Forensic anthropology in the 1990s.

Forensic anthropology has undergone considerable change over the past 10 years. Today it is utilized by most law enforcement, coroner, and medical examiner systems. The techniques for determination of age at death, sex, race, and stature from skeletal remains have been modified and greatly expanded. The role of the forensic anthropologist within a medicolegal context is much broader than in previous years. In addition to establishing individual identity, forensic anthropologists are now consulted for trauma analysis, facial reconstruction, photographic superimposition, determination of time interval since death, and crime-scene recovery. Not all physical anthropologists are forensic anthropologists. Qualified individuals are certified, through rigorous examination, by the American Board of Forensic Anthropology. A list of board-certified forensic anthropologists may be obtained through the American Academy of Forensic Sciences.     

Shark‐Inflicted Trauma: A Case Study of Unidentified Remains Recovered from the Gulf of Mexico*

Abstract: Here, we present a case of an unidentified male whose remains, except for the right arm, were recovered from the Gulf of Mexico 10 years prior to osteological analysis by forensic anthropologists. After the poorly preserved soft tissue was removed and the bones cleaned, forensic analysis revealed an unusual series of hard tissue trauma later attributed by a shark expert as shark scavenging and/or predation. Identified were five unique hard tissue trauma patterns that are bite mark artifacts produced by sharks: punctures without fractures, punctures with associated fractures, striations with bone shaving, overlapping striations, and incised bone gouges. The cooperation among experts provided a comprehensive death case analysis and a better understanding of shark‐inflicted trauma on human skeletal remains.     

Dietary health does affect histological age assessment: an evaluation of the Stout and Paine (1992) age estimation equation using secondary osteons from the rib

An age at death estimation equation that uses rib histological variables presented by Stout and Paine was used to evaluate a skeletal population of individuals with a known age at death and cause of death from either malnutrition or the niacin deficiency disease pellagra. The sample was comprised of 26 autopsied black South Africans. Histological analysis of mounted thin sections involved the microscopic measurement of cortical area and a count of the number of intact and fragmentary secondary osteons for the entire cross-section of the rib. Rib osteon population density values were then calculated for each case. It was found that this equation under-aged individuals on average by 29.2 years. Overall, secondary osteon size and Haversian canals tended to be larger than expected, while cortical bone area was less when compared with a control population. The implications of these findings are critical given that many of the skeletal remains examined by forensic anthropologists come from marginalized backgrounds, including malnutrition. This research suggests that measurements based on healthy cases may not be useful in an analysis of individuals with poor diet and health. It is argued that new standards for histological age assessment methods need to be created that account for variation in the health status of individuals examined by forensic anthropologists.     

Patterns of Trauma Induced by Motorboat and Ferry Propellers as Illustrated by Three Known Cases from Rhode Island*

Abstract: Understanding patterns of trauma is important to determining cause and manner of death. A thorough evaluation of taphonomy, trauma, and bone fracture mechanisms is necessary to reconstruct the circumstances of the death. This study examines the skeletal trauma caused by boat propeller strikes in terms of wound characteristics and location based on three cases from Rhode Island. These case studies review the traumatic characteristics caused by propeller injuries and highlight the anatomic regions most likely to sustain skeletal trauma. With this information, investigators may be able to identify propeller trauma even in severely decomposed remains. The discussion of boat propeller trauma also raises issues regarding how forensic anthropologists and forensic pathologists classify trauma (specifically blunt force vs. sharp) and highlights semantic issues arising in trauma classification. The study also discusses why these propeller cases should be classified as blunt trauma rather than sharp or chop/hack trauma. Ultimately, the authors urge consistency and communication between pathologist and forensic anthropologists performing trauma analyses.     

The reliability of forensic osteology--a case in point. Case study

The medico-legal investigation of skeletons is a trans-disciplinary effort by forensic scientists as well as physical anthropologists. The advent of DNA extraction and amplification from bones and teeth has led to the assumption that morphological assessment of skeletal remains might soon become obsolete. But despite the introduction and success of molecular biology, the analysis of skeletal biology will remain an integral part of the identification process. This is due to the fact, that the skeletal record allows relatively fast and accurate inferences about the identity of the victim. Moreover, a standard biological profile may be established to effectively narrow the police investigator's search parameters. The following study demonstrates how skeletal biology may collaborate in the forensic investigation and support DNA fingerprinting evidence.In this case, the information gained from standard morphological methods about the unknown person's sex, age and heritage immediately led the police to suspect, that the remains were that of a young man from Vietnam, who had been missing for 2.5 years. The investigation then quickly shifted to prove the victim's identity via DNA extraction and mtDNA sequence analysis and biostatistical calculations involving questions of kinship [4].     

