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Age-at-death estimation is an important issue in forensic medicine and anthropology. Initially, methods relied on morphological criteria, but with the advancement of radiology, new techniques such as morphological studies on multi-slice computed tomography (CT) reconstructions have emerged. Recent studies have shown promising results by investigating the correlation between age and bone mineral density (BMD). However, there is currently a lack of data on post-mortem CTs (PMCT) involving decomposed bodies, and limited information exists regarding changes in Hounsfield Units measurement in a post-mortem context. In light of these gaps, our study aimed to examine the relationship between age at death and pubic and ilium BMD using a sample of forensic bodies. We also aimed to determine whether post-mortem processes, such as putrefaction, could interfere with this correlation. Our retrospective analysis encompassed 637 PMCTs conducted before medicolegal autopsies at Tours University Hospital. Utilizing simple and multiple linear regressions, we investigated the correlation between age and pubic and ilium BMD, as well as the relationship between BMD and the radiologic alteration index (RAI), a scale employed to quantify the degree of putrefaction. Our findings indicate promising outcomes in age-at-death estimation using pubic and/or ilium BMD for bodies exhibiting no or moderate decomposition (RAI < 80), particularly among individuals under 40 years old. However, for highly decomposed corpses (RAI ≥ 80), the presence of gas infiltration significantly influences the BMD of both the ilium and pubis. Consequently, we advocate for the incorporation of the RAI score into the age estimation equation to enhance the accuracy of our results in such cases. Further investigation involving a larger cohort of decomposed bodies could facilitate refinement and validation of our method within this specific population. [ABSTRACT FROM AUTHOR]

Necrophagous flies, particularly blowflies, serve as vital indicators in forensic entomology and ecological studies, contributing to minimum postmortem interval estimations and environmental monitoring. The study investigates variations in the predominant cuticular hydrocarbons (CHCs) viz. n-C25, n-C27, n-C28, and n-C29 of empty puparia of Calliphora vicina Robineau-Desvoidy, 1830, (Diptera: Calliphoridae) across diverse environmental conditions, including burial, above-ground and indoor settings, over 90 days. Notable trends include a significant decrease in n-C25 concentrations in buried and above-ground conditions over time, while n-C27 concentrations decline in buried and above-ground conditions but remain stable indoors. Burial conditions show significant declines in n-C27 and n-C29 concentrations over time, indicating environmental influences. Conversely, above-ground conditions exhibit uniform declines in all hydrocarbons. Indoor conditions remain relatively stable, with weak correlations between weathering time and CHC concentrations. Additionally, machine learning techniques, specifically Extreme Gradient Boosting (XGBoost), are employed for age estimation of empty puparia, yielding accurate predictions across different outdoor and indoor conditions. These findings highlight the subtle responses of CHC profiles to environmental stimuli, underscoring the importance of considering environmental factors in forensic entomology and ecological research. The study advances the understanding of insect remnant degradation processes and their forensic implications. Furthermore, integrating machine learning with entomological expertise offers standardized methodologies for age determination, enhancing the reliability of entomological evidence in legal contexts and paving the way for future research and development. [ABSTRACT FROM AUTHOR]

Cranial vault thickness (CVT) and its variations provide valuable insights into an individual's biological attributes such as age and sex. This paper aimed to assess the correlations of CVT with age and sex and develop a regression model for age estimation in Malaysian subadults using computed tomography (CT) images. A total of 521 CT images (male/female: 279/242; age range: 0–20 years; Malay/Chinese/Indian: 221/145/155) were included in the study. Correlations of CVT measurements with age and sex, including frontal bone thickness (FBT), occipital bone thickness (OBT), left parietal bone thickness (LPBT), and right parietal bone thickness (RPBT) were assessed and regression formulae were developed for age estimation in subadults. A significant correlation between CVT measurements and age was demonstrated (p < 0.001). Age estimation was most accurate in the younger age group (< 2 years) at frontal and occipital, and accuracy decreases in the older age groups. Additionally, sexual dimorphism was evident in the frontal and parietal bone thickness within the age range of 3–6 years and 16–20 years, respectively. In conclusion, the findings suggested CVT measurements could be used to corroborate other age estimation methods for subadults. [ABSTRACT FROM AUTHOR]

Introduction: Evaluation of the eruption of mandibular third molars in orthopantomograms (OPGs) is a method of forensic age assessment. The objective of our study was to provide valid reference data for this trait within a population of black South Africans. The study was guided by the criteria for reference studies in age assessment. Materials and methods: A study population from Pretoria, South Africa comprising 670 OPGs obtained from 338 black females and 332 black males aged between 15.00 and 25.97 years was analysed. All OPGs were performed for medical indication during the period from 2011 to 2022 and were retrospectively evaluated. From the 670 OPGs, a total of 1021 mandibular third molars were evaluated. The assessment of mandibular third molars was conducted using the staging scale presented by Olze et al. in 2012. Two experienced dentists evaluated the OPGs independently of each other. If the two examiners diverged in their assessments, a consensus stage was assigned. Results: As expected, the mean, median and minimal age increased with higher stages for both teeth and both sexes. The minimum age recorded for stage D, indicating complete tooth eruption, was 15.79 years in females and 16.62 years in males. Conclusion: As it is the case for previous reference studies in other countries, placing exclusive reliance on the evaluation of mandibular third molar eruption is inadequate for ascertaining the age of majority among Black South Africans. Future studies need to examine if our results are transferable to other countries in Sub-Saharan Africa. [ABSTRACT FROM AUTHOR]

Sex and chronological age estimation are crucial in forensic investigations and research on individual identification. Although manual methods for sex and age estimation have been proposed, these processes are labor-intensive, time-consuming, and error-prone. The purpose of this study was to estimate sex and chronological age from panoramic radiographs automatically and robustly using a multi-task deep learning network (ForensicNet). ForensicNet consists of a backbone and both sex and age attention branches to learn anatomical context features of sex and chronological age from panoramic radiographs and enables the multi-task estimation of sex and chronological age in an end-to-end manner. To mitigate bias in the data distribution, our dataset was built using 13,200 images with 100 images for each sex and age range of 15–80 years. The ForensicNet with EfficientNet-B3 exhibited superior estimation performance with mean absolute errors of 2.93 ± 2.61 years and a coefficient of determination of 0.957 for chronological age, and achieved accuracy, specificity, and sensitivity values of 0.992, 0.993, and 0.990, respectively, for sex prediction. The network demonstrated that the proposed sex and age attention branches with a convolutional block attention module significantly improved the estimation performance for both sex and chronological age from panoramic radiographs of elderly patients. Consequently, we expect that ForensicNet will contribute to the automatic and accurate estimation of both sex and chronological age from panoramic radiographs. [ABSTRACT FROM AUTHOR]

Age estimation methods through the metric study of bones in non-adult individuals are a frequently used tool in Biological and Forensic Anthropology; however, few methodological validation studies are available, given the limited number of adequate samples for their study. In this context, the aim of this work is to test the effectiveness of the methods of Fazekas and Kósa (1978), Saunders et al. (1993), Rissech and Black (2007) and Cardoso et al. (2017), for age estimation through the measurements of the scapula, in the non-adult osteological collection of the Anthropology Laboratory of the University of Granada (Spain), one of the most representative identified non-adult collections internationally. For this purpose, nine different measurements of the scapula were taken from a total of 149 non-adult individuals, applying the regression formulae proposed by the different authors and verifying their accuracy. Results have shown the existence of significant differences between the real age and the estimated age in almost all the tested methods, with degrees of error that differ from those obtained in the original study populations. Based on the obtained results, it is recommended to use the method of Cardoso et al. in postnatal individuals, always adjusting the degrees of error with those obtained in this work. [ABSTRACT FROM AUTHOR]

