A methodological approach to age estimation of the intra-puparial period of the forensically relevant blow fly Calliphora vicina via Fourier transform infrared spectroscopy.

Estimating the age of immature blow flies is of great importance for forensic entomology. However, no gold-standard technique for an accurate determination of the intra-puparial age has yet been established. Fourier transform infrared (FTIR) spectroscopy is a method to (bio-)chemically characterise material based on the absorbance of electromagnetic energy by functional groups of molecules. In recent years, it also has become a powerful tool in forensic and life sciences, as it is a fast and cost-effective way to characterise all kinds of material and biological traces. This study is the first to collect developmental reference data on the changes in absorption spectra during the intra-puparial period of the forensically important blow fly Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae). Calliphora vicina was reared at constant 20°C and 25°C and specimens were killed every other day throughout their intra-puparial development. In order to investigate which part yields the highest detectable differences in absorption spectra throughout the intra-puparial development, each specimen was divided into two different subsamples: the pupal body and the former cuticle of the third instar, that is, the puparium. Absorption spectra were collected with a FTIR spectrometer coupled to an attenuated total reflection (ATR) unit. Classification accuracies of different wavenumber regions with two machine learning models, i.e., random forests (RF) and support vector machines (SVMs), were tested. The best age predictions for both temperature settings and machine learning models were obtained by using the full spectral range from 3700 to 600 cm-1. While SVMs resulted in better accuracies for C. vicina reared at 20°C, RFs performed almost as good as SVMs for data obtained from 25°C. In terms of sample type, the pupal body gave smoother spectra and usually better classification accuracies than the puparia. This study shows that FTIR spectroscopy is a promising technique in forensic entomology to support the estimation of the minimum post-mortem interval (PMImin), by estimating the age of a given insect specimen.

Age estimation using medial clavicle by histomorphometry method with artificial intelligence: A review.

This review research critically assesses the evolving landscape of age estimation methodologies, with a particular focus on the innovative integration of histomorphometry and artificial intelligence (AI) in the analysis of the medial clavicle. The medial clavicle emerges as a crucial skeletal feature for predicting age, offering valuable insights into the morphological changes occurring throughout an individual's lifespan. Through an in-depth exploration of histological complexities, including variations in osteons, trabecular structures, and cortical thickness, this review elucidates their utility as viable indicators for age-related evaluations. This framework is augmented by the incorporation of AI technology, which enables automatic picture identification, feature extraction, and complicated pattern analysis. Our review of previous research highlights the promise of AI in improving prediction models for nuanced age estimates, highlighting the importance of large-scale, diversified datasets and thorough cross-validation. This thorough study, which addresses ethical concerns as well as the influence of population-specific characteristics, moves the debate around age estimate ahead, presenting insights with consequences for forensic anthropology, clinical diagnoses, and future research avenues.

Artificial intelligence and dental age estimation: development and validation of an automated stage allocation technique on all mandibular tooth types in panoramic radiographs.

Age estimation in forensic odontology is mainly based on the development of permanent teeth. To register the developmental status of an examined tooth, staging techniques were developed. However, due to inappropriate calibration, uncertainties during stage allocation, and lack of experience, non-uniformity in stage allocation exists between expert observers. As a consequence, related age estimation results are inconsistent. An automated staging technique applicable to all tooth types can overcome this drawback.This study aimed to establish an integrated automated technique to stage the development of all mandibular tooth types and to compare their staging performances.Calibrated observers staged FDI teeth 31, 33, 34, 37 and 38 according to a ten-stage modified Demirjian staging technique. According to a standardised bounding box around each examined tooth, the retrospectively collected panoramic radiographs were cropped using Photoshop CC 2021® software (Adobe®, version 23.0). A gold standard set of 1639 radiographs were selected (n31 = 259, n33 = 282, n34 = 308, n37 = 390, n38 = 400) and input into a convolutional neural network (CNN) trained for optimal staging accuracy. The performance evaluation of the network was conducted in a five-fold cross-validation scheme. In each fold, the entire dataset was split into a training and a test set in a non-overlapping fashion between the folds (i.e., 80% and 20% of the dataset, respectively). Staging performances were calculated per tooth type and overall (accuracy, mean absolute difference, linearly weighted Cohen's Kappa and intra-class correlation coefficient). Overall, these metrics equalled 0.53, 0.71, 0.71, and 0.89, respectively. All staging performance indices were best for 37 and worst for 31. The highest number of misclassified stages were associated to adjacent stages. Most misclassifications were observed in all available stages of 31.Our findings suggest that the developmental status of mandibular molars can be taken into account in an automated approach for age estimation, while taking incisors into account may hinder age estimation.

