人骨骼及牙齿DNA提取方法的比较和优化

目的人骨骼和牙齿DNA提取方法的比较和优化。方法收集18份不同个体的长骨、30颗磨牙和同一个体2根股骨、8颗磨牙。利用TissueLyser-Ⅱ组织破碎仪和PreFiler Express BTA^TM法医DNA提取试剂盒（BTA法）,应用Automate Express^TM自动化法医DNA提取系统提取DNA,进行STR分型,与脱钙法进行比较,并进行实验条件优化。结果用TissueLyser-Ⅱ结合BTA法,约2.5h即可完成骨骼和牙齿的DNA提取,分型成功率分别为94.4%和96.7%。与脱钙法比较,两种方法获得DNA质量浓度和检出率比较接近（P〈0.05）,但BTA法在操作过程方面更具优势。最佳样本量为100mg,消化时间为2h。结论采用TissueLyser-Ⅱ组织破碎仪结合BTA法对骨骼和牙齿进行DNA提取和分型检验,能满足实际检案的要求,可在法医学实践中选择使用。     

Clinically Detectable Dental Identifiers Observed in Intra‐oral Photographs and Extra‐oral Radiographs,Validated for Human Identification Purposes

Screening the prevalence and pattern of dental identifiers contributes toward the process of human identification. This research investigated the uniqueness of clinical dental identifiers in photographs and radiographs. Panoramic and lateral cephalometric radiographs and five intra‐oral photographs of 1727 subjects were used. In a target set, two observers examined different subjects. In a subset, both observers examined the same subjects (source set). The distance between source and target subjects was quantified for each identifier. The percentage of subjects in the target set being at least as close as the correct subject was assessed. The number of molars (34.6%), missing teeth (42%), and displaced teeth (59.9%) were the most unique identifiers in photographs and panoramic and lateral cephalometric radiographs, respectively. The pattern of rotated teeth (14.9%) was the most unique in photographs, while displaced teeth was in panoramic (37.6%) and lateral cephalometric (54.8%) radiographs. Morphological identifiers were the most unique, highlighting their importance for human identifications.     

Isolation of tooth pulp cells for sex chromatin studies in experimental dehydrated and cremated remains.

In experiments designed to assess sex chromatin in artificially mummified and heated pulp tissue, a method was devised that successfully separates cells while minimizing nuclear damage. Sex chromatin (both Barr bodies and F-bodies) is shown to preserve in dehydrated human pulps up to one year. Human pulp tissue retains sex diagnostic characteristics when heated to 100 degrees C for up to 1 h. Parallel experiments on extracted teeth from young pigs reveals comparable tissue preservation. Heat penetration is retarded, however, in unextracted pig teeth in fleshed jaws such that temperatures could be raised to 300 degrees C for longer than 1 h. Heat penetration into fleshed material was further tested by the insertion of thermocouple probes to assess the temperature attained within the pulp chamber. At chamber temperatures up to 75 degrees C sex diagnosis in human pulps from extracted teeth was still possible. In outdoor incineration of fleshed pigs' heads in an open fire, 75 degrees C in the pulp chamber was reached at a fire temperature within the range 500-700 degrees C. The implications of these findings for forensic situations are described.     

The Use of Radio-Frequency Identification Tags for Labeling Dentures—Scanning Properties

Abstract:  The inclusion of radio-frequency identification (RFID)-tags within dental prostheses has been suggested as means of effectively labeling such devices and permitting rapid and reliable identification of the wearer. Previous studies have suggested that patients will accept denture labeling and recognize the need for such systems. However, they demand systems that are aesthetic, durable, and secure. One concern over the use of RFID-tags is that they could be scanned by third parties without the patient's knowledge. This study categorizes the scanning patterns of RFID-tags both in vitro and in vivo to provide data for patients for the consent process and for forensic dentists to ensure that they are scanning prostheses optimally. The results demonstrate that the RFID chips can only be read when the interrogator is in close proximity to the denture and thus should alleviate any concerns over privacy issues. However, evidence obtained from both the literature and experiments suggests that authorities must agree upon a unified standard for chip and reader specifications and protocols in order to avoid cases in which RFID-tags may fail to be read by an incompatible reader.     

