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Skeletal remains encountered frequently in forensic applications are a challenging specimen, since their DNA is usually degraded due to harsh conditions, limiting the utilization of skeletal DNA. Forensic scientists have tried various methods to extract DNA from skeletal remains of low quantity and poor quality or improve detecting technology for more information from compromised DNA. Compared with traditional capillary electrophoresis (CE), massively parallel sequencing (MPS) is more sensitive to shorter fragments, able to detect allele sequences for variations from core motif or flanking regions, and able to detect more markers with a higher discrimination power. In this study, short tandem repeats (STR) and single nucleotide polymorphisms (SNP) from 35 human skeletons were genotyped by MPS platform, and CE method was also used to perform STR genotyping. The results indicated that the detection rates reached 100.00% in 16 of 35 samples with MPS method, while the same 100.00% was reached in only 9 samples with CE. The success rates of MPS were also higher than that of CE method in shared 21 loci (excluding Y-indel, DYS391, and SE33), especially in loci detected by MPS method only. Besides, all SNPs (124 and 90 SNPs in males and females) were detected in 18 samples of 35 samples by MPS method. Some intra-allelic sequence variants were observed in eight loci (D21S11, D8S1179, D5S2800, D3S1358, vWA, D2S1338, D1S1656, D12S391) using MPS technology. Interestingly, there is a sample showing genotyping disagreement in FGA locus. The clone sequencing verified that a "T" deletion discovered in flanking sequence of FGA led to wrong genotyping on Ampliseq Converge. Our results indicated that MPS could be adopted in qualified labs as a supplementary when the DNA of skeletal remains are hard to identify. [ABSTRACT FROM AUTHOR]

Before amalgamating published isotope data, comparability should be demonstrated. This paper compares carbon and oxygen isotopic compositions of 30 enamel samples measured by two laboratories. The aims were to see what, if any, isotopic variation was observed, to determine the causes as needed and to correct if possible. Bioapatite was acidified at 90°C in 2006 and at 26°C in 2017, while δ values were corrected via one‐point normalization in 2006 and by two‐point normalization in 2017. One case (of the 30) produced different δ values between the analysis dates, suggesting contamination. Repeated carbon isotope ratio measurements were not meaningfully different. Repeated oxygen isotope ratio measurements were significantly different, even following correction for acid‐carbonate fractionation at different temperatures and the renormalization of 2017 δ values using one point; however, differences were not meaningful for interpretations. Results were used to calculate real interpretative differences (RIDs) for comparing enamel bioapatite as 0.6‰ for δ13C values and as 1.6‰ for δ18O values. [ABSTRACT FROM AUTHOR]

When deaths occur in water, soft tissue decomposes after a temperature‐dependent period, making DNA identification dependent on bone. This study examined the effects of water on bone DNA purity and quantity, and determined the best of three extraction methods for isolating DNA. The organic phenol–chloroform method consistently extracted DNA approximating the accepted 260/280 purity value (~1.8); ChargeSwitch® gDNA Plant Kit and DNeasy Blood and Tissue Kit produced fair and unacceptable values, respectively. The purity value for humerus and rib samples was consistent across accumulated degree days (ADD). Significant differences in quantification among extraction methods and between bone types were identified. Ribs and ChargeSwitch® gDNA Plant Kit samples produced the lowest mean Ct values of the bone types and the extraction methods, respectively. Therefore, this study proposes that magnetic bead technology extraction methods and ribs be considered when processing bones that have been submerged in water for any length of time. [ABSTRACT FROM AUTHOR]

This study introduces a newly developed in‐house SNaPshot single‐base extension (SBE) multiplex assay for forensic single nucleotide polymorphism (SNP) genotyping of fresh and degraded samples. The assay was validated with fresh blood samples from four different populations. In addition, altogether 24 samples from skeletal remains were analyzed with the multiplex. Full SNP profiles could be obtained from 14 specimens, while ten remains showed partial SNP profiles. Minor allele frequencies (MAF) of bone samples and different populations were compared and used for association of skeletal remains with a certain population. The results reveal that the SNPs of the bone samples are genetically close to the Pathan population. The findings show that the new multiplex system can be utilized for SNP genotyping of degraded and forensic relevant skeletal material, enabling to provide additional investigative leads in criminal cases. [ABSTRACT FROM AUTHOR]

