•DNA quantity and STR typing success of 48 bone types in three WWII skeletons differed by anatomical region but not by skeleton.•Metatarsals, metacarpals, and petrous bones outperformed all other ...bone types in DNA quantity and quality.•The ten highest-yielding bone types differed from skeleton to skeleton.•The third metacarpal yielding the highest DNA concentration in all three skeletons.•The data obtained have implications for skeletal sampling from old and highly degraded skeletal remains.
DNA yield varies by anatomical region, and the selection of bone types that yield maximum recovery of DNA is important to maximize the success of human identification of skeletal remains. The goal of our study was to explore inter- and intra-individual variation in DNA content by measuring nuclear DNA quantity and quality and autosomal STR typing success to determine the most promising skeletal elements for bone sampling. To exclude the influence of taphonomic issues as much as possible, three complete male skeletons from a single Second World War mass grave were examined and all representative skeletal element types of the human body were analyzed. Forty-eight different types of bones from the head, torso, arm, leg, hand, and foot were sampled from each skeleton, 144 bones altogether. The samples were cleaned, and half a gram of bone powder was decalcified using a full demineralization extraction method. The DNA was purified in a Biorobot EZ1 (Qiagen). DNA content and rates of DNA degradation were determined with the PowerQuant (Promega), and the Investigator ESSplex SE QS (Qiagen) was used for STR typing. The highest-yielding bones mostly produced the most complete STR profiles. Among the skeletal elements containing on average the most DNA and producing the most complete profiles in all three skeletons examined were metacarpals, metatarsals, and the petrous portion of the temporal bone. Metatarsals and metacarpals can easily be sampled without using a saw, thus reducing potential DNA contamination. Skeletons from the Second World War can be used as a model for poorly preserved skeletal remains, and the results of the investigation can be applied for genetic identification of highly degraded skeletal remains in routine forensic casework. Although the research was limited to only three skeletons found in a unique mass grave, the data obtained could contribute to sampling strategies for identifying old skeletal remains. More Second World War skeletons will be analyzed in the future to investigate inter-bone variation in the preservation of DNA.
Objectives
The aims of the study are to investigate the effects of latitude, settlement type, age, and sex on the prevalence of vitamin D deficiency disease in the Roman Empire using human skeletal ...remains from cemetery sites (1st to 6th cent. AD).
Materials and Methods
Data from 2,787 individuals (1,143 subadults, 1,644 adults) from 18 cemeteries associated with 15 different settlements in the Mediterranean and north‐western Europe were analyzed. Vitamin D deficiency disease (rickets, osteomalacia) was identified using standard paleopathological criteria. Multivariate statistical analysis was used analyze the effects of the variables of interest on the prevalence of vitamin D deficiency.
Results
The overall prevalence of rickets in subadults (<20 years) was 5.7%, and 3.2% of adults showed osteomalacia and/or residual rickets. There was a positive association between rickets in subadults and latitude, with numerous cases of rickets among infants. There was no general association with sex or settlement type, although an elevated prevalence of rickets was observed at a cemetery associated with a settlement (Ostia, Italy), which had multi‐storey buildings.
Discussion
The association of rickets with latitude may reflect care practices that, in more northerly locations where solar radiation is less intense, placed infants at increased risk of insufficient sunlight exposure to permit adequate vitamin D biosynthesis. The elevated level of vitamin D deficiency at Ostia may reflect, at least in part, the lack of sunlight due to dense occupation of multi‐storey blocks that prevented direct sunlight from reaching living quarters and the streets between these closely spaced buildings.
We examined fragmentary human skeletal remains from Escalon Cave near Surigao City, northeastern Mindanao, the Philippines, with respect to the morphology of bones and teeth, radiocarbon dating, and ...mitochondrial DNA (mtDNA) haplogroup. These remains contained parts of the left temporal bone, the right humerus, the right femur, the upper right first and second premolars, and the first molar. They are presumed to belong to an early-middle adult male, whom we named the Escalon Man. Using the femur sample, we estimated the 14C date of the individual at 2692 ± 39 years BP (uncalibrated). His stature, estimated from the maximum femur length, was about 170 cm—this makes him much taller than the Aeta and the Mamanwa, who are indigenous negrito hunter-gatherers of the Philippines. The femur is sturdy and shows well-developed muscle markings. Numerous narrow grooves on the humerus were found in close proximity to each other, but we could not prove whether these are artificial cut marks made by humans with sharp-edged tools or scratches from some non-human entity. The mesiodistal crown diameters of the molar and premolars are large and resemble those of native Oceanians. Analysis of mtDNA haplogroup was carried out using a DNA sample extracted from a molar. A next-generation sequencer was used to determine the nucleotide sequences of the mtDNA genome. The results indicated that the Escalon Man belongs to the haplogroup E1a1a, which is known to be the marker of Austronesian-speaking agriculturist populations that originated in Taiwan and spread southwards through the Philippines to the Western Pacific, since about 4000 BP. Thus, the Escalon Man was likely a member of the late Neolithic or early Metal Age agriculturist peoples who settled in northeastern Mindanao and who may be the ancestors of the territory’s present-day occupants, such as the Manobo.