Variations in cuticular hydrocarbons of Calliphora vicina (Diptera: Calliphoridae) empty puparia: Insights for estimating late postmortem intervals.

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.

Forensic age estimation of adolescents using computed tomography of the clavicles.

In forensic age estimation, CT imaging of the clavicles is used to determine an age over completed 21 years. If ossification of the medial clavicular epiphysis is complete, young men are assumed to be over 21 years of age. The aim of this study is to check the statistical parameters (specificity, predictive probability) for the characteristic "completed ossification of the medial clavicles". 285 male patients who, for various reasons, received a chest CT at the Medical Center of the University of Freiburg between 1st December 2019 and 6th December 2022 were screened for the study, of whom 203 patients were included in the study. The stage of clavicular ossification was classified as stage 1 - 5 according to Schmeling. While 70 out of 71 patients under 21 years of age were correctly estimated to be under 21 years of age, there was one patient whose ossification on one side was classified as stage 4 and who would therefore have been estimated to be over 21 years of age. If only subjects whose ossification stage was the same on both sides are included, the specificity of the test method is 100% and the positive predictive probability is 100%. If patients for whom only one side is stage 4 are also included, the specificity is 98.6%. Thus, only the complete and symmetrical ossification of both clavicles (stage 4 according to the Schmeling classification) in a standardised thin-layer CT can be classified as a reliable indicator of an age over 21 years in young men. In the case of asymmetric ossification of the medial clavicles (stage 4 is not reached on one side), false positive evaluations and the incorrect assumption of an age over 21 years can occur.

Forensic age estimation by MRI of the knee - comparison of two classifications for ossification stages in a German population.

AIM AND OBJECTIVES: In forensic age estimation e.g. for judicial proceedings surpassed age thresholds can be legally relevant. To examine age related differences in skeletal development the recommendations by the Study Group on Forensic Age Diagnostics (AGFAD) are based on ionizing radiation (among others orthopantomograms, plain x-rays of the hand). Vieth et al. and Ottow et al. proposed MRI-classifications for the epiphyseal-diaphyseal fusion of the knee joint to define different age groups in healthy volunteers. The aim of the present study was to directly compare these two classifications in a large German patient population. MATERIALS AND METHODS: MRI of the knee joint of 900 patients (405 female, 495 male) from 10 to 28 years of age were retrospectively analyzed. Acquired T1-weighted turbo spin-echo sequence (TSE) and T2-weighted sequence with fat suppression by turbo inversion recovery magnitude (TIRM) were analyzed for the two classifications. The different bony fusion stages of the two classifications were determined and the corresponding chronological ages assigned. Differences between the sexes were analyzed. Intra- and inter-observer agreements were determined using Cohen's kappa. RESULTS: With the classification of Ottow et al. it was possible to determine completion of the 18th and 21st year of life in both sexes. With the classification of Vieth et al. completion of the 18th year of life for female patients and the 14th and 21st year of life in both sexes could be determined. The intra- and inter-observer agreement levels were very good (κ > 0.82). CONCLUSION: In the large German patient cohort of this study it was possible to determine the 18th year of life with for both sexes with the classification of Ottow et al. and for female patients with the classification of Vieth et al. It was also possible to determine the 21st year of life for all bones with the classification of Ottow et al. and for the distal femur with the classification of Vieth et al.