Abstract: The postmortem interval (PMI) of skeletal remains is a crucial piece of information that can help establish the time dimension in criminal cases. Unfortunately, the accurate and reliable determination of PMI from bone continues to evade forensic investigators despite concerted efforts over the past decades to develop suitable qualitative and quantitative methods. A relatively new PMI method based on the analysis of citrate content of bone was developed by Schwarcz et al. The main objective of our research was to determine whether this work could be externally validated. Thirty‐one bone samples were obtained from the Forensic Anthropology Center, University of Tennessee, Knoxville, and the Onondaga County Medical Examiner's Office. Results from analyzing samples with PMI greater than 2 years suggest that the hypothetical relationship between the citrate content of bone and PMI is much weaker than reported. It was also observed that the average absolute error between the PMI value estimated using the equation proposed by Schwarcz et al. and the actual (“true”) PMI of the sample was negative indicating an underestimation in PMI. These findings are identical to those reported by Kanz et al. Despite these results this method may still serve as a technique to sort ancient from more recent skeletal cases, after further, similar validation studies have been conducted. [ABSTRACT FROM AUTHOR]

The floods in Bosnia and Herzegovina in May 2014 caused landslides all over the country. In the small village of Šerići, near the town of Zenica, a landslide destroyed the local cemetery, relocated graves, and commingled skeletal remains. As the use of other physical methods of identification (facial recognition, fingerprint analysis, dental analysis, etc.) was not possible, DNA analysis was applied. DNA was isolated from 20 skeletal remains (bone and tooth samples) and six reference samples (blood from living relatives) and amplified using PowerPlex ® Fusion and PowerPlex®Y23 kits. DNA profiles were generated for all reference samples and 17 skeletal remains. A statistical analysis (calculation of paternity, maternity, and sibling indexes and matching probabilities) resulted in 10 positive identifications. In this study, 5 individuals were identified based on one reference sample. This has once again demonstrated the significance of DNA analysis in resolving the most complicated cases, such as the identification of commingled human skeletal remains. [ABSTRACT FROM AUTHOR]

The main idea behind age assessment in adults is related to the analysis of the physiological degeneration of particular skeletal structures with age. The main issues with these procedures are due to the fact that they have not been tested on different modern populations and in different taphonomic contexts and that they tend to underestimate the age of older individuals. The purpose of this study was to test the applicability and the reliability of these methods on a contemporary population of skeletal remains of 145 elderly individuals of known sex and age. The results show that, due to taphonomic influences, some skeletal sites showed a lower survival. Therefore, the methods with the highest percentage of applicability were Lovejoy (89.6%) and Rougé-Maillart (81.3%), followed by Suchey-Brooks (59.3%), and those with the lowest percentage of applicability were Beauthier (26.2%) and Iscan (22.7%). In addition, this research has shown how for older adults the study of both acetabulum and auricular surface may be more reliable for aging. This is also in accordance with the fact that auricular surface and the acetabulum are the areas more frequently surviving taphonomic insult. [ABSTRACT FROM AUTHOR]

Present study represents a novel approach of PCR ready-human DNA extraction method from skeletal remains using copper nanoparticles (CuNPs) for personnel identification. To achieve rapid, cost effective, sensitive and non-hazardous method for DNA extraction we utilized CuNPs synthesized using microwave. The applicability of this approach was first tested in blood samples and afterwards, this system was extended to skeletal remains' samples also. This method yields good quality DNA that are ready for PCR reactions from small quantities of blood and skeletal remains. Consequently, even small quantities of nanoparticles could be potentially utilized for a highly efficient isolation of DNA from skeletal remains as well as from ancient archaeological samples. The present method has the advantages that it is quick with high yield, inexpensive, robust, environment friendly and does not require use of hazardous organic solvents. [ABSTRACT FROM AUTHOR]

We employed our previously developed 27-plex ancestry-informative single nucleotide polymorphism (SNP) panel to infer the ancestral components of bone remains of a possible foreign pilot found in south-western China. For ancestry assignment of this unknown individual, we first obtained the 27-SNP genotype of the individual. Then, based on a reference database of 3081 individuals from 33 populations, we calculated the match probability and likelihood ratio using the self-developed software program Forensic Intelligence. Inferred ancestral components of this individual were calculated by structure at K = 3. A complete profile was obtained for the individual using our multiplexed SNP assay. The European population was within one order of magnitude of the highest likelihood. The major ancestral component of this individual was 97.6% European. [ABSTRACT FROM AUTHOR]

