeding around the left kidney. C Disjunction between the L2 and L3 vertebrae (►) after removal of the hematoma and the intervertebral mass
Histopathology of the retroperitoneum. Tumor proliferation (A and B), composed of diffuse spindle cells that infiltrate the muscle (*). Spindle cells have a sparse cytoplasm and a large oblong nucleus, which is sometimes nucleolated. The collagenous stroma is the site of inflammatory cells, mainly lymphocytes. This proliferation infiltrates the 3 tunics of the aorta (C and D), causing a clear rupture of the aorta (¤). HES = hematoxylin–eosin-safran staining
Histopathology of the subcutaneous lesion on the posterior side of the right forearm (A and B) and the periorbital fat (C). In the hypodermis, proliferative conjunctival spindle cells are seen (¤), among which capillaries with thickened walls are present (►). The epidermis and dermis are normal. In the periorbital fat, no recent hemorrhage is observed but again increased connective tissue (*) with capillaries with thickened walls, cytosteatonecrosis lesions, and siderophages are seen. HES = hematoxylin–eosin-safran staining
All unexpected deaths of children require an autopsy to determine the cause of death. In cases of aortic rupture, the immediate cause of death is easily identified at autopsy. Although the majority of aortic ruptures are caused by high-energy trauma, other causes should not be missed. We present and discuss the case of a 29-month-old child who died suddenly at home. Her recent medical history and the ecchymotic lesions observed on external examination of the body appeared potentially suspicious of physical abuse. The autopsy concluded that death was due to complete rupture of the abdominal aorta with associated vertebral disjunction. At first glance, the overall forensic picture could suggest a traumatic death. However, careful inspection of the retroperitoneum revealed a discrete atypical mass of infiltrative tissue within the hematoma. Histopathological examinations confirmed tumor proliferation of the soft tissues, triggering vascular and spinal injuries. Other paraneoplastic elements or metastases were ultimately revealed (orbital and subcutaneous). Overall, this was a rare and fatal case of abdominal aortic rupture induced by tumors. Due to the mechanisms and the forces needed to cause vertebral dislocations and aortic rupture, the combination of the two is highly suggestive of child abuse when an accidental traumatic history is absent or inconsistent with the injuries. Nevertheless, this case illustrates the importance of a systematic and rigorous forensic examination, rather than ignoring other possible diagnoses.
Positive (left) and negative (right) photographs of the Turin Shroud (TS), depicting the frontal and dorsal image of a male human body (photo from, ©Vernon Miller, 1978)
Experimental findings regarding the complex “blood stains attributed to injuries caused by the crown of thorns”, in comparison with the corresponding “blood stains” on a the Turin Shroud (TS) (photo from, ©Vernon Miller, 1978); b simulation 1a: liquid/moist blood from antemortem scalp wounds is covering the hair and the face; c simulation 1b: blood from antemortem scalp wounds is covering the hair and the face, blood is coagulating and/or drying and blood components are mobilised by postmortem washing; (d) simulation 2: as simulation 1b but with additional oiling of face and hairline. The findings in d seem to be most similar to the findings on the TS (a)
Experimental findings regarding the complex “serum ring around the lance wound”: dried fluid spots after application of small samples of whole blood (a), vital whole blood on vital serum (b), postmortem whole blood on postmortem serum (c) and samples of postmortem blood on postmortem pleural effusion (d) onto cotton cloths. The findings in b–d are very similar; visually, a “serum ring” can not be distinguished from a “pleural effusion ring”
Experiments regarding the complex “belt of blood”. Experimental provocation of a belt of blood: a simulation of a flow of blood or bloodied water from a chest wound by application of blood to the right lateral chest wall (according to the assumed location of the lance wound); b rotation of the body to the left side; c tucking the cloth under the body; d wrapping the cloth around the body
Experiments regarding the complex “belt of blood”. Experimental findings on the cloth after the provocation of a belt of blood as illustrated in Fig. 4: a using undiluted blood and b, c using watery dilutions of blood (1:4 and 1:10, respectively); the findings appear very similar to the belt of blood on the TS (d) (photo from, ©Vernon Miller, 1978), especially when diluted blood samples were used
The Turin Shroud (TS) is a Christian relic interpreted to be the burial cloth of Jesus of Nazareth. It exhibits red discolorations that have been interpreted as blood stains and that are the subjects of a highly controversial discussion. We conducted experiments to identify theoretically possible explanations for the stains attributed to the crown of thorns, the lance wound and the belt of blood. In the experiments with a focus on the stains attributed to the crown of thorns, a very similar stain pattern as on the TS could be provoked by simulating the following sequence of events: blood from antemortem scalp wounds is covering hair and face; blood is coagulating and/or drying; blood components are mobilised by postmortem washing and oiling. A stain pattern very similar to the belt of blood on the TS was successfully provoked by simulating the following sequence of events: The body is lying in a supine position, blood or bloodied water flowing from a wound at the right lateral chest wall; the body is rotated to the left side; the Shroud is tucked under the back; the body is rotated back to a supine position and laid onto the Shroud. The so-called serum ring surrounding the stain attributed to the lance wound could be reproduced by sequential application of serum and whole blood samples or of pleural effusion and whole blood samples onto cotton cloth. It is obvious that any attempt to interpret the assumed blood stain pattern on the TS has serious limitations. Nevertheless, it seems remarkable that we were able to reproduce findings that appear to be very similar to stains on the TS.
