Archival, paleopathological and aDNA-based techniques in leprosy research and the case of Father Petrus Donders at the Leprosarium 'Batavia', Suriname.

OBJECTIVE: We assessed whether Petrus Donders (died 1887), a Dutch priest who for 27 years cared for people with leprosy in the leprosarium Batavia, Suriname, had evidence of Mycobacterium (M.) leprae infection. A positive finding of M. leprae ancient (a)DNA would contribute to the origin of leprosy in Suriname. MATERIALS: Skeletal remains of Father Petrus Donders; two additional skeletons excavated from the Batavia cemetery were used as controls. METHODS: Archival research, paleopathological evaluation and aDNA-based testing of skeletal remains. RESULTS: Neither archives nor inspection of Donders skeletal remains revealed evidence of leprosy, and aDNA-based testing for M. leprae was negative. We detected M. leprae aDNA by RLEP PCR in one control skeleton, which also displayed pathological lesions compatible with leprosy. The M. leprae aDNA was genotyped by Sanger sequencing as SNP type 4; the skeleton displayed mitochondrial haplogroup L3. CONCLUSION: We found no evidence that Donders contracted leprosy despite years of intense leprosy contact, but we successfully isolated an archaeological M. leprae aDNA sample from a control skeleton from South America. SIGNIFICANCE: We successfully genotyped recovered aDNA to a M. leprae strain that likely originated in West Africa. The detected human mitochondrial haplogroup L3 is also associated with this geographical region. This suggests that slave trade contributed to leprosy in Suriname. LIMITATIONS: A limited number of skeletons was examined. SUGGESTIONS FOR FURTHER RESEARCH: Broader review of skeletal collections is advised to expand on diversity of the M. leprae aDNA database.

DNA identification of human remains in Disaster Victim Identification (DVI): An efficient sampling method for muscle, bone, bone marrow and teeth.

In disaster victim identification (DVI), DNA profiling is considered to be one of the most reliable and efficient means to identify bodies or separated body parts. This requires a post mortem DNA sample, and an ante mortem DNA sample of the presumed victim or their biological relative(s). Usually the collection of an adequate ante mortem sample is technically simple, but the acquisition of a good quality post mortem sample under unfavourable DVI circumstances is complicated due to the variable degree of preservation of the human remains and the high risk of DNA (cross) contamination. This paper provides the community with an efficient method to collect post-mortem DNA samples from muscle, bone, bone marrow and teeth, with a minimal risk of contamination. Our method has been applied in a recent, challenging DVI operation (i.e. the identification of the 298 victims of the MH17 airplane crash in 2014). 98,2% of the collected PM samples provided the DVI team with highly informative DNA genotyping results without the risk of contamination and consequent mistyping the victim's DNA. Moreover, the method is easy, cheap and quick. This paper provides the DVI community with a step-wise instructions with recommendations for the type of tissue to be sampled and the site of excision (preferably the upper leg). Although initially designed for DVI purposes, the method is also suited for the identification of individual victims.

Classification criteria for diffuse idiopathic skeletal hyperostosis: a lack of consensus.

Objectives: DISH is a condition characterized by flowing ossifications of the spine with or without ossifications of entheses elsewhere in the body. Studies on the prevalence and pathogenesis of DISH use a variety of partly overlapping combinations of classification criteria, making meaningful comparisons across the literature difficult. The aim of this study was to systematically summarize the available criteria to support the development of a more uniform set of diagnostic/classification criteria. Methods: A search was performed in Pubmed, Embase, Cochrane Library and Web of Science using the term DISH and its synonyms. Articles were included when two independent observers agreed that the articles proposed a new set of classification criteria for DISH. All retrieved articles were evaluated for methodological quality, and the presented criteria were extracted. Results: A total of 24 articles met the inclusion criteria. In all articles, spinal hyperostosis was required for the diagnosis of DISH. Peripheral, extraspinal manifestations were included as a (co-)requirement for the diagnosis DISH in five articles. Most discrepancies revolved around the threshold for the number of vertebral bodies affected and to defining different developmental phases of DISH. More than half of the retrieved articles described a dichotomous set of criteria and did not consider the progressive character of DISH. Conclusion: This systematic review summarizes the available different classification criteria for DISH, which highlights the lack of consensus on the diagnosis of (early) DISH. Consensus criteria, including consecutive phases of new bone formation that characterize DISH, can be developed based upon established diagnostic/classification criteria.