Various staging scales have been proposed for the assessment of the visibility of the periodontal ligament space of mandibular third molars on dental panoramic radiographs (PANs) for forensic age assessment in living individuals. However, up to now, there has been no systematic comparison between these staging scales available. We directly compared the 2010 staging scale proposed by Olze et al. with the 2017 staging by Lucas et al. and the 2020 staging by Guo et al. in a German study population. We evaluated 233 PANs from 115 females and 118 males aged 20.0 to 40.9 years using three independent examiners, with one examiner conducting two assessments. We examined the correlation between age and stage, as well as the inter- and intra-rater reliabilities. While the point estimates for the correlation coefficient and the reliability measures were lowest for the Guo scale and highest for the Olze scale, confidence intervals showed a large overlap, particularly for the scales of Olze et al. and Lucas et al. The correlation coefficients between stage and age were consistently lower in females than in males across all methods. In summary, we showed that the staging scales of Olze et al. and Lucas et al. were very similar. The Olze method showed higher point estimates across all analyses, and because there are more reference data available for this method, we argue that it should be preferred as the method of choice for further studies in the field. However, Guo method could be considered for instances, in which the inter-radicular periodontal ligament is not evaluable. [ABSTRACT FROM AUTHOR]

Age assessment of migrants is crucial, particularly for unaccompanied foreign minors, a population facing legal, social, and humanitarian challenges. Despite existing guidelines, there is no unified protocol in Europe for age assessment. The Forensic Anthropology Society of Europe (FASE) conducted a comprehensive questionnaire to understand age estimation practices in Europe. The questionnaire had sections focusing on the professional background of respondents, annual assessment numbers, requesting parties and reasons, types of examinations conducted (e.g., physical, radiological), followed protocols, age estimation methods, and questions on how age estimates are reported. The questionnaire's findings reveal extensive engagement of the forensic community in age assessment in the living, emphasizing multidisciplinary approaches. However, there seems to be an incomplete appreciation of AGFAD guidelines. Commonalities exist in examination methodologies and imaging tests. However, discrepancies emerged among respondents regarding sexual maturity assessment and reporting assessment results. Given the increasing importance of age assessment, especially for migrant child protection, the study stresses the need for a unified protocol across European countries. This can only be achieved if EU Member States wholeheartedly embrace the fundamental principles outlined in EU Directives and conduct medical age assessments aligned with recognized standards such as the AGFAD guidelines. [ABSTRACT FROM AUTHOR]

Insects, especially blow flies, are forensically relevant to determine the minimal postmortem interval (PMImin), based on the fact that they are usually the first colonisers of a body. By estimating the age of immature blow flies, interferences can be made about the time since death. Whilst morphological parameters are valuable for age estimation of blow fly larvae, gene expression profiling is more applicable for blow fly pupae. Here, the age-dependent changes in the gene expression levels during the development are analysed. 28 temperature-independent markers have already been described for the age estimation of pupae of the forensically important blow fly Calliphora vicina and are analysed by RT-qPCR. To allow simultaneous analysis of these age markers, a multiplex assay was developed in the present study. After reverse transcription, the markers are analysed simultaneously in an endpoint PCR and subsequently separated by capillary electrophoresis (CE). This method is highly attractive due to its quick and easy procedure and interpretation. The present age prediction tool was adapted and validated. The multiplex PCR assay reproduced the same expression profiles as the RT-qPCR assay based on the same markers. The statistical evaluation shows that the new assay has a lower precision but a better trueness for age determination compared to the RT-qPCR assay. Since the new assay is also qualified to estimate the age of C. vicina pupae and is practical, cost-effective and, even more importantly, time-saving, it is attractive for use in forensic casework. [ABSTRACT FROM AUTHOR]

Purpose: To investigate prediction of age older than 18 years in sub-adults using tooth tissue volumes from MRI segmentation of the entire 1st and 2nd molars, and to establish a model for combining information from two different molars. Materials and methods: We acquired T2 weighted MRIs of 99 volunteers with a 1.5-T scanner. Segmentation was performed using SliceOmatic (Tomovision©). Linear regression was used to analyse the association between mathematical transformation outcomes of tissue volumes, age, and sex. Performance of different outcomes and tooth combinations were assessed based on the p-value of the age variable, common, or separate for each sex, depending on the selected model. The predictive probability of being older than 18 years was obtained by a Bayesian approach using information from the 1st and 2nd molars both separately and combined. Results: 1st molars from 87 participants, and 2nd molars from 93 participants were included. The age range was 14-24 years with a median age of 18 years. The transformation outcome (high signal soft tissue + low signal soft tissue)/total had the strongest statistical association with age for the lower right 1st (p= 7.1*10-4 for males) and 2nd molar (p=9.44×10-7 for males and p=7.4×10-10 for females). Combining the lower right 1st and 2nd molar in males did not increase the prediction performance compared to using the best tooth alone. Conclusion: MRI segmentation of the lower right 1st and 2nd molar might prove useful in the prediction of age older than 18 years in sub-adults. We provided a statistical framework to combine the information from two molars. [ABSTRACT FROM AUTHOR]

This study proposes an assessment of the accuracy of the Fazekas and Kósa and Nagaoka methods by measuring the squamosal and petrous portions of the temporal bone, whose application in the Mediterranean population is not recommended. Therefore, our proposal is a new formula to estimate the age of skeletal remains from individuals at 5 months gestational age to 1.5 postnatal years with the temporal bone. The proposed equation was calculated on a Mediterranean sample identified from the cemetery of San José, Granada (n = 109). The mathematical model used is the exponential regression of the estimated age for each measure and sex, and both in combination, using an inverse calibration and cross-validation model. In addition, the estimation errors and the percentage of individuals within a 95% confidence interval were calculated. The lateral development of the skull, especially the growth of the length of the petrous portion, showed the greatest accuracy, while its counterpart, the width of the pars petrosa, showed the lowest accuracy, so its use is discouraged. The positive results from this paper should be useful in both forensic and bioarchaeological contexts. [ABSTRACT FROM AUTHOR]

In forensic investigations, age estimation is vital for determining whether a suspect is under or over the legally defined adult age. With breakthroughs in RNA sequencing technology, small noncoding RNAs have provided new ways to solve problems related to the age estimation of trace or aged samples, owing to their small molecular weight and better stability. In our previous study, we had applied miRNAs for the age estimation of bloodstains; however, further improvement of the existing model is needed. PIWI-interacting RNAs (PiRNAs), which are 24–32 nt noncoding small RNA molecules involved in the PIWI-piRNA pathway, play an important role in the aging process. In this study, we explored the possibility of simultaneously analyzing piRNAs and miRNAs for better age estimation purpose. Through massively parallel sequencing, five age-related piRNAs were identified in blood samples that had been stored for eight years. Further real-time PCR analysis revealed that two piRNAs (piR-000753 and piR-020548) showed relatively higher efficiency in age estimation. Additionally, two age-related miRNAs (miR-324-3p and miR-330-5p) were used to build the estimation model. Among all algorithms tested, gradient boosting showed the lowest mean absolute error (MAE) and root mean square error (RMSE) values (3.171 and 4.403 years, respectively) for the validation dataset (n = 110). The errors of the model were less than 5 years and 10 years for 81.82% and 96.36% of the samples, respectively. The results suggest that the combined use of piRNA and miRNA markers may increase the accuracy of age estimation, and our new model has great potential for application in forensic casework. [ABSTRACT FROM AUTHOR]