Gonial Angle in Forensic Anthropology to Determine Age and Gender: A Population-Specific Analysis.

Background The study highlights the gonial angle as a key craniofacial landmark for age and gender determination in forensic cases. It emphasizes population-specific analysis, enhancing precision by recognizing variations between populations. By clarifying the gonial angle's forensic use, the study offers clear guidelines, improving forensic practices. Moreover, the gonial angle and age and gender correlations are thoroughly examined, offering important information on their forensic relevance. The results highlight how crucial population-specific research is to improving the precision and dependability of forensic age and gender estimation techniques, which advances forensic anthropology and supports forensic investigations around the globe. Aim and objective The purpose of this study is to assess the accuracy of age and gender estimates using gonial angles. The objectives of this research are to evaluate the precision of age and gender estimates utilizing the gonial angle. Materials and methods This present study comprises two groups based on age groups: Group I belongs to 51 to 60 years of age, and Group II belongs to 61 to 70 years of age. Making use of G-Power software (version 3.1.9.4, Düsseldorf, Germany), the sample size was determined. The calculation ensured 95% statistical power at a significance level (alpha error probability) of 0.05. To achieve sufficient statistical power, a total of 1000 samples were included, with a projected required sample size of 92. A total of 1000 samples, consisting of 500 male and 500 female panoramic radiographs, were meticulously selected for the study. The samples picked were within the age range of 51 to 70 years. Orthopantomograms were determined using Planmeca software (Planmeca Romexis®, Version 6.0, USA Inc.). Descriptive statistics, including prediction classification analysis of age and gender, were conducted using SPSS Statistics version 16.0 (SPSS Inc., Released 2007, SPSS for Windows, Version 16.0, Chicago, SPSS Inc.). Results According to this study, the mean gonial angle of males aged 51 to 60 years is larger (124.7370 degrees) than that of females (119.6371 degrees). The female group's mean estimates are more accurate, as seen by the smaller standard error (0.20844) compared to the male group's (0.60998). A statistically significant difference in mean gonial angles between the genders is evident, with males having a larger gonial angle (p-value <0.001). In the age range of 61 to 70 years, the mean gonial angle of females is higher (128.4322 degrees) than that of males (124.0529 degrees). In this instance, the male group's standard error is smaller (0.14968) than the female group's (0.30028), indicating more accurate mean estimates. Once more, a statistically significant difference is indicated by a p-value of less than 0.001, with females having a larger gonial angle than males. Conclusion Our study revealed that the gonial angle of the mandible can be considered a reliable parameter for gender identification. The study's limitation is its inability to reliably identify gender in the subadult population and in cases of edentulousness. An orthopantomogram is a trustworthy and accurate method for taking the different measurements needed to identify the gender of a particular mandible.

Cerebrovascular lesion loads and accelerated brain aging: insights into the cognitive spectrum.

Introduction: White matter hyperintensities (WMHs) and cerebral microbleeds are widespread among aging population and linked with cognitive deficits in mild cognitive impairment (MCI), vascular MCI (V-MCI), and Alzheimer's disease without (AD) or with a vascular component (V-AD). In this study, we aimed to investigate the association between brain age, which reflects global brain health, and cerebrovascular lesion load in the context of pathological aging in diverse forms of clinically-defined neurodegenerative conditions. Methods: We computed brain-predicted age difference (brain-PAD: predicted brain age minus chronological age) in the Comprehensive Assessment of Neurodegeneration and Dementia cohort of the Canadian Consortium on Neurodegeneration in Aging including 70 cognitively intact elderly (CIE), 173 MCI, 88 V-MCI, 50 AD, and 47 V-AD using T1-weighted magnetic resonance imaging (MRI) scans. We used a well-established automated methodology that leveraged fluid attenuated inversion recovery MRIs for precise quantification of WMH burden. Additionally, cerebral microbleeds were detected utilizing a validated segmentation tool based on the ResNet50 network, utilizing routine T1-weighted, T2-weighted, and T2* MRI scans. Results: The mean brain-PAD in the CIE cohort was around zero, whereas the four categories showed a significantly higher mean brain-PAD compared to CIE, except MCI group. A notable association trend between brain-PAD and WMH loads was observed in aging and across the spectrum of cognitive impairment due to AD, but not between brain-PAD and microbleed loads. Discussion: WMHs were associated with faster brain aging and should be considered as a risk factor which imperils brain health in aging and exacerbate brain abnormalities in the context of neurodegeneration of presumed AD origin. Our findings underscore the significance of novel research endeavors aimed at elucidating the etiology, prevention, and treatment of WMH in the area of brain aging.

