Investigation of post mortem brain, rectal and forehead temperature relations.

It is well known that magnetic resonance (MR) imaging is temperature sensitive, which is highly relevant for post mortem examinations. Therefore, the determination of the exact temperature of the investigated body site, e.g. the brain, is crucial. However, direct temperature measurements are invasive and inconvenient. Thus, in view of post mortem MR imaging of the brain, this study aims at investigating the relation between the brain and the forehead temperature for modelling the brain temperature based on the non-invasive forehead temperature. In addition, the brain temperature will be compared to the rectal temperature. Brain temperature profiles measured in the longitudinal fissure between the brain hemispheres, as well as rectal and forehead temperature profiles of 16 deceased were acquired continuously. Linear mixed, linear, quadratic and cubic models were fitted to the relation between the longitudinal fissure and the forehead and between the longitudinal fissure and the rectal temperature, respectively. Highest adjusted R2 values were found between the longitudinal fissure and the forehead temperature, as well as between the longitudinal fissure and the rectal temperature using a linear mixed model including the sex, environmental temperature and humidity as fixed effects. The results indicate that the forehead, as well as the rectal temperature, can be used to model the brain temperature measured in the longitudinal fissure. Comparable fit results were observed for the longitudinal fissure-forehead temperature relation and for the longitudinal fissure-rectal temperature relation. Combined with the fact that the forehead temperature overcomes the problem of measurement invasiveness, the results suggest using the forehead temperature for modelling the brain temperature in the longitudinal fissure.

Fully automated radiologic identification focusing on the sternal bone.

A crucial task in forensic investigations is the identification of unknown deceased. In general, secure identification methods rely on a comparison of ante mortem (AM) with post mortem (PM) data. However, available morphologic approaches are often dependent on the expertise and experience of the examiner, and often lack standardisation and statistical evidence. The objective of this study was therefore to overcome the current challenges via developing a fully automated radiologic identification (autoRADid) method based on the sternal bone. An anonymised AM data set consisting of 91 chest computed tomography (CT) scans, as well as an anonymised PM data set of 42 chest CT scans were included in this work. Out of the 91 available AM CT data sets, 42 AM scans corresponded to the 42 PM CT scans. For the fully automated identification analysis, a custom-made python pipeline was developed, which automatically registers AM data to the PM data in question using a two-step registration method. To evaluate the registration procedure and subsequent identification success, the image similarity measures Jaccard Coefficient, Dice Coefficient, and Mutual Information were computed. The highest value for each metric was retrieved in order to analyse the correspondence between AM and PM data. For all three similarity measures, 38 out of the 42 cases were matched correctly. This corresponds to an accuracy of 91.2%. The four unsuccessful cases incorporated surgical interventions taking place between the AM and the PM CT acquisition or poor CT scan quality preventing robust registration results. To conclude, the presented autoRADid method seems to be a promising fully automated tool for a reliable and facile identification of unknown deceased. A final pipeline combining all three similarity measures is open source and publicly available for efficient future identifications of unknown deceased.

A nontuberculous mycobacterium could solve the mystery of the lady from the Franciscan church in Basel, Switzerland.

BACKGROUND: In 1975, the mummified body of a female has been found in the Franciscan church in Basel, Switzerland. Molecular and genealogic analyses unveiled her identity as Anna Catharina Bischoff (ACB), a member of the upper class of post-reformed Basel, who died at the age of 68 years, in 1787. The reason behind her death is still a mystery, especially that toxicological analyses revealed high levels of mercury, a common treatment against infections at that time, in different body organs. The computed tomography (CT) and histological analysis showed bone lesions in the femurs, the rib cage, and the skull, which refers to a potential syphilis case. RESULTS: Although we could not detect any molecular signs of the syphilis-causing pathogen Treponema pallidum subsp. pallidum, we realized high prevalence of a nontuberculous mycobacterium (NTM) species in brain tissue sample. The genome analysis of this NTM displayed richness of virulence genes and toxins, and similarity to other infectious NTM, known to infect immunocompromised patients. In addition, it displayed potential resistance to mercury compounds, which might indicate a selective advantage against the applied treatment. This suggests that ACB might have suffered from an atypical mycobacteriosis during her life, which could explain the mummy's bone lesion and high mercury concentrations. CONCLUSIONS: The study of this mummy exemplifies the importance of employing differential diagnostic approaches in paleopathological analysis, by combining classical anthropological, radiological, histological, and toxicological observations with molecular analysis. It represents a proof-of-concept for the discovery of not-yet-described ancient pathogens in well-preserved specimens, using de novo metagenomic assembly.