The skeletal remains of eight Australian Aboriginals with healed depressed skull fractures were examined. Male:female ratio 5:3; age range 20‐60 yrs. Burial dates by 14C dating in three cases were 500 years BP (n = 2) and 1300 BP. There were 13 healed depressed skull fractures manifested by shallow indentations of cortical bone and thinning of diploe, with no significant disturbance of the inner skull tables. Nine (69%) were located within 35 mm of the sagittal suture/midline. These lesions represent another acquired feature that might be helpful in suggesting that a skull is from a tribal Aboriginal individual and may be particularly useful if the remains are represented by only fragments of calvarium. While obviously not a finding specific to this population, these healed injuries would be consistent with the possible results of certain types of conflict behavior reported in traditional Aboriginal groups that involved formalized inflicted blunt head trauma. [ABSTRACT FROM AUTHOR]

Pelvic ring fractures are associated with high rates of mortality and thus can provide key information about circumstances surrounding death. These injuries can be particularly informative in skeletonized remains, yet difficult to diagnose and interpret. This study adapted a clinical system of classifying pelvic ring fractures according to their resultant degree of pelvic stability for application to gross human skeletal remains. The modified Tile criteria were applied to the skeletal remains of 22 individuals from the Cleveland Museum of Natural History and Universidad Nacional Autónoma de México that displayed evidence of pelvic injury. Because these categories are tied directly to clinical assessments concerning the severity and treatment of injuries, this approach can aid in the identification of manner and cause of death, as well as interpretations of possible mechanisms of injury, such as those typical in car-to-pedestrian and motor vehicle accidents. [ABSTRACT FROM AUTHOR]

A recently introduced method to determine the postmortem interval ( PMI) based on quantification of the citrate content in bone was applied on the temporal bones and femora of 20 individuals buried in wooden coffins ( WO) and body bags ( BB), respectively. Concerning known vs. calculated PMI, a significant difference between the temporal and the femur bone samples of the same individuals was observed in the BB group ( p = 0.012). In contrast, differences were insignificant for the WO group ( p = 0.400). Moreover, similar levels of underestimation of PMIs resulted from the analysis of the femora for both burial groups ( p = 0.247). Also, there was consistently less citrate preserved in the flat temporal bones as compared to the femora, indicating that the cortical layer of the long bones should be preferentially employed for citrate-based PMI estimations. The results call for additional research on subsurface-buried and surface-deposited remains to enhance the accuracy of the published PMI equation. [ABSTRACT FROM AUTHOR]

Determination of sex constitutes the most important element during the identification process of human skeletal remains. Several sex-specific features of human skeleton have been exploited for sex determination with varying reliability. This study aims to obtain sexual dimorphic standards for ulnae of the north Indian population. Eight measurements were obtained on a sample of 106 ulnae (males-80, females-26) in the age range of 25-65 years. The sexual dimorphism index and demarking points were calculated for all the variables. The data were then subjected to stepwise and direct discriminant function analysis. The best discriminator of sex was the maximum length (84.9%) followed by radial notch width (84%). In stepwise analysis, these two variables were selected and provided an accuracy of 88.7% (M-87.5%, F-92.3%). The proximal end provided a classification rate of 81.1% (M-80%, F-84.6%) with selection of the notch length and olecranon width. [ABSTRACT FROM AUTHOR]

The estimation of the postmortem interval (PMI) is still one of the most challenging issues in forensic investigations, especially in cases in which advanced transformative phenomena have taken place. The dating of skeletal remains is even more difficult and sometimes only a rough determination of the PMI is possible. Recent studies suggest that plant analysis can provide a reliable estimation for skeletal remains dating, when traditional techniques are not applicable. Forensic Botany is a relatively recent discipline that includes many subdisciplines such as Palynology, Anatomy, Dendrochronology, Limnology, Systematic, Ecology, and Molecular Biology. In a recent study, Cardoso et al. ( Int J Legal Med 2010;124:451) used botanical evidence for the first time to establish the PMI of human skeletal remains found in a forested area of northern Portugal from the growth rate of mosses and shrub roots. The present paper deals with a case in which the study of the growth rate of the bryophyte Leptodyctium riparium (H edw.) Warnst, was used in estimating the PMI of some human skeletal remains that were found in a wooded area near Perugia, in Central Italy. [ABSTRACT FROM AUTHOR]

The Armed Forces DNA Identification Laboratory reports the mitochondrial DNA (mt DNA) sequences of over 800 skeletal samples a year for the Joint POW/ MIA Accounting Command-Central Identification Laboratory. These sequences are generated from degraded skeletal remains that are presumed to belong to U.S. service members missing from past military conflicts. In the laboratory, it is possible to control for contamination of remains; however, in the field, it can be difficult to prevent modern DNA from being transferred to skeletal elements and being carried forward through the analysis process. Four such cases are described here along with the controls in place in the laboratory to eliminate the possibility of the exogenous DNA being reported as authentic. In each case, the controls implemented by the laboratories prevented the false reporting of contaminant exogenous DNA from remains that were either faunal or human, but lacked endogenous DNA. [ABSTRACT FROM AUTHOR]

