The influence of cooling on biomechanical time since death estimations using ovine brain tissue.

The significance of biomechanical analyses for forensic time since death estimations has recently been demonstrated. Previous biomechanical analyses successfully discriminated post-mortem brain tissue from tissue with a post-mortem interval of at least one day when held at 20 °C. However, the practical utility of such analyses beyond day one at 20 °C was limited. This study investigates the storage, loss, and complex shear modulus of various brain regions in sheep stored at 4 °C in 24-hour intervals over four days post-mortem using rheometry tests. The aim is to identify the critical biomechanical tissue property values to predict post-mortem time and assess the temperature sensitivity of the rheometry method by comparing results to recent findings at 20 °C. Thirty sheep brains were examined, including the frontal lobe, parietal lobe, anterior and posterior deep brain, superior colliculi, pons, medulla, and cerebellum. Rheometry tests were conducted, and receiver operator characteristic analyses were employed to establish cut-off values. At 4 °C storage, all investigated biomechanical properties of the examined brain regions remained stable for at least one day post-mortem. Using cerebellar samples stored at 4 °C, a post-mortem interval of at least two days could be determined with excellent diagnostic ability. Complex shear modulus values below 1435 Pa or storage modulus values below 1313 Pa allowed prediction of two or more days post-mortem. Comparisons between 4 °C and 20 °C revealed brain region-specific results. For instance, the complex shear moduli of the anterior deep brain at 4 °C were significantly higher on all individual testing days when compared to 20 °C. In contrast, the combined medulla and pons samples were similar on each day. Rheometry testing of brain tissue consistently stored at 4 °C since death proved valuable for forensic time since death estimations starting from two days after death.

Microstructural analysis on the innervation of the anterior, medial, and lateral human hip capsule: Preliminary evidence on its neuromechanical contribution.

OBJECTIVE: Capsular repair aims to minimize damage to the hip joint capsular complex (HJCC) and subsequent dislocation risk following total hip arthroplasty (THA). Numerous explanations for its success have been advocated, including neuromuscular feedback loops originating from within the intact HJCC. This research investigates the hypothesis that the HJCC contributes to hip joint stability by analyzing HJCC innervation. METHOD: Twenty-nine samples from the anterior, medial, and lateral aspects of the midportion HJCC of 29 individuals were investigated stereologically and immunohistochemically to identify encapsulated mechanoreceptors according to a modified Freeman and Wyke classification, totaling 11,745 sections. Consecutive slices were observed to determine the nerve course within the HJCC. RESULTS: Few encapsulated mechanoreceptors were found in the HJCC subregions and overlying tissues across the cohort studied. Of regions studied, no significant regional differences in the density of mechanoreceptors were found. No significant difference in mechanoreceptor density was found between sides (left, 10.2×10-4/mm3, 4.0×10-4 - 19.0×10-4/mm3; right 12.9×10-4/mm3, 5.0×10-4 - 22.0×10-4/mm3; mean, 95% confidence intervals) sexes (female 10.4×10-4/mm3, 4.0×10-4 - 18.0×10-4/mm3; male 11.6×10-4/mm3, 5.0×10-4 - 20.0×10-4/mm3; mean, 95% confidence intervals), nor in correlation with age demographics. Myelinated nerves coursed consistently within the HJCC in various orientations. CONCLUSION: Sparse mechanoreceptor density suggests that the HJCC contributes to a limited extent to hip joint stabilization. HJCC nerve terminals may potentially contribute to neuromuscular feedback loops with associated muscles to mediate joint stability in tandem with the active and passive components of the joint.

The use of brain tissue mechanics for time since death estimations.

