Echocardiographic age and sex estimation using artificial intelligence predicts overall survival

The continued development in methylome analysis has enabled a more precise assessment of DNA methylation, but treatment of target tissue prior to analysis may affect DNA analysis. Prediction of age based on methylation levels in the genome (DNAmAge) has gained much interest in disease predisposition (biological age estimation), but also in chronological donor age estimation in crime case samples. Various epigenetic clocks were designed to predict the age. However, it remains unknown how the storage of the tissues affects the DNAmAge estimation. In this study, we investigated the storage method impact of DNAmAge by the comparing the DNAmAge of the two commonly used storage methods, freezing and formalin-fixation and paraffin-embedding (FFPE) to DNAmAge of fresh tissue. This was carried out by comparing paired heart tissue samples of fresh tissue, samples stored by freezing and FFPE to chronological age and whole blood samples from the same individuals. Illumina EPIC beadchip array was used for methylation analysis and the DNAmAge was evaluated with the following epigenetic clocks: Horvath, Hannum, Levine, Horvath skin+blood clock (Horvath2), PedBE, Wu, BLUP, EN, and TL. We observed differences in DNAmAge among the storage conditions. FFPE samples showed a lower DNAmAge compared to that of frozen and fresh samples. Additionally, the DNAmAge of the heart tissue was lower than that of the whole blood and the chronological age. This highlights caution when evaluating DNAmAge for FFPE samples as the results were underestimated compared with fresh and frozen tissue samples. Furthermore, the study also emphasizes the need for a DNAmAge model based on heart tissue samples for an accurate age estimation.

DNA methylation markers in combination with skeletal and dental ages to improve age estimation in children

To develop and validate formulas for age and sex estimation based on the pulp cavity volume of teeth using cone beam CT. The sample was composed of 116 cone beam CT scans from Brazilian individuals of both sexes, ranging in age from 13 to 70 years. A total of 232 teeth (upper central incisors and canines) were evaluated. Two calibrated examiners determined pulp cavity volumes using the ITK-SNAP software. Pearson's correlation test was