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.     

Mitochondrial DNA typing--a new level for solving identification problems in forensic medical expert identification of unidentified remains of victims of terrorist acts in Moscow and the armed conflict in the Chechen Republic

Two large-scale episodes described in this paper reflect the first in Russia use of molecular genetic matrilinear markers (analysis of polymorphism of sequences of amplified fragments of mitochondrial DNA hypervariable locuses) in solution of a complex identification problem: forensic medical identification of unidentified fragments of victims of explosions of houses in Moscow in September, 1999, and of soldiers dead in the war conflict in the Chechen Republic in 1994-1996. The results of this work and methodological experience gained in it essentially extend the potentialities of expert studies as regards forensic medical identification of victims of large scale disasters, terroristic acts, and war conflicts.     

Teeth as a source of DNA for forensic identification of human remains: A Review

Teeth and bones are frequently the only sources of DNA available for identification of degraded or fragmented human remains. The unique composition of teeth and their location in the jawbone provide additional protection to DNA compared to bones making them a preferred source of DNA in many cases. Despite this, post-mortem changes in the structure and composition of teeth, and the location and diagenesis of DNA within them are poorly understood. This review summarises current knowledge of tooth morphology with respect to DNA content and preservation, and discusses the way in which post-mortem changes will affect the recovery of DNA from teeth under a range of commonly used extraction protocols. We highlight the benefits and pitfalls of using specific tooth tissues for DNA extraction and make recommendations for tooth selection and sampling that will maximise DNA typing success. A comprehensive understanding of tooth structure and an appreciation of the relationship between DNA and mineralized tissues in post-mortem teeth are critical for optimal sample selection. More informed sampling methods that target specific tooth tissues will increase the likelihood of successful genetic analysis and allow for efficient and timely missing persons case work and disaster victim identification response.     

Utilization of complex technologies of molecular genetics for forensic expert identification of unidentified remains of victims of terrorist acts in Moscow in 1999

This paper presents the first experience gained in Russia in expert identification practice with complex utilization of technologies of molecular genetic individualization of biological objects for solving identification tasks requiring the maximum complete armory of mutually supplementing means. This is particularly important for forensic expert personality identification in cases with many deaths, when indirect identification has to be resorted to, consisting in the use of biological samples from relatives of the victims as the identifying objects. Methodological approaches and concepts of expert studies, used for identification of the remains of people who died in terroristic acts committed in Moscow in September, 1999, are discussed. The results of this study and methodological experience notably extend the potentialities of expert evaluation as regards forensic medical identification of victims of overall disasters, terroristic acts, and war conflicts.     

Polymorphism of structural genes of human mitochondrial DNA for forensic medical personal identification

An experimental study is described, which deals with polymorphism of nucleotide sequences of three structural genes of mitochondrial genome, i.e. of the 3d, small 4th, and 6th subunits of the NADN dehydrogenase complex (ND3, ND4L and ND6) sampled from Russian population. The genetic primary structure was analyzed in 63 unrelated individuals. The investigated locuses were shown to possess a pronounced polymorphism. A total of 19 polymorphic positions were detected in the ND3, ND4L and ND6 gene region within the studied sampling. Besides, a possibility is demonstrated in the paper that the mtDNA structural genes can be used as additional identification markers in the forensic experimental typing of the mtDNA control region.     

The possible use of the dielectric indices of human hair in forensic medical expertise]

Method of measuring dielectric parameters of human hair was developed. It was shown that values of relative dielectric permeability of hair may serve as objective criteria in the course of determination of their similarity.     

Using electrophoresis for the identification of subjects in forensic medical expertise

It was proven as advisable to apply the electrophoretic method in sophisticated expertise cases for the purpose of defining the nature of objects in examinations of material evidence.     

Recognition of cemetery remains in the forensic setting

Cemetery remains exposed through vandalism or natural phenomena are frequently brought to the attention of law enforcement agents or medical examiners. Although it is often difficult to distinguish cemetery remains from those of medicolegal significance, clues to their origin may exist. Characteristics consistent with cemetery remains include physical characteristics associated with the embalming process. Characteristics indicative of cemetery remains include functional or ornamental artifacts associated with the coffin, devices used in embalming the body, and elevated levels of embalming chemicals in the soft tissue.     

Ascertainment of the cause and tempo of death in forensic medical practice

In order to develop a method of ascertaining a cause of death and of evaluating its tempo the authors undertook a histological study and a study of a dehydration degree in 76 cases of death due to the below reasons: mechanical trauma, mechanical asphyxia, burn shock, and carbon monoxide intoxication. Acute emphysema of lungs, lack of edemas in lungs and brain, desquamation of pulmonary macrophages, hyperemia of renal capillaries, primary urine in some glomerules, prevalence of ischemic changes with karyopyknosis in medulla and hypohydration of brain were typical of fast death. Prevalence of dystelectasis in lungs, emptying of pulmonary capillaries, evident edema of lungs and brain, desquamation of pulmonary macrophages, lack of primary urine in glomerular capsules, acute circulatory disorders of renal hemodynamics with sweating of fibrin and erythrocyte glomerules into cavities, necrotic nephrosis, pigment cylinders in renal tubules, thrombi in vessels, prevalence of chromatolysis, karyolysis and cytolysis in medulla, pronounced glial reaction as well as pronounced edema of brain and its hyperhydration were typical of slow death.     