Computed tomography osteoabsorptiometry for imaging of degenerative disc disease.

BACKGROUND: Lower back pain is a common condition with significant morbidity and economic impact. The pathophysiology is poorly understood but is in part attributable to degenerative disc disease (DDD). The healthy intervertebral disc ensures spine functionality by transferring the perceived load to the caudally adjacent vertebrae. The exposure to recurring mechanical load is mirrored in the mineralization pattern of the subchondral bone plate (SBP), where increased bone density is a sign of repetitive localized high stress. Computed tomography -osteoabsorptiometry (CT-OAM) is a technique based on conventional CT scans that displays the mineral density distribution in the SBP as a surface-color map. The objective of this study was to measure and analyze the SBP mineral density patterns of healthy lumbar intervertebral disc (IVDs) and those suffering DDD using CT-OAM densitograms. These findings should provide in vitro insight into the long-term morphological properties of the IVD and how these differ in the state of disc degeneration. METHODS: The CT-data sets of spines from 17 healthy individuals and 18 patients displaying DDD in the lumbar spine were acquired. Individual vertebrae of both cohorts were 3D reconstructed, processed using image analysis software, and compared to one another. Maximum intensity projection of the subchondral mineralization provided surface densitograms of the SBP. The relative calcium concentration, the local maxima of mineralization, and a mean surface projection of level-defined SBPs were calculated from the densitogram and statistically compared. RESULTS: The inferior SBP, adjacent to degenerating disc, display an 18-41 % higher relative calcium concentration than their healthy counterparts. In the opposing superior SBPs the relative calcium content is significantly increased. Whereas it is reasonably consistent for L1-L3 (L1: 132 %, L2: 127 %, L3: 120 %), the increase grows in caudal direction (L4: 131 %, L5: 148 %, S1: 152 %). Furthermore, a change in the areal distribution of excessive mineralization can be differentiated between healthy and diseased motion segments. CONCLUSIONS: The acquired data provide in vitro proof of the mechanical and anatomical properties of the SBP in relation to the state of disc degeneration. In conjunction with the diagnostic use of CT-osteoabsorptiometry, our data provide a basis for a non-invasive and sensitive technique that correlates with disc functionality. This could be promising in various cases, from early identification of early stages of DDD, tracking disease progression, and assessing the repercussions of surgical procedures or experimental therapies.

Post mortem brain temperature and its influence on quantitative MRI of the brain.

OBJECTIVE: MRI temperature sensitivity presents a major issue in in situ post mortem MRI (PMMRI), as the tissue temperatures differ from living persons due to passive cooling of the deceased. This study aims at computing brain temperature effects on the MRI parameters to correct for temperature in PMMRI, laying the foundation for future projects on post mortem validation of in vivo MRI techniques. MATERIALS AND METHODS: Brain MRI parameters were assessed in vivo and in situ post mortem using a 3 T MRI scanner. Post mortem brain temperature was measured in situ transethmoidally. The temperature effect was computed by fitting a linear model to the MRI parameters and the corresponding brain temperature. RESULTS: Linear positive temperature correlations were observed for T1, T2* and mean diffusivity in all tissue types. A significant negative correlation was observed for T2 in white matter. Fractional anisotropy revealed significant correlations in all gray matter regions except for the thalamus. DISCUSSION: The linear models will allow to correct for temperature in post mortem MRI. Comparing in vivo to post mortem conditions, the mean diffusivity, in contrast to T1 and T2, revealed additional effects besides temperature, such as cessation of perfusion and active diffusion.

Tissue sample analysis for post mortem determination of brain edema.