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.     

Current practices by forensic anthropologists in adult skeletal age estimation

When determining an age estimate from adult skeletal remains, forensic anthropologists face a series of methodological choices. These decisions, such as which skeletal region to evaluate, which methods to apply, what statistical information to use, and how to combine information from multiple methods, ultimately impacts the final reported age estimate. In this study, a questionnaire was administered to 145 forensic anthropologists, documenting current trends in adult age at death estimation procedures used throughout the field. Results indicate that the Suchey-Brooks pubic symphysis method (1990) remains the most highly favored aging technique, with cranial sutures and dental wear being the least preferred, regardless of experience. The majority of respondents stated that they vary their skeletal age estimate process case-by-case and ultimately present to officials both a narrow and broad possible age range. Overall, respondents displayed a very high degree of variation in how they generate their age estimates, and indicated that experience and expertise play a large role in skeletal age estimates.     

The Effects of Different Maceration Techniques on Nuclear DNA Amplification Using Human Bone

Abstract: Forensic anthropologists routinely macerate human bone for the purposes of identity and trauma analysis, but the heat and chemical treatments used can destroy genetic evidence. As a follow‐up to a previous study on nuclear DNA recovery that used pig ribs, this study utilizes human skeletal remains treated with various bone maceration techniques for nuclear DNA amplification using the standard Combined DNA Index System (CODIS) markers. DNA was extracted from 18 samples of human lower leg bones subjected to nine chemical and heat maceration techniques. Genotyping was carried out using the AmpF?STR^® COfiler^® and AmpF?STR^® Profiler Plus^® ID kits. Results showed that heat treatments via microwave or Biz/Na₂CO₃ in sub‐boiling water efficiently macerate bone and produce amplifiable nuclear DNA for genetic analysis. Long‐term use of chemicals such as hydrogen peroxide is discouraged as it results in poor bone quality and has deleterious effects on DNA amplification.     

Simple and highly effective DNA extraction methods from old skeletal remains using silica columns

The recovery of DNA data from old skeletal remains is often difficult due to degraded and very low yield of extracted DNA and the presence of PCR inhibitors. Herein, we compared several silica-based DNA extraction methods from artificially degraded DNA, DNA with PCR inhibitors and DNA from old skeletal remains using quantitative real-time PCR. We present a modified large-scale silica-based extraction combined with complete demineralization, that enables maximum DNA recovery and efficient elimination of PCR inhibitors. This is performed with high concentration of EDTA solution for demineralization of bone powder followed by QIAamp^® spin columns and buffers from the QIAquick^® PCR purification kit. We have successfully used this modified technique to perform STR analysis for 55-year-old skeletal remains. The results of this study will contribute to solve the forensic cases dealing with skeletal remains.     

A multidisciplinary investigation of chronic animal abuse: Collaboration between veterinary forensics and forensic anthropology

An unknown juvenile female mixed breed dog was found non‐ambulatory on a dead‐end street in an urban setting adjacent to a public park. During initial veterinary examination, she was assessed to have untreatable injuries and was humanely euthanized. The forensic veterinarian requested consultation from a forensic anthropologist to assist with documenting antemortem skeletal trauma. Analyses of skeletal tissues indicated numerous injuries in various stages of healing diagnostic of non‐accidental injuries. Veterinary forensic cases may benefit from collaborative analysis of bony remains by forensic anthropologists.     

