Application of mtDNA sequence analysis in forensic casework for the identification of human remains

In four forensic cases of unidentified skeletal remains investigated in the last year, we were able to attach three to missing persons. In one case we could show that the discovered bone sample did not fit to a missing child. The method for mitochondrial DNA analysis for the routine identification of skeletal remains was established in our institute by typing bone samples of defined age obtained from Frankfurt's cemetery. Reproducible results were obtained for bones up to 75 years old. For analysis the bone samples were pulverised to fine powder, decalcified and DNA was extracted. From the DNA we amplified a 404-bp fragment from HV-1 and a 379-bp fragment from HV-2 of the mtDNA control region. After sequencing of the PCR products, the results were compared to the Anderson reference sequence and to putative maternal relatives.     

Anthropological and radiographic comparison of vertebrae for identification of decomposed human remains

This case study demonstrates the importance of involving an anthropologist in forensic situations with decomposed remains. Anthropological consultation was used in conjunction with the comparison of antemortem and postmortem radiographs to establish positive identification of unknown, decomposed remains. The remains had no traditional identifying features such as fingerprints or dental. Through anthropological analysis, it was determined the decedent was male, between 20 and 23 years at time of death and c. 5'2' tall. This information allowed for a presumptive identification and a request for antemortem radiographs. The missing person was identified comparing the spinous processes of the cervical and thoracic vertebrae between ante- and postmortem radiographs.     

论失踪损害赔偿之诉的法律适用——以马航客机失踪事件切入

自然人的失踪通常都会给其近亲属造成严重的精神损害和一定的财产损害,依据我国法律现有规定及其解释,在失踪是因可归责于他人的事由所致时,失踪者近亲属应独立享有对相关责任人的损害赔偿请求权。肯定此类损害赔偿请求权,不仅符合分配正义原则,而且能更好地实现权益保护与行为自由的平衡。在单纯的失踪损害赔偿之诉中,失踪者近亲属可以请求赔偿精神损害、寻亲费用和扶养费损失等,在失踪者已被宣告死亡时,还可请求死亡赔偿金。     

Victim of a dictatorial regime: identification of Mr. Roberto Gomensoro Josman

Forensic cases are ideal to test osteological techniques developed by physical anthropologists. Forensic anthropology is a scientific discipline that applies population-based standards to individual skeletal remains. Many complex techniques are used in an attempt to make a positive identification. Several of these techniques, specifically digital video superimposition and DNA, were used to identify the victim in this case. The purpose of this paper is to describe anthropological techniques used to identify the remains of an unknown person who was later identified as Mr. Roberto Gomensoro Josman, the victim of a Uruguayan dictatorial regime. Mr. Gomensoro Josman disappeared after authorities of the Uruguayan dictatorial government (1973-1984) arrested him. Six days later an unknown body was found floating in Lake Rincon del Bonete. The corpse was found tied with wire and weighted with three large stones used to keep the body submerged. An autopsy was performed and the body was buried as an unknown person in the grave identified as number 10936 of Tacuarembo Cemetery. On December 2002 the Peace and Justice Service asked the local judge to authorize the exhumation of the remains. The exhumed body was headless. An investigation revealed that the local medical examiner who had autopsied the remains on March 1973 had retained the victim's skull in his office. Osteological analysis indicated the victim was a white male in his 20s. Four good quality photographs of Mr. Gomensoro who was known to be missing were compared with the skull. To confirm the identification from the video a DNA analysis was carried out comparing the victim with relatives. DNA typing confirmed the results of the earlier identification.     

Investigation of Different Ways in Which the CODIS 7.0 May be Used in Mass Disaster Identification

DNA analysis is a key method for the identification of human remains in mass disasters. Reference samples from relatives may be used to identify missing persons by kinship analysis. Different methods of applying the CODIS in disaster victim identification (DVI) were investigated. Two searches were evaluated: (i) relating family relatives to a pedigree tree (FPT) and (ii) relating unidentified human remains to a pedigree tree (UPT). A joint pedigree likelihood ratio (JPLR) and rank were calculated for each search. Both searches were similar in average JPLR and rank. In exceptional cases, namely the existence of a mutation different from the CODIS model, a nonbiological father, a mistake in STR, or incorrect profile association, the UPT search returned one true rank, whereas the FPT search returned no results. This paper suggests a novel strategy to overcome these limitations and increase efficiency in conducting identification of mass disaster victims.     