The aim of this article is to provide a scientific and statistical basis to identify the murder weapon in stabbing cases from the geometric characteristics of the stab-marks left on human ribs. For this purpose, a quantitative predictive model is developed, based on geometric measurements of the stab-mark and its location along the rib. A general method based on Bayesian inference and probabilities is used for the model development, rather than a deterministic model given its inability in certain occasions to identify the murder weapon. Following the process explained in this article to collect the stab-mark information required, the complete probabilistic model exposed attained a high accuracy in the identification of the murder weapon between two macroscopically identical blades with a microscopic alteration in one of them (more than 90% of correct identification is achieved).
The structure and growth cycle of human hair
The process of hair analysis from crime scene to laboratory in forensic genetics. This figure illustrates the analyses that are possible from various aspects, such as morphology, mitochondrial DNA, nuclear DNA, DNA methylation, and protein when a hair is found at the crime scene
Hair is one of the most common pieces of biological evidence found at a crime scene and plays an essential role in forensic investigation. Hairs, especially non-follicular hairs, are usually found at various crime scenes, either by natural shedding or by forcible shedding. However, the genetic material in hairs is usually highly degraded, which makes forensic analysis difficult. As a result, the value of hair has not been fully exploited in forensic investigations and trials. In recent years, with advances in molecular biology, forensic analysis of hair has achieved remarkable strides and provided crucial clues in numerous cases. This article reviews recent developments in DNA and protein analysis of hair and attempts to provide a comprehensive solution to improve forensic hair analysis.
The structure-of-interest (SOI), defined as the sternoclavicular joints, together with their contributing portions of the sternum and the medial clavicles
Schematic workflow diagram of the proposed medial clavicular epiphyseal cartilage localization. 1. Annotation: CT images are manually annotated with two-dimensional ground-truth bounding boxes in axial slices around the structure-of-interest (SOI). The SOI is an easy-to-identify proxy structure for the actual medial clavicular epiphyseal cartilage. 2. RetinaNet Training: A RetinaNet is trained to detect the SOI in axial slices and predict bounding boxes. 3: Localization in CT scan: The SOI can be localized in an unknown CT scan of the upper body. For this purpose, the trained RetinaNet is applied to each slice in a CT scan and all positive detections are collected. Afterwards, the center of the bounding box which corresponds to the best detection (highest classification score) is used as the predicted location for the SOI
(A–F) Bounding boxes around the SOI in different CT scans after preprocessing. The SOI is defined as the sternoclavicular joints, together with their contributing portions of the sternum and the medial clavicles. In addition, (A) illustrates the 4 bounding box location parameters x, y, width (w), and height (h)
The left panels show axial CT slices with ground-truth bounding boxes around the SOI (yellow boxes) and detections (if predicted by network) (red boxes). The right panels show the central sagittal slice of the respective CT. The position of the axial slice in the left panel is indicated by the dashed blue line in the right panel. The red area in the right panel indicates the positions of all axial slices which contain the SOI and have ground-truth bounding boxes annotated. The heatmaps next to the right panels show the classification score returned by the RetinaNet for each axial slice (light orange = 1.0; black = 0.0). Detections made by the RetinaNet are true positive, if the axial slice has a ground-truth bounding box (red area) and the classification score is > 0.05 (e.g., light orange). A Shows an example of a true-positive localization of the SOI; i.e., the highest classification score was returned for a slice which indeed contains the SOI. B shows an example of a false-negative localization; i.e., the RetinaNet returned only classification scores < 0.05 even though the SOI is present in one or more slices
Background Deep learning is a promising technique to improve radiological age assessment. However, expensive manual annotation by experts poses a bottleneck for creating large datasets to appropriately train deep neural networks. We propose an object detection approach to automatically annotate the medial clavicular epiphyseal cartilages in computed tomography (CT) scans. Methods The sternoclavicular joints were selected as structure-of-interest (SOI) in chest CT scans and served as an easy-to-identify proxy for the actual medial clavicular epiphyseal cartilages. CT slices containing the SOI were manually annotated with bounding boxes around the SOI. All slices in the training set were used to train the object detection network RetinaNet. Afterwards, the network was applied individually to all slices of the test scans for SOI detection. Bounding box and slice position of the detection with the highest classification score were used as the location estimate for the medial clavicular epiphyseal cartilages inside the CT scan. Results From 100 CT scans of 82 patients, 29,656 slices were used for training and 30,846 slices from 110 CT scans of 110 different patients for testing the object detection network. The location estimate from the deep learning approach for the SOI was in a correct slice in 97/110 (88%), misplaced by one slice in 5/110 (5%), and missing in 8/110 (7%) test scans. No estimate was misplaced by more than one slice. Conclusions We demonstrated a robust automated approach for annotating the medial clavicular epiphyseal cartilages. This enables training and testing of deep neural networks for age assessment.