[Identifying victims of a disaster]. / Identificatie van slachtoffers van een ramp.

Identifying the victims of a disaster is important for the next of kin, to issue a death certificate and, if necessary, for forensic investigations. In the Netherlands victims are identified by the Dutch disaster victim identification team, which is part of the national forensic investigation team ('Landelijk Team Forensische Opsporing'). Ante-mortem data are collected during the identification process; these include the victim's specific medical characteristics and the DNA profile of the victim and their family members. The victim's own doctor can play an important role in the ante-mortem investigation because of his or her knowledge of their personal medical details, and of the possible availability of samples for establishing a DNA profile. The ante-mortem data are then compared with post-mortem data. For a definitive identification at least 1 primary identification characteristic has to be established from the physical remains - dermatoglyphics, the DNA profile or the dental status.

Are the alleged remains of Johann Sebastian Bach authentic?

A skeleton alleged to be that of Johann Sebastian Bach (1685-1750) was exhumed from a graveyard in Leipzig, Germany, in 1894, but its authenticity is not established. In 1895, anatomist Wilhelm His concluded from his examination of the skeleton and reconstruction of the face that it most likely belonged to Bach. In 1949, surgeon Wolfgang Rosenthal noticed exostoses on the skeleton and on x-rays of 11 living organists and proposed a condition, Organistenkrankheit, which he interpreted as evidence that the skeleton was Bach's. However, our critical assessment of the remains analysis raises doubts: the localisation of the grave was dubious, and the methods used by His to reconstruct the face are controversial. Also, our study of the pelvic x-rays of 12 living professional organists failed to find evidence for the existence of Organistenkrankheit. We believe it is unlikely that the skeleton is that of Bach; techniques such as DNA analysis might help resolve the question but, to date, church authorities have not approved their use on the skeleton.

Femur, rib, and tooth sample collection for DNA analysis in disaster victim identification (DVI) : a method to minimize contamination risk.

Although much literature is available on DNA extraction from tissue samples to obtain the best possible genotyping results, to the best of our knowledge no written recommendations exist on how to excise or extract bone and tooth samples from a victim to facilitate this. Because the possibility of cross-contamination is high, especially when excising numerous samples under disaster conditions, it is important to minimize this risk and to keep samples in optimum condition. In this paper a standard operating procedure is proposed for collection of femur, rib, and tooth samples to aid victim identification both after mass disasters and in (single) forensic investigations.

Variables affecting the probability of complete fusion of the medial clavicular epiphysis.

In this study, we have combined data on clavicle fusion from different studies and applied a binomial logistic regression analysis. As such, we aimed to assess whether or not variables such as sex, socioeconomic status, and ethnicity influence the probability of having mature, i.e., completely fused clavicles at a given age. We further explored whether the method of clavicle examination, i.e., diagnosis from either a dry bone specimen, an examination of X-rays, or an examination of computed tomography scans, affects the probability of being diagnosed with mature clavicles. It appeared that only ethnicity did not significantly affect this probability. Finally, we illustrated how the logit model may be used to predict the probability of being diagnosed with mature clavicles.

Cross-sectional anthropometric study of the external ear.

This cross-sectional anthropometric study was aimed at estimating expansion of the external ear during adult life, in order to evaluate the extent to which anatomical features appearing in earprints may vary with time. A review of the literature was provided. Data extracted from photographed ears of 1353 subjects were analyzed. The effect of age on auricle length, earlobe length, and auricle width was explored using univariate analyses of variance. The regression coefficients of age on these dimensions were, respectively, 0.178, 0.115, and 0.073 mm/yr for males, and 0.162, 0.100, and 0.073 mm/year for females (p=0.000). Regression coefficients of age corrected for stature were assumed to be less accurate. Anthelix prominence and helix width were analyzed using data of 175 subjects, and appeared unaffected by age. As lobe expansion appeared to exceed the estimated cartilage expansion, it was assumed that particularly the imprint of the lobe would be less stable with time.

Histological age prediction from the femur in a contemporary Dutch sample. The decrease of nonremodeled bone in the anterior cortex.