Genetic Identification of Communist Crimes’ Victims (1944–1956) Based on the Analysis of One of Many Mass Graves Discovered on the Powazki Military Cemetery in Warsaw,Poland

As the result of the communist terror in Poland, during years 1944–1956 more than 50,000 people died. Their bodies were buried secretly, and most places are still unknown. The research presents the results of identification of people buried in one of many mass graves, which were found at the cemetery Pow?zki Military in Warsaw, Poland. Exhumation revealed the remains of eight people, among which seven were identified genetically. Well‐preserved molars were used for the study. Reference material was collected from the closest living relatives. In one case, an exhumation of victim's parents had to be performed. DNA from swabs was extracted with a PrepFiler^® BTA Forensic DNA Extraction Kit and organic method. Autosomal, Y‐STR amplification, and mtDNA sequencing were performed. The biostatistical calculations resulted in LR values from 1608 to 928 × 10¹⁸. So far, remains of more than 50 victims were identified.     

Choosing relatives for DNA identification of missing persons

DNA-based analysis is integral to missing person identification cases. When direct references are not available, indirect relative references can be used to identify missing persons by kinship analysis. Generally, more reference relatives render greater accuracy of identification. However, it is costly to type multiple references. Thus, at times, decisions may need to be made on which relatives to type. In this study, pedigrees for 37 common reference scenarios with 13 CODIS STRs were simulated to rank the information content of different combinations of relatives. The results confirm that first-order relatives (parents and fullsibs) are the most preferred relatives to identify missing persons; fullsibs are also informative. Less genetic dependence between references provides a higher on average likelihood ratio. Distant relatives may not be helpful solely by autosomal markers. But lineage-based Y chromosome and mitochondrial DNA markers can increase the likelihood ratio or serve as filters to exclude putative relationships.     

A multifaceted,multijurisdictional, multiagency,and multidisciplinary approach to investigating unidentified and missing persons cases in Australia

Currently, there are approximately 750 unidentified human remains and 2500 long-term missing persons in Australia. The Australian Federal Police National DNA Program for Unidentified and Missing Persons (Program) is using a multifaceted, multijurisdictional, multiagency, and multidisciplinary approach in a dedicated effort to identify these unknown deceased persons, scientifically link them to known missing persons, and provide answers to their families. The nationally coordinated Program provides its police, forensic, and coronial stakeholders with a suite of contemporary forensic technologies, databases, and experts to forensically examine the skeletonised remains and recover post-mortem data for comparison to the available ante-mortem data for each missing person. Through a number of physical and virtual public outreach activities, families with missing relatives have been encouraged to provide vital ante-mortem forensic information, records, and samples to aid the identification process. To date, this unique Program has assisted to resolve a number of unidentified and missing persons cases from both historical and contemporary contexts, using a combination of genetic and non-genetic techniques, and local and national databases. The centralisation of Program capabilities, expertise, and resources to conduct this type of unique and challenging casework is proving to be the most effective and efficient way to generate investigative leads, identify human remains, and resolve long-term missing persons cases in Australia.     

DNA identification of skeletal remains by investigator’s intuition

In June 2006 a decapitated woman was found in a parking area of the motorway in the area of Prato (Florence). Since the body was beheaded and no victim’s documents or objects were present at the crime scene, identification at that time was impossible. However, DNA profile from woman’s bones were collected. In the same year (2003), a mother had reported her daughter's disappearance but the two events were not related at that time. About ten years later the mother’s DNA profile was finally acquired for a genetic identification of another girl’s body found in the Ferrara area. These genetic profiles were completely discordant. All these genetic comparisons were carried out on behalf of the prosecutors of the cities involved in the findings of the bodies and in the disappearance complaints, but due to the lack of a database the events remained disconnected. In January 2017, the head of the scientific police of Prato who had followed the investigation and questioned the mother of the missing girl found about ten years later, suggested to the magistrate to order the comparison of the mother's DNA with the genetic profile of the bones found in 2006. This comparison finally allowed the identification of the missing daughter.This story highlights the importance of having forensic DNA database to search for missing persons and how the investigator's intuition can play a key role in resolving criminal cases. In fact, databases of unknown bodies and relatives of missing persons were created in Italy as a part of national DNA database just at the beginning of 2018.     

DNA profiles from biological trace material contained in underwear and toothbrushes

There are special cases where it is extremely difficult to find a close relative for obtaining reference samples, such as in adoption cases or foreigners without relatives in the country. In these cases, it is possible to use personal objects from the missing person. We developed a protocol for obtaining DNA profiles from toothbrushes and underwear and evaluated two DNA extraction methods (Silica and magnetic particles), comparing the DNA quantity and the quality of the obtained profiles. Results showed that for underwear, silica in suspension recovered higher DNA quantity but magnetic particles (PrepFiler®) generated more complete profiles. For toothbrushes PrepFiler® recovered a higher DNA quantity and generated more complete DNA profiles. Statistical analyses showed there were no significant differences between both methods. Higher DNA quantities were recovered from underwear samples, however toothbrushes generated higher quality in DNA profiles, without obtaining mixed profiles, concluding that toothbrushes are a better item than underwear. Considering that silica in suspension method is three-times cheaper than PrepFiler®, we propose it is possible to successfully obtain DNA profiles from personal objects using a low-cost DNA extract method.     