Measuring the distance of the wisdom teeth from the occlusal plane as forensic-odontological method for chronological age determination

As part of an epidemiological study on tooth eruption it was attempted to define easily accessible criteria for age estimation in living human beings based on the distance of the wisdom teeth from the occlusal plane. 253 cast models of 149 female and 104 male patients were used for evaluation. Due to the great variability observed in the eruption of the wisdom teeth, only a small fraction of the total number of 10,924 cast models was suitable for investigation. Only teeth with the axis being perpendicular to the plane of occlusion during eruption according to radiological and clinical findings were included. After defining these inclusion criteria, the distance of the wisdom teeth from the plane of occlusion was determined and correlated to the age of the subjects. The results confirmed the known large variability concerning wisdom tooth eruption and did not show any reliable relation between age and distance values. Nevertheless, a remarkable finding that may be relevant for forensic odontostomatology was seen in one case where the lower wisdom teeth were taller than the occlusal plane, although the female patient was younger than 14 years. The upper molars were retained and root development was incomplete.     

Determination of season at death using dental cementum increment analysis

Dental cementum anchors teeth into their sockets via the periodontal ligament. In mammals, dental cementum is laid down in alternating opaque and translucent bands representing winter (dormant) and summer (growth) seasons. Each pair of bands represents 1 year of life. Estimates of age at death based on counting pairs of bands are highly accurate. If the timing of the transition between winter and summer bands could be identified in humans, dental cementum increment analysis could be used to specify the season at death. This pilot project attempts to determine when these transitions occur and thus provide forensic anthropologists with a technique that would refine our estimates of postmortem interval. Extracted teeth were obtained from a local oral surgeon, embedded, sectioned, ground, polished, and examined under transmitted polarized light. The outermost increment was identified and measured along with other randomly chosen like bands. A transition from translucent to opaque bands was observed in teeth extracted in early October, while teeth extracted in early April exhibited nascent translucent bands. Further, significant correlations were observed between band thickness and number of days into either season, suggesting that band width increases as either season progresses. In this blind study, extracted teeth were effectively sorted into the spring/summer or fall/winter season 99% of the time.     

自动荧光分析法检验陈旧骨骼和牙齿的vWA和LPL位点

为鉴定陈旧骨骼和牙齿的尸源 ,对DNA的提取方法进行了研究,并建立了vWA和LPL位点的自动荧光分析技术,该法简便、快速、有效、可对存放2、3、4、7年的陈旧骨骼和牙齿成功地进行检验,在检验中能直接确定样品的等位基因     

Is re-creation of human identity possible using tooth prints? An experimental study to aid in identification

Enamel in teeth, as documented is the hardest substance in the entire human body. Tooth prints are the enamel rod end patterns on the tooth surface. Could this fact be a boon in utilizing it for an individual's identification in the hands of forensic experts? A study has been attempted towards the same. These tooth prints were recorded for 60 extracted tooth specimens after acid etching using a cellophane tape. Subsequently a digital image of the print was obtained which was subjected to biometric conversion using Verifinger standard SDK version 6.0 software followed by the use of Automated Fingerprint Identification System (AFIS) software for comparison of the tooth prints. In results we observed that tooth prints were composed of varied patterns and sub-patterns. A comparison was made between the tooth prints of different and the same individual and also between different classes of teeth (incisors, canines, premolars and molars). None of the patterns exhibited intra- and inter-individual similarity. Nor did any particular class of the tooth could be preferentially used over the other for an individual identification. This field demands a need for further exploration towards the use of tooth prints for establishing an individual's identity.     

