Personal identification from human remains by mitochondrial DNA sequencing

The authors report four cases in which severely damaged human remains were identified by mitochondrial DNA (mtDNA) sequencing. Degraded DNA was extracted from highly adipoceratous tissues using the phenol-chloroform method and polymerase chain reaction amplified for sequencing of two hypervariable regions, hypervariable region 1 and hypervariable region 2, of mitochondrial DNA. They also sequenced these regions of blood samples that were obtained from the presumptive mother or sister of the human remains. The sequencing results were compared with each other and with the Anderson's sequence. It was concluded from the sequence data that a lower part of a body in case 1 and some organs in case 2 were from the same woman, and a human head in case 3 and a female body in case 4 were from the relative of a presumptive mother and a sister, respectively.     

Distribution of digoxin, digitoxin and their cardioactive metabolites in human heart and kidney tissue. A postmortem study

A method was developed for the specific determination of digoxin and digitoxin, as well as their semisynthetic derivatives and dependent cardioactive metabolites, in autopsy samples of heart and kidney. A collective of six patients on long-term treatment with therapeutic doses of beta-acetyldigoxin had a mean myocardial digoxin content of 46.1 +/- 25.0 ng/g (SD); kidney: 50.3 +/- 30.3 ng/g. Digoxigenin bisdigitoxoside represented the second most important metabolite in heart and kidney; digoxigenin monodigitoxoside and digoxigenin follow, respectively. In a collective of seven patients on maintenance treatment with digitoxin, the mean tissue levels were higher but the metabolic pattern was similar (myocardial digitoxin content: 78.9 +/- 38.4 ng/g, renal content: 104.1 +/- 44.1 ng/g). The amount of digoxin formed by hydroxylation under long-term treatment with digitoxin in heart and kidney were approximately 10 ng/g. A case of digoxin intoxication differed both in the tissue content and in the metabolic distribution.     

The identification of a dismembered human body: a multidisciplinary approach

A criminal case was directed to a multidisciplinary forensic team for identification, concerning a victim whose head, having two gunshot wounds, had been separated by a sharp instrument and was recovered 6 months later. The purpose of this research was to determine the sex and age of the victim for human identification. Primarily, macroscopic examination of the skull, tooth, and DNA analysis was conducted for sex determination. A rough assessment of age was made from the skull based on anthropological findings, however a more definitive result of age estimation was determined utilizing dental morphology. The dental data showed an age range of 32-37 from the mineral examination and the formulation of microscopic measurements. The results obtained from the skull and dental analysis matched with the physical characteristics of the victim's body, the known personal data of this person, and with the superposition of the photos gathered by a formal request. Besides, the result of DNA profiling of the victim showed male gender and direct relationship with the victim's presumed wife and daughter. Generally, research on human identification consists of sex and age determination. The sex characteristics can be precisely proved from DNA tests. However, age can be estimated by skeletal, and dental analysis. In this case the performed sex and age analysis lead the research to the selective matching of the missing person's identity.     

Elucidation of a poisoning case from the analysis of formalin in which brain tissue was preserved

The toxicological analysis of post-mortem materials often help determine the cause of death. When tissues have been fixed in formalin it may be possible to extract several drugs from the tissue and/or the preserving fluid. An unusual case of multiple poisoning is discussed from the aspect of laboratory analysis.     

Successful extraction of human genomic DNA from serum and its application to forensic identification

We report here on the successful extraction of human genomic DNA from a serum sample in a forensic case. The extracted DNA was successfully used for the identification of remains presumably immersed for more than three weeks for which the only comparison sample was a 250-microL serum aliquot kept frozen in a laboratory. The analysis made it possible to identify a second victim as the daughter of the first.     

Evaluation and identification of dismembered human remains.

On occasion, pathologists are confronted with cases involving dismembered human remains. Such cases present unique and interesting problems of postmortem identification. Four cases involving dismembered human remains are presented here and practical suggestions on how identification can be achieved are provided.     

Dual examinations for identification of urine as being of human origin and for DNA-typing from small stains of human urine

Concurrent methods for identification of urine as being of human origin, and for DNA-typing from small stains of human urine were examined. A urine stain was extracted with phosphate-buffered saline (PBS), and the extract was filtered using a Centricon-100 device. The filtrate was subjected to electrospray ionization liquid chromatography-mass spectrometry (ESI-LC-MS) for identification of human urine and a DNA-typing sample was obtained by dialfiltration of the residue using a DNA purification kit. After the purified residue was treated with an AmpflSTR Profiler PCR amplification kit, the DNA-types were analyzed by capillary electrophoresis using a Genetic Analyzer. It was possible to identify a urine stain as being of human origin, and complete DNA profiles could be successfully obtained from a urine stain which had been created by 50 microL of female urine. Serial analyses of urine stains found at a crime scene provide effective information for forensic investigation. This method is recommended for stain identification and for DNA-typing from a urine stain.     