Box plots between age and each of the seven acetabular variables (V1–V7) to illustrate the relationship between the stage of each variable (horizontal axis) and age (vertical axis) comparing Portuguese (blue) and North American (green) males. The central line in the boxes indicates the median; the bottom and top of the box represent the first and third quartiles, respectively; and the ends of the whiskers correspond to the minimum and maximum values
Box plots between age and each of the seven acetabular variables (V1–V7) to illustrate the relationship between the stage of each variable (horizontal axis) and age (vertical axis) comparing Portuguese (blue) and North American (green) females. The central line in the boxes indicates the median; the bottom and top of the box represent the first and third quartiles, respectively; and the ends of the whiskers correspond to the minimum and maximum values
Since investigation of the timing of the skeletal traits among the acetabula of different populations is lacking, this study aims to evaluate the relevance of geographical origin in the acetabulum aging process and in the usability of the SanMillán-Rissech aging method. The acetabula of 826 European North Americans derived from the Bass Collection (USA) have been analyzed and compared with 611 Portuguese acetabula from the Luis Lopes Collection (Portugal) applying the most updated acetabular age estimation technique (2017). After evaluating and comparing the acetabular aging rates between both populations by Mann–Whitney U tests, the inaccuracy values (bias and absolute error) were analyzed and compared using population-specific reference samples and using references differing in geographical origin by Wilcoxon tests. In general terms, the North Americans age faster than the Portuguese, especially the females, reaching the consecutive acetabular stages at younger ages. Regarding the SanMillán-Rissech method accuracy, using population-specific reference samples produces, as a general rule, better outcomes. In addition, an exhaustive meta-analysis of inaccuracy values has demonstrated that this method provides better estimation values than pubic symphysis and auricular surfaces regardless of the geographic coherence of the reference sample. These inter-population skeletal differences are derived from different factors than age, highlighting the impact of both biological and social background on age estimation. A thorough analysis of the skeletal age-based timing becomes essential to understanding, deciphering and being able to minimize bias and potential inaccuracy or even counteract them when applying the age estimation methods to different populations.
Example of costal neck measurements developed by Partido-Navadijo et al. [18]: a) total length of the neck, from the head to the tubercle (TLN), length of the segment head + neck (LSHN), length of the segment neck + tubercle (LSNT); b) cranio-caudal diameter of the neck in the centre point (CCDN); c) anteroposterior diameter of the neck in the centre point (APDN)
Example of costal neck measurements developed by Partido-Navadijo et al. [18]: a) length of the head (LH); b) length of the tubercle (LT); c) total length of the rib (TLR) (R1 only)
Example of an ossified sternal chondrocostal cartilage fragment (limited by the dotted lines) and visualization of the ridge (square and arrow) that allows obtaining the total length of the first rib (TLR) measurement
Sexual dimorphism is the basis for sex estimation research in skeletal remains, helping the positive identification of individuals in forensic backgrounds. In this regard, it has been proved that the metrical analysis of the costal neck of the first four ribs in Spanish contemporary adult skeletons shows high sexual dimorphism, especially in the first rib. However, the validation of any method developed in identified skeletal collections must be compulsorily reproduced in other skeletonized sets to verify either its potential application in individuals from other locations, or the existence of possible biases associated with inter-population variation. Due to the geographical and socioeconomic proximity between Spain and Portugal, this paper aims to check the utility of the discriminant functions designed by Partido-Navadijo et al. (2021) in the two skeletal collections of the University of Coimbra (Portugal). Results show the utility of these discriminant functions in Portuguese population, with frequencies of cases correctly assigned reaching up to 93.6% in the Coimbra Identified Skeletal Collection (CISC); and up to 92.6% in the 21st Century Identified Skeletal Collection (CEI/XXI). However, two particularities between the Spanish and Portuguese osteological assemblages can be highlighted: the possible existence of secular changes in relation to the CISC collection (1904–1938); and a reversal in the degree of costal dimorphism, being the third and fourth ribs of the CEI/XXI collection more dimorphic than the first ones.
The illustration of the proposed method
The accuracy of age estimation in the test set. a The differences of absolute error between age groups and significant differences were noticed between every two groups. The lines represent the mean absolute error (symbol in the middle) with SD. b Violin diagram of errors between age groups. The red dotted lines represent the median; the blue dotted lines range from the first quartile to the last quartile. The outer shape of the violin is the kernel density estimate. c The confusion matrices for multi-group classification in the test sets. d A comparison of higher error and lower error in different age groups
The accuracy of sex estimation in the test set. a The confusion matrices for sex estimation in the test sets. b The ROC curves of the DASE-Net for sex estimation
Heatmaps of different age groups and the whole test set. The red indicates high attention and the blue indicates low attention. The chronological age and the prediction error are shown in white color. The last row shows the average heatmaps of all test samples
The heatmaps of different ages of three subjects. The red indicates high attention and the blue indicates low attention. The chronological age and the prediction error are shown in white color
Teeth-based age and sex estimation is an important task in mass disasters, criminal scenes, and archeology. Although various methods have been proposed, most of them are subjective and influenced by observers’ experiences. In this study, we aimed to develop a deep learning model for automatic dental age and sex estimation from orthopantomograms (OPGs) and compare to manual methods. A large dataset of 15,195 OPGs (age range, 16 ~ 50 years; mean age, 29.65 years ± 9.36 [SD]; 10,218 females) was used to train and test a hybrid deep learning model which is a combination of convolutional neural network and transformer model. The final performance of this model was evaluated on additional independent 100 OPGs and compared to the manual method for external validation. In the test of 1413 OPGs, the mean absolute error (MAE) of age estimation was 2.61 years by this model. The accuracy and the area under the receiver operating characteristic curve (AUC) of sex estimation were 95.54% and 0.984. The heatmap indicated that the crown and pulp chamber of premolars and molars contain the most age-related information. In the additional independent 100 OPGs, this model achieved an MAE of 3.28 years for males and 3.79 years for females. The accuracy of this model was much higher than that of the manual models. Therefore, this model has the potential to assist radiologists in automated age and sex estimation.
