Internal validation of gestational age estimation algorithms in health-care databases using pregnancies conceived through fertility procedures.

Fertility procedures recorded in health-care databases can be used to estimate the start of pregnancy, which can serve as a reference standard to validate gestational age estimates based on International Classification of Diseases codes. In a cohort of 17 398 US MarketScan pregnancies (2011-2020) in which conception was achieved via fertility procedures, we estimated gestational age at the end of pregnancy using algorithms based on (1) time (days) since the fertility procedure (the reference standard); (2) International Classification of Diseases, Ninth Revision (ICD-9)/International Classification of Diseases, Tenth Revision (ICD-10) (before/after October 2015) codes indicating gestational length recorded at the end of pregnancy (method A); and (3) ICD-10 end-of-pregnancy codes enhanced with Z3A codes denoting specific gestation weeks recorded at prenatal visits (method B). We calculated the proportion of pregnancies with an estimated gestational age falling within 14 days ($\pm$14 days) of the reference standard. Method A accuracy was similar for ICD-9 and ICD-10 codes. After 2015, method B was more accurate than method A: For term births, within-14-day agreement was 90.8% for method A and 98.7% for method B. Corresponding estimates were 70.1% and 95.6% for preterm births; 35.3% and 92.6% for stillbirths; 54.3% and 64.2% for spontaneous abortions; and 16.7% and 84.6% for elective terminations. ICD-10-based algorithms that incorporate Z3A codes improve the accuracy of gestational age estimation in health-care databases, especially for preterm births and non-live births.

Epigenetic Age Estimation by Detecting DNA Methylation Status in Buccal Swabs.

The analysis of DNA methylation (DNAm) levels at specific CpG sites represents one of the most promising molecular techniques for estimating an individual's age. To date, a considerable number of studies have reported the development of age prediction models on the basis of DNAm in body fluids, with only a few utilizing buccal swabs. The objective of this study was to identify age-dependent methylation CpG sites in three different genes (HOXC4, TRIM59, and ELOVL2) in buccal swab samples from the Chinese Han population. A total of 461 buccal swabs, with an age range of 0.4-80.8 years, were divided into a training set (n = 325) and a validation set (n = 136). Samples were analyzed by pyrosequencing in order to identify age-related genes with correlation coefficient. A random forest regression model was ultimately proposed, including eight CpGs in three genes, with a mean absolute error (MAE) of 2.119 years. The model performs independent validation set with an MAE of 4.391 years. Our findings illustrate that buccal swabs present a suitable alternative to biological traces for age prediction based on DNAm pattern using pyrosequencing and random forest regression, offering the additional advantage of being collected noninvasively.

A semen-specific deoxyribonucleic acid methylation model for epigenetic age estimation and its robustness under environmental challenges.

In forensic investigations, semen samples are a common form of biological evidence, especially in cases involving sexual assault. Therefore, accurately estimating the age of an individual is crucial in criminal cases. This study presents a novel age estimation model based on semen-specific CpG methylation patterns. A multiplex panel was developed, consisting of 12 CpG sites (PARP14, C5orf25, cg23488376, MXRA5, PFKFB3, DLL1, NOX4, cg12837463, TTC7B, KCNA7, NKX2-1, and SYNE4), which exhibit strong correlations with age. Additionally, this study investigates the resilience of these methylation markers under simulated environmental challenges. We collected ejaculate samples from a diverse cohort of 115 male individuals, aged 20-71 years, who underwent deoxyribonucleic acid extraction and bisulfite conversion. Methylation levels of the selected CpG sites were assessed using a SNaPshot assay, which revealed significant correlations with chronological age. We developed and validated two robust age estimation models through stepwise and enter regression analyses, achieving reliable accuracy with mean absolute errors ranging from 3.81 to 4.1 years. Additionally, the study also investigated the robustness of semen stains under diverse environmental conditions, including fabric type, washing, hematin exposure, and UV-C light. The selected methylation markers demonstrated remarkable resilience despite the challenges posed by washing procedures and environmental exposure, confirming their potential for age estimation in forensic genetics. This research presents successful age estimation models, emphasizing the strong correlations between methylation levels and chronological age. The proposed methodology's accuracy is affirmed through model validation on an independent test set, while also highlighting the resilience of semen stains on fabrics under varying storage and washing conditions.

Does third molar agenesis influence the second lower molar mineralization?

Different studies have established that the mineralization stages of the second mandibular molar can be used in forensic age estimation. Nowadays, the estimate's accuracy is an ethical concern, producing as few false positives (individuals incorrectly classified as older than a determined threshold) and false negatives (individuals incorrectly classified as younger than a determined threshold) as possible. Some have hypothesized that changes in teeth number may influence tooth mineralization, altering the age estimate process. This paper analyzes whether third molar agenesis affects the second mandibular molar mineralization time frame. To do so, 355 orthopantomograms were evaluated for third molar agenesis, and the second mandibular molar mineralization stage was assessed using the Demirjian stages. Student's t-test was used to compare the difference in the mean age at which the various stages of 37 mineralization were reached in the groups with and without third molar agenesis. The level of statistical significance was set at 5%. The results pointed to a delay in second mandibular molar mineralization in the case of agenesis, suggesting the need to consider this when estimating age using dental techniques.