Time since death estimation is a vital part of forensic pathology. Despite the known tissue degradation after death, the efficacy of using biomechanical tissue properties to estimate time since death remains unexplored. Here, eight brain tissue localizations were sampled from the frontal lobe, parietal lobe, anterior and posterior deep brain, superior colliculi, pons, medulla, and cerebellum of 30 sheep; were then stored at 20 °C; and subsequently subjected to rheometry tests on days zero to four after death. Overall, the measured tissue storage modulus, loss modulus, and complex shear modulus decreased after death for all of the tested regions in a site-specific manner. Day zero to day one changes were the only 24-h interval, for which statistically significant differences in tissue mechanical moduli were observed for some of the tested brain regions. Based on receiver operator characteristic analyses between day zero and the pooled data of days one to four, a post mortem interval of at least 1 day can be determined with a sensitivity of 90%, a specificity of 92%, and a positive likelihood ratio of 10.8 using a complex shear modulus cut-off value of 1461 Pa for cerebellar samples. In summary, biomechanical properties of brain tissue can discriminate between fresh and at least 1-day-old samples stored at 20 °C with high diagnostic accuracy. This supports the possible value of biomechanical analyses for forensic time since death estimations. A striking advantage over established methods to estimate the time since death is its usability in cases of disintegrated bodies, e.g. when just the head is found.

A disaster victim identification workshop focused on forensic odontology using embalmed human remains.

A high number of victims of mass casualty incidences are identified through their teeth. While forensic odontologists need to have a complex skillset during a disaster victim identification (DVI) response, hands-on training opportunities are rare. In countries with very limited forensic casework, such as New Zealand, many forensic odontologists find it difficult to achieve the number of annual forensic dental identifications required to maintain their credentialling. This report details the development of a hands-on forensic odontology-focused DVI workshop using human Crosado-embalmed remains. Anonymous participant evaluations, including five-point Likert and open-ended items, were performed in both years the workshop was held. A total of 10 and 17 participants, predominantly dentists, attended the workshop in 2020 and 2021, respectively. Participant feedback was extremely positive. Likert items were statistically similar between participants in both years. Open-ended items revealed positive feedback regarding the use of cadaveric remains, the gained hands-on experience, or the teamwork aspect. Participants who attended the workshop in both years commented on the positive aspect of repetition to cement their skills. As areas of improvement, participants named (for example) time management and the number of portable X-ray devices, leading to changes that were implemented in 2021. Moreover, the participants expressed interest to further their skills on decomposed, burnt, and fragmented human remains, which for ethical reasons has yet to be implemented. The DVI workshop described here, using embalmed human remains, provides an opportunity to add dental identifications toward annual credentialling requirements for forensic odontologists. Participants rated the course to be excellent overall and highly relevant for their role. For future workshops, there is an interest to include further aspects of the DVI response such as fingerprinting or police work as well as remains, which are altered due to natural or physical reasons.

Density of TMEM119-positive microglial cells in postmortem cerebrospinal fluid as a surrogate marker for assessing complex neuropathological processes in the CNS.

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

Forensic biomarkers of lethal traumatic brain injury.

Traumatic brain injury (TBI) is a major cause of death and its accurate diagnosis is an important concern of daily forensic practice. However, it can be challenging to diagnose TBI in cases where macroscopic signs of the traumatic head impact are lacking and little is known about the circumstances of death. In recent years, several post-mortem studies investigated the possible use of biomarkers for providing objective evidence for TBIs as the cause of death or to estimate the survival time and time since death of the deceased. This work systematically reviewed the available scientific literature on TBI-related biomarkers to be used for forensic purposes. Post-mortem TBI-related biomarkers are an emerging and promising resource to provide objective evidence for cause of death determinations as well as survival time and potentially even time since death estimations. This literature review of forensically used TBI-biomarkers revealed that current markers have low specificity for TBIs and only provide limited information with regards to survival time estimations and time since death estimations. Overall, TBI fatality-related biomarkers are largely unexplored in compartments that are easily accessible during autopsies such as urine and vitreous humor. Future research on forensic biomarkers requires a strict distinction of TBI fatalities from control groups, sufficient sample sizes, combinations of currently established biomarkers, and novel approaches such as metabolomics and mi-RNAs.

Fat Is Consistently Present within the Plantar Muscular Space of the Human Foot-An Anatomical Study.