A new excavation of the Iyai rock-shelter site has uncovered more than a dozen human skeletal remains from the Initial Jomon period. We describe here an almost complete female skeleton (Iyai 1), and ...examine this in the context of morphological variation in Jomon females, especially those of the Initial Jomon period. Two radiocarbon dates based on the Iyai 1 skeletal samples show a calibrated date c. 8300–8200 calBP, belonging to the later part of the Initial Jomon period. The Iyai 1 skeleton was found in a burial pit with an unusual body placement. Although it was a flexed burial with the both arms and limbs flexed, the upper and lower body was disconnected at the waist and the two portions placed on top of each other: the upper body was laid on its abdomen with the head faced into the pelvis, and the lower body was found under the upper trunk in a supine position. The individual is a young female, 146 cm in height using Sasou–Fujii method, and the age at death is estimated as c. 20–40 years. The neurocranium is long (cranial index = 79.9) and high (length–height index = 77.1), the face is wide and low (upper-facial index = 43.4). The dentition shows nearly horizontal heavy wear but no dental caries with an edge-to-edge bite occlusion. The postcranial long bones are gracile, although they exhibit clear musculature impressions in some attachment areas. No paleopathological bone modification was found. The talus exhibits a ‘squatting’ facet at the anterior contact of the talocrural joint. The stable isotopic signature of Iyai 1 falls among those of the inland Initial Jomon people, indicating that her proteins mainly came from C3 plants and terrestrial animals.
Las enfermedades dentales son frecuentemente reportadas en los restos humanos antiguos y su estudio brinda información acerca de la paleodieta. En esta investigación se evaluó la relación entre las ...enfermedades dentales y el perfil dietético de un grupo de individuos de estatus social bajo que habitó el sitio arqueológico Agua Caliente, Costa Rica, entre los años 900-1550 d.C. La muestra de estudio estuvo integrada por 24 individuos: 5 juveniles (sexo indeterminado) y 19 adultos (4 femeninas, 5 masculinos y 10 de sexo indeterminado). La dentición se analizó de manera macroscópica para identificar cinco enfermedades: caries, cálculos dentales, enfermedad periodontal, lesiones periapicales y pérdida ante mortem de piezas dentales. Las enfermedades más frecuentes fueron las caries y los cálculos dentales. Probablemente, estas dos afecciones estuvieron involucradas en la formación de las otras enfermedades identificadas. Los resultados obtenidos indicaron que los carbohidratos y las proteínas formaron parte de la paleodieta de estos individuos, así como otros alimentos. Estas fuentes dietéticas fueron consistentes con el registro arqueológico, ya que se evidenciaron en los restos paleobotánicos y de fauna, las estrategias de subsistencia y la tecnología empleada para la obtención y procesamiento de estos alimentos. Los hallazgos de este estudio ofrecen un primer acercamiento de la asociación entre las enfermedades dentales y la paleodieta de los habitantes de Agua Caliente, pero es necesario desarrollar más investigaciones, aplicar métodos complementarios (análisis de isótopos y cálculos dentales) y analizar colecciones osteológicas humanas de mayor tamaño.
Retrieving DNA from highly degraded human skeletal remains is still a challenge due to low concentration and fragmentation, which makes it difficult to extract and purify. Recent works showed that ...silica‐based methods allow better DNA recovery and this fact may be attributed to the type of bones and the quality of the preserved tissue. However, more systematic studies are needed to evaluate the efficiency of the different silica‐based extraction methods considering the type of bones. The main goal of the present study is to establish the best extraction method and the type of bone that can maximize the recovery of PCR‐amplifiable DNA and the subsequent retrieval of mitochondrial and nuclear genetic information. Five individuals were selected from an archaeological site located in Catalonia–Spain dating from 5th to 11th centuries AD. For each individual, five samples from different skeletal regions were collected: petrous bone, pulp cavity and cementum of tooth, and rib and limb bones. Four extraction methods were tested, three silica‐based (silica in‐suspension, HE column and XS plasma column) and the classical method based on phenol–chloroform. Silica in‐suspension method from petrous bone and pulp cavity showed the best results. However, the remains preservation will ultimately be the key to the molecular result success.