Mandible is the hardest and most durable bone of the skull exhibiting a high degree of sexual dimorphism. Especially ramus of mandible is subjected to greater stress than any other bone of the skull because of the process of mastication. This study has been performed to establish the osteometric standards for practical use in forensic context over Indian population using mandibular ramus. The sample consists of 116 mandibles of Northern Indian population (M:F; 92:24, mean age 37.4 years), collected from the Department of Forensic Medicine, IMS, BHU, Varanasi. Osteometric informations about five metric parameters (coronoid height, projective height, condylar height, and maximum breadth and minimum breadth of ramus) were taken with sliding calipers. These parameters were subjected to different discriminant function analysis using SPSS 16.0. All parameters showed significant sexual dimorphism ( p < 0.001 in all cases) with an overall accuracy of 80.2%, and coronoid height was the single best parameter providing an accuracy of 74.1%. [ABSTRACT FROM AUTHOR]

The identification of missing casualties of the Korean War (1950-1953) has been performed using mitochondrial DNA (mtDNA) profiles, but recent advances in DNA extraction techniques and approaches using smaller amplicons have significantly increased the possibility of obtaining DNA profiles from highly degraded skeletal remains. Therefore, 21 skeletal remains of Korean War victims and 24 samples from biological relatives of the supposed victims were selected based on circumstantial evidence and/or mtDNA-matching results and were analyzed to confirm the alleged relationship. Cumulative likelihood ratios were obtained from autosomal short tandem repeat, Y-chromosomal STR, and mtDNA-genotyping results, and mainly confirmed the alleged relationship with values over 10. The present analysis emphasizes the value of mini- and Y-STR systems as well as an efficient DNA extraction method in DNA testing for the identification of old skeletal remains. [ABSTRACT FROM AUTHOR]

Bones fluoresce when exposed to certain wavelengths of shortwave light, and this property can be useful in locating and sorting skeletal remains in forensic contexts. The proteins in bone collagen are largely responsible for its fluorescent properties, but these proteins degrade and denature over time. This study examined the fluorescence of bones from four temporal groups (recent, semi-recent, ancient, and historic) ranging from 0 to 1064 years before present. Specimens were photographed under 490 nm wavelength light, and fluorescence was quantified by converting intensity to a gray scale value based on the RGB color model using ImageJ® software. Significant ( p < 0.05) differences were found in mean fluorescence between all four temporal groups, and a 0.324 coefficient of correlation indicates a significant (inverse) relationship between fluorescence and time. Bone fluorescence decreases with time, but some fluorescence is retained even in older samples. Fluorescence can therefore be reliably used in many modern skeletal remains searches. [ABSTRACT FROM AUTHOR]

Burned skeletal material is often very fragile and at high risk for fragmentation during packaging and transportation. One method that has been suggested to protect bones in these cases is to carefully wrap them in aluminum foil. Traces of aluminum, however, are known to transfer from foil packaging materials to food products. If such transfer occurs between aluminum foil and bones, it could interfere with subsequent chemical, elemental and isotopic analyses, which are becoming more common in forensic anthropological investigations. This study examined aluminum levels in bones prior to and following the use of aluminum foil packaging and storage for a 6-week period. Results indicate no significant change in the detected levels of aluminum ( p > 0.05), even when packaged in compromised foil and exposed to elevated temperatures. Aluminum foil can therefore continue to be recommended as a packaging medium without affecting subsequent chemical examinations. [ABSTRACT FROM AUTHOR]

The Fosse Ardeatine massacre was a mass execution carried out in Rome on March 24, 1944 by Nazi German occupation troops during the Second World War as a reprisal for a partisan attack conducted on the previous day in central Rome. The 335 civilians were taken to the “Cave Ardeatine” and they were shot. Only 323 corpses out of 335 have been identified. The aim of this work is the genetic and anthropological analysis of the remains exhumed from grave number 329 of Fosse Ardeatine’s Shrine to assess their identity. So far, such remains have been supposed to belong to MM but mitochondrial analysis excluded a biological relationship to two living maternal relatives. Our analysis indicated that remains recovered in grave number 329 do not belong to MM. This result suggests that genetic analysis of the remains should be also applied to the other 12 unknown corpses to elucidate their identity. [ABSTRACT FROM AUTHOR]