Background and Objectives: The foot comprises of active contractile and passive connective tissue components, which help maintain stability and facilitate movement during gait. The role of age- or pathology-related degeneration and the presence of fat within muscles in foot function and pain remains unclear. The existence of fat has to date not been quantified or compared between individuals according to age, sex, side or subregion. Materials and Methods: 18 cadaveric feet (mean age 79 years) were sectioned sagittally and photographed bilaterally. Fat in the plantar muscular space of the foot (PMSF) was quantified through the previously validated manual fat quantification method, which involved observing photographs of each section and identifying regions using OsiriX. Fat volume and percentage was calculated using a modified Cavalieri's method. Results: All feet had fat located within the PMSF, averaging 25.8% (range, 16.5-39.4%) of the total PMSF volume. The presence of fat was further confirmed with plastination and confocal microscopy. Conclusions: These findings suggest that fat within the PMSF is a consistent but highly variable finding in elderly cohorts. Fat within the foot muscles may need to be considered a norm when comparing healthy and non-healthy subjects, and for therapeutic interventions to the foot. Further work is required to understand in detail the morphological and mechanical presence of fat in the foot, and compare these findings with pathological cohorts, such as sarcopenia. Additionally, future work should investigate if fat may compensate for the degeneration of the intrinsic muscles of the foot, with implications for both the use of orthotics and pain management.

Preliminary observations of the sequence of damage in excised human juvenile cranial bone at speeds equivalent to falls from 1.6 m.

There is much debate within the forensic community around the indications that suggest a head injury sustained by a child resulted from abusive head trauma, rather than from accidental causes, especially when a fall from low height is the explanation given by a caregiver. To better understand this problem, finite element models of the paediatric head have been and continue to be developed. These models require material models that fit the behaviour of paediatric head tissues under dynamic loading conditions. Currently, the highest loading rate for which skull data exists is 2.81 ms-1. This study improves on this by providing preliminary experimental data for a loading rate of 5.65 ± 0.14 ms-1, equivalent to a fall of 1.6 m. Eleven specimens of paediatric cranial bone (frontal, occipital, parietal and temporal) from seven donors (age range 3 weeks to 18 years) were tested in three-point bending with an impactor of radius 2 mm. It was found that prompt brittle fracture with virtually no bending occurs in all specimens but those aged 3 weeks old, where bending preceded brittle fracture. The maximum impact force increased with age (or thickness) and was higher in occipital bone. Energy absorbed to failure followed a similar trend, with values 0.11 and 0.35 mJ/mm3 for age 3 weeks, agreeing with previously published static tests, increasing with age up to 9 mJ/mm3 for 18-year-old occipital bone. The preliminary data provided here can help analysts improve paediatric head finite element models that can be used to provide better predictions of the nature of head injuries from both a biomechanical and forensic point of view.

GFAP positivity in neurons following traumatic brain injuries.

Glial fibrillary acidic protein (GFAP) is a well-established astrocytic biomarker for the diagnosis, monitoring and outcome prediction of traumatic brain injury (TBI). Few studies stated an accumulation of neuronal GFAP that was observed in various brain pathologies, including traumatic brain injuries. As the neuronal immunopositivity for GFAP in Alzheimer patients was shown to cross-react with non-GFAP epitopes, the neuronal immunopositivity for GFAP in TBI patients should be challenged. In this study, cerebral and cerebellar tissues of 52 TBI fatalities and 17 controls were screened for immunopositivity for GFAP in neurons by means of immunohistochemistry and immunofluorescence. The results revealed that neuronal immunopositivity for GFAP is most likely a staining artefact as negative controls also revealed neuronal GFAP staining. However, the phenomenon was twice as frequent for TBI fatalities compared to non-TBI control cases (12 vs. 6%). Neuronal GFAP staining was observed in the pericontusional zone and the ipsilateral hippocampus, but was absent in the contralateral cortex of TBI cases. Immunopositivity for GFAP was significantly correlated with the survival time (r = 0.306, P = 0.015), but no correlations were found with age at death, sex nor the post-mortem interval in TBI fatalities. This study provides evidence that the TBI-associated neuronal immunopositivity for GFAP is indeed a staining artefact. However, an absence post-traumatic neuronal GFAP cannot readily be assumed. Regardless of the particular mechanism, this study revealed that the artefact/potential neuronal immunopositivity for GFAP is a global, rather than a regional brain phenomenon and might be useful for minimum TBI survival time determinations, if certain exclusion criteria are strictly respected.

Ultrasound in legal medicine-a missed opportunity or simply too late? A narrative review of ultrasonic applications in forensic contexts.