Accuracy of automated forensic dental age estimation lab (F-DentEst Lab) on large Malaysian dataset.

When a disaster occurs, the authority must prioritise two things. First, the search and rescue of lives, and second, the identification and management of deceased individuals. However, with thousands of dead bodies to be individually identified in mass disasters, forensic teams face challenges such as long working hours resulting in a delayed identification process and a public health concern caused by the decomposition of the body. Using dental panoramic imaging, teeth have been used in forensics as a physical marker to estimate the age of an individual. Traditionally, dental age estimation has been performed manually by experts. Although the procedure is fairly simple, the large number of victims and the limited amount of time available to complete the assessment during large-scale disasters make forensic work even more challenging. The emergence of artificial intelligence (AI) in the fields of medicine and dentistry has led to the suggestion of automating the current process as an alternative to the conventional method. This study aims to test the accuracy and performance of the developed deep convolutional neural network system for age estimation in large, out-of-sample Malaysian children dataset using digital dental panoramic imaging. Forensic Dental Estimation Lab (F-DentEst Lab) is a computer application developed to perform the dental age estimation digitally. The introduction of this system is to improve the conventional method of age estimation that significantly increase the efficiency of the age estimation process based on the AI approach. A total number of one-thousand-eight-hundred-and-ninety-two digital dental panoramic images were retrospectively collected to test the F-DentEst Lab. Data training, validation, and testing have been conducted in the early stage of the development of F-DentEst Lab, where the allocation involved 80 % training and the remaining 20 % for testing. The methodology was comprised of four major steps: image preprocessing, which adheres to the inclusion criteria for panoramic dental imaging, segmentation, and classification of mandibular premolars using the Dynamic Programming-Active Contour (DP-AC) method and Deep Convolutional Neural Network (DCNN), respectively, and statistical analysis. The suggested DCNN approach underestimated chronological age with a small ME of 0.03 and 0.05 for females and males, respectively.

Leverage Effective Deep Learning Searching Method for Forensic Age Estimation.

Dental age estimation is extensively employed in forensic medicine practice. However, the accuracy of conventional methods fails to satisfy the need for precision, particularly when estimating the age of adults. Herein, we propose an approach for age estimation utilizing orthopantomograms (OPGs). We propose a new dental dataset comprising OPGs of 27,957 individuals (16,383 females and 11,574 males), covering an age range from newborn to 93 years. The age annotations were meticulously verified using ID card details. Considering the distinct nature of dental data, we analyzed various neural network components to accurately estimate age, such as optimal network depth, convolution kernel size, multi-branch architecture, and early layer feature reuse. Building upon the exploration of distinctive characteristics, we further employed the widely recognized method to identify models for dental age prediction. Consequently, we discovered two sets of models: one exhibiting superior performance, and the other being lightweight. The proposed approaches, namely AGENet and AGE-SPOS, demonstrated remarkable superiority and effectiveness in our experimental results. The proposed models, AGENet and AGE-SPOS, showed exceptional effectiveness in our experiments. AGENet outperformed other CNN models significantly by achieving outstanding results. Compared to Inception-v4, with the mean absolute error (MAE) of 1.70 and 20.46 B FLOPs, our AGENet reduced the FLOPs by 2.7×. The lightweight model, AGE-SPOS, achieved an MAE of 1.80 years with only 0.95 B FLOPs, surpassing MobileNetV2 by 0.18 years while utilizing fewer computational operations. In summary, we employed an effective DNN searching method for forensic age estimation, and our methodology and findings hold significant implications for age estimation with oral imaging.