The post mortem evaluation of a brain edema is routinely performed by pathologists based on the macroscopic signs during autopsy. This method represents the current gold standard, but is subjective and observer dependent. Therefore, three post mortem evaluation methods of brain samples were analyzed in this work: histology, wet-dry weight and normalized cerebral weight, which was described in 2020 by Bauer et al. Tissue samples from six different regions of 34 brains were collected and examined both by rating of histological slides and by measuring the water content by using a drying oven. The rating of the histological slides, stained with hematoxylin and eosin, was performed by two pathologists independently. For the water content, the wet weight and the dry weight of each sample were set in relation. The normalized cerebral weight was calculated by dividing the brain weight by the brain volume, which were both determined during autopsy and in computed tomography images, respectively. A fair to moderate interrater agreement was obtained for the histologic evaluation and a significant correlation was present between one rater and the wet-dry weight and the normalized cerebral weight method. When classifying according to the gold standard, a significant difference was detected between the edematous and nonedematous cases by using the wet-dry weight method in the cerebral cortex and by using the normalized cerebral weight method. However, the significant correlations and group differences were limited to the aforementioned results. In conclusion, both the histological and the wet-dry weight method show limited benefits for the classification of brain edema and the histology analysis is highly observer dependent. The normalized cerebral weight method, however, reveals a significant effect between the edematous and nonedematous cases when classifying according to the gold standard. Therefore, we suggest to apply this method for the assessment of brain edema since it is objective and rater independent. Nevertheless, the exact evaluation of brain edema remains a challenging task, especially due to the continuous transition between no edema and edema.

Histomorphological assessment of isolated abdominal organs after targeted perfusion with the contrast agent Angiofil® in postmortem computed tomography angiography.

The current literature about histological alterations and artefacts associated with postmortem computed tomography angiography (PMCTA) is scarce and limited to qualitative research. The aim of this study was to describe and quantify histological alterations and artefacts in histologic specimens of the inner organs. The spleen, the liver, the kidneys and the pancreas were eviscerated in the course of standard forensic autopsy, followed by manual arterial and venous perfusion of the organs with the lipophilic contrast agent Angiofil®. Computed tomography (CT) of the organ packages was executed to evaluate if the perfusion with Angiofil® was sufficient. The CT scans revealed a good delineation of the vessel trees up to the organ periphery, suggesting a sufficient perfusion. Tissue samples were taken before and after perfusion, stained with hematoxylin-eosin (H&E) and examined by using microscopy. Native and perfused specimens were assessed on the basis of a qualitative rating scale in regard to the integrity of the cells, the tissue architecture and induced alterations. Subsequently the observed differences before and after perfusion with Angiofil® were analyzed quantitatively. While the liver, spleen and kidneys could be evaluated, further microscopic examinations of the pancreatic tissue were not possible due to vast autolytic changes. On the cellular level no changes in the liver, spleen and kidneys were observed after administering Angiofil®. While the tissue architecture of these organs remained intact, a significant blood volume displacement occurred. As a consequence, tissue with hypoemic or normal blood state demonstrated congestion. Further, optically empty spaces emerged regularly in the vessels and the parenchyma of each organ as a surrogate for the lipophilic contrast agent. With a few exceptions an adequate perfusion up to the microscopic level was observed. Following the results of this investigation, CT-controlled manual injection of Angiofil® into isolated organs may be an alternative to the generalized in situ application of contrast agents in the course of PMCTA. It is of eminent importance to know the artefacts induced by an Angiofil® injection in order to avoid a misinterpretation or a masking of findings. Thus, a collection of native tissue specimens has to be done before contrast agent application.

Postmortem CT Angiography Compared with Autopsy: A Forensic Multicenter Study.