OBJECTIVES: Conventional autopsies remain the gold standard of postmortem healthcare quality assurance and help gathering extended knowledge on diseases. In answer to constantly declining autopsy rates non- or minimally invasive autopsy methods were introduced. Ultrasound is a well-established tool for imaging commonly used in clinical practice. This narrative review aims to summarize the current literature regarding the feasibility and validity of ultrasound in a forensic context. MATERIAL AND METHODS: A PubMed database search was carried out. Abstracts were scanned for pre-defined ex- and inclusion criteria, followed by a snowball search procedure applied to the primarily included articles. RESULTS: Forty-five publications met our inclusion criteria. The selected articles concern the feasibility of ultrasound in pre- or postmortem settings, forensic age estimation, and minimally invasive approaches. For imaging, ultrasound was deemed a reliable tool for the examination of epiphyses und superficial wounds, with limitations regarding internal organs and image quality due to postmortem changes. Ultrasound-guided minimally invasive approaches yielded higher success rates for adequate tissue sampling. Many investigations were carried out in low- and middle-income countries focusing on infectious diseases. CONCLUSION: Ultrasound seems a promising but underutilized imaging tool in legal medicine to date. Promising approaches on its feasibility have been conducted. Especially for minimally invasive methods, ultrasound offered significant improvements on qualified biopsy sampling and thus appropriate diagnostics. Moreover, ultrasonic evaluation of epiphyses for age estimation offered valuable results. Nevertheless, further assessment of ultrasonic feasibility in forensic contexts is needed.

Anatomical structures at risk using different approaches for sacrospinous ligament fixation.

For 50 years now, sacrospinous ligament fixation (SSLF) has been used to treat pelvic organ prolapse consequent on altered integrity of the pelvic myofascial structures. It is usually performed vaginally, but it has recently been performed laparoscopically through either an anterior or a posterior approach, with the broad ligament as a landmark to differentiate the two. In the present study, these two laparoscopic approaches were assessed using Thiel-embalmed cadavers. The anterior and posterior approaches were compared in terms of the closest distance to anatomical structures at risk, including pelvic viscera, the obturator nerve, and vascular structures. The posterior approach was more often closer to the investigated vessels and the rectum. The obturator nerve and the ureter were close to both the anterior and posterior approaches. The urinary bladder was closer using the anterior approach. From an anatomical standpoint, therefore, the anterior laparoscopic approach for SSLF is more likely to cause injury to the urinary bladder, whereas the posterior approach is more prone to causing rectal and vessel injuries. This study illustrates, from a basic science perspective, the importance of combining fascia research, novel endoscopic or minimally invasive surgical exposures informed by anatomy, and contemporary trends in gynecology in order to improve patient outcomes. Clin. Anat. 33:522-529, 2020. © 2019 Wiley Periodicals, Inc.

What is Considered a Variation of Biomechanical Parameters in Tensile Tests of Collagen-Rich Human Soft Tissues? - Critical Considerations Using the Human Cranial Dura Mater as a Representative Morpho-Mechanic Model.

Background and Objectives: Profound knowledge on the load-dependent behavior of human soft tissues is required for the development of suitable replacements as well as for realistic computer simulations. Regarding the former, e.g., the anisotropy of a particular biological tissue has to be represented with site- and direction-dependent particular mechanical values. Contrary to this concept of consistent mechanical properties of a defined soft tissue, mechanical parameters of soft tissues scatter considerably when being determined in tensile tests. In spite of numerous measures taken to standardize the mechanical testing of soft tissues, several setup- and tissue-related factors remain to influence the mechanical parameters of human soft tissues to a yet unknown extent. It is to date unclear if measurement extremes should be considered a variation or whether these data have to be deemed incorrect measurement outliers. This given study aimed to determine mechanical parameters of the human cranial dura mater as a model for human soft tissues using a highly standardized protocol and based on this, critically evaluate the definition for the term mechanical "variation" of human soft tissue. Materials and Methods: A total of 124 human dura mater samples with an age range of 3 weeks to 94 years were uniformly retrieved, osmotically adapted and mechanically tested using customized 3D-printed equipment in a quasi-static tensile testing setup. Scanning electron microscopy of 14 samples was conducted to relate the mechanical parameters to morphological features of the dura mater. Results: The here obtained mechanical parameters were scattered (elastic modulus = 46.06 MPa, interquartile range = 33.78 MPa; ultimate tensile strength = 5.56 MPa, interquartile range = 4.09 MPa; strain at maximum force = 16.58%, interquartile range = 4.81%). Scanning electron microscopy revealed a multi-layered nature of the dura mater with varying fiber directions between its outer and inner surface. Conclusions: It is concluded that mechanical parameters of soft tissues such as human dura mater are highly variable even if a highly standardized testing setup is involved. The tissue structure and composition appeared to be the main contributor to the scatter of the mechanical parameters. In consequence, mechanical variation of soft tissues can be defined as the extremes of a biomechanical parameter due to an uncontrollable change in tissue structure and/or the respective testing setup.