Between February 1999 and May 2005, 116 DNA extractions were completed on skeletal remains from routine casework. Overall, at least a partial mitochondrial DNA (mtDNA) profile was obtained on 83.6% of samples. Skeletal remains fell into two general categories: (1) samples for body identifications submitted by law enforcement and (2) samples submitted to answer historical or family identity questions. Body identification cases were more likely to yield full mtDNA profiles, whereas historical cases were more likely to result in partial profiles. Overall, the ability to obtain a full or partial profile primarily reflects the difference in the average age and condition of the samples in these two categories and thus, difference in the quantity and quality of the DNA. Cremated remains were uniformly unsuccessful, whereas infant/fetal remains were uniformly successful. Heteroplasmy in skeletal remains was observed at a rate similar to that in hair (∼10%). For body identification cases, skeletal remains had the same mtDNA profile as the accompanying reference sample in 50% of cases. [ABSTRACT FROM AUTHOR]

When conventional methods of identification, such as visual recognition and dental comparison, cannot be used to identify a deceased person, it becomes necessary to consider alternative methods. The presence of an orthopedic implant in a body may assist identification if ante-mortem medical records are available for comparison. Another method of identification involves comparison of ante-mortem and postmortem radiographs. Eight cases are reported from Forensic Science SA where the presence of orthopedic implants and/or ante-mortem radiographs were used to try to establish identification. In six cases, positive identification was established, and in two cases with upper limb orthopedic implants, the bones remained unidentified. Manufacturers were unable to provide any information about the distribution and use of the implants that could be of use with identification, as there are no requirements in Australia for individual medical implants to be tracked. Such a system has the potential to aid postmortem identification if serial codes were etched onto implants that could then be traced to manufacturers, surgeons, and recipients of these devices. [ABSTRACT FROM AUTHOR]

Abstract: We present individual body identification efforts, to identify skeletal remains and relatives of missing persons of an explosion took place inside one of the munitions recesses of the Armoured Brigade of the Galapagos Armoured Cavalry, in the city of Riobamba, Ecuador, on Wednesday, November 20, 2002. Nineteen samples of bone remains and two tissue samples (a blood stain on a piece of fabric) from the zero zone were analysed. DNA extraction was made by Isoamilic Phenol–Chloroform–Alcohol, and proteinase K. We increased PCR cycles to identify DNA from bones to 35 cycles in some cases. An ABI 310 sequencer was used. Determination of the fragment size and the allelic designation of the different loci was carried out by comparison with the allelic ladders of the PowerPlex©16 kit and Gene Scan Analysis Software® programme. Five possible family groups were established and were compared with the profiles found. Classical Bayesian methods were used to calculate the Likelihood Ratio and it was possible to identify five different genetic profiles in our country. This paper is important because is a novel experience for our forensic services, because this was the first time DNA had been used as an identification method in disasters, and it was validated by Ecuadorian justice like a very effective method. [Copyright &y& Elsevier]

Abstract: Twenty-six bone DNA identification cases are described. The postmortem periods of the studied remains ranged from three days to over 30 years, and the locations where the remains were found varied resulting in a variety of postmortem conditions. Nuclear DNA typing using an AmpFLSTR Profiler kit and mitochondrial DNA (mtDNA) typing of hypervariable regions 1 and 2 (HV1 and HV2) in a control region were performed both with decalcified and non-treated bone powder samples. Decalcification was shown to improve the success of DNA typing. The nucleotide sequences of the HV1 and HV2 regions were successfully determined in all cases examined. Nuclear DNA typing was very successful, more than half of the loci were typed during multiple amplifications (10 loci in one reaction) in 23 cases. Polymerase chain reaction (PCR) inhibition was observed in five cases including three samples that were found buried in soil. This inhibitory effect was identified as the result of unbalanced multiple PCR during the profiler test. These results revealed that DNA typing targeting nuclear DNA is a potentially powerful tool for bone identification. [Copyright &y& Elsevier]

Abstract: Estimation of stature is one of the important initial steps during forensic analysis of human skeletal remains. The aim of the present study was to derive a linear regression formula for estimating stature of adult South Indian males from the length of the sternum. The study included 35 male sternums of South Indian origin dissected from cadavers during medico-legal autopsies. The linear regression equation [Stature=117.784+(3.429×Sternal length)] was derived to estimate the stature from the length of the sternum. The correlation coefficient was 0.638. The standard error of the estimate was 5.64cm. This preliminary study concludes that the length of the sternum can be used as a tool for stature estimation in adult South Indian males. [Copyright &y& Elsevier]