Introduction Fatal intoxications, both accidental and intentional, are a global issue. In the Western world, intoxications with pharmaceuticals dominate, but in other parts of the world, other substances are more common. In a forensic setting, elemental intoxications are of great importance when investigating both accidental, suicidal, and homicidal deaths. The current study presents normal postmortem reference concentrations of 68 elements in femoral blood and urine. In addition, possible sources of error such as contamination from sample tubes, preservative potassium fluoride (KF) solution, and storage time are evaluated. Methods Paired femoral blood and urine samples from 120 cases of death by suicidal hanging in Sweden were collected. Additionally, multiple batches of sample tubes and multiple batches of KF solution were also analyzed. Concentrations of elements were determined by double focusing sector field ICP-MS. Results Key descriptive statistics for 68 elements are provided in blood and urine. Contamination from sample tubes was minor compared to the overall mean elemental concentrations in both blood and urine. KF solution contained a large assortment of elements, but the overall contribution is relatively minor for most elements given the small amounts of solution added to samples. There were significant differences for 22 elements in blood and 17 elements in urine between samples with short and long storage time. Conclusion The present study provides an important tool when evaluating postmortem elemental concentrations. It fills a needed gap between large antemortem population studies and postmortem case reports or small case series of elemental intoxications.
Age distribution for every third molar (18, 28, 38, 48) based on the three-stage eruption scoring system in males
Age distribution for every third molar (18, 28, 38, 48) based on the three-stage eruption scoring system in females
Percent of individuals classified as 18 years and older for each phase
The biological aspects of determining the dental age of subadults represent an important interdisciplinary scientific link with applications in criminal law and in forensic anthropology and dentistry. In criminal procedural law, it is necessary to determine the exact age of an undocumented person in view of the application of the provisions on juvenile offenders and minor victims. Chronological age can be estimated from the development of the third molars, as these are the only teeth that develop at the age of 18. The aim of this study was to verify the applicability of the Gambier method based on the eruption of the third permanent molars in the mandible and maxilla, to contribute to forensic age assessment. The analyzed group that met the criteria consisted of 811 orthopantomograms (OPGs) (339 females and 472 males) between the ages of 13 and 25 years. The OPGs were retrospectively analyzed according to the method of Gambier et al. (Int J Legal Med 133:625–632, 29), which refers to the eruption stages of the third molar. Differences between eruption stages of maxillary and mandibular third molars were statistically significant in both biological sexes. Intersexual differences in mean age were significant only at stage 3 for any M3 tooth and at stage 1 for mandibular M3. There were no statistically significant differences between the left and right mandibular and maxillary third molars, respectively. Differences between mandibular and maxillary M3 were significant only for stage 1 in males on the left side and for stage 2 in both sexes and sides. The method used allowed the best classification of individuals into minor and adult groups (based on phase D—90.41% of individuals, based on the third stage of the mandibular left third molar—86.29%). Based on the results obtained, we can assume that the method cannot be used alone in the initial examination of living individuals, since all four third molars must be assessed and there are no additional findings from practice.
Joachim Gerchow, editor-in-chief of the Zeitschrift für Rechtsmedizin—Journal of Legal Medicine from 1970 to 1990 (photo reproduction courtesy of Hogrefe Verlag)
Countries of origin of the authors. The country of the first author determined the country of origin. The numbers in brackets indicate the number of articles written by authors from the respective countries. Other countries are USA (17), Spain (13), Belgium (12), CSSR (9), Bulgaria (7), Norway (7), Yugoslavia (3), USSR (3), UK (3) and Qatar (2). Brazil, China, France, India, the Netherlands and Thailand are represented with one article each
Number of traffic fatalities in West Germany from 1950 to 1990.
Source of data: Statistisches Bundesamt [18]
From volume 67 (1970) onwards, the journal appeared under the new bilingual title Zeitschrift für Rechtsmedizin—Journal of Legal Medicine. The editorial board was expanded and internationalised. From 1970 to 1990, 1416 articles were published in 36 volumes. 1036 articles were in German and 380 in English. The authors of 411 articles came from non-German-speaking countries. Compared to the periods under review in the first two parts of our article series, the proportion of papers on forensic genetics increased significantly between 1970 and 1990, with a small increase in publications on the identification of unknown dead bodies. An opposite trend was observed in the articles on forensic psychiatry and psychology, sexual medicine and social medicine. This development reflects a further sharpening of the discipline’s profile.