Difficulties of sex determination from forensic bone degraded DNA: A comparison of three methods

Sex determination is of paramount importance in forensic anthropology. Numerous anthropological methods have been described, including visual assessments and various measurements of bones. Nevertheless, whatever the method used, the percentage of correct classification of a single bone usually varies between 80% and 95%, due to significant intra- and inter-population variations, and sometimes variations coming from secular trends. DNA is increasingly used in a forensic context. But forensic DNA extraction from bone raises several issues, because the samples are very often badly altered and/or in very small quantity. Nuclear DNA is difficult to get from degraded samples, according to low copy number, at least in comparison with mitochondrial DNA. In a forensic context (as in a paeleoanthropological context) DNA sex determination is usually complicated by the weak amount of DNA, the degraded nature of nucleic acids, the presence of enzymatic inhibitors in DNA extracts, the possible faint amplification of Y band and the risk of contamination during either excavation or manipulation of samples.The aim of this work was to compare three methods of DNA sex determination from bones: procedure #1 using a single PCR amplification, procedure #2 using a double PCR amplification, and procedure #3 adding bleaching for decontamination of the bone, instead of simply rubbing the bone. These processes were applied to samples of bones (49 samples coming from 39 individuals) that were in various states of post mortem alteration.The main results are the following. (i) No DNA could be extracted from three skulls (parietal bones, mastoid process), the compact bone of one rib, and the diaphysis of one femur; (ii) there was a contamination in three skulls; and (iii) the Y band did not appear in two male cases, with one of the three procedures (male tibia, procedure #2) and with procedures #2 and #3 (male femur).This study emphasises the main issue while working with altered bones: the impossibility to extract DNA in some cases, and, worth of all, the contamination of the sample or the faint amplification of Y band which leads to a wrong sex answer. Multiple and significant precautions have to be taken to avoid such difficulties.     

CT技术在法医人类学研究中应用的进展

近年来,将计算机断层扫描(CT)等影像学新技术应用于法医人类学个体识别的研究受到国内外法医人类学家的重视。CT具有清晰快速的成像能力以及强大的后处理功能,无需骨骼前处理就能提供高分辨率模型图像,对法医人类学具有重要的研究和应用价值。本文就CT技术在法医人类学研究中应用的进展进行综述,以期为相关研究和实践提供参考和借鉴。     

The effects of skeletal preparation techniques on DNA from human and non-human bone

The forensic pathologist increasingly relies on the forensic anthropologist to be the consulting expert in human identification. Likewise, if identification is not possible from visual inspection of skeletal remains, the forensic biologist may be called upon to conduct DNA analysis. The possibility of downstream DNA testing needs to be considered when skeletal preparation techniques are employed to deflesh human remains, as they have the potential to strongly impact genetic analyses and subsequent identification. In this study, three cleaning techniques, boiling bone in water, in bleach, and in powdered detergent/sodium carbonate, were tested for their effect on nuclear and mtDNA recovery from a variety of human and non-human bones. A statistically significant reduction in DNA yields occurred in non-human bones cleaned with bleach, and DNA degradation was apparent electrophoretically. The human bones also showed much lower yields from bleach cleaning, while the detergent/carbonate method allowed the largest segments of DNA to be amplified, indicating it may have a less degradative effect on bone DNA than either of the other cleaning processes.     

The Use of an Alternate Light Source for Detecting Bones Underwater

When searching underwater crime scenes or disaster scenes for fragmentary human remains, it may be advantageous for forensic divers to be able to detect the presence of bones and teeth among other marine materials (such as shells and rocks). In terrestrial environments, this can typically be accomplished by visual and instrumental methods, but underwater conditions make it difficult to employ detection and sorting techniques in these environments. This study investigates fluorescence of bones and teeth and other marine materials using a submersible alternate light source (ALS) and concludes that an ALS can be a useful tool for detecting bones and teeth in underwater searches as well in terrestrial searches and laboratory environments. The results could impact the methods and equipment used by forensic divers and forensic anthropologists when searching for skeletal remains, potentially increasing the quantity and efficiency of forensic evidence recovered.     

Identification of the source of ivory idol by DNA analysis

In this study, we describe a forensic case dealing with the identification of the source of the processed ivory object by DNA analysis. Two pieces of Lord Krishna's idols from a shop were confiscated by an investigating agency of the Indian government and forwarded to us to identify the source of its origin. We succeeded in isolating DNA from both processed ivory idols by using the phenol/chloroform DNA extraction method. The extracted DNA was subjected to PCR amplification using an elephant-specific mitochondrial DNA (mtDNA) D-loop marker. DNA sequence analysis of the amplified fragment of mtDNA D-loop region confirmed that the idols were consistent with Asian elephant with 99% similarity.     