The Cayor Atlas: a black Senegalese dental atlas for permanent tooth formation.

INTRODUCTION: Many aspects of tooth development have been documented, particularly in Caucasian populations. However, dental development has not been extensively studied in West Africa. OBJECTIVE: The present study was designed to provide information on the sequences of tooth calcification in West African black Senegalese children and to compare the results with those of other populations, notably the London Atlas. METHODS: A total of 556 orthopantomograms (OPGs) from 289 males and 266 females with a mean age of 11.34 ± 3.84 years were analyzed. Demirjian A-H staging was applied to record the stages of tooth development. Tables of tooth development stages for each tooth were generated separately for age cohorts and by sex. The most common stage of tooth formation (modal) was the characteristic age stage of development. Differences between boys and girls and between maxillary and mandibular teeth were also analyzed using chi-squares. Accuracy was assessed by comparing the age estimated by the Dental Development Atlas for this population (Cayor Atlas) and the London Atlas tooth with chronological age using the Bland-Altman test. RESULTS: There was no significant difference in tooth development between girls and boys, p > 0.05. Maxillary teeth had similar dental development to mandibular teeth, p > 0.05. The Pearson correlation test showed a strong correlation between chronological age and the age estimated by the Cayor atlas, p < 0.001. The Bland-Altman test also showed greater accuracy than the London Atlas. CONCLUSION: These results show dental calcification sequences different from those of the London Atlas Tooth and the Witts Atlas (Atlas of Black South African Subjects).

Charting the growth through intelligence: A SWOC analysis on AI-assisted radiologic bone age estimation.

Bone age estimation (BAE) is based on skeletal maturity and degenerative process of the skeleton. The clinical importance of BAE is in understanding the pediatric and growth-related disorders; whereas medicolegally it is important in determining criminal responsibility and establishing identification. Artificial Intelligence (AI) has been used in the field of the field of medicine and specifically in diagnostics using medical images. AI can greatly benefit the BAE techniques by decreasing the intra observer and inter observer variability as well as by reducing the analytical time. The AI techniques rely on object identification, feature extraction and segregation. Bone age assessment is the classical example where the concepts of AI such as object recognition and segregation can be used effectively. The paper describes various AI based algorithms developed for the purpose of radiologic BAE and the performances of the models. In the current paper we have also carried out qualitative analysis using Strength, Weakness, Opportunities and Challenges (SWOC) to examine critical factors that contribute to the application of AI in BAE. To best of our knowledge, the SWOC analysis is being carried out for the first time to assess the applicability of AI in BAE. Based on the SWOC analysis we have provided strategies for successful implementation of AI in BAE in forensic and medicolegal context.

Radiological assessment of periodontal ligament space visibility on third molars for forensic age assessment - a comparison study of three different staging scales.

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.

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.

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.

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.

Computed tomography study of cranial vault thickness in Malaysian subadult population.

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.

Forensic age estimation of the knee by post-mortem DR, CT, and MR imaging: a comparative study.

It is believed by many that reference data for age estimation purposes must be imaging-modality specific. A study from our department has however proven otherwise. We therefore found it interesting to investigate this further by looking at the level of agreement between different imaging modalities. The aim of this study was to investigate the level of agreement between the three radiological modalities, computed tomography (CT), magnetic resonance imaging (MRI), and digital radiography (DR), in assessing the ossification of the epiphyses of the knee. A total of 34 deceased individuals of 10-25 years of age, brought in for a medicolegal autopsy at our department, were scanned by CT, MRI, and DR. The ossification stages of the three bones of the right knee, distal femoral, proximal tibial, and proximal fibular epiphysis were assessed using the established combined staging method by Schmeling et al. and Kellinghaus et al. Analysis of the results by Cohen's weighted kappa showed a good agreement between CT and DR (K = 0.61-0.70), and MRI and DR (K = 0.68-0.79) but only moderate agreement between CT and MRI (K = 0.55-0.57). This leads us to conclude that different radiological images cannot be used interchangeably for age estimation purposes, so reference material needs to be imaging-modality specific. However, to make a more general conclusion research on a larger population is needed.

Novel ratio-expressions of genes enables estimation of wound age in contused skeletal muscle.

Given that combination with multiple biomarkers may well raise the predictive value of wound age, it appears critically essential to identify new features under the limited cost. For this purpose, the present study explored whether the gene expression ratios provide unique time information as an additional indicator for wound age estimation not requiring the detection of new biomarkers and allowing full use of the available data. The expression levels of four wound-healing genes (Arid5a, Ier3, Stom, and Lcp1) were detected by real-time polymerase chain reaction, and a total of six expression ratios were calculated among these four genes. The results showed that the expression levels of four genes and six ratios of expression changed time-dependent during wound repair. The six expression ratios provided additional temporal information, distinct from the four genes analyzed separately by principal component analysis. The overall performance metrics for cross-validation and external validation of four typical prediction models were improved when six ratios of expression were added as additional input variables. Overall, expression ratios among genes provide temporal information and have excellent potential as predictive markers for wound age estimation. Combining the expression levels of genes with ratio-expression of genes may allow for more accurate estimates of the time of injury.