Fixed human tissues: a resource for the identification of individuals

Polymorphic genetic loci of the deoxyribonucleic acid (DNA) present in formalin-fixed, paraffin-embedded tissues were successfully analyzed by utilizing the polymerase chain reaction. Using this analysis, with three different polymorphic loci [human leucocyte antigen (HLA) DQ alpha, low-density lipoprotein receptor, and parathyroid hormone], fixed tissues representing 14 different individuals were genotyped and could be distinguished from each other. The techniques were further applied to the fixed autopsy tissues of a man in which a question of paternity arose postmortem. Since many individuals have surgical procedures or autopsy, these readily available fixed tissues represent an additional resource for the identification of individuals.     

Radiographic human identification using bones of the hand: a validation study

The 1993 Supreme Court case Daubert v. Merrell-Dow Pharmaceuticals, Inc. underscores the importance of validating forensic science techniques. This research examines the validity of using posterior-anterior radiographs of the hand to make positive identifications of unknown human remains. Furthermore, this study was constructed to satisfy the requirements of Daubert's guidelines of scientific validity by establishing a standard methodology for hand radiograph analysis, testing the technique, and noting rates of error. This validation study required twelve participant examiners from the forensic science community, working independently, to attempt to match 10 simulated postmortem radiographs of skeletonized hands to 40 simulated antemortem radiographs of fleshed cadaver hands. The overall accuracy rate of the twelve examiners was 95%, while their collective sensitivity and specificity were 95% and 92%, respectively. However, the accuracy of each examiner was related to the amount of radiological training and experience of the observer. Six Ph.D. forensic anthropologists and four experienced forensic anthropology graduate students correctly identified all the matches. Participant examiners noted bone morphology, trabecular patterns of the proximal and middle phalanges, and distinctive radiopaque and radiolucent features as the anatomical features that aided the identification process. The hand can be an important skeletal element for radiographic positive identification because it contains 27 individual bones for comparative analysis.     

Zinc test as a new tool for identification of human seminal stains

An extremely simple qualitative method for identification of seminal stains based on high levels of zinc in human semen is described. It uses reaction of 1-(2-pyridylazo)-2-naphthol with zinc to develop a deep red color. The data are presented on the sensitivity, stability and specificity of the present method. We can recommend it for identification of human semen especially in old or denatured samples.     

Rapid and sensitive identification of human blood by an ELISA-ABC method using a biotinylated antibody against human HbA0

A direct enzyme-linked immunosorbent assay (ELISA) using an avidin-biotin complex (ABC) system for the identification of human blood is described. In this ELISA-ABC method, in which biotin-labeled goat IgG antibody against human HbA0 was used, it was possible clearly to distinguish human blood from the blood of other species, including that of Japanese monkeys. It took about 3 h to obtain the results. Human Hb concentrations ranging from 22 ng to 169 micrograms produced a positive reaction, and the minimum detection limit in terms of the highest possible dilution of human blood was 1:640,000.     

马和人股骨碎片组织形态学结构特征及其鉴别

目的研究成体马和成年人的股骨组织形态学结构,分析其组织学特征,建立人与马之间的种属鉴别方法。方法取来源于尸体检验的成人右股骨中段约4cm,同时收集马的右腿股骨,取中段约4cm,脱钙后制作成厚约25μm的骨组织切片,在光学显微镜下观察,将显微镜下的图像录入电脑,选取25项指标进行逐步判别分析。结果马和人在哈氏系统数量、哈氏系统直径等13个指标上具有显著差异,用这些指标建立种属判别数学模型,经盲测检验,二者的判别率可达99.6%。结论马和人的股骨组织学结构具有明显的种属特征,建立的判别方程可以有效地鉴别马和人的股骨碎片。     

Retrieval of analytical data and substance identification in systematic toxicological analysis by the mean list length approach

Chromatographic techniques are basic tools in systematic toxicological analysis. Extensive data bases with retention parameters of known drugs to aid in the identification of substances found are available or in preparation. For a search in such a data base the computer is indispenable. The commonly used window search has some disadvantages which can be overcome by a search based on the statistical concept, the mean list length. The latter retrieval system gives for each candidate in the identification process a probability value. It is shown that these probability values are highly influenced by the reproducibility of the retention parameters of the analytical systems used. Explanations for these phenomena are given.     