In forensic medicine practice, age estimation—both in living and deceased individuals—can be requested due to legal requirements. Radiologic methods, such as X-rays, for the estimation of bone age have been discussed, and ethical concerns have been raised. Given these factors, radiologic methods that reduce radiation exposure have gained importance and have become one of the research topics in forensic medicine. In this study, the MR images of the ankles of patients aged between 8 and 25 years, obtained with a 3.0 T MR scanner, were evaluated retrospectively according to the staging method defined by Vieth et al. In the study, the ankle MR images of 201 cases (83 females and 118 males) with sagittal T1-weighted turbo spin echo and T2-weighted short tau inversion recovery sequences were evaluated independently by two observers. According to the results of our study, the intra- and inter-observer agreements are at a very good level for both the distal tibial and calcaneal epiphyses. All the cases detected as stages 2, 3, and 4 in both sexes for both the distal tibial and the calcaneal epiphyses have been determined to be under the age of 18 years. According to the data obtained from our study, we consider that stage 5 for males and stage 6 for both sexes in the distal tibial epiphysis and stage 6 for males in the calcaneal epiphysis can be used to estimate the age of 15 years. As far as we know, our study is the first to evaluate ankle MR images with the method defined by Vieth et al. Further studies should be conducted to evaluate the validity of the procedure. [ABSTRACT FROM AUTHOR]

This article is a retrospective analysis of postmortem computed tomography scans of ossification stages of the anterior and posterior intra-occipital sutures, the anterior arch of the atlas, and the neurocentral junction of the axis. We also analyzed the development of secondary ossification centers in the proximal humeral, femoral, and tibial epiphyses, and the distal femoral and tibial epiphyses. Additionally, the development of primary ossification centers in the wrist and metacarpals, and maxillary and mandibular deciduous tooth maturation. A total of 58 cadavers (35 males, 23 females), whose age ranged from 3rd month of pregnancy to 14 years, were analyzed. The results of this study show that analysis of synchondrosis closure, primary, and secondary ossification center development and deciduous tooth changes are a good tool for age estimation in subadults group (fetuses, newborns, infants, and children). The results of the study in a Polish population are consistent with those reported by other authors. [ABSTRACT FROM AUTHOR]

This study sought to develop an age-estimation formula to evaluate the extent of median palatine suture (MP) closure using postmortem computed tomographic (PMCT) images. The PMCT images of 634 Japanese subjects (mean age, 54.5 years; standard deviation [SD], 23.2 years) with known age and sex were examined. The degree of suture closure of the MP, anterior median palatine suture (AMP), and posterior median palatine suture (PMP) was measured and scored (suture closure score, SCS), and a single linear regression analysis was conducted with age at death. On the analysis, SCS of MP, AMP, and PMP showed a significant correlation with age (p < 0.001). The correlation coefficient of MP was higher (0.760, male; 0.803, female; and 0.779, total) than that of AMP (0.726, male; 0.745, female; and 0.735, total) or PMP (0.457, male; 0.630, female; and 0.549, total). The regression formula and standard error of estimation (SEE) of MP were calculated as Age = 100.95 × SCS + 20.51 (SEE 14.87 years) for male subjects, Age = 91.93 × SCS + 26.65 (SEE 14.12 years) for female subjects, and Age = 95.17 × SCS + 24.09 (SEE 14.59 years) for the total, respectively. In addition, another 50 Japanese subjects were randomly selected to validate the age-estimation formula. In this validation, the actual age of 36 subjects (72%) was within the estimated age ± SEE. This study showed that the age estimation formula using PMCT images of MPs was potentially useful for estimating the age of unidentified corpses. [ABSTRACT FROM AUTHOR]

When investigating a death, post-mortem identification provides with results of great legal and humanitarian significance. The effectiveness of the methods used to estimate age depends on the reference population, considering variables such as sex and ancestry. The aim of this study was to validate the Iscan method to estimate age in a Spanish forensic population, comparing the estimates obtained in dry bones and 3D reconstructions created with a surface scanner. We carried out a cross-sectional study on 109 autopsied corpses (67% male), scanning the sternal end of the right fourth rib in a 3D mesh, using an EinScan-Pro® surface scanner (precision: 0.05 mm). Two observers estimated the phases in dry bones and 3D images according to the Iscan method and to the sex of the subject. The mean age was 57.73 years (SD = 19.12 years;18–93 years). The intra-observer agreement was almost perfect in bones (κ = 0.877–0.960) and 3D images (κ = 0.954), while the inter-observer agreement was almost perfect in bones (κ = 0.813) and substantial in 3D images (κ = 0.727). The correlation with the Iscan phases was very strong in bones (Rho = 0.794–0.820; p < 0.001) and strong in 3D images (Rho = 0.690–0.691; p < 0.001). Both sex-adjusted linear regression models were significant (dry bones: R2 = 0.65; SEE = ± 11.264 years; 3D images: R2 = 0.50; SEE = ± 13.537 years) from phase 4 onwards. An overestimation of age was observed in the first phases, and an underestimation in the later ones. Virtual analysis using a surface scanner in the fourth rib is a valid means of estimating age. However, the error values and confidence intervals were considerable, so the joint use of different methods and anatomical sites is recommended. [ABSTRACT FROM AUTHOR]

Purpose: Our aim was to investigate tissue volumes measured by MRI segmentation of the entire 3rd molar for prediction of a sub-adult being older than 18 years. Material and method: We used a 1.5-T MR scanner with a customized high-resolution single T2 sequence acquisition with 0.37 mm iso-voxels. Two dental cotton rolls drawn with water stabilized the bite and delineated teeth from oral air. Segmentation of the different tooth tissue volumes was performed using SliceOmatic (Tomovision©). Linear regression was used to analyze the association between mathematical transformation outcomes of the tissue volumes, age, and sex. Performance of different transformation outcomes and tooth combinations were assessed based on the p value of the age variable, combined or separated for each sex depending on the selected model. The predictive probability of being older than 18 years was obtained by a Bayesian approach. Results: We included 67 volunteers (F/M: 45/22), range 14–24 years, median age 18 years. The transformation outcome (pulp + predentine)/total volume for upper 3rd molars had the strongest association with age (p = 3.4 × 10−9). Conclusion: MRI segmentation of tooth tissue volumes might prove useful in the prediction of age older than 18 years in sub-adults. [ABSTRACT FROM AUTHOR]

Age-at-death estimation is of great relevance for the identification of unknown deceased individuals. In skeletonised corpses, teeth and bones are theoretically available for age estimation, but in many cases, only single bones or even only bone fragments are available for examination. In these cases, conventional morphological methods may not be applicable, and the application of molecular methods may be considered. Protein-based molecular methods based on the D-aspartic acid (D-Asp) or pentosidine (Pen) content have already been successfully applied to bone samples. However, the impact of the analysed type of bone has not yet been systematically investigated, and it is still unclear whether data from samples of one skeletal region (e.g. skull) can also be used for age estimation for samples of other regions (e.g. femur). To address this question, D-Asp and Pen were analysed in bone samples from three skeletal regions (skull, clavicle, and rib), each from the same individual. Differences between the bone types were tested by t-test, and correlation coefficients (ρ) were calculated according to Spearman. In all types of bone, an age-dependent accumulation of D-Asp and Pen was observed. However, both parameters (D-Asp and Pen) exhibited significant differences between bone samples from different anatomical regions. These differences can be explained by differences in structure and metabolism in the examined bone types and have to be addressed in age estimation based on D-Asp and Pen. In future studies, bone type-specific training and test data have to be collected, and bone type-specific models have to be established. [ABSTRACT FROM AUTHOR]