Establishment of a multiplex PCR-CE assay for the simultaneous and rapid analysis of age markers for Calliphora vicina pupae.

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.

Age estimation of burnt human remains through DNA methylation analysis.

The identification of human fire victims is a challenging task in forensic medicine. The heat-induced alterations of biological tissues can make the conventional anthropological analyses difficult. Even if the DNA profile of the victim is achieved, it is possible that no match can be found in a forensic DNA database, thus hindering positive identification. In such cases, any information useful to nail down a possible identity should be collected, such as DNA methylation analysis which could provide useful investigative leads. In the present study, five age-related epigenetic markers (ELOVL2, FHL2, KLF14, C1orf132, and TRIM59) were initially analysed in blood samples of 72 living Italian individuals of known age, using a Single Base Extension (SBE) assay. An age prediction model was built by multiple linear regression including all the markers (Mean Absolute Error, MAE: 3.15 years). This model was tested on 29 blood samples collected during autopsies from burnt human remains, already identified through DNA analysis, providing a MAE of 6.92 years. The model allowed a correct prediction in 79.3% of the cases (95% prediction interval), while six cases were associated with inaccurate predictions (min-max prediction error: 9.8-37.3 years). Among the different sample variables considered to explain these results, only the DNA degradation index was a relevant factor affecting the reliability of the predictions. In conclusion, the SBE typing of blood from burnt remains proved to be a reliable tool to estimate chronological age of most of the samples, also in consideration of its cost-effectiveness and the availability of CE sequencers in every forensic genetics laboratory.

Application of Vieth staging in forensic age estimation in the living using MRI of the distal radial epiphysis.

Forensic age estimation is crucial in various legal and civil contexts, particularly in regions experiencing significant migration and inadequate birth registration systems. This study evaluates the applicability of the Vieth staging system for forensic age estimation in the living using MRI of the distal radial epiphysis. A retrospective analysis was conducted on 620 left wrist MRI scans from individuals aged 9.92 to 29.58 years. The study demonstrated high intra- and inter-observer agreement values (κ = 0.974 and κ = 0.961), confirming the method's reliability. Spearman's rank correlation analysis showed significant positive correlations between age and ossification stage for both sexes. The minimum ages observed for males were 9.92 years at stage 2, 15.00 years at stage 3, 15.00 years at stage 4, 17.00 years at stage 5, and 20.00 years at stage 6. For females, the minimum ages were 10.08 years at stage 2, 12.33 years at stage 3, 14.25 years at stage 4, 16.33 years at stage 5, and 18.42 years at stage 6. The study supports the applicability of the Vieth methodology for forensic age estimation in the living and suggests that MRI could be a non-invasive and potentially effective tool for determining critical age thresholds in forensic contexts. Further research is recommended to refine these methods and explore their applicability across different populations.

Studying eruption patterns of mandibular third molars for forensic age assessment: Introducing current reference data from a population of black South Africans.

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.

Automatic and robust estimation of sex and chronological age from panoramic radiographs using a multi-task deep learning network: a study on a South Korean population.

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.

Development of a screening system of gene sets for estimating the time of early skeletal muscle injury based on second-generation sequencing technology.

The present study is aimed to address the challenge of wound age estimation in forensic science by identifying reliable genetic markers using low-cost and high-precision second-generation sequencing technology. A total of 54 Sprague-Dawley rats were randomly assigned to a control group or injury groups, with injury groups being further divided into time points (4 h, 8 h, 12 h, 16 h, 20 h, 24 h, 28 h, and 32 h after injury, n = 6) to establish rat skeletal muscle contusion models. Gene expression data were obtained using second-generation sequencing technology, and differential gene expression analysis, weighted gene co-expression network analysis (WGCNA) and time-dependent expression trend analysis were performed. A total of six sets of biomarkers were obtained: differentially expressed genes at adjacent time points (127 genes), co-expressed genes most associated with wound age (213 genes), hub genes exhibiting time-dependent expression (264 genes), and sets of transcription factors (TF) corresponding to the above sets of genes (74, 87, and 99 genes, respectively). Then, random forest (RF), support vector machine (SVM) and multilayer perceptron (MLP), were constructed for wound age estimation from the above gene sets. The results estimated by transcription factors were all superior to the corresponding hub genes, with the transcription factor group of WGCNA performed the best, with average accuracy rates of 96% for three models' internal testing, and 91.7% for the highest external validation. This study demonstrates the advantages of the indicator screening system based on second-generation sequencing technology and transcription factor level for wound age estimation.