This paper presents an uncomplicated and minimally invasive method for age-at-death determination in a contemporary Dutch (West European) population, by modifying the approach of assessment based on the age-related remodeling of bone tissue. In contrast to the usual "osteon count," a "non-remodeled tissue count" is undertaken. To optimize the method, proper zeroing of the polarization filter set of the microscope is essential. Instructions for setting the filters are given. A sample of femoral shaft segments totaling 162 individuals with ages ranging from 15 to 96 years is analyzed. Subperiosteal quantitative assessments are recorded at the most anterior point of the femoral shaft and also at points 25 degrees to the left and to the right of that point. Interobserver agreement in the assessments shows an acceptable degree of correlation. Bone remodeling with age does not progress in a linear, but in a curvilinear manner. Dependence of predicted age on nonremodeled surface counts in the analyzed areas of the anterior cortex of the femur appears to be significant. A set of regression equations is given. Sex can be ignored in age prediction. The small but statistically significant dependence of predicted age on cadaver length corresponds with the present strong secular increase in stature in the Netherlands. A concise catalogue with micrograph examples for every 10-year period in life is available upon request.

Improving the visibility of tooth cementum annulations by adjustment of the cutting angle of microscopic sections.

Age at death assessments by counting tooth cementum annulations (TCAs) in unstained undecalcified microscopic ground sections of (single rooted) teeth is, amongst others, problematic because of the unclear distinction between the bright and darker annulations. Counting is hampered by optical superimposition of the tangentially positioned layers of cementum in the section since 'regular transverse sections' run perpendicular to the axis of a cone-shaped root with its yearly deposited cone-shaped layers of cementum. This study demonstrates that to improve the visibility of the annulations, the cutting angle should be perpendicular to the exterior of a root, not perpendicular to its axis. The site where the cut hits the root perpendicular should show the best possible distinction between the TCAs. Here, superimposition of the now vertical positioned layers within the section will result in increased contrast between bright and darker layers. A procedure for such preparation is given.

Individualization of earprints : Variation in prints of monozygotic twins.

This article presents a study of the variation in the appearance of earprints from six pairs of monozygotic (MZ) twins, applying different methods with varying degrees of objectivity. By analyzing prints of MZ twins, we purposely select pairs of individuals for whom the degree of interindividual variation is low. We first provide an account of the corresponding characteristics and the differences that occur between prints from the individual members. We evaluate differences in both content (i.e., presence, shape, and intensity of imprinted features) and geometry (i.e., position of imprinted features). Differences are visualized by way of digital overlays. Differences in geometry are further evaluated by performing a hierarchical cluster analysis using variables derived from the position of seemingly corresponding features. As a next step, we explore a method to fully automatically analyze prints. In this approach, image regions are selected and matched automatically. A suggestion on how to apply this method to calculate the evidential value of an earprint is provided. To our knowledge, this is the first time that any method capable of fully automated earprint comparisons has been presented in the literature.

Earprints in forensic investigations.

This article provides an overview of the theoretical and practical issues surrounding the use of earprints in forensic research. In the first section, we provide a limited account of the history of earprints in forensic investigations and their use as evidence in court. Criticism of the use of earprints for individualization is addressed and fundamental questions that require attention are summarized. In the second section, we summarize the results of various studies that we have performed on earprint variation. Potential causes of intraindividual variation in earprint appearance are, for instance, variations in the way the prints are lifted, variations in the time the donor spends listening, and variations in the force that is applied during listening. Concerning the latter, we have explored various factors that could influence applied force. We have also studied changes in the morphology of live ears during adulthood.

Exploring the effect of the occurrence of sound on force applied by the ear when listening at a surface.

In this article, we explore the effect of the occurrence of a target sound on the force that is applied by the ear when listening at a surface because differences in applied force induce variation in carprints of a single ear. Forty two subjects each listened four times at a surface. During two of these listening efforts there was silence. While the subjects were listening, we measured the amount of force that was applied to the surface. To explore the effect of the occurrence of a target sound upon applied force, we applied a mixed model analysis of variance. The force applied by the ear appeared to be uncorrelated to presence or absence of a target sound. This lack of association appeared not to be the result of potential confounders. Only repetition (i.e., the position of a listening effort in the series of four), appeared to significantly affect applied force; this force being generally lower during a first listening effort.

Exploratory study on classification and individualisation of earprints.

The FearID research project is aimed at the individualisation of earprints for the purpose of forensic research. The study presented here was carried out within the framework of this project. It intends to combine a review of what is known from literature on the classification and individualisation of earprints with results from a preliminary study of earprints. Possibilities for, and limitations to, the use of earprints in forensic investigation are addressed. Differences between eliminating a suspect, placing a suspect at a crime scene, and linking crimes by prints left at different scenes are considered.