Immunohistochemical proof of amelogenin in teeth--a contribution to the evaluation of the age in the identification of unknown corpses

When a corpse is found that cannot be identified, one of the forensic tasks is to determine how old the person was when she or he died. To solve this frequently asked question in forensic practice, the enamel protein amelogenin was marked immunohistochemically in 249 extracted teeth. Amelogenin is already produced during prenatal development and is gradually used up throughout life into old age. Hence, the demonstrability decreases with age. The extent of the labelling can be quantified by measuring the mean optical density (MOD). While it is not possible to determine the age on the basis of the mean optical density measurements alone, logistic regression of the MOD together with dichotomisation of the teeth according to age allows statements as to whether the person was younger or older than 30 years.     

The diversity of dental patterns in the orthopantomography and its significance in human identification

The primary aim of this study was to evaluate the utility of orthopantomography for human identification. Three hundred orthopantomograms were randomly selected from those stored at Dental Hospital of Yonsei University in Seoul. Dental patterns observed in the orthopantomograms were converted into eight codes and their diversity was calculated. The diversity of dental patterns in the orthopantomogram data was 99.92% for full dentition and the diversity of mandible (99.28%) was slightly higher value than that of maxilla (98.22%). In the case of molars and premolars in both jaws, the diversity values ranged from 92.13% to 96.04%. It was founded that orthopantomography is a valuable means of personal identification not only based on full dentition but also when only the posterior teeth are available. The present study indicates that orthopantomography is excellent means of forensic identification.     

Studying survivors of near-lethal suicide attempts as a proxy for completed suicide in prisons

Sex determination is a necessary step in the investigation of unidentified human remains from a forensic context. Teeth, as one of the strongest tissues in the human body, can be used for this purpose. Most studies of sexual dimorphism in teeth are based on the traditional mesiodistal and buccolingual crown measurements. The purpose of this study is to examine the degree of sexual dimorphism in permanent molars of modern Greeks using crown and cervical diagonal diameters, and to evaluate their applicability in sex determination. A total of 344 permanent molars in 107 individuals (53 male and 54 female) from the Athens Collection were examined. Crown and cervical diagonal diameters of both maxillary and mandibular molars were measured. It was found that males have larger molars than females and in 19 out of 24 dimensions measured male molars exceeded female molars significantly (P<0.05). The most dimorphic molars are the maxillary second molar, and the mandibular second and first molars. Although other molars were also sexually dimorphic they did not have a statistically significant difference in all dimensions. Cervical diagonal diameters have found to be more sexually diamorphic than crown diagonal diameters. In discriminant function analysis the variables entered more frequently were the cervical diagonal diameters mainly of mandibular molars. Classification accuracy was found to be 93% for the total sample, 77.4% for upper jaw, and 88.4% for the lower jaw. Accuracy rates were higher for cervical than crown diagonal diameters. The data generated from the present study suggest that this metric method can be useful and reliable for sex determination, especially when the traditional dental measurements are not applicable.     

Accuracy of BR Regression Equation for Impacted Teeth in Age Estimation in Haryana Population of India

Abstract:  Dental cementum is a vital tissue that demonstrates continuous apposition throughout the life of a tooth. Positive correlation has been reported between the coronal displacement of cementum and age in impacted teeth and BR regression equation has been proposed. The present study is an attempt to calculate accuracy of BR regression equation in Haryana population of India. The study was conducted on 20 impacted mandibular third molars extracted from healthy patients, aged between 22 and 32 years. The bucco-lingual ground sections were prepared and the distance between the edges of enamel and cementum were measured with micrometer attached to a light microscope and BR regression equation was applied for age estimation. Although the estimated age with BR regression equation was less than chronological age it was not statistically significant. Hence, the BR regression equation can be used in medicolegal as well as in forensic odontology for age estimation.     