Next-generation sequencing and single-cell RNA sequencing (scRNA-seq) technologies have advanced rapidly in recent years. scRNA-seq reveals the unique gene expression of each cell type, providing directions for exploring cell heterogeneity, cell type–specific responses to injury/disease, and the mechanisms underlying these processes. The development of sequencing technology and improved sequencing throughput have brought about a revolution in single-cell transcriptome study, bringing great benefits to the fields of medicine and biomedical science. From our perspective, certain issues in forensic medicine may potentially be addressed using single-cell transcriptome studies; however, this powerful technique has not yet attracted sufficient attention in forensic medicine-associated research. Therefore, examining and reviewing the latest developments and applications of single-cell transcriptome studies, we present our views on the future directions of forensic research using this technology, aiming to expand the frontiers of forensic science. [ABSTRACT FROM AUTHOR]

The acetabulum presents as a well-preserved evidence, resistant to taphonomic degradation changes and can thus aid in the age estimation process. A CT-based examination of the acetabulum can further help simplify the process of age estimation by overcoming the time-consuming process of maceration and by doing away with the interference resulting from tissue remnants. The aim of the present study was to evaluate the role of the acetabulum for age estimation in an Indian population through a CT-based examination, using principal component analysis and regression models. CT images of 400 individuals aged 10 years and above were evaluated according to the features defined in the San-Millán-Rissech method of age estimation. Five of the seven morphological features defined by San-Millán-Rissech were appreciable on CT scans, and, to enable further statistical analysis, a cumulative score was computed using these five features. A significant correlation of 0.835 and 0.830 for the right and left acetabulum, respectively, was obtained between computed cumulative scores and chronological age of individuals. No significant sex differences were observed in the scoring of different age-related morphological changes. Regression models were generated using individual features and cumulative scores. Regression models derived using the cumulative score yielded inaccuracy values of 9.67 years for the right acetabulum and 9.15 years for the left acetabulum. Inaccuracy and bias values were computed for each individual feature, as well as for each decade, using mean point ages established within the original study. Amongst the various features, acetabular rim porosity was seen to have the lowest values of inaccuracy (11.50 years) and bias (2.32 years) and activity on outer edge of acetabular fossa the highest (inaccuracy and bias values of 22.36 years and 21.50 years, respectively). Taking into consideration this differential contribution towards age estimation, weighted coefficients and mean point ages for different morphological features were determined using principal component analysis. Subsequently, summary age models were generated from the obtained weighted coefficients and mean age values. Summary age models derived in the present study yield lower estimates of inaccuracy of 7.60 years for the right acetabulum and 7.82 years for the left acetabulum. While regression models derived in the present study allow for age estimation using even a single appreciable feature, summary age models take into account the contribution of each feature and generate more accurate estimates of age. Both statistical computations yield reduced error rates and thus can render greater applicability to the acetabulum in forensic age estimation. [ABSTRACT FROM AUTHOR]

Numerous intrinsic and extrinsic factors influence bone remodelling rates and have shown to affect the accuracy of histological aging methods. The present study investigates the rib cortex from two Mediterranean skeletal collections exploring the development of population-specific standards for histomorphometric age-at-death estimation. Eighty-eight standard ribs from two samples, Cretans and Greek-Cypriots, were processed histologically. Thirteen raw and composite histomorphometric parameters were assessed and observer error tested. The correlation between age and the parameters and the differences between sex and population subsamples were explored through group comparisons and analysis of covariance. General linear models assessed through data fit indicators and cross-validation were generated from the total dataset, and by sex and population subsamples. Most of the histological variables showed a statistically significant correlation with age with some differences observed by sex and by sample. From the twelve models generated, the optimal model for the whole sample included osteon population density (OPD), osteon perimeter, and osteon circularity producing an error of 10.71 years. When sex and samples were separated, the best model selected included OPD and osteon perimeter producing an error of 8.07 years for Greek-Cypriots. This research demonstrates the feasibility of quantitative bone histology to estimate age, obtaining errors rates in accordance with macroscopic ageing techniques. Sex and sample population differences need further investigation and inter-population variation in remodelling rates is suggested. Moreover, this study contributes to the creation of population-specific standards for Cretans and Greek-Cypriots. [ABSTRACT FROM AUTHOR]

Identification of the living and the dead individual is essential in routine forensic dental examinations. Age determination can be of great value in forensic odontology, not only in identifying bodies but also in relation to crime. When subjects have extensive changes that external features provide no information, teeth are often the only means of identification. Several procedures for age-at-death estimation in adults have been introduced. Two of them, cementum annulation and dentin translucency, are frequently used as a single dental indicator. Cementum annulation refers to an alternating dark and light band; each pair of it represents 1 year. Meanwhile, dentin translucency is the other dental physiological process that begins in the second or third decade of life and progresses with age. There are still few studies that compared both methods and their accuracy in estimating adult age at death. Therefore, this study aims to test and compare cementum annulation and dentin translucency accuracy by performing a systematic search on five online databases (Pubmed, Scopus, Ebsco, ScienceDirect, and Wiley). All the research articles must be published in the last 10 years, and the full paper must be available in English. Out of the total 1178 literature, 28 studies were recruited for qualitative analysis and 23 studies for meta-analysis. The results show that dentin translucency age estimation is more accurate than the cementum annulation method in the entire population. It is recommended to use the cementum annulation method for younger adults (15–44 years) and the dentin translucency method for the older ones (≥ 45 years). [ABSTRACT FROM AUTHOR]

Age estimation is an essential element in the field of forensics and is also of judicial interest in forensic medicine of the living. Despite all the methods, age estimation is often imprecise with many biases, especially in mature individuals. The main objective of our study is to assess the correlation between age and computed tomography bone mineral density of the medial end of the clavicle. A sample of 180 chest multi-slice computed tomography scans without contrast injection performed in living individuals, with suspected respiratory infection, aged 15 to 100 years was used. Bone density measurement was performed on the medial meta-epiphyseal region of the clavicles. A significant negative correlation was found between bone density and age of individuals (p-value < 0.05). The mean absolute error was calculated for men at 13.4 years and 13.1 years for women, which was associated with an absence of bias. Good precision of the estimate for both sexes was also calculated on a subgroup of individuals whose age was greater than 40 years, with average absolute errors of about 12 years, which was associated with an absence of relative error. In view of our preliminary results, the study of bone density of the medial end of the clavicle could be of interest in the estimation of age. Several studies will be necessary to evaluate the reproducibility of these results on independent bone samples and in the estimation of age in the living individual. [ABSTRACT FROM AUTHOR]

Forensic entomology is primarily concerned with the estimation of time since death and involves determination of the age of immature insects colonising decomposing remains. Accurate age determination of puparia is usually accomplished by dissection, which means destructive sampling of evidence. As part of improving abilities to correctly identify species and developmental age, it is highly desirable to have available non-destructive methods. In this study, we acquired external hyperspectral imaging (HSI) data (77 spectral bands, 389-892 nm) from the dorsal and ventral sides of individual puparia of two species of blowfly (Diptera: Calliphoridae), Calliphora dubia Macquart 1855 and Chrysomya rufifacies Macquart 1842. Puparia were dissected to determine the presence/absence of eight internal morphological development characteristics (legs, wings, labella, abdominal segments, antennae, thoracic bristles, orbital/facial bristles and eye colour and arista). Based on linear discriminant analysis and independent validation of HSI data, reflectance features from puparia could be used to successfully (1) distinguish the two species (classification accuracy = 92.5 %), (2) differentiate dorsal and ventral sides of puparia (classification accuracy C. dubia = 81.5 %; Ch. rufifacies = 89.2 %) and (3) predict the presence of these morphological characteristics and therefore the developmental stage of puparia (average classification accuracy using dorsal imaging: C. dubia = 90.3 %; Ch. rufifacies = 94.0 %). The analytical approach presented here provides proof of concept for a direct puparial age relationship (i.e. days since the onset of pupation) between external puparial reflectance features and internal morphological development. Furthermore, this approach establishes the potential for further refinement by using a non-invasive technique to determine the age and developmental stage of blowflies of forensic importance.