Ultrastructural characteristic of gunpowder charge remains in forensic medical examination of gunshot bullet injuries

Particles of 3 different marks of gunpowder, withdrawing from experimental gunshot injuries, were examined with scanning electronic microscopy. Features of ultrastructure, making possible to differentiate gunpowder of studied marks, were detected.     

300例心源性猝死案例的法医学鉴定分析

目的探讨心源性猝死案例的病因、诱发因素及法医病理学特点,以期为心源性猝死案件的法医病理诊断提供参考。方法收集法医病理检案中的心源性猝死案例,对其年龄、性别、死亡时间、死因、诱因及场所等流行病学特点进行回顾分析。结果 300例心源性猝死案例中男性多于女性,18~45岁发病最多;病因包括冠心病(129例),心肌病(78例),心肌炎(36例)等;应激为SCD最常见的诱发因素(157例)。结论 1应激和冠心病是心源性猝死的重要危险因素。2心源性猝死的法医学鉴定除了要进行系统的尸检及病理学检查外,还要对死者的相关资料如:基础疾病、诱因、死亡时间、死亡经过等进行全面系统的分析以明确死因。     

Effects of processing techniques on the forensic DNA analysis of human skeletal remains

Human remains processed by forensic anthropologists may potentially be used for genetic analysis. Therefore, the condition of the deoxyribonucleic acid (DNA) in processed remains may become an issue for future analysis. Processing techniques employed by anthropologists are highly variable and scanning electron microscopy reveals significant alterations to the bone surface depending upon the technique used. Such damage to the bone indicates differences may exist in quality and quantity of DNA extracted. This study assessed how five processing procedures used by major forensic anthropology laboratories around the country affects the amounts of DNA extracted from human rib bones and the subsequent DNA analysis. The DNA was analyzed using the short tandem repeat (STR) locus CSF1PO and amelogenin. The findings indicate processing procedures used by forensic anthropologists do not adversely affect DNA analysis but prolonged exposure to heat during processing may decrease the yield of information from the DNA.     

必须重视法医的命案现场勘验

命案现场勘验是法医的一项重要工作任务。但近几年没有得到足够的重视，甚至有的认为法医就是检验尸体，现场勘验可有可无，从而导致临案法医的作用被弱化和检案水平得不到提高。笔者就此结合案例进行相关探讨。     

必须重视法医的命案现场勘验

命案现场勘验是法医的一项重要工作任务。但近几年没有得到足够的重视 ,甚至有的认为法医就是检验尸体 ,现场勘验可有可无 ,从而导致临案法医的作用被弱化和检案水平得不到提高。笔者就此结合案例进行相关探讨。案例介绍【案例 1】　某日下午 ,本县一招待所客房内一 80岁老妪死于床上 ,刑技人员处理现场后即将尸体交由法医检验 ,发现 :( 1)死者颈部有 4道勒索 (第 1条为长条毛巾 ,其他 3条为长短不一的布条 ) ,都在颈前部打有死结 ,其索道间绞夹着头发和衣领等 ,勒索相应处软组织出血 ;( 2 )颜面部青紫肿胀 ,左耳道出血 ,心血呈暗红色流动性…     

The use of moxibustion scars in the identification of individuals in forensic medicine

The use of moxibustion today does not only take place in Eastern Asia, but recently also in Europe and the United States. Moxibustion scars are often seen in autopsy cases in Japan. However, no reports of the use of moxibustion in autopsy cases have been reported. This paper reports on an autopsy case in which moxibustion scars were used to extrapolate on the past symptoms of the deceased, date of therapy and the location of the moxibustion clinic which administered the moxibustion.     

Validation of the barcoding gene COI for use in forensic genetic species identification

The application of forensics to wildlife crime investigation routinely involves genetic species identification based on DNA sequence similarity. This work can be hindered by a lack of authenticated reference DNA sequence data resulting in weak matches between evidence and reference samples. The introduction of DNA barcoding has highlighted the expanding use of the mtDNA gene, cytochrome c oxidase I (COI), as a genetic marker for species identification. Here, we assess the COI gene for use in forensic analysis following published human validation guidelines. Validation experiments investigated reproducibility, heteroplasmy, mixed DNA, DNA template concentration, chemical treatments, substrate variation, environmental conditions and thermocycling parameters. Sequence similarity searches using both GenBank BLASTn and BOLD search engines indicated that the COI gene consistently identifies species where authenticated reference sequence data exists. Where misidentification occurred the cause was attributable to either erroneous reference sequences from published data, or lack of primer specificity. Although amplification failure was observed under certain sample treatments, there was no evidence of environmentally induced sequence mutation in those sequences that were generated. A simulated case study compared the performance of COI and cytochrome b mtDNA genes. Findings are discussed in relation to the utility of the COI gene in forensic species identification.     

Role of orthopedic implants and bone morphology in the identification of human remains

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.