Although the primary objective of forensic DNA analyses of unidentified human remains is positive identification, cases involving historical or archaeological skeletal remains often lack reference ...samples for comparison. Massively parallel sequencing (MPS) offers an opportunity to provide biometric data in such cases, and these cases provide valuable data on the feasibility of applying MPS for characterization of modern forensic casework samples. In this study, MPS was used to characterize 140-year-old human skeletal remains discovered at a historical site in Deadwood, South Dakota, United States. The remains were in an unmarked grave and there were no records or other metadata available regarding the identity of the individual. Due to the high throughput of MPS, a variety of biometric markers could be typed using a single sample.
Using MPS and suitable forensic genetic markers, more relevant information could be obtained from a limited quantity and quality sample. Results were obtained for 25/26 Y-STRs, 34/34 Y SNPs, 166/166 ancestry-informative SNPs, 24/24 phenotype-informative SNPs, 102/102 human identity SNPs, 27/29 autosomal STRs (plus amelogenin), and 4/8 X-STRs (as well as ten regions of mtDNA). The Y-chromosome (Y-STR, Y-SNP) and mtDNA profiles of the unidentified skeletal remains are consistent with the R1b and H1 haplogroups, respectively. Both of these haplogroups are the most common haplogroups in Western Europe. Ancestry-informative SNP analysis also supported European ancestry. The genetic results are consistent with anthropological findings that the remains belong to a male of European ancestry (Caucasian). Phenotype-informative SNP data provided strong support that the individual had light red hair and brown eyes.
This study is among the first to genetically characterize historical human remains with forensic genetic marker kits specifically designed for MPS. The outcome demonstrates that substantially more genetic information can be obtained from the same initial quantities of DNA as that of current CE-based analyses.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
•An integrated multidisciplinary approach to resolve long-standing unidentified cases.•Challenges and considerations to manage unidentified human remains.•Humanitarian forensic capacity in ...Cyprus.•Lessons learned in complex cases of skeletal analysis and identification.
The Committee on Missing Persons in Cyprus (CMP) is a bicommunal committee established in 1981, tasked to determine the fate of 2002 individuals who went missing during the intercommunal fighting of 1963−64 and the events of 1974. The CMP operates strictly within a humanitarian framework, using a multidisciplinary approach to conclude individual identifications of remains exhumed throughout the island, where all information obtained from different phases of the CMP Project is integrated and assessed in a comprehensive manner. By 2017, although over 1000 sets of remains were recovered and either identified or resolved by the CMP, 137 challenging cases remained unidentified at the CMP Anthropological Laboratory. To resolve these cases, different strategies were adopted where the investigatory component was enhanced through the implementation of new data mining approaches, and the genetic-related data were revised and updated through the adoption of new DNA technologies and the improvement of the Family Reference Samples Database. These new approaches resulted in a dramatic reduction of the number of unidentified cases (by over 70 %) as well as the timeframe required for future identifications. These approaches could serve as an example in other humanitarian contexts facing similar challenges as they can have a profound impact on the families of missing persons.
•STR loci were amplified from single-source DNA obtained from bone samples.•Peaks caused by co-amplified microbial DNA may have an impact on STR profile interpretation.•This study informs on observed ...positions of microbial peaks in human STR profiles obtained with different multiplex STR kits.•We present an approach for distinguishing microbial peaks from human alleles.
STR-based DNA analysis is still the main tool for human DNA identification in most forensic DNA laboratories. DNA typing of aged human skeletal elements faces well-known interpretation challenges characteristic of degraded and low copy number DNA samples. Analyzing tens of thousands of human bone and teeth samples, we found that the occasional presence of artefactual peaks of presumed microbial origin adds another layer of complexity to the interpretation of STR profiles. In this paper, we present our approach and suggest guidelines for identifying and distinguishing non-human peaks, developed over the last 18 years. Additionally, we report a compendium of artefact peaks of presumed microbial origin recorded in human STR profiles obtained from bone and teeth samples, originating from Iraq, Chile, Maldives, Brazil and Western Balkans. Our experience has shown that these artefacts are not uncommon in bone STR testing, suggesting the possibility of occurrence in other forensic contexts, particularly trace DNA samples. Raising awareness among the forensic DNA community and accounting for this phenomenon is important for accurate STR interpretation.
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