Age estimation is a key factor for identification procedure in forensic context. Based on anthropological findings, degenerative changes of the sternal extremity of the 4th rib are currently used for age estimation. These have been adapted to post-mortem computed tomography (PMCT). The aim of this study was to validate a post-mortem computed tomography method based on a revision of the Iscan's method on a French sample. A total of 250 PMCT (aged from 18–98 years (IQR 36–68 years, median 51 years); 68 (27%) females) from the Medicolegal Institute of Paris (MLIP) were analyzed by two radiologists. The sternal extremity of 4th right rib was scored using method adapted from Iscan et al. Weighted κ was used to evaluate intra- and inter-observer reliability and Spearman correlation was performed to evaluate relationship between age and score. Confidence intervals for individual prediction of age based on 4th rib score and sex were computed with bootstrapping. The intra-observer reliability and inter-observer reliability were almost perfect (weighted κ = 0.85 [95%CI: 0.78–0.93] and 0.82 [95%CI 0.70–0.96] respectively). We confirmed a high correlation between the 4th rib score and subject age (rho = 0.72, p < 0.001), although the confidence intervals for individual age prediction were large, spanning over several decades. This study confirms the high reliability of Iscan method applied to PMCT for age estimation, although future multimodal age prediction techniques may help reducing the span of confidence intervals for individual age estimation. Trial registration: INDS 0,509,211,020, October 2020, retrospectively registered. [ABSTRACT FROM AUTHOR]

Objectives: To assess the performance of knee MRI for forensic age prediction and classification for 12-, 14-, 16-, and 18-year thresholds. Methods: The ossification stages of distal femoral epiphyses and proximal tibial epiphyses were assessed using an integrated staging system by Schmeling et al. and Kellinghaus et al. for knee 3.0T MRI with T1-weighted turbo spin-echo (T1-TSE) in sagittal orientation among 852 Chinese Han individuals (483 males and 369 females) aged 7–30 years. Regression models for age prediction were constructed and their performances were evaluated based on mean absolute deviation (MAD) values. In addition, the performances of age classification were assessed using receiver operating characteristic (ROC) analyses. Results: The intra- and inter-observer agreement levels were very good (κ > 0.80). The complete fusion of those two types of epiphyses took place before 18.0 years in our study participants. The minimum MAD values were 2.51 years (distal femur) and 2.69 years (proximal tibia) in males, and 2.75 years (distal femur) and 2.87 years (proximal tibia) in females. The specificity values of constructed prediction models were all above 90% for the 12-, 14-, and 16-year thresholds, compared to the 74.8–84.6% for the 18-year threshold. Better performances of age prediction and classification were observed in males by distal femoral epiphyses. Conclusions: Ossification stages via 3.0T MRI of the knee with T1-TSE sequence using an integrated staging system could be a reliable noninvasive method for age prediction or for age classification for 12-, 14-, and 16-year thresholds, especially in males by distal femoral epiphyses. However, assessments based on the full bony fusion of the distal femoral epiphysis and proximal tibial epiphysis seemed not reliable for age classification for the 18-year threshold in the Chinese Han population. [ABSTRACT FROM AUTHOR]

Objectives: To validate the fitness of the age estimation model in Black Americans, which was previously and solely established for the Chinese population based on pulp chamber volume of the first molars from cone beam computed tomography (CBCT), and to establish a new age estimation model for Black Americans. Materials and methods: A total of 203 subjects with CBCT scans, including 119 Chinese and 84 Black Americans, were retrospectively identified. The age range of subjects was between 11 and 87 years. For both populations, automated 3D pulp chamber segmentation of the first molars was performed by deep learning, followed by volume calculation and age estimation by a logarithmic regression model, which was established in a prior study solely on Chinese population. Additionally, a separate logarithmic regression analysis was carried out on Black Americans. The performance of age estimation was assessed by the mean absolute error (MAE), root mean square error (RMSE), Wilcoxon signed rank test, and coefficient of determination (R2) between the actual and estimated human ages. Results: When applying the age estimation model established in the prior study, MAE = 7.994 years and RMSE = 10.065 years were observed in the Chinese population, while MAE = 14.049 years and RMSE = 17.866 years were observed in Black Americans. The new age estimation model established for Black Americans was AGE = 89.752 − 21.176 × lnV (V = pulp chamber volume), with MAE = 7.930 years, RMSE = 10.664 years, and coefficient of determination (R2) = 0.600. Conclusions: Population-specific age estimation is needed when applied in Black Americans and Chinese people based on pulp chamber volume of the first molars from CBCT. [ABSTRACT FROM AUTHOR]

Assessment of third molar mineralization is a basic forensic odontological method available for age estimation in children and adolescents. Due to their protracted development into the late adolescence, their development has become the subject of interest and acquired great forensic importance. It is clearly evident in the literature that impaction factor has an influence on the rate of maturation of the third molars. In this regard, the present study was undertaken to test the relationship between impaction and its concomitant effect on the mineralization of the third molar and forensic age estimation. A total of 520 digital orthopantomograms (OPG) of 260 males and 260 females of south Indian origin were assessed retrospectively. Maturation of the third molars (I3M) was assessed using the method of Cameriere et al. (Int J Leg Med 122 (6):493–497, 2008). Impaction status was determined according to the position of lower right and left third molars in relation to the long axis of the second molars. Out of the total sample, 68.1% of lower left third molars and 71.2% of lower right molars were impacted. Statistical measures for I3M were calculated in the males and females for both impacted and non-impacted third molars. It was ascertained that the mean age and I3M values were higher in impacted third molars, indicating the slower rate of maturation than non-impacted ones. The sensitivity and specificity of the test (I3M < 0.08) was 96.2% and 72.4% for non-impacted third molars and 46.2% and 87% for impacted third molars, respectively. Our findings indicate that the cut-off value of I3M < 0.08 resulted in greater number of age misclassifications (false negatives) for impacted molars, highlighting the need for new cut-off value of I3M to meet the legal standards. It was concluded that the mineralization of impacted third molars was slower compared to non-impacted ones in south Indian males and females. Further studies are warranted to validate these findings among a larger and more diverse sample. [ABSTRACT FROM AUTHOR]