A biomechanical comparison between human calvarial bone and a skull simulant considering the role of attached periosteum and dura mater.

PURPOSE: Current forensic analysis of blunt force trauma relies on the use of cadaveric or animal tissues, posing ethical and reproducibility concerns. Artificial substitutes may help overcome such issues. However, existing substitutes exhibit poor anatomic and mechanical biofidelity, especially in the choice of skull simulant material. Progress has been made in identifying materials that have similar mechanical properties to the human skull bone, with the potential to behave similarly in mechanical loading. AIMS: To compare the biomechanical properties of the human calvarial bone with an epoxy resin-based simulant material. Data collected was also used to analyse the effect of periosteal attachment on the mechanical properties of skull bone compared with that of the counterpart samples. METHODS: Fifty-six human skull bone specimens were prepared from two cadaveric heads. Half of these specimens were removed of periosteum and dura mater as the PR (periosteum removed) group, whereas periosteum was left attached in the PA (periosteum attached) group. Duplicates of the bone specimens were fabricated out of an epoxy resin and paired in corresponding PR and PA groups. The specimens were loaded under three-point bending tests until fracture with image-based deformation detection. RESULTS: Comparison of the epoxy resin and skull specimens yielded similarity for both the PR and PA groups, being closer to the PA group (bending modulus resin PR 2665 MPa vs. skull PR 1979 MPa, resin PA 3165 MPa vs. skull PA 3330 MPa; maximum force resin PR 574 N vs. skull PR 728 N, resin PA 580 N vs. skull PA 1034 N; strain at maximum force resin PR 2.7% vs. skull PR 5.1%, resin PA 2.3% vs. skull PA 3.5%, deflection at maximum force resin PR 0.5 mm vs. skull PR 0.8 mm, resin PA 0.5 mm vs. skull PA 1.0 mm). Bending strength was significantly lower in the resin groups (resin PR 43 MPa vs. skull PR 55 MPa, resin PA 44 MPa vs. skull PA 75 MPa). Moreover, the correlations of the mechanical data exhibited closer accordance of the PR group with the epoxy resin compared with the PA group with the epoxy resin. CONCLUSIONS: The load-deformation properties of the epoxy resin samples assessed in this study fell within a closer range to the skull specimens with PR  than with PA. Moreover, the values obtained for the resin fall within the reference range for skull tissues in the literature suggesting that the proposed epoxy resin may provide a usable artificial substitute for PA but does not totally represent the human skull in its complex anatomical structure.

Tissue biomechanics of the human head are altered by Thiel embalming, restricting its use for biomechanical validation.

Thiel embalming is a well-known method of anatomical fixation giving lifelike optical and haptic tissue properties. Beyond these characteristics, Thiel embalming may also be a promising method to provide lifelike tissues for validation purposes of human head biomechanics. Recent investigations using Thiel-embalmed human tissues of the upper and lower limb yielded contradicting biomechanical results on fixation-induced changes in the tissues' load-deformation behavior. It is to date unclear if Thiel embalming may have a softening or stiffening effect on human soft tissues or no global effect on biomechanics compared to the fresh state, with the latter being the most desirable outcome. The given study aimed at assessing the effects of Thiel embalming on the uniaxial tensile properties of human head soft tissues. Age-matched fresh and Thiel-embalmed dura mater, temporalis muscle, temporalis muscle fascia, and scalp samples were examined. Dura, fascia, and scalp samples showed significantly different elastic moduli compared to fresh tissues (all P < 0.01). The observed ultimate tensile strength supports the theory of an increased collagen crosslinking of the embalmed tissues when compared to the fresh state. Thiel-embalmed muscles failed any tensile testing approach as a result of the muscles dissolving due to the embalming. Furthermore, collagen integrity seems altered in scanning electron microscopy by the Thiel embalming, limiting their use for ultrastructural failure analyses. Thiel-embalmed soft tissues may consequently not serve to reflect the biomechanical properties of the human head. Consequently, the application of Thiel embalming should be limited to preliminary tests for biomechanical purposes. Clin. Anat. 32:903-913, 2019. © 2019 Wiley Periodicals, Inc.