Age estimation in Indian soldiers: Maxillary anterior tooth pulp/tooth volume ratio analysis with cone beam computed tomography.

Background: The objective of this study was to investigate the utility of Cone Beam Computed Tomography (CBCT)-based pulp tooth volume- ratio of maxillary anterior teeth for accurate age estimation. The project aimed to utilize the HOROS software for image analysis and develop prediction models using regression analysis. Methods: 1800 male patients in the age group of 20 to 40 years were selected, and maxillary anterior teeth were picked. High-resolution CBCT scans were collected, and image analysis in terms of pulp volume (PV), tooth volume (TV), and pulp-volume-to-tooth-volume ratio (PV/TV) was calculated using HOROS software. Simple linear regression analysis was used to develop prediction models correlating the PV/TV with chronological age. Results: PV/TV of all teeth ranged between 0.073 and 0.214. Pearson correlation coefficient was used to evaluate the correlation between the chronological age and the PV/TV. It shows a statistically significant (positive) but low correlation between age and PV/TV 13 and 22 (combined), respectively, and the highest Pearson correlation (0.849) for maxillary canine (13). This study presents four models for age estimation with maximum standard error ranging between 3.5 and 4.3 and an accuracy of 96%. Conclusion: This study illustrates the effectiveness of CBCT-based PV/TV of maxillary anterior teeth for age assessment. Accurate prediction models were constructed by using regression analysis and the HOROS software. These findings enhance the study of forensic odontology and have potential applications in forensic investigations, archaeological research, and legal-age assessment. Further research is necessary to validate and refine the prediction models, expanding their applicability to larger and more diverse population samples.

A novel technique for estimating age and demography of long-lived seabirds (genus Pterodroma) using an epigenetic clock for Gould's petrel (Pterodroma leucoptera).

Understanding the demography of wildlife populations is a key component for ecological research, and where necessary, supporting the conservation and management of long-lived animals. However, many animals lack phenological changes with which to determine individual age; therefore, gathering this fundamental information presents difficulties. More so for species that are rare, highly mobile, migratory and those that reside in inaccessible habitats. Until recently, the primary method to measure demography is through labour intensive mark-recapture approaches, necessitating decades of effort for long-lived species. Gadfly petrels (genus: Pterodroma) are one such taxa that are overrepresented with threatened and declining species, and for which numerous aspects of their ecology present challenges for research, monitoring and recovery efforts. To overcome some of these challenges, we developed the first DNA methylation (DNAm) demography technique to estimate the age of petrels, using the epigenetic clock of Gould's petrels (Pterodroma leucoptera). We collected reference blood samples from known-aged Gould's petrels at a long-term monitored population on Cabbage Tree Island, Australia. Epigenetic ages were successfully estimated for 121 individuals ranging in age from zero (fledgling) to 30 years of age, showing a mean error of 2.24 ± 0.17 years between the estimated and real age across the population. This is the first development of an epigenetic clock using multiplex PCR sequencing in a bird. This method enables demography to be measured with relative accuracy in a single sampling trip. This technique can provide information for emerging demographic risks that can mask declines in long-lived seabird populations and be applied to other Pterodroma populations.

Enhancing ECG-based heart age: impact of acquisition parameters and generalization strategies for varying signal morphologies and corruptions.