Introduction: The estimation of age-at-death of unidentified cadavers is a central aspect of the identification process. With increasing age, the incidence of glomerulosclerosis and the thickness of the carotid wall have been observed to also increase. This correlation has been demonstrated in various international histological studies. The aim of our study was to assess whether these correlations also apply to a Western European population. Methodology: In this retrospective observational study, kidney and common carotid artery samples from 216 cases autopsied at the Institute of Legal Medicine at the Justus-Liebig University in Giessen, Germany, were examined. Only cases with available tissue samples from both body sides were included. Exclusion criteria were poor sample quality and an age younger than 21 years. After histological processing, the tissue samples were assessed and digitally evaluated. Regression and classification analyses were used to investigate the correlation between age-at-death and intima-media thickness and age-at-death and the incidence of renal glomerular sclerosis. Results: Of the 216 autopsy cases, 183 were included for evaluation. Analysis of the carotid artery segments showed a strong correlation (Pearson correlation coefficient r = 0.887) between the intima-media-complex thickness and chronological age. Classification of the glomerulosclerotic incidence showed a correlation of 37.7–43.1% with the predicted age group. Discussion: Both the intima-media thickness and the proportion of sclerotic glomeruli can be used to estimate age in Western European cadavers. On the basis of these results, both methods are suited to supplement other already established methods for age-at-death estimation in the identification of an unknown cadaver. [ABSTRACT FROM AUTHOR]

Third molars are one of the few biological markers available for age estimation in juveniles, especially for the estimation of the 18-year-age threshold. Literature has indicated that impaction has an influence on the development of third molars, which could possibly result in age misclassifications. The present study is intended to identify an alternative cut-off value of the third molar maturity index (I3M) in impacted mandibular third molars and also to evaluate its applicability in estimating the major (≥ 18 years)/minor (< 18 years) status. A total of 1330 digital orthopantomograms (OPGs) of 665 male and 665 female south Indian adolescents aged from 15 to 22 years were collected and assessed. Eight hundred forty OPGs (63.1%) represented test sample, and 490 OPGs (36.9%) represented validation sample. I3M was measured for the total sample. Logistic regression, receiver operating characteristic (ROC) curve analysis, and Youden's index were used to test the performance of the method in the test sample. An alternative cut-off value of I3M < 0.17 was established for the highest value of the Youden's index of 0.598 for both sexes. When tested in validation sample, it has resulted in sensitivity and specificity values of 0.91 (95% CI; 0.86–0.95) and 0.90 (95% CI; 0.78–0.91) in males and 0.86 (95% CI; 0.80–0.92) and 0.90 (95% CI; 0.83–0.95) in females. In conclusion, cut-off value of I3M < 0.17 could accurately discriminate adults from minors with impacted mandibular third molars. However, more work is needed to be done among a more diverse sample to confirm these findings. [ABSTRACT FROM AUTHOR]

Estimation of age at death is important in forensic investigations of unknown remains. There have been several reports on applying the degree of osteophyte formation—an age-related change in the vertebral body—for age estimation; however, this method is not yet established. This study investigated a method for age estimation of modern Japanese individuals using osteophytes measured on CT images. The sample included 250 cadavers (125 males) aged 20–95 years. The degree of osteophyte formation was evaluated as score O (0–5 points), and the degree of fusion of the osteophytes between the upper and lower vertebrae was evaluated as score B (0–2 points). Age estimation equations were developed using regression analyses with seven variables, determined by scores O and B, and the equation with the smallest standard error of estimate (SEE) was obtained when the number of vertebrae with score O ≥ 2 was used as the explanatory variable. Age estimation with SEE of about 10 years was possible even when partial vertebrae with a high degree of osteophyte formation were used, showing its potential for practical application. The cutoff value for age estimation was established using the receiver operating characteristic curve analysis, wherein good results were obtained for all variables (area under the curve ≥ 0.8). The combination of the estimation equation and the cutoff value can narrow the range of age estimates. [ABSTRACT FROM AUTHOR]

Bloodstains are frequently encountered at crime scenes and they provide important evidence about the incident, such as information about the victim or suspect and the time of death or other events. Efforts have been made to identify the age of the bloodstain's donor through genomic approaches, but there are some limitations, such as the availability of databases and the quality dependence of DNA. There is a need for the development of a tool that can obtain information at once from a small blood sample. The aim of this study is to identify bloodstain metabolite candidates that can be used to determine donor age. We prepared bloodstain samples and analyzed metabolites using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Eighteen molecular features (MFs) were selected as candidates using volcano plots and multivariate analysis. Based on the MS/MS spectrum of the MFs, the following nine metabolites were identified from the METaboliteLINk database: Δ2-cis eicosenoic acid, ergothioneine, adenosine 5′-monophosphate, benzaldehyde, phenacylamine, myristic acid ethyl ester, p-coumaric acid, niacinamide, and N-arachidonoyl-l-alanine. These nine age markers at high or low abundances could be used to estimate the age of a bloodstain's donor. This study was the first to develop metabolite age markers that can be used to analyze crime scene bloodstains. [ABSTRACT FROM AUTHOR]

Estimating the age of the developmental stages of the blow fly Calliphora vicina (Diptera: Calliphoridae) is of forensic relevance for the determination of the minimum post-mortem interval (PMImin). Fly eggs and larvae can be aged using anatomical and morphological characters and their modification during development. However, such methods can only hardly be applied for aging fly pupae. Previous study described age estimation of C. vicina pupae using gene expression, but just when reared at constant temperatures, but fluctuating temperatures represent a more realistic scenario at a crime scene. Therefore, age-dependent gene expression of C. vicina pupae were compared at 3 fluctuating and 3 constant temperatures, the latter representing the mean values of the fluctuating profiles. The chosen marker genes showed uniform expression patterns during metamorphosis of C. vicina pupae bred at different temperature conditions (constant or fluctuating) but the same mean temperature (e.g. constant 10 °C vs. fluctuating 5–15 °C). We present an R-based statistical tool, which enables estimation of the age of the examined pupa based on the analysed gene expression data. [ABSTRACT FROM AUTHOR]

Forensic age estimation in living individuals is mainly based on radiological features, but direct radiography and computed tomography lead to a rise in ethical concerns due to radiation exposure. Thus, the contribution of magnetic resonance imaging (MRI) to age estimation of living individuals is a subject of ongoing research. In the current study, MRIs of shoulder were retrospectively collected from a modern Chinese Han population and data from 835 individuals (599 males and 236 females) in the age group 12 to 30 years were obtained. A staging technique based on (Schmidt et al. Int J Legal Med 121(4):321-324, 2007) and (Kellinghaus et al. Int J Legal Med 124(4):321–325, 2010) was used and all images were evaluated with T1-wieghted turbo spin echo (T1-TSE) sequence and T2-weighed fat suppression (T2-FS) sequence. One-sided images were assessed because data from both sides were considered coincidental, as no significant differences were found (P > 0.05). Two MRI sequences were evaluated separately and subsequently compared. Regression models and supportive vector classification (SVC) models were established accordingly. The intraobserver and interobserver agreement levels were good. Compared with T1-TSE sequence, the R2 values of T2-FS sequence were generally higher, while the mean absolute deviation (MAD) values were slightly lower. For T2-FS sequence, the MAD value was 1.49 years in males and 2.19 years in females. With two MRI sequences incorporated, the SVC model obtained with 85.7% correctly classified minors and 96.2% correctly classified adults in males, while 83.3% and 98.0% respectively in females. In conclusion, T2-FS sequence may slightly outperform the T1-TSE sequence in shoulder MRI analysis for age estimation, while shoulder MRIs could be a reliable prediction indicator for the 18-year threshold and two MRI sequences incorporated are encouraged. [ABSTRACT FROM AUTHOR]