Frequency and intensity of pulmonary bone marrow and fat embolism due to manual or automated chest compressions during cardiopulmonary resuscitation.

Iatrogenic consequences of cardiopulmonary resuscitation (CPR) include sternal or rib fractures, pulmonary bone marrow embolisms (BME) and fat embolisms (FE). This report aimed to analyze the frequency and intensity of pulmonary BME and FE in fatal cases receiving final CPR efforts with the use of automated chest compression devices (ACCD) or manual chest compressions (mCC). The study cohort (all cardiac causes of death, no ante-mortem fractures) consisted of 15 cases for each group 'ACCD', 'mCC' and 'no CPR'. Lung tissue samples were retrieved and stained with hematoxylin eosin (n = 4 each) and Sudan III (n = 2 each). Evaluation was conducted microscopically for any existence of BME or FE, the frequency of BME-positive vessels, vessel size for BME and the graduation according to Falzi for FE. The data were compared statistically using non-parametric analyses. All groups were matched except for CPR duration (ACCD > mCC) but this time interval was linked to the existence of pulmonary BME (p = 0.031). Both entities occur in less than 25% of all cases following unsuccessful CPR. BME was only detectable in CPR cases, but was similar between ACCD and mCC cases for BME frequency (p = 0.666), BME intensity (p = 0.857) and the size of BME-affected pulmonary vessels (p = 0.075). If any, only mild pulmonary FE (grade I) was diagnosed without differences in the CPR method (p = 0.624). There was a significant correlation between existence of BME and FE (p = 0.043). Given the frequency, intensity and size of pulmonary BME and FE following CPR, these conditions may unlikely be considered as causative for death in case of initial survival but can be found in lower frequencies in autopsy histology.

Subthalamic nucleus volumes are highly consistent but decrease age-dependently-a combined magnetic resonance imaging and stereology approach in humans.

The subthalamic nucleus (STN) is a main target structure of deep brain stimulation (DBS) in idiopathic Parkinson's disease. Nevertheless, there is an ongoing discussion regarding human STN volumes and neuron count, which could potentially have an impact on STN-DBS. Moreover, a suspected functional subdivision forms the basis of the tripartite hypothesis, which has not yet been morphologically substantiated. In this study, it was aimed to investigate the human STN by means of combined magnetic resonance imaging (MRI) and stereology. STN volumes were obtained from 14 individuals (ranging from 65 to 96 years, 25 hemispheres) in 3 T MRI and in luxol-stained histology slices. Neuron number and cell densities were investigated stereologically over the entire STN and in pre-defined subregions in anti-human neuronal protein HuC/D-stained slices. STN volumes measured with MRI were smaller than in stereology but appeared to be highly consistent, measuring on average 99 ± 6 mm3 (MRI) and 132 ± 20 mm3 (stereology). The neuron count was 431,088 ± 72,172. Both STN volumes and cell count decreased age-dependently. Neuron density was different for the dorsal, medial and ventral subregion with significantly higher values ventrally than dorsally. Small variations in STN volumes in both MRI and stereology contradict previous findings of large variations in STN size. Age-dependent decreases in STN volumes and neuron numbers might influence the efficacy of STN-DBS in a geriatric population. Though the study is limited in sample size, site-dependent differences for the STN subregions form a morphological basis for the tripartite theory. Hum Brain Mapp 38:909-922, 2017. © 2016 Wiley Periodicals, Inc.

Neuroforensomics: metabolites as valuable biomarkers in cerebrospinal fluid of lethal traumatic brain injuries.