Sudden death cases in the young population remain without a conclusive cause of decease in almost 40% of cases. In these situations, cardiac arrhythmia of genetic origin is suspected as the most plausible cause of death. Molecular autopsy may reveal a genetic defect in up to 20% of families. Most than 80% of rare variants remain classified with an ambiguous role, impeding a useful clinical translation. Our aim was to update rare variants originally classified as of unknown significance to clarify their role. Our cohort included fifty-one post-mortem samples of young cases who died suddenly and without a definite cause of death. Five years ago, molecular autopsy identified at least one rare genetic alteration classified then as ambiguous following the American College of Medical Genetics and Genomics’ recommendations. We have reclassified the same rare variants including novel data. About 10% of ambiguous variants change to benign/likely benign mainly because of improved population frequencies. Excluding cases who died before one year of age, almost 21% of rare ambiguous variants change to benign/likely benign. This fact makes it important to discard these rare variants as a cause of sudden unexplained death, avoiding anxiety in relatives’ carriers. Twenty-five percent of the remaining variants show a tendency to suspicious deleterious role, highlighting clinical follow-up of carriers. Periodical reclassification of rare variants originally classified as ambiguous is crucial, at least updating frequencies every 5 years. This action aids to increase accuracy to enable and conclude a cause of death as well as translation into the clinic.
D-aspartic acid (D-Asp) content (as ln [(1 + D/L)/(1 – D/L)]; D= D-aspartic acid; L= L-aspartic acid) in total protein (TP) of female (◇) and male (■) bone samples, related to the age at death: a skull; n = 50 (21 females, 29 males), b clavicle; n = 50 (21 females, 29 males), and c rib; n = 46 (18 females, 28 males)
D-aspartic acid (D-Asp) content (as ln [(1 + D/L)/(1 – D/L)]; D= D-aspartic acid; L= L-aspartic acid) in ln [(1 + D/L)/(1 – D/L)] in non-collagenous total protein (NCP) of female (○) and male (▲) bone samples, related to the age at death: a skull; n = 49 (20 females, 29 males), b clavicle; n = 46 (20 females, 26 males), and c rib; n = 47 (20 females, 27 males)
Pentosidine (Pen) concentration [nmol/mg] of female (ж) and male (●) bone samples, related to the age at death: a skull; n = 45 (20 females, 25 males), b clavicle; n = 43 (18 females, 25 males), and c rib; n = 37 (13 females, 24 males)
Age-at-death estimation is of great relevance for the identification of unknown deceased individuals. In skeletonised corpses, teeth and bones are theoretically available for age estimation, but in many cases, only single bones or even only bone fragments are available for examination. In these cases, conventional morphological methods may not be applicable, and the application of molecular methods may be considered. Protein-based molecular methods based on the D-aspartic acid (D-Asp) or pentosidine (Pen) content have already been successfully applied to bone samples. However, the impact of the analysed type of bone has not yet been systematically investigated, and it is still unclear whether data from samples of one skeletal region (e.g. skull) can also be used for age estimation for samples of other regions (e.g. femur). To address this question, D-Asp and Pen were analysed in bone samples from three skeletal regions (skull, clavicle, and rib), each from the same individual. Differences between the bone types were tested by t-test, and correlation coefficients ( ρ ) were calculated according to Spearman. In all types of bone, an age-dependent accumulation of D-Asp and Pen was observed. However, both parameters (D-Asp and Pen) exhibited significant differences between bone samples from different anatomical regions. These differences can be explained by differences in structure and metabolism in the examined bone types and have to be addressed in age estimation based on D-Asp and Pen. In future studies, bone type-specific training and test data have to be collected, and bone type-specific models have to be established.
Informed consent is based on basic ethical principles that should be considered when conducting biomedical and behavioral research involving human subjects. These principles—respect, beneficence, and justice—form the foundations of informed consent which in itself is grounded on three fundamental elements: information, comprehension, and voluntary participation. While informed consent has focused on human subjects and research, the practice has been adopted willingly in the forensic science arena primarily to acquire reference samples from family members to assist in identifying missing persons. With advances in molecular biology technologies, data mining, and access to metadata, it is important to assess whether the past informed consent process and in particular associated risks are concomitant with these increased capabilities. Given the state-of-the-art, areas in which informed consent may need to be modified and augmented are as follows: reference samples from family members in missing persons or unidentified human remains cases; targeted analysis of an individual(s) during forensic genetic genealogy cases to reduce an investigative burden; donors who provide their samples for validation studies (to include population studies and entry into databases that would be applied to forensic statistical calculations) to support implementation of procedures and operations of the forensic laboratory; family members that may contribute samples or obtain genetic information from a molecular autopsy; and use of medical and other acquired samples that could be informative for identification purposes. The informed consent process should cover (1) purpose for collection of samples; (2) process to analyze the samples (to include type of data); (3) benefits (to donor, target, family, community, etc. as applicable); (4) risks (to donor, target, family, community, etc. as applicable); (5) access to data/reports by the donor; (6) sample disposition; (7) removal of data process (i.e., expungement); (8) process to ask questions/assessment of comprehension; (9) follow-up processes; and (10) voluntary, signed, and dated consent. Issues surrounding these topics are discussed with an emphasis on addressing risk factors. Addressing informed consent will allow human subjects to make decisions voluntarily and with autonomy as well as secure the use of samples for intended use.