Identification of Missing Norwegian World War II Soldiers,in Karelia Russia

This article presents the multidisciplinary effort in trying to identify the skeletal remains of 100 Norwegian soldiers serving in the German army, killed in Karelia Russia in 1944, from the recovery of the remains through the final identification using DNA. Of the 150 bone samples sent for DNA testing, 93 DNA profiles were obtained relating to 57 unique individuals. The relatives could not be directly contacted as the soldiers were considered as traitors to Norway; therefore, only 45 reference samples, relating to 42 cases of the missing, were donated. DNA matches for 14 soldiers and 12 additional body part re‐associations for these individuals were found. Another 24 bone samples were re‐associated with 16 individuals, but no familial match was found. More than six decades after the end of WWII, DNA analysis can significantly contribute to the identification of the remains.     

The collapse of an Italian cemetery into the sea: Forensic approach to human remains identification

On February 22nd, 2021, a landslide on the Italian coast caused the collapse of an old cemetery. About 370 coffins tumbled and more than 200 fell into the sea. 333 groups of unidentified human remains were found: 140 decomposed bodies and 193 bags of commingled skeletal remains. The Medical Staff of Legal and Forensic Medicine was involved for analyzing the remains in order to identify and bury them. The remains involved belonged to people who died between the end of the XIX century and 2017; all were interesting by advanced transformative phenomena. For the identifications, new forms, based on the Interpol DVI ones, were created. Information was collected by relatives through a specific antemortem form. Relatives’ information and post-mortem data were compared: 19 body were identified thanks to secondary methods (like object in the bury, dresses, medical devices). 147 bone samples (long bones and teeth) were collected for the genetic analysis. Among the 77 relatives eligible for a genetic comparison, 66 gave consent to DNA swab for collection and genetic typing. Currently, after 48 samples DNA analysis (STRs and Y-polymorphism) 12 remains were identified, 21 presented a profile suitable for comparison but without attribution, and 7 did not return a comparable profile caused by stochastic effects. 31 subjects have been identified and the genetics analysis are still in progress. The Cemetery collapse shows that every disaster requires a tailored approach.     

DNA Typing for the Identification of Old Skeletal Remains from Korean War Victims*

Abstract: 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 10⁵. 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.     

A Retrospective Review of Forensic Odontology Reports Written by the Joint POW/MIA Accounting Command Central Identification Laboratory for Remains Identified from the Korean War

As of August 2014, the Joint POW/MIA Accounting Command has identified the remains of 1980 previously unknown U.S. service members; 280 were from the Korean War. To determine the accuracy and completeness of the available antemortem (AM) dental records, a review of the AM/postmortem (AM/PM) dental record comparisons from 233 Forensic Odontology Reports written in support of remains identified from the Korean War was performed. Seventy‐two AM/PM comparisons resulted in exact dental chartings while 161 contained discrepancies which were explainable. Explainable discrepancies include undocumented treatment (103), incorrectly charted third molars as missing (82), differing opinions of specific molars present/missing (20), and erroneous treatment documentation and/or misidentification of teeth present/missing (22, other than molars). Reassessment has revealed varying levels of completeness for our available AM dental records, the need to thoroughly review our computerized comparisons, adjust our comparisons to include molar pattern variations/third molars, and updating our database comparison program.     

DNA Identification of Commingled Human Remains from the Cemetery Relocated by Flooding in Central Bosnia and Herzegovina

The floods in Bosnia and Herzegovina in May 2014 caused landslides all over the country. In the small village of ?eri?i, 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.     

Palynology and mycology as biological evidence in a homicide case

Criminal cases are studied from several disciplines to link a suspect with a criminal act. In this case, a man was reported missing in a coastal area in Buenos Aires Province, Argentina. The victim's relatives pointed to a possible suspect, and the local police carried out the investigation. We contributed to this research by applying palynological and mycological techniques. Palynomorphs and fungal spores offer valuable trace evidence, as they can be easily transferred between objects and crime scenes due to their minute size and persist on them for a long time. The victim was found 25 days later, lying on sandy soil, which partially covered the body, 35 km from where the suspect was arrested. Comparative samples were collected from the crime scene and the suspect's home and belongings (clothes, footwear, and seized vehicle). The palynological associations obtained from the crime scene and the defendant's belongings were dominated by diatoms and acritarchs (Acantomorphitae), all elements of marine origin, and a high CFU number of Bipolaris cynodontis, which allowed the defendant's clothing to be related to the place of corpse discovery. Soil from the defendant's home had an entirely continental composition, and the fungal biota was characteristic of prairie areas which were different from those of the crime scene.