Purpose To determine if postmortem computed tomography (CT) and postmortem CT angiography help to detect more lesions than autopsy in postmortem examinations, to evaluate the strengths and weaknesses of each method, and to define their indications. Materials and Methods Postmortem CT angiography was performed on 500 human corpses and followed by conventional autopsy. Nine centers were involved. All CT images were read by an experienced team including one forensic pathologist and one radiologist, blinded to the autopsy results. All findings were recorded for each method and categorized by anatomic structure (bone, organ parenchyma, soft tissue, and vascular) and relative importance in the forensic case (essential, useful, and unimportant). Results Among 18 654 findings, autopsies helped to identify 61.3% (11 433 of 18 654), postmortem CT helped to identify 76.0% (14 179 of 18 654), and postmortem CT angiography helped to identify 89.9% (16 780 of 18 654; P < .001). Postmortem CT angiography was superior to autopsy, especially at helping to identify essential skeletal lesions (96.1% [625 of 650] vs 65.4% [425 of 650], respectively; P < .001) and vascular lesions (93.5% [938 of 1003] vs 65.3% [655 of 1003], respectively; P < .001). Among the forensically essential findings, 23.4% (1029 of 4393) were not detected at autopsy, while only 9.7% (428 of 4393) were missed at postmortem CT angiography (P < .001). The best results were obtained when postmortem CT angiography was combined with autopsy. Conclusion Postmortem CT and postmortem CT angiography and autopsy each detect important lesions not detected by the other method. More lesions were identified by combining postmortem CT angiography and autopsy, which may increase the quality of postmortem diagnosis. Online supplemental material is available for this article.

Identification of deceased based on sternal bone computed tomography features.

Identification of deceased with unclear identity is a common problem in forensic science, whereby radiologic comparison can be applied as method for identification. As this comparison is mostly made on a visual basis, it is highly dependent on the examiner and often lacks standardized procedures and statistical support. The aim of this study therefore was to develop a reproducible and examiner independent method for radiologic identification (RADid) based on morphometric and morphological features of sternal bone computed tomography (CT). Furthermore, the feasibility of an automated comparison of a post-mortem (PM) case against a database of ante-mortem (AM) cases was evaluated. 44 in situ PM CT scans of sternal bone and their corresponding AM CT scans were analysed and reproducible features were selected based on intra- and interrater reliability assessments. The selected features were further tested by contrasting AM and PM data. Moreover, an automated identity evaluation was developed by calculating the number of matching features between each PM case and an AM database consisting of 94 cases including the 44 with corresponding PM scans. Several features showed to be reliable according to their resulting correlation coefficient values (greater or equal to 0.60). The suitability and stability of these features was confirmed by contrasting AM and PM CT scans. Finally, the automated comparison was successful in 76.7% of the cases, whereby an unambiguous identification was possible in 65.1%. The present study reflects the benefits of a standardized and statistically established identification method and demonstrates the high potential of the sternal bone as a suitable structure for RADid.

Evidence of neurofibromatosis type 1 in a multi-morbid Inca child mummy: A paleoradiological investigation using computed tomography.

OBJECTIVE: In this study, an Inca bundle was examined using computed tomography (CT). The primary aim was to determine the preservation status of bony and soft tissues, the sex, the age at the time of death, possible indicators for disease or even the cause of death, as well as the kind of mummification. A secondary aim was to obtain a brief overview of the wrapping in order to gain additional information on the cultural background. MATERIALS AND METHODS: The bundle belongs to the Museum of Cultures in Basel, Switzerland, and was bought in Munich, Germany, in 1921. Radiocarbon dating of the superficial textile yielded a calibrated age between 1480 and 1650 AD. The mummy was investigated using multi-slice CT with slice thickness of 0.75 mm and 110 kilovolt. For standardized assessment of soft tissue preservation, a recently developed checklist was applied. RESULTS: CT revealed the mummy of a seven to nine year old boy with superior preservation of bony and soft tissues allowing detailed assessment. Indicators of neurofibromatosis type 1 (paravertebral and cutaneous neurofibromas, a breast neurofibroma, sphenoid wing dysplasia), Chagas disease (dilatation of the esophagus, stomach, rectum, and large amounts of feces), and lung infection (pleural adherence, calcifications), probably due to tuberculosis, were found. Furthermore, signs of peri-mortem violence (transection of the chest and a defect in the abdominal wall) were detected. CT images revealed a carefully performed wrapping. CONCLUSION: CT examination of the Inca bundle proved to be an important non-destructive examination method. Standardized assessment, especially of the soft tissue structures, allowed for diagnoses of several diseases, indicating a multi-morbid child at the time of death. The careful wrapping pointed to a ceremonial burial. Within the cultural background, the signs of fatal violence were discussed as a possible result of war, murder, accident, or human sacrifice.

[The problem of highlights in forensic photography]. / Zur Problematik der Glanzlichtbildung bei der Fotodokumentation im Obduktionssaal.