Exemplary impression of the black and white graphic novel-style graphics of the game
Serious games (computer-based learning games) are increasingly used in medical education at various levels, as user access is independent of location and time and promotes non-linear learning. In legal medicine, interactive digital media are still scarce. The freely accessible online serious game “Adventure Legal Medicine” was developed as part of the “Hamburg Open Online University”. The goal was to teach the basics of forensic casework in a point-and-click adventure setting consisting of five cases. During development, 40 medical students were asked to evaluate the game anonymously. The System Usability Scale (SUS) resulted in a mean score of 86.7 (SD 8.3), which corresponds to above-average usability. Further specific evaluations revealed a good to very good rating of the game with no differences in terms of gender ( p = 0.214), first-year versus advanced students ( p = 0.393) and students who never/rarely or sometimes/often played computer games ( p = 0.780). Since there are only a few digital media so far that allow curricular integration into undergraduate teaching in legal medicine, this serious game represents a possibility to integrate digital media into both face-to-face teaching and distance learning and to use it as a supplement to the medical school’s own teaching offer, encouraging users to actively engage with the subject.
Location of the burial site and the bone fragments analyzed in this study. a Location of Qingdao City, where the murder case took place. The base map was obtained from the USGS National Map Viewer, public domain ( b The burial site in the suspect’s yard. c Nineteen bone fragment samples excavated at the burial site, and the test numbers are marked at the bottom of each sample
Spectrum of A14 obtained by the MALDI-TOF mass spectrometer. Five peptide markers assigned A14 to Homo sapiens are colored in red
The genetic profile of A14 by outgroup f3 and PCA. a The top 14 populations (including the putative parents) sharing the highest amount of genetic drift with A14 measured by f3 (Mbuti; X, A14). b Principal component analysis of A14 and the putative parents projected onto present-day Asian populations
Pairwise mismatch rate (PMR) between A14 and the putative parents. Baseline was generated by PMR calculated within northern Han individuals (n = 10)
Bones are one of the most common biological types of evidence in forensic cases. Discriminating human bones from irrelevant species is important for the identification of victims; however, the highly degraded bones could be undiagnostic morphologically and difficult to analyze with standard DNA profiling approaches. The same challenge also exists in archaeological studies. Here, we present an initial study of an analytical strategy that involves zooarchaeology by mass spectrometry (ZooMS) and ancient DNA methods. Through the combined strategy, we managed to identify the only biological evidence of a two-decades-old murder case — a small piece of human bone out of 19 bone fragments — and confirmed the kinship between the victim and the putative parents through joint application of next-generation sequencing (NGS) and Sanger sequencing methods. ZooMS effectively screened out the target human bone while ancient DNA methods improve the DNA yields. The combined strategy in this case outperforms the standard DNA profiling approach with shorter time, less cost, as well as higher reliability for the genetic identification results. Highlights • The first application of zooarchaeology by mass spectrometry technique in the forensic case for screening out human bones from bone fragment mixtures. • Application of ancient DNA technique to recover the highly degraded DNA sequence from the challenging sample that failed standard DNA profiling approaches. • A fast, sensitive, and low-cost strategy that combines the strengths of protein analysis and DNA analysis for kinship identification in forensic research.
In times of peace and except for terrorist attacks, fatalities by explosions are rare. Fireworks have deadly potential, especially self-made or illegally acquired devices. The use of professional pyrotechnics by untrained persons poses a life-threatening hazard. We present a case of devastating blunt force and blast injuries to the head and chest of a young man. After ignition of a display shell (syn. a real shell or mortar shell) without the use of a launching pipe, the device hit the man’s face, nearly simultaneously followed by the explosion of the burst charge. The autopsy revealed injuries to the face and forehead as well as extensive tissue structure damage and a massive contusion with a bloody edema of the lungs. Autopsy results are supplemented with CT imaging and 3D reconstruction of the fractured mid face, as well as histological and toxicological examinations. This case of a misused display shell demonstrates both its devastating destructive potential and the corresponding and rarely observed injury pattern.