Methods for physical stabilization of ashed teeth in incinerated remains

Methods for physically stabilizing the extremely fragile ashed teeth that are often encountered in incinerated human remains were investigated. Results of a questionnaire sent to forensic anthropologists and forensic odontologists disclosed that, for these two groups, the most popular methods currently used are impregnation with a solution of polyvinyl acetate or application of cyanoacrylate cement, respectively. In addition, extracted human teeth were incinerated in the laboratory and impregnated with commercially available preparations of either cyanoacrylate cement, clear acrylic spray paint, hair spray, spray furniture varnish, clear fingernail polish, quick-setting epoxy cement, Duco household cement, polyvinyl acetate polymer in acetone, or self-curing clear dental acrylic resin. Every substance tested successfully stabilized the incinerated teeth. Clear acrylic spray paint was judged the most efficacious overall because of its ease of application, availability, inexpensiveness, and rapidity of setting.     

Forensic human identification: Investigation into tooth morphotype and DNA extraction methods from teeth

When a body is decomposed, hard tissues such as teeth may provide the only DNA source for human identification. There is currently no consensus as to the best DNA extraction method, and there is a lack of empirical data regarding tooth morphotype and condition that may impact DNA recovery. Therefore, this study sought to investigate which variables significantly improved DNA concentration, integrity and profiling success. A total of 52 human teeth were assessed, representing all tooth morphotypes from three deceased individuals. DNA was extracted using both the QIAamp® DNA Investigator Kit and the phenol-chloroform method. DNA concentration and degradation index were assessed using real time PCR, prior to conventional DNA profiling. Contrary to international guidelines promoting the use of molars, DNA profiling from molars was the least successful, with premolars, followed by canines, performing the best. The presence of fillings reduced the DNA quantity and quality obtained and may explain the poor performance of molars. DNA from the maxillae were significantly less degraded when the QIAamp® was used, although this did not influence DNA profiling success. A significant increase in DNA concentration, integrity and profiling success was observed in diseased teeth (periodontitis) compared to those without disease. This may be due to increased white blood cell presence at the site. There was no significant difference in DNA profiling success between the two DNA extraction methods. However, different teeth yielded failed DNA profiles for each extraction method, suggesting that repeated attempts, using alternative DNA extraction methods, is recommended. The recovery of additional DNA profiling information from degraded samples may help to ultimately reduce the burden of unidentified human remains.     

利用AutoMate Express~(TM)自动化法医DNA提取系统提取骨骼及牙齿DNA

目的探讨建立骨骼及牙齿DNA自动化提取的新方法。方法将33份骨骼及15份牙齿样本分别用冷冻研磨和手工处理两种方法研磨成粉,采用AutoMate ExpressTM自动化法医DNA提取系统提取DNA并定量。结果 AutoMate ExpressTM自动化法医DNA提取系统能够在3h左右完成骨骼、牙齿DNA的提取,两种方法处理的骨骼样本所得DNA质量浓度差异无统计学意义。冷冻研磨处理的骨骼和牙齿样本均获得了较好的STR分型结果,且牙齿样本所得DNA质量浓度高于手工提取所得。结论应用AutoMate ExpressTM自动化法医DNA提取系统是自动化提取骨骼、牙齿DNA的一种新方法,可应用于法医实际案件检验。     

The Potential of X‐Ray Diffraction in the Analysis of Burned Remains from Forensic Contexts*

Abstract: In view of the difficulties in extracting quantitative information from burned bone, we suggest a new and accurate method of determining the temperature and duration of burning of human remains in forensic contexts. Application of the powder X‐ray diffraction approach to a sample of human bone and teeth allowed their microstructural behavior, as a function of temperature (200–1000°C) and duration of burning (0, 18, 36, and 60 min), to be predicted. The experimental results from the 57 human bone sections and 12 molar teeth determined that the growth of hydroxylapatite crystallites is a direct and predictable function of the applied temperature, which follows a nonlinear logistic relationship. This will allow the forensic investigator to acquire useful information about the equilibrium temperature brought about by the burning process and to suggest a reasonable duration of fire exposure.     