Use of bleach to eliminate contaminating DNA from the surface of bones and teeth

The extraction of DNA from archaeological or forensic skeletal remains can provide quite powerful data for analysis, but is plagued by a unique set of methodological problems. One of the most important methodological problems to overcome in such analyses is the presence of modern contamination on the surfaces of bones and teeth, which can lead to false positives and erroneous results unless it is removed before DNA extraction is initiated. Ancient DNA (aDNA) researchers and forensic scientists have employed a number of techniques to minimize such contamination. One such technique is the use of bleach (sodium hypochlorite--NaOCl) to "destroy" contaminating DNA. However, a consensus on the optimum concentration of sodium hypochlorite to be used and the amount of time the bone or tooth should be exposed to it has not emerged. The present study systematically approaches the issue by introducing contamination to ancient bones (from approximately 500 BP) and determining which of several sodium hypochlorite treatments best eliminates surface contamination. The elimination of surface contamination from bone requires immersion in at least 3.0% (w/v) sodium hypochlorite (approximately equal parts of commercial bleach and water) for at least 15 min. Endogenous DNA proved to be quite stable to even extreme sodium hypochlorite treatments (6% for 21 h), suggesting that DNA adsorbs to hydroxyapatite in the bone and that this process facilitates the preservation of DNA in ancient skeletal remains.     

A minimally destructive technique for sampling dentin powder for mitochondrial DNA testing

The purpose of this paper is to present the horizontal sectioning technique used by odontologists at the Central Identification Laboratory to sample dentin for mtDNA analysis. From the perspective of DNA testing, anthropologists and odontologists at the Central Identification Laboratory work with ancient remains. In many instances, the lack of comprehensive antemortem records, the potential for fragmentation and commingling, and environmental exposure makes the use of traditional forensic identification techniques difficult or impossible. Teeth are highly resistant to environmental degradation and are an excellent source of mitochondrial DNA (mtDNA). This technique is simple, quick, and relatively conservative, allowing for preservation of the majority of the external portion of the tooth structure.     

Guidelines for the recognition of cemetery remains in Greece

Forensic pathologists frequently consult anthropologists for the identification of skeletonized human remains. These remains may be the result of criminal activity or remains that were unearthed because of erosion, or during construction projects. In some cases, human remains that had been previously buried in a cemetery may be the subject of a forensic investigation. Early recognition of cemetery remains prevents unnecessary efforts and conserves precious resources. One of the key characteristics of cemetery remains is the presence of embalmed tissue. However, there are countries where embalming is not a common practice, and other clues must be sought for identifying previously buried remains. Current funerary customs in Greece and, in particular, the tradition of exhumations result in a large number of misplaced human remains. The present study presents examples of cemetery remains from Greece and offers guidelines for recognizing changes on skeletal remains that may be indicative of a cemetery origin. Location of discovery, condition of the remains, and the types of associated artifacts are all factors that aid forensic anthropologists in identifying cemetery remains.     

Optimization and validation of a fully automated silica-coated magnetic beads purification technology in forensics

Automated procedures for forensic DNA analyses are essential not only for large-throughput sample preparation, but are also needed to avoid errors during routine sample preparation. The most critical stage in PCR-based forensic analysis is DNA isolation, which should yield as much highly purified DNA as possible. The extraction method used consists of pre-treatment of stains and samples, cell lysis using chaotropic reagents, binding of the DNA to silica-coated magnetic particles, followed by elution of the DNA. Our work focuses mainly on sample preparation, obtaining the maximum possible amount of biological material from forensic samples, and the following cell lysis, to create a simple standardized lysis protocol suitable for nearly all forensic material. After optimization and validation, the M-48 BioRobot((R)) workstation has been used for more than 20,000 routine lab samples. There has been no evidence of cross contamination. Resulting DNA from as small as three nuclear cells yield reliable complete STR amplification profiles. The DNA remains stable after 2 years of storage.     

Effect of Temperature on Bone Tissue: Histological Changes

The analysis of burned human remains has been of great interest among forensic anthropologists largely due to the difficulty that their recovery, classification, reconstruction, and identification present. The main purpose of this analysis is to present histological methodology for the interpretation of bones altered by thermal processes. We include analyses of the microscopic changes among bones exposed to different temperatures, with the goal of establishing categories of histological morphology in relation to fire temperature. Samples of bone (ilium) were exposed systematically to controlled temperatures. Analysis of the resulting histological changes has allowed the formation of a clear four‐stage classification of the alterations observed. This classification should prove useful in assessing bone changes in relation to temperature of exposure, particularly in cases where this temperature was previously not known.