Electrocardiogram (ECG) is a non-invasive approach to capture the overall electrical activity produced by the contraction and relaxation of the cardiac muscles. It has been established in the literature that the difference between ECG-derived age and chronological age represents a general measure of cardiovascular health. Elevated ECG-derived age strongly correlates with cardiovascular conditions (e.g., atherosclerotic cardiovascular disease). However, the neural networks for ECG age estimation are yet to be thoroughly evaluated from the perspective of ECG acquisition parameters. Additionally, deep learning systems for ECG analysis encounter challenges in generalizing across diverse ECG morphologies in various ethnic groups and are susceptible to errors with signals that exhibit random or systematic distortions To address these challenges, we perform a comprehensive empirical study to determine the threshold for the sampling rate and duration of ECG signals while considering their impact on the computational cost of the neural networks. To tackle the concern of ECG waveform variability in different populations, we evaluate the feasibility of utilizing pre-trained and fine-tuned networks to estimate ECG age in different ethnic groups. Additionally, we empirically demonstrate that finetuning is an environmentally sustainable way to train neural networks, and it significantly decreases the ECG instances required (by more than 100 × ) for attaining performance similar to the networks trained from random weight initialization on a complete dataset. Finally, we systematically evaluate augmentation schemes for ECG signals in the context of age estimation and introduce a random cropping scheme that provides best-in-class performance while using shorter-duration ECG signals. The results also show that random cropping enables the networks to perform well with systematic and random ECG signal corruptions.

Reliability of a forensic odontology method for age-at-death estimation in adults: A Mexican case study.

This study aimed to evaluate the reliability of an age estimation method based on the pulp/tooth area ratio by assessing intra- and inter-examiner agreement across five observers at different intervals. Using the same X-ray device and technical parameters, 96 digital periapical X-ray images of upper and lower canines were obtained from 28 deceased people in Central America, whose age at death ranged from 19 to 49 years. Excellent and good agreement of results were achieved, and there were no statistically significant differences. The R2 value for upper teeth (54.0%) was higher than the R2 value for lower teeth (45.7%). The highest intraclass correlation coefficient value was 0.995 (0.993-0.997) and the lowest 0.798 (0.545-0.895). Inter-examiner agreement was high with values of 0.975 (0.965-0.983) and 0.927 (0.879-0.955). This method is adequate for assessing age in missing and unidentified people, including victims of mass disasters.

Comparison of the Cameriere's third molar maturity index and Olze et al.'s stages of radiographic visibility of the root pulp in a Turkish population.

Purpose: The purpose of this study was to compare the Cameriere's third molar maturity index and Olze et al.'s stages of radiographic visibility of the root pulp in estimating the age of maturity in the Turkish population. The age of majority, which is legally significant, marks the transition from childhood to adulthood. In Turkey, the age of majority is set at 18 years. As the third molars continue to develop at this age, they can serve as an indicator of dental age. Materials and methods: A total of 705 panoramic radiographs obtained from individuals aged 15 to 22 years, including children and adults, were included in this study. The left mandibular third molars were evaluated on panoramic radiographs using Cameriere's third molar maturity index and Olze's method of radiographic root pulp visibility (RPV) stages. Minimum and maximum values were noted for each stage, and a median with upper and lower quartiles, as well as mean and standard deviation were calculated. Sensitivity and specificity values were calculated. Results: In males, Cameriere's third molar maturity index demonstrated a sensitivity of 0.77% and specificity of 0.96%, while in females, it showed a sensitivity of 0.57% and specificity of 0.92%. Regarding Olze et al.'s stage 0, the sensitivity and specificity values were 0.86% and 0.79% in males, and 0.85% and 0.75% in females, respectively. Conclusion: Although both methods can be used to distinguish individuals below or above the age of 18, the cut-off value suggested by Cameriere's method resulted in a higher rate of type 2 error (false negativity). Therefore, the method proposed by Olze et al., based on the radiographic visibility of the root pulp, can be employed to differentiate between adults and minors in the Turkish population.

Intrapuparial stage aging and PMI estimation based on the developmental transcriptomes of forensically important Aldrichina grahami (Diptera: Calliphoridae) gene expression.