Response received from the participants regarding a the types of face mask used, and b no. of times the mask is worn before its disposal c duration of wearing masks per day
Amount of DNA extracted from different types of face masks with respect to different portions of masks and duration of wearing of the face-masks (n = 50)
Quality of STR profiles obtained from the DNA yield from different face mask types
After the onset of COVID-19 pandemic, a sharp surge in the usage of the face-masks throughout the globe has been observed. Pre-experiment survey of 252 individuals indicated a higher use of cotton-make masks (41%), followed by N-95 make (31%), and surgical disposable masks (26%). It was also further revealed that a higher fraction of individuals wear a face-mask more than 3 times (37%) before its disposal. In order to assess the potential usability of different mask types as forensic DNA evidence, a study was conducted on 50 healthy individuals. DNA content of different fractions such as the portion of mask covering the mouth region and the ear-piece showed a good source of host DNA. Though no statistically significant difference (P < 0.05) was found in the DNA quantity obtained from different face mask types, an increasing trend was obtained in the order: cloth make type (7.031 ± 0.31 ng), N-95 make (4.711 ± 0.15 ng), and surgical disposable type (2.17 ± 0.13 ng). The time of wearing of a face-mask showed a positive correlation with the yield of DNA irrespective of the face-mask type used. Samples retrieved from both the portions covering the mouth area and the ear-piece showed a good source of genomic DNA yielding an average of 4.82 ± 0.11 ng and 4.44 ± 0.10 ng of DNA, respectively. Irrespective of the face-mask types, number of reuse, and the portion of the mask, 66.66–96.11% of samples showed a complete autosomal STR DNA profile. This suggests that if a face-mask is found at the crime scene, it should be collected and preserved as a potential source of DNA evidence for routine forensic DNA analysis.
The pepper spray launcher “Jet Protector JPX®” [13]
Skin wound caused by the second pepper spray shot
Appearance of the scar about half a year after the incidence
Initial dispersion of a jet from a cartridge-powered irritant pistol. Muzzle on the left side (extract from a high-speed video performed by the group of forensic physics/ballistics at the ballistic laboratory of the legal medicine Bern under supervision of Beat P. Kneubuehl)
Pepper spray launchers are more precise and wind stable compared to conventional pepper sprays and are commonly used as a self-defensive tool. With the advanced potential, they may also harbour a greater risk for injuries, especially if they are not used within the suggested safety distance. If the shooting distance is below 1.5 m, energy densities may exceed the threshold energy density for the penetration of skin leading to skin laceration. We present a case where a man is hit by the liquid jet of a JPX Jet Protector® with an estimated shooting distance of 0.3 m. The man suffered from a bleeding skin laceration, which had to be sewed in the hospital. This case report furthermore outlines the potentially dangerous effect of pepper spray launchers and thereby their role in forensic investigations.
Chrysomya megacephala development from newly hatched larvae until pupation (in hours) reared on AlP with their mean larval length in control and dosage groups
The weight of Chrysomya megacephala larvae in dosage groups and in the control group until pupation (in hours). The larvae in groups B and D were the heaviest of the groups, and group D retained its heavy weight until pupation
Change in larval length in Chrysomya rufifacies dosage groups and the control group until pupation (in hours)
Comparison of the weight of Chrysomya rufifacies larvae in dosage groups and in the control group until pupation (in hours). Group D larvae reached peak weight at 114 h, whilst the other groups did not
Chrysomya megacephala and C. rufifacies are globally predominant species of blowflies that are amongst the primary colonisers of decomposing carcasses. The development of blowflies has been studied extensively to estimate the minimum postmortem interval (PMImin). This article evaluates the effect of aluminium phosphide (AlP) on the larval morphometry and life cycles of C. megacephala and C. rufifacies. The rabbits were given different dosages of AlP and their livers were excised to rear C. megacephala and C. rufifacies. Through multiple-regression analysis, we assessed the overall impact of varying dosages of AlP on larvae of both species. AlP accelerated development until pupation, whilst the time until emergence remained the same. At higher concentrations, AlP had a positive effect on the development of these species. Since both species are abundant, forensic entomology studies examining the effect of AlP on morphometry and developmental stages would provide valuable information to investigators assessing PMImin.
The classification system by Vieth et al. [14] applied to the DFE. Schematic drawings and case examples from our study cohort
Box-and-whisker plot diagram showing the age ranges obtained for all five ossification stages (stages 2–6) and for both sexes. Whiskers indicate the minimum and maximum ages, unless otherwise indicated by stars (statistical outliers)
The age of majority, which corresponds to the age of 18 years in most European countries, plays a crucial role for a large number of legal decisions. Accordingly, an increasing number of requests by authorities to forensic age estimation experts comprise the question of whether the age of 18 years has been reached by an individual. In recent years, novel study data suggested that magnetic resonance imaging (MRI) of the knee might likewise allow for the determination of majority beyond reasonable doubt. However, the data basis, especially concerning the distal femoral epiphysis (DFE), is still poor. For this reason, 392 routine MRI cases of the knee (204 males and 188 females of a Western Caucasian population, aged between 12 and 25 years) were retrospectively analyzed. T1-weighted and water-selective fat-saturated PD/T2-weighted sequences, generated at 1.5 and 3.0 T clinical MR scanners, were available. Ossification stages of the DFE were determined by means of the classification system by Vieth et al. (Eur Radiol 2018; 28:3255–3262). Both the intra-observer agreement and inter-observer agreement were found to be “very good” (κ = 0.899 and κ = 0.830). The present study confirmed that MRI of the DFE is suitable to determine majority in both sexes when stage 6 is present as the study revealed minimum ages above the age of 18 years for this stage (20.40 years in males and 20.60 years in females). Accordingly, the data represent a strong support for the so far existing database. Hence, the investigation of the knee using routine MRI appears to become a realistic alternative for forensic age estimation practice in the near future.