A problem of forensic photography, especially when using direct flashlight, is the appearance of highlights on wet or shiny surfaces because of light reflections. Highlights can cause considerable difficulties if they interfere with relevant findings, because in a flare image information is reduced. Forensic literature describes several procedures to prevent the formation of highlights, which we compared to each other using flashlight in order to find the most effective and practicable method for reducing reflections. First, bounce flash photography can completely prevent flashlight flares on shiny surfaces such as putrefied bodies. Second, powdering such surfaces with rice powder can eliminate disruptive flares even when using direct flashlight but causes visible artefacts when using higher magnifications. As our study showed, bounce flash photography is an easy way to reduce highlights on photographs without the application of complex techniques or methods that may even alter relevant findings.

[Photodocumentation during autopsy]. / Fotodokumentation im Obduktionssaal--Untersuchungen zur Hintergrundgestaltung.

Photography during autopsy is an important part of forensic imaging and essential for the documentation of autopsy findings. A forensic photograph mainly serves the purpose of providing evidence and should be authentic. But aesthetic artwork may improve both the attractiveness and acceptance of autopsies and thus help to increase the autopsy rate. Different materials were compared to each other as photographic backdrop, particularly with regard to reflection and contrast, stability of exposure measurement and monitoring of colour fidelity. The photofoil 9010 Dove Grey proved to be a material of outstanding quality. In practical application, the foil was scratch-resistant and easy to clean. Furthermore, backdrop boards with a gap, e.g. for head and neck, allow special perspectives without a distracting background.

[Semen analysis using methods of molecular genetics--a case report]. / Anwendung des molekulargenetischen Spermanachweises--ein Fallbeispiel.

The presented case report describes the exclusion of the suspect of a sexual offence by means of methods of molecular genetics. Pretests for prostate-specific antigen performed at the beginning of the investigation and cytological sperm tests were negative. Nevertheless, by combining the methods of differential lysis and DNA quantification a small number of spermatozoa could be demonstrated in the trace evidence. Subsequently, the profile of the woman's boyfriend, with whom she had had unprotected vaginal intercourse two days before the incident, could be detected in the fraction of the hard lysis.

Mitochondrial D-loop (CA)n repeat length heteroplasmy: frequency in a German population sample and inheritance studies in two pedigrees.

Sequence analysis of the human mitochondrial genome (mtDNA) has proven to be a valuable tool in forensic identity testing and the analysis of crime scene stains. In contrast to the very expensive sequencing technique, typing of different length variants can greatly facilitate screening of a large number of traces for their relevance during casework. Within the mitochondrial control region, a dinucleotide (CA)( n ) repeat locus is present. To assess the discrimination power of this marker, we have determined (CA)( n ) allele distribution and the frequency of heteroplasmy in a population sample of 2,458 Germans. The inclination to develop heteroplasmic mixtures (CA)( n )/(CA)( n-1) was positively correlated with the number of CA repeats in the mtDNA. In addition, we have studied the inheritance patterns of (CA)( n ) repeat sequence heteroplasmy in two pedigrees. In one pedigree, we also found a length heteroplasmy in the homopolymeric C-tract (nt 303-309). Our data show stable inheritance of heteroplasmy within the homopolymeric C-stretch, but rather unstable inheritance regarding the (CA)( n ) repeat locus.

Forensic mass screening using mtDNA.

At the forensic autopsy of a sexual murder victim, some trace hairs, possibly belonging to the perpetrator, were saved. Initially, the analysis of a pubic hair shaft only revealed the presence of the mitochondrial (mt) DNA haplotype profile consisting of the (CA)(6) allele and the complete hypervariable region 1 (HV1) and 2 (HV2) sequence. Later, typing of some further telogene trace hairs, which had been stored for several years, yielded a nuclear short tandem repeat (STR) profile. We used both the mtDNA haplotype and the STR profile to start a DNA mass screening project involving 2,335 male citizens of the relevant communities. MtDNA screening was carried out by using the CA repeat amplification in combination with an SNP typing procedure based on the restriction site analysis of amplified d-loop sequences. The aim of our paper is to put mass screening with mtDNA up for discussion.

Cell line DNA typing in forensic genetics--the necessity of reliable standards.