Background: The expression profiles of differentially expressed genes (DEGs) during pupal development have been demonstrated to be vital in age estimation of forensic entomological study. Here, using forensically important Aldrichina grahami (Diptera: Calliphoridae), we aimed to explore the potential of intrapuparial stage aging and postmortem interval (PMI) estimation based on characterization of successive developmental transcriptomes and gene expression patterns. Methods: We collected A. grahami pupae at 11 successive intrapuparial stages at 20 °C and used the RNA-seq technique to build the transcriptome profiles of their intrapuparial stages. The DEGs were identified during the different intrapuparial stages using comparative transcriptome analysis. The selected marker DEGs were classified and clustered for intrapuparial stage aging and PMI estimation and then further verified for transcriptome data validation. Ultimately, we categorized the overall gene expression levels as the dependent variable and the age of intrapuparial A. grahami as the independent variable to conduct nonlinear regression analysis. Results: We redefined the intrapuparial stages of A. grahami into five key successive substages (I, II, III, IV, and V), based on the overall gene expression patterns during pupal development. We screened 99 specific time-dependent expressed genes (stage-specific DEGs) to determine the different intrapuparial stages based on comparison of the gene expression levels during the 11 developmental intrapuparial stages of A. grahami. We observed that 55 DEGs showed persistent upregulation during the development of intrapuparial A. grahami. We then selected four DEGs (act79b, act88f, up and ninac) which presented consistent upregulation using RT-qPCR (quantitative real-time PCR) analysis, along with consideration of the maximum fold changes during the pupal development. We conducted nonlinear regression analysis to simulate the calculations of the relationships between the expression levels of the four selected DEGs and the developmental time of intrapuparial A. grahami and constructed fitting curves. The curves demonstrated that act79b and ninac showed continuous relatively increasing levels. Conclusions: This study redefined the intrapuparial stages of A. grahami based on expression profiles of developmental transcriptomes for the first time. The stage-specific DEGs and those with consistent tendencies of expression were found to have potential in age estimation of intrapuparial A. grahami and could be supplementary to a more accurate prediction of PMI.

Age Estimation by Machine Learning and CT-Multiplanar Reformation of Cranial Sutures in Northern Chinese Han Adults.

OBJECTIVES: To establish age estimation models of northern Chinese Han adults using cranial suture images obtained by CT and multiplanar reformation (MPR), and to explore the applicability of cranial suture closure rule in age estimation of northern Chinese Han population. METHODS: The head CT samples of 132 northern Chinese Han adults aged 29-80 years were retrospectively collected. Volume reconstruction (VR) and MPR were performed on the skull, and 160 cranial suture tomography images were generated for each sample. Then the MPR images of cranial sutures were scored according to the closure grading criteria, and the mean closure grades of sagittal suture, coronal sutures (both left and right) and lambdoid sutures (both left and right) were calculated respectively. Finally taking the above grades as independent variables, the linear regression model and four machine learning models for age estimation (gradient boosting regression, support vector regression, decision tree regression and Bayesian ridge regression) were established for northern Chinese Han adults age estimation. The accuracy of each model was evaluated. RESULTS: Each cranial suture closure grade was positively correlated with age and the correlation of sagittal suture was the highest. All four machine learning models had higher age estimation accuracy than linear regression model. The support vector regression model had the highest accuracy among the machine learning models with a mean absolute error of 9.542 years. CONCLUSIONS: The combination of skull CT-MPR and machine learning model can be used for age estimation in northern Chinese Han adults, but it is still necessary to combine with other adult age estimation indicators in forensic practice.

Dental Age Estimation in Northern Chinese Han Children and Adolescents Using Demirjian's Method Combined with Machine Learning Algorithms.