Estimation of age of majority has important applications in the forensic daily practice because of the increasing demand for age estimates of unaccompanied minors. Diagnostic accuracy of I3M to assess legal adult age of 18 years has been already tested in several specific population samples. The aim of this work was to compare the available data about sensitivity and specificity of the third molar maturity index (I3M) from five different regional groups' radiographic samples to study possible ethnical difference. For this purpose, a sample of 6157 orthopantomograms (OPGs), coming from 15 countries, was analysed. Data about sensitivity and specificity were pooled using a bivariate modelling approach. The one-way MANOVA analysis was applied to assess the likelihood that sensitivity and specificity of the five regional groups are sampled from the same population. The result of the one-way MANOVA showed that both sensitivity and specificity did not depend from the regional groups. The obtained sensitivity was 84% (95% CI: 82% and 85%), and its specificity was 94% (95% CI: 93% and 95%). Based on these results, the I3M was a useful statistical tool to identify whether a subject has reached the legal age of 18 years old, regardless of his or her ethnicity. [ABSTRACT FROM AUTHOR]

Introduction: The acetabulum has been reported as a reliable age estimation marker. However, analyzing its morphological changes can be challenging using computed tomography (CT) imaging. Newly introduced global illumination rendering (GIR) applied to CT can improve the visualization of the fine details and thus the method's performance. This study aimed to analyze age estimation using morphological features of the acetabulum using GIR applied to CT. Methods: We collected 200 postmortem CT scans. A segmentation of the acetabular joint was initially done. Then, three-dimensional (3D) reconstruction of the images was performed using GIR. These images were saved and then analyzed by two operators based on the three morphological criteria described in the Rougé-Maillart method. Reproducibility was assessed by intraclass correlation (ICC). Age estimation was assessed by multiple linear regression. Results: The sample was composed of 155 males and 45 females, with a mean age of 50 ± 18.3 years old. We observed high agreement in both the inter-observer and intra-observer reproducibility for the three variables (ICC of 75.6 to 90.8% and 89.3 to 95.8%, respectively) and the total score (ICC of 93.5% and 95%, respectively). The three variables, as well as the total score, were significantly correlated with age groups. The total score showed a prediction rate higher than 85% for ages under 40 and over 70 years old. We identified three models with two validated models with an adjusted R2 of 85.6% and 84.8%, respectively; a standard error of 0.688 and 0.706, respectively; and a good correlation of all variables and no inter-correlation. The first validated model included the three morphological criteria scores, and the second model was based on the total score. Conclusion: GIR applied to CT provides photorealistic images that can be useful for forensic imaging intended for age estimation based on morphological methods. [ABSTRACT FROM AUTHOR]

Evaluation of secondary dentin formation is generally suitable for age assessment. We investigated the potential of modern magnetic resonance imaging (MRI) technology to visualize the dental pulp in direct comparison with cone beam computed tomography (CBCT). To this end, we examined 32 extracted human teeth (teeth 11–48 [FDI]) using 9.4-T ultrashort echo time (UTE)-MRI and CBCT (methods). 3D reconstruction was performed via both manual and semi-automatic segmentation (settings) for both methods in two runs by one examiner. Nine teeth were also examined by a second examiner. We evaluated the agreement between examiners, scan methods, and settings. CBCT was able to determine the pulp volume for all teeth. This was not possible for two teeth on MRI due to MRI artifacts. The mean pulp volume estimated by CBCT was consistently higher (~ 43%) with greater variability. With lower variability in its measurements, evaluation of pulp volume using the MRI method exhibited greater sensitivity to differences between settings (p = 0.016) and between examiners (p = 0.009). The interactions of single-rooted teeth and multi-rooted teeth and method or setting were not found to be significant. For examiner agreement, the mean pulp volumes were similar with overlapping measurements (ICC > 0.995). Suitable for use in age assessment is 9.4-T UTE-MRI with good reliability and lower variation than CBCT. For MRI, manual segmentation is necessary due to a more detailed representation of the interior of the pulp cavity. Since determination of pulp volume is expected to be systematically larger using CBCT, method-specific reference values are indispensable for practical age assessment procedures. The results should be verified under in vivo conditions in the future. [ABSTRACT FROM AUTHOR]

Age estimation is an important challenge in many fields, including immigrant identification, legal requirements, and clinical treatments. Deep learning techniques have been applied for age estimation recently but lacking performance comparison between manual and machine learning methods based on a large sample of dental orthopantomograms (OPGs). In total, we collected 10,257 orthopantomograms for the study. We derived logistic regression linear models for each legal age threshold (14, 16, and 18 years old) for manual method and developed the end-to-end convolutional neural network (CNN) which classified the dental age directly to compare with the manual method. Both methods are based on left mandibular eight permanent teeth or the third molar separately. Our results show that compared with the manual methods (92.5%, 91.3%, and 91.8% for age thresholds of 14, 16, and 18, respectively), the end-to-end CNN models perform better (95.9%, 95.4%, and 92.3% for age thresholds of 14, 16, and 18, respectively). This work proves that CNN models can surpass humans in age classification, and the features extracted by machines may be different from that defined by human. [ABSTRACT FROM AUTHOR]

Age at death estimation in cases of human skeletal finds is an important task in forensic medicine as well as in anthropology. In forensic medicine, methods based on "molecular clocks" in dental tissues and bone play an increasing role. The question, whether these methods are applicable also in cases with post-depositional intervals far beyond the forensically relevant period, was investigated for two "protein clocks", the accumulation of D-aspartic acid (D-Asp) and the accumulation of pentosidine (Pen) in dentine. Eight teeth of skeletons from different burial sites in Austria and with post-depositional intervals between c. 1216 and c. 8775 years were analysed. The results of age at death estimation based on D-Asp and Pen in dentine were compared to that derived from a classical morphological examination. Age at death estimation based on D-Asp resulted consistently in false high values. This finding can be explained by a post-mortem accumulation of D-Asp that may be enhanced by protein degradation. In contrast, the Pen-based age estimates fitted well with the morphological age diagnoses. The described effect of post-mortem protein degradation is negligible in forensically relevant time horizons, but not for post-depositional intervals of thousands of years. That means that the "D-Asp clock" loses its functionality with increasing post-depositional intervals, whereas Pen seems to be very stable. The "Pen-clock" may have the potential to become an interesting supplement to the existing repertoire of methods even in cases with extremely long post-depositional intervals. Further investigations have to test this hypothesis. [ABSTRACT FROM AUTHOR]

Since forensic age estimation is not a valid medical indication, research on the use of nonionizing methods is increasing. Ultrasonography is a radiological approach that protects patients from radiation exposure and offers special convenience to them. In this study, ultrasonography was used for age estimation by investigating the degree of ossification of the distal radial epiphysis. Its applicability on the Turkish population was investigated. The left wrist of 688 (322 males, 366 females) patients between the ages of 9 and 25 years was prospectively evaluated by ultrasonography. The intra- and interobserver reliabilities in evaluating the distal radial epiphysis and Cohen's kappa statistics show that the interobserver error was very low, and the kappa value was found to be 0.919. Stage 3 and 4 ossification of the distal radial epiphysis was first detected at age 14.3 and 15.3 years in males and 12.7 and 14.8 years in females, respectively. The data obtained may help determine legally critical age limits of 14 and 15. Although it does not seem useful for the age of 18, ultrasonography may be recommended in selected cases as a fast, inexpensive, frequently reproducible radiological method without concern about radiation and without a predictable health risk. [ABSTRACT FROM AUTHOR]