过敏性猝死者咽喉和胃肠组织内P物质的表达

目的观察P物质在过敏性猝死人体咽喉和胃肠组织的表达,并探讨其在法医学鉴定中的作用和意义。方法应用免疫组化方法对15例过敏性猝死者咽喉和胃肠组织的P物质表达进行研究,同时以冠心病猝死者作为对照;应用计算机图像分析系统对免疫组化染色结果进行分析,计算阳性指数(PI)。结果与对照组相比,实验组咽喉和胃肠组织P物质表达明显增加(P<0.001)。结论咽喉和胃肠组织的P物质表达可作为过敏性猝死的法医学鉴定的形态学依据和参考指标。     

Value of human lip lines and nail striations in identification

The pattern of the lips and the longitudinal striation of the lower concave surface of the nails of fingers and toes can be resource of identification. The types of the lip lines are individual and probably constant for some years. We demonstrated that the pattern of all of 20 human nails are over the space of one year the same and found no interindividual correspondence. The investigation rate for examination amount to 36% for the nails of the fingers and 55% for suchlike toes.     

Confidence and accuracy of eyewitness identification from lineups

Recent studies of confidence-accuracy correlations in eyewitness-identification experiments have produced highly variable results (Deffenbacher, 1980; Wells & Murray, 1984) with some correlations near 0 (Malpass & Devine, 1981) and others as high as 0.5 (Brigham et al., 1982). Much of this variability may be accounted for by differences, in the design of the research. Specifically, lineup experiments in which the to-be-identified person (target) is always present tend to produce higher confidence-accuracy correlations than studies also providing blank or target-absent lineups replacing the target with a lookalike. A reanalysis of previously collected data (Lindsay & Wells, 1980; Wallbridge & Lindsay, 1982) revealed that lineup foils (nontargets) were identified with significantly less confidence than targets or lookalikes but that targets and lookalikes were identified with equal confidence. Two experiments were conducted to test the hypothesis that confidence of identification is a function of the similarity of the identified person to the original target. The first experiment using nonstudent subjects (n=53) and multiple targets (n=14) to increase generalizability, demonstrated that the person in the lineup who most closely resembled the target was identified with the highest mean level of confidence. The second experiment examined confidence in identification following a crime staged for 260 introductory psychology students. The results indicated that the similarity of foils to the target (established by independent ratings) was predictive of both the frequency and confidence of identification of faces.     

Psychedelic fungus (Psilocybe sp.) authentication in a case of illegal drug traffic: sporological,molecular analysis and identification of the psychoactive substance

In nature, there are >200 species of fungi with hallucinogenic properties. These fungi are classified as Psilocybe, Gymnopilus, and Panaeolus which contain active principles with hallucinogenic properties such as ibotenic acid, psilocybin, psilocin, or baeocystin. In Chile, fungi seizures are mainly of mature specimens or spores. However, clandestine laboratories have been found that process fungus samples at the mycelium stage. In this transient stage of growth (mycelium), traditional taxonomic identification is not feasible, making it necessary to develop a new method of study.Currently, DNA analysis is the only reliable method that can be used as an identification tool for the purposes of supporting evidence, due to the high variability of DNA between species. One way to identify the species of a distinctive DNA fragment is to study PCR products analyzed by real time PCR and sequencing. One of the most popular sequencing methods of forensic interest at the generic and intra-generic levels in plants is internal transcribed spacer (ITS). With real time PCR it is possible to distinguish PCR products by differential analysis of their melting temperature (Tm) curves.This paper describes morphological, chemical, and genetic analysis of mycelia of psychedelic fungi collected from a clandestine laboratory. The fungus species were identified using scanning electron microscopy (SEM), mass spectrometry, HRM analysis, and ITS sequencing. The sporological studies showed a generally smooth surface and oval shape, with maximum length 10.1?μm and width 6.4?μm. The alkaloid Psilocyn was identified by mass spectrometry, while HRM analysis and ITS sequencing identified the species as Psilocybe cubensis. A genetic match was confirmed between the HRM curves obtained from the mycelia (evidence) and biological tissue extracted from the fruiting bodies. Mycelia recovered from the evidence and fruiting bodies (control) were genetically indistinguishable.