OBJECTIVES: To investigate the application value of combining the Demirjian's method with machine learning algorithms for dental age estimation in northern Chinese Han children and adolescents. METHODS: Oral panoramic images of 10 256 Han individuals aged 5 to 24 years in northern China were collected. The development of eight permanent teeth in the left mandibular was classified into different stages using the Demirjian's method. Various machine learning algorithms, including support vector regression (SVR), gradient boosting regression (GBR), linear regression (LR), random forest regression (RFR), and decision tree regression (DTR) were employed. Age estimation models were constructed based on total, female, and male samples respectively using these algorithms. The fitting performance of different machine learning algorithms in these three groups was evaluated. RESULTS: SVR demonstrated superior estimation efficiency among all machine learning models in both total and female samples, while GBR showed the best performance in male samples. The mean absolute error (MAE) of the optimal age estimation model was 1.246 3, 1.281 8 and 1.153 8 years in the total, female and male samples, respectively. The optimal age estimation model exhibited varying levels of accuracy across different age ranges, which provided relatively accurate age estimations in individuals under 18 years old. CONCLUSIONS: The machine learning model developed in this study exhibits good age estimation efficiency in northern Chinese Han children and adolescents. However, its performance is not ideal when applied to adult population. To improve the accuracy in age estimation, the other variables can be considered.

Adults Ischium Age Estimation Based on Deep Learning and 3D CT Reconstruction.

OBJECTIVES: To develop a deep learning model for automated age estimation based on 3D CT reconstructed images of Han population in western China, and evaluate its feasibility and reliability. METHODS: The retrospective pelvic CT imaging data of 1 200 samples (600 males and 600 females) aged 20.0 to 80.0 years in western China were collected and reconstructed into 3D virtual bone models. The images of the ischial tuberosity feature region were extracted to create sex-specific and left/right site-specific sample libraries. Using the ResNet34 model, 500 samples of different sexes were randomly selected as training and verification set, the remaining samples were used as testing set. Initialization and transfer learning were used to train images that distinguish sex and left/right site. Mean absolute error (MAE) and root mean square error (RMSE) were used as primary indicators to evaluate the model. RESULTS: Prediction results varied between sexes, with bilateral models outperformed left/right unilateral ones, and transfer learning models showed superior performance over initial models. In the prediction results of bilateral transfer learning models, the male MAE was 7.74 years and RMSE was 9.73 years, the female MAE was 6.27 years and RMSE was 7.82 years, and the mixed sexes MAE was 6.64 years and RMSE was 8.43 years. CONCLUSIONS: The skeletal age estimation model, utilizing ischial tuberosity images of Han population in western China and employing the ResNet34 combined with transfer learning, can effectively estimate adult ischium age.

Adolescents and Children Age Estimation Using Machine Learning Based on Pulp and Tooth Volumes on CBCT Images.

OBJECTIVES: To estimate adolescents and children age using stepwise regression and machine learning methods based on the pulp and tooth volumes of the left maxillary central incisor and cuspid on cone beam computed tomography (CBCT) images, and to compare and analyze the estimation results. METHODS: A total of 498 Shanghai Han adolescents and children CBCT images of the oral and maxillofacial regions were collected. The pulp and tooth volumes of the left maxillary central incisor and cuspid were measured and calculated. Three machine learning algorithms (K-nearest neighbor, ridge regression, and decision tree) and stepwise regression were used to establish four age estimation models. The coefficient of determination, mean error, root mean square error, mean square error and mean absolute error were computed and compared. A correlation heatmap was drawn to visualize and the monotonic relationship between parameters was visually analyzed. RESULTS: The K-nearest neighbor model (R2=0.779) and the ridge regression model (R2=0.729) outperformed stepwise regression (R2=0.617), while the decision tree model (R2=0.494) showed poor fitting. The correlation heatmap demonstrated a monotonically negative correlation between age and the parameters including pulp volume, the ratio of pulp volume to hard tissue volume, and the ratio of pulp volume to tooth volume. CONCLUSIONS: Pulp volume and pulp volume proportion are closely related to age. The application of CBCT-based machine learning methods can provide more accurate age estimation results, which lays a foundation for further CBCT-based deep learning dental age estimation research.