The evaluation of epiphyseal areas by magnetic resonance imaging (MRI) for forensic age estimation is an important supportive diagnostic method to prevent repeated radiation exposure without a valid medical reason. There are still not enough individuals being analyzed with MRI for age estimation. The aim of this study was to investigate the utility of T1-weighted turbo spin echo (T1-TSE) MRI sequences in determining the degree of ossification of the distal femoral and proximal tibial epiphyses in a Turkish population. In this study, images from 649 patients (335 males and 314 females) aged 10–30 years were retrospectively evaluated with sagittal T1-weighted turbo spin echo (T1-TSE) MRI sequences of the knee. Proximal tibial and distal femoral epiphysis were scored by two different observers twice using the combined staging system described by Schmeling and Kellinghaus. Spearman's rank correlation analysis indicated a significant positive relationship between age and ossification stages of the distal femoral and proximal tibial epiphyses (p < 0.001). The intra- and inter-observer reliabilities in evaluating the femur and tibia were separately determined and gave promising results and Cohen's kappa statistics ranged from κ = 0.886 and κ = 0.961. The minimal ages of patients with stage 4 ossification were 15.1 years for females and 15.8 years for males for the distal tibial epiphysis and 15.4 years for females and 17 years for males for the distal femoral epiphysis. This study show that (T1-TSE) MRI and the applicability and Schmeling and Kellinghaus staging method of the knee can be performed for living 14- to 17-year-old individuals in need of a supportive noninvasive method for estimating forensic age. [ABSTRACT FROM AUTHOR]

Age estimation is a crucial element of forensic medicine to assess the chronological age of living individuals without or lacking valid legal documentation. Methods used in practice are labor-intensive, subjective, and frequently comprise radiation exposure. Recently, also non-invasive methods using magnetic resonance imaging (MRI) have evaluated and confirmed a correlation between growth plate ossification in long bones and the chronological age of young subjects. However, automated and user-independent approaches are required to perform reliable assessments on large datasets. The aim of this study was to develop a fully automated and computer-based method for age estimation based on 3D knee MRIs using machine learning. The proposed solution is based on three parts: image-preprocessing, bone segmentation, and age estimation. A total of 185 coronal and 404 sagittal MR volumes from Caucasian male subjects in the age range of 13 and 21 years were available. The best result of the fivefold cross-validation was a mean absolute error of 0.67 ± 0.49 years in age regression and an accuracy of 90.9%, a sensitivity of 88.6%, and a specificity of 94.2% in classification (18-year age limit) using a combination of convolutional neural networks and tree-based machine learning algorithms. The potential of deep learning for age estimation is reflected in the results and can be further improved if it is trained on even larger and more diverse datasets. [ABSTRACT FROM AUTHOR]

The Bayesian approach is being a fundamental tool in forensic and legal field where inferences and decisions are made. In this study, a full Bayesian calibration model was developed to make probabilistic inferences about age estimation in a reference sample of 891 periapical X-rays of upper and lower canines. These teeth belonged to both deceased and living adult subjects, aged between 20 and 86 years, coming from five different countries (Turkey, Italy, Portugal, Japan and Mexico). For this purpose, the narrowing of pulp chamber due to the apposition of secondary dentine was analysed by means of the pulp/tooth area ratio. To determine the agreement of the method, intra- and inter-observer differences for measuring process were calculated by means of the intraclass correlation coefficient (ICC) analysis. Observer error tests showed excellent agreement between observers and between repeated assessments. According to the results of the ANCOVA, neither nationality nor sex was associated to the secondary dentine apposition while it is associated with individual's age. The results of the present study indicated that the concept of probability is intrinsically linked to the assessment of age in a forensic context, and the Bayesian approach could be considered a robust tool to overtake the bias generated by traditional regression models, thus helping the decision-making process in a legal framework. [ABSTRACT FROM AUTHOR]

The present study correlated the mineralization of third molars to chronological age using a modified classification based on Demirjian's stages in a Brazilian subpopulation and compared with the original classification. A total of 1082 patients with age ranging from 6 to 26 years were included in the sample, with at least one third molar on panoramic radiographs. The third molars were classified according to the original Demirjian classification (8 stages) and a new model based on the Demirjian method, where the original stages were grouped into four stages: AB—enamel mineralization; CD—crown dentin mineralization; EFG—root formation; and H—complete development. Statistical analyses were performed by Kruskal-Wallis/Dunn tests (α = 0.05) and the multinomial logistic regression model. Data were analyzed according to percentiles for the probability of an individual being over 18 years old. The mean ages of the stages in both classifications did not present a significant difference between superior and inferior arches (p < 0.05). The differences in mean ages between all the stages of mineralization were statistically significant (p < 0.001) only for the 4-stage classification. Males attained root formation and complete formation earlier than females (p < 0.05) in the 4-stage classification. The modified classification system showed dependence between chronological age and mineralization stages of third molars, simplifying the age estimation process. At stage H, females present a 95.7% chance of being over 18, while for males, this probability is 89.6%. This modified classification system simplifies the dental age estimation process based on third molars and can be used as a reference for future studies. [ABSTRACT FROM AUTHOR]

Dental age estimation in living individuals is one of the most frequent requests undertaken by forensic odontologists. The aim of this study was to estimate the dental age by pulp/tooth volume ratio, as measured on cone beam computed tomography (CBCT) images, in a Spanish population. This study included 313 teeth from 107 adult individuals, 56 females and 51 males with a mean age of 44 ± 14 years. The statistical analysis of the results took account of clustering (multiple teeth in individuals). Linear regression models were constructed on the relationship between pulp/tooth volume ratio and chronological age for each tooth type. The highest coefficient of determination (R2) value was provided by the upper incisors (36.6%), and the difference between chronological and estimated age was less than 5 years in 31.3% of the sample and less than 10 years for 65.7%. CBCT is an accurate imaging technique to measure dental volume with a relatively low radiation dose, and it can be used to assess dental age in living adult individuals. Volumetric changes in the pulp cavity with increasing age proved valuable to estimate dental age in this Spanish population. [ABSTRACT FROM AUTHOR]

Age estimation based on the analysis of DNA methylation patterns has become a focus of forensic research within the past few years. However, there is little data available regarding postmortem DNA methylation analysis yet, and literature mainly encompasses analysis of blood from corpses without any signs of decomposition. It is not entirely clear yet which other types of specimen are suitable for postmortem epigenetic age estimation, and if advanced decomposition may affect methylation patterns of CpG sites. In living persons, buccal swabs are an easily accessible source of DNA for epigenetic age estimation. In this work, the applicability of this approach (buccal swabs as source of DNA) under different postmortem conditions was tested. Methylation levels of PDE4C were investigated in buccal swab samples collected from 73 corpses (0–90 years old; mean: 51.2) in different stages of decomposition. Moreover, buccal swab samples from 142 living individuals (0–89 years old; mean 41.2) were analysed. As expected, methylation levels exhibited a high correlation with age in living individuals (training set: r2 = 0.87, validation set: r2 = 0.85). This was also the case in postmortem samples (r2 = 0.90), independent of the state of decomposition. Only in advanced putrified cases with extremely low DNA amounts, epigenetic age estimation was not possible. In conclusion, buccal swabs are a suitable and easy to collect source for DNA methylation analysis as long as sufficient amounts of DNA are present. [ABSTRACT FROM AUTHOR]