Radiographic evaluation of teeth subjected to high temperatures: experimental study to aid identification processes

The radiographic evaluation of dental remains represents a significant aspect in the forensic identification process, particularly after an exposure to fire. The aim of this "in vitro" study was to evaluate the radiographic features of unrestored, endodontically treated and restored teeth after exposure to an experimental range of high temperatures. Ninety human teeth were divided into two groups: (1) unrestored teeth, as a control group and (2) teeth endodontically treated (condensation technique) and restored with amalgam or composite fillings. Before testing the high temperatures, periapical radiographs of all teeth were performed. The tests of exposure to heat were carried out in an oven for six different temperatures (200, 400, 600, 800, 1000 and 1100 degrees C (392, 752, 1112, 1472, 1832, 2012 degrees F)). After each exposure, periapical radiographs of all the teeth were taken. The radiographic appearance of all the teeth before and after the thermal stresses were evaluated and the differences were recorded. The results of the radiographic examination showed that a number of significant radiographic details were conserved: the composite fillings were in place maintaining the shape till 600 degrees C (1112 degrees F), the amalgam fillings were in place maintaining the shape till 1000 degrees C (1832 degrees F) and the endodontic treatments were recognisable till 1100 degrees C (2012 degrees F).     

Age estimation of unaccompanied minors. Part II. Dental aspects

Recent years have brought a worldwide increase in cross-border migration due to a globalized economy and ongoing belligerent conflicts. As a result, the percentage of foreigners among the general population has steadily increased not only in Germany, but also in other countries. This trend has triggered a growing demand for forensic medicine to assess the age of adolescents and young adults. The individuals examined here are unaccompanied minors without valid identification documents who do not know their age or else are suspected of not giving their correct age. The mineralization of third molars is the main criterion for dental age estimation of living subjects in the relevant age group. To date insufficient knowledge has been obtained about how ethnic origin can influence tooth mineralization. This, however, constitutes a restraint on the reliability of age estimates and hence on the forensic value of information essential to legal security. A comparative study was conducted to present comparative data on third molar mineralization in a Caucasoid, Mongoloid and African sample. In conclusion, forensic age estimates of living subjects would be more powerful tools if population-specific standards were applied to evaluations of wisdom tooth mineralization. Since the mineralization of third molars is usually completed by the age of 19 or 20 years, this feature cannot be used to ascertain whether a person has attained the forensically relevant age of 21 years. The question was whether determination based on an orthopantomogram of a combination of features relevant to dental age estimation of adults supplies forensically useful information for ascertaining whether a person has attained 21 years of age. The features considered include the DMFT index of all permanent teeth, the DMFT index of all permanent teeth excluding third molars and the DFT index of third molars projecting beyond the occlusal plane. It can be concluded that an evaluation of the variations of the DMFT index does not by itself yield sufficient data to determine with the accuracy required in criminal proceedings whether a person has attained 21 years of age. An additional X-ray examination of the medial clavicular epiphyseal cartilage is, therefore, strongly recommended when assessing whether a person is over 21.     

Study of Short- and Long-Term Storage of Teeth and Its Influence on DNA

Abstract:  DNA degradation can interfere with the resolution of forensic cases. Allelic dropout often reduces the opportunity for adequate comparisons between degraded and reference samples. This study analyzed DNA degradation in 24 extracted teeth after storage at room temperature for 0, 2, 5, and 10 years. DNA concentration, quantified by dot-blot hybridization, declined significantly for the first 2 years, but there was no significant further degradation from the second to the tenth year of storage. COfiler™ analysis was used and the allelic dropout ratio for the amelogenin locus relative to CSF1PO locus was also estimated. Statistically significant differences were found between fresh teeth and teeth from the 2- and 5-year groups but not from the 10-year group. Under our storage conditions most of the DNA degradation occurred during the first 2 years. Further research is needed to control for individual and external factors that could affect DNA.