The incorporation of reference DNA is crucial to the validation of any DNA typing protocol. This paper aims to provide a panel of reference DNAs for actual forensic profiling strategies, i.e. autosomal and gonosomal STR typing as well as mtDNA sequencing. We have characterised three human lymphoid cell lines, GM9947, GM9948 and GM3657, and considered 58 autosomal and gonosomal microsatellites as well as the mitochondrial control region sequence. Well-established markers and STRs recently developed for forensic use were involved. K562 DNA samples which we purchased from two different suppliers were also analysed. They revealed conflicting results with regard to the ChrX STR marker genotype. Hence, we suggest that K562 is no longer used for the calibration of profiling techniques. Our investigation establishes a panel of one female and two male DNA samples as an STR allelic ladder calibration tool and offers information on six alleles of each autosome (AS) marker, three alleles of each X chromosome (ChrX) marker and two alleles of each ChrY marker. In addition, sequences of the mitochondrial control region of the three DNAs are communicated in order to provide sequencing quality control.

Mitochondrial diversity of a northeast German population sample.

Mitochondrial DNA sequences of the hypervariable regions HV I and HV II were analyzed in 300 unrelated individuals born and living in the northeast corner of Germany (Western Pomerania) to generate a database for forensic identification purposes in this region. Sequence polymorphism were detected using PCR and direct sequencing analysis. A total of 242 different haplotypes were found as determined by 147 variable positions. The most frequent haplotype (263G, 315.1C) was found in 10 individuals and is also the most common sequence in Europe. Three other haplotypes were shared by 5 individuals, 2 sequences by 4, 8 haplotypes by 3, 15 sequences by 2 persons, and 213 sequences were unique. The genetic diversity was estimated to be 0.99 and the probability of two random individuals showing identical mitochondrial DNA (mtDNA) haplotypes is 0.6%. A comparison with other studies from Germany showed only little differences in the distribution of haplogroups. Nevertheless, one frequent haplotype in northeast Germany (five unrelated individuals) could only rarely be found in other German and European regions. Our results may indicate that despite a high admixture proportion in the German population some regions could demonstrate certain characteristic features.

Variability of mitochondrial DNA in a population sample from Iceland.

Over the past decade investigations of human mitochondrial DNA (mtDNA) have considerably contributed to our knowledge about human evolution and migration. The genome of the Icelandic population is of special interest since Iceland has been genetically isolated for centuries. The sequence of the hypervariable regions HVS-I and HVS-II of the mtDNA control region was generated for 100 Icelandic individuals. A total of 75 different mtDNA sequences were observed, of which 19 sequences were shared by more than one individual, 16 sequences were shared by two individuals and two sequences were shared by three individuals; the most frequent haplotype (16129 A, 16239 T, 00263 G and 00315.1 C) was found six times. Both the genetic diversity (0.9925+/-0.0031) and the average number of pairwise nucleotide differences (7.371) were comparable with most of the other European populations. However, we found a smaller number of distinct mitochondrial lineages, suggesting that founder effects and genetic drift may have exerted a visible influence on the Icelandic genetic diversity. We compared these data with 1400 other European sequences from the D-Loop-BASE database. The paper discusses the evolutionary relationship between Icelandic and Central European mtDNA under due consideration of the historical context. Finally, our study has been aimed at increasing the number of mtDNA sequences available throughout the world and contributing to human genome investigations.

The impact of mtDNA analysis between positions nt8306 and nt9021 for forensic casework.

Identification in forensic casework by mtDNA sequencing is predominantly done by sequencing the non-coding control region (HVI and HVII). In an attempt to further increase the discriminatory power of mtDNA analysis, we sequenced a coding region between nt8306 and nt9021 to identify additional polymorphisms in a group of 61 unrelated German individuals who had mtDNA profiles that occurred more than two times each, as well as a control group of 119 unrelated Germans whose profiles occurred one or two times each. Within these 180 individuals, 38 different polymorphisms in this region were observed; 64.4% of these individuals displayed the Cambridge reference sequence profile plus A8860G. For 28 individuals with the two most common profiles, A263G-315.1insC (N = 18) and A263G-309.1insC-315.1insC (N = 10), additional polymorphisms in this coding region permitted further discrimination of 56 and 40% of the individuals, respectively.