Traumatic brain injury (TBI) is a ubiquitous, common sequela of accidents with an annual prevalence of several million cases worldwide. In forensic pathology, structural proteins of the cellular compartments of the CNS in serum and cerebrospinal fluid (CSF) have been predominantly used so far as markers of an acute trauma reaction for the biochemical assessment of neuropathological changes after TBI. The analysis of endogenous metabolites offers an innovative approach that has not yet been considered widely in the assessment of causes and circumstances of death, for example after TBI. The present study, therefore, addresses the question whether the detection of metabolites by liquid-chromatography-mass spectrometry (LC/MS) analysis in post mortem CSF is suitable to identify TBI and to distinguish it from acute cardiovascular control fatalities (CVF). Metabolite analysis of 60 CSF samples collected during autopsies was performed using high resolution (HR)-LC/MS. Subsequent statistical and graphical evaluation as well as the calculation of a TBI/CVF quotient yielded promising results: numerous metabolites were identified that showed significant concentration differences in the post mortem CSF for lethal acute TBI (survival times up to 90 min) compared to CVF. For the first time, this forensic study provides an evaluation of a new generation of biomarkers for diagnosing TBI in the differentiation to other causes of death, here CVF, as surrogate markers for the post mortem assessment of complex neuropathological processes in the CNS ("neuroforensomics").

On a potential morpho-mechanical link between the gluteus maximus muscle and pelvic floor tissues.

Stress urinary incontinence presents a condition not only found in female elderlies, but also in young athletes participating in high-impact sports such as volleyball or trampolining. Repeated jumps appear to be a predisposing factor. Yet the pathophysiology remains incompletely elucidated to date; especially with regard to the influence of the surrounding buttock tissues including gluteus maximus. The present study assessed the morpho-mechanical link between gluteus maximus and the pelvic floor female bodies. 25 pelves obtained from Thiel embalmed females were studied in a supine position. Strands of tissues connecting gluteus maximus with the pelvic floor obtained from 20 sides were assessed mechanically. Plastinates were evaluated to verify the dissection findings. In total, 49 hemipelves were included for data acquisition. The fascia of gluteus maximus yielded connections to the subcutaneous tissues, the fascia of the external anal sphincter and that of obturator internus and to the fascia of the urogenital diaphragm. The connection between gluteus maximus and the urogenital diaphragm withstood an average force of 23.6 ± 17.3 N. Cramér φ analyses demonstrated that the connections of the fasciae connecting gluteus maximus with its surroundings were consistent in the horizontal and sagittal planes, respectively. In conclusion, gluteus maximus is morphologically densely linked to the pelvic floor via strands of connective tissues investing the adjacent muscles. Though gluteus maximus has also been reported to facilitate urinary continence, the here presented morpho-mechanical link suggests that it may also have the potential to contribute to urinary stress incontinence. Future research combining clinical imaging with in-situ testing may help substantiate the potential influence from a clinical perspective.

Sample size considerations in soft tissue biomechanics.

Biomechanical experiments help link tissue morphology with load-deformation characteristics. A tissue-dependent minimum sample number is indispensable to obtain accurate material properties. Stress-strain properties were retrieved from human dura mater and scalp skin, exemplifying two distinct soft tissues. Minimum sample sizes necessary for a stable estimation of material properties were obtained in a simulation study. One-thousand random samples were sequentially drawn for calculating the point at which a majority of the estimators settled within a corridor of stability at given tolerance levels around a 'complete' reference for the mean, median and coefficient of variation. Stable estimations of means and medians can be achieved below sample sizes of 30 at a ± 20%-tolerance within 80%-conformity for scalp skin and dura. Lower tolerance levels or higher conformity dramatically increase the required sample size. Conformity was barely ever reached for the coefficient of variation. The parameter type appears decisive for achieving conformity. STATEMENT OF SIGNIFICANCE: Biomechanical trials utilizing human tissues are needed to obtain material properties for surgical repair, tissue engineering and modeling purposes. Linking tissue mechanics with morphology helps elucidate form-function relationships, the 'morpho-mechanical link'. For material properties to be accurate, it is vital to examine a minimum number of samples. This number may vary between tissues, and the effects of intrinsic tissue characteristics on data accuracy are unclear to date. This study used data obtained from human dura and skin to compute minimum sample sizes required for estimating material properties at a stable level. It was shown that stable estimations are possible at a ± 20%-tolerance within 80%-conformity below sample sizes of 30. Higher accuracy warrants much higher sample sizes for most material properties.