One of the most important pieces of evidence to find the suspect of a sexual assault are semen stains. It has been observed that suspects do not use condoms in many cases and transfer seminal fluids over the body of the victim, to their clothes, to bed linen, or to other objects in the environment. However, in many cases, it has been seen that the suspects or the victims tend to clean their bodies and clothes because of psychological conditions such as fear, panic, and anxiety. This study aims to determine whether human identification can be accomplished from washed semen stains by evaluating the quantity of extracted DNA obtained from washed clothes. In this study, both cotton- and nylon-blended fabrics were stained with semen samples taken from 10 volunteers and washed with detergent at 40 °C, 60 °C, and 90 °C. After stains were made re-visible using an ultraviolet (UV) light source, DNA was extracted using two different extraction methods (phenol–chloroform and spin-column). As a result, DNA extraction methods, washing conditions, and obtained DNA quantity from different washed fabrics were compared. It was shown that DNA could be obtained from all samples and that these DNA amounts could be used to perform identification. This study gives hope to the victims who have not been able to go to the judicial units immediately after the incident for fear or any other reasons and who think that they cannot get results because they have washed the laundry.
β-APP expression in rat brainstem (IHC × 200). a The control group did not show expression of β-APP, b–d the TAI group showed strong expression of β-APP, e–g the RGD group showed expression of β-APP but at lower levels than in the TAI group
The ultrastructure of microtubules and microfilaments in rat brainstem at 24 h postinjury. a–c Myelinated nerve fibers and myelin lamellar structures were severely separated in some axons (asterisk). The microtubules and microfilaments were disorganized, disintegrated, and even completely absent (triangle), and only residual axolemma can be observed in some axons. d–f Unmyelinated nerve fibers showed more severe cytoskeletal damage (triangle). The extent of axonal injury in the RGD group was significantly milder than in the TAI group
Immunoblot analysis of α-tubulin expression in rat brainstem. α-Tubulin expression in brainstems was lower in the TAI group than in the controls and was higher in the RGD group than in the TAI group
Immunoblot analysis of NF-L expression in rat brainstem. NF-L expression was increased in TAI and was lower in the RGD group than in the TAI group
The expression of β-integrin in the pons of forensic cases. a The pons of an individual who died from coronary heart disease showed expression of β1-integrin. b The pons of an individual who died from TAI showed stronger, more centralized β1-integrin expression
Traumatic axonal injury (TAI) accounts for a large proportion of the mortality of traumatic brain injury (TBI). The diagnosis of TAI is currently of limited use for medicolegal purposes. It is known that axons in TAI are diffusely damaged by secondary processes other than direct head injury. However, the physiopathological mechanism of TAI is still elusive. The present study used RGD peptide, an antagonist of the mechanotransduction protein integrin, to explore the role of integrin-transmitted mechanical signalling in the pathogenesis of rat TAI. The rats were subjected to a linearly accelerating load, and changes in beta-amyloid precursor protein (β-APP) expression, skeleton ultrastructure, skeleton protein neurofilament light (NF-L), and α-tubulin in the brainstem were observed, indicating that RGD could relieve the severity of axonal injury in TAI rats. In addition, the expression of β-integrin was stronger and centralized in the brainstem of the deceased died from TAI compared to other nonviolent causes. This study examined the pathophysiology and biomechanics of TAI and assessed the role of integrin in the injury of microtubules and neurofilaments in TAI. Thus, we propose that integrin-mediated cytoskeletal injury plays an important role in TAI and that integrin has the potential as a biomarker for TAI.
PRISMA chart
Immunohistochemical marker detection time
Proteins and mRNA detection time
The capability of discriminating between a vital and a post-mortem injury has always been a central theme in forensic pathology, particularly when the corpse is an advanced state of decomposition. Post-mortem decay of the body can mask or disrupt the classical features of a skin lesion, making it difficult to establish the cause and manner of death. Taphonomically challenging situations pose several interpretative issues of skin lesions which need to be addressed with scientifically recent methods that are still limited in the forensic literature. For that reason, the present research aims at resuming what is currently available in the attempt to provide some insight regarding this topic. This review considers only original researches, in which the markers of vitality were studied a significant amount of time after death, in order to test post-mortem persistency of these markers over time. A number of 132 original articles and reviews were considered, and the most significant results are resumed in an overview table and in two intuitive figures. Though many researchers tried to establish the vitality of lesions in specimen, few analysed samples from bodies when a significant degree of putrefaction or burning had occurred. The most significant marker proved to be GPA, which sowed a satisfying persistence over time (up to 6 months in air putrefaction and 15 days in water). However, what clearly emerged is that further studies are needed to address the challenges of taphonomically transformed specimen and to possibly neutralize the variability of experimental conditions, which affect the reproducibility of results. In conclusion, this study could be a starting point for providing food for thoughts about the most useful markers to search for in unusually tricky autopsy cases.
Top-cited authors
Andreas Schmeling
  • Universitätsklinikum Münster
Heidi Pfeiffer
  • University of Münster
Thomas Bajanowski
  • University Hospital Essen
Volker Vieth
  • Mathias-Spital Rheine - Stiftung privaten Rechts
Cristian Palmiere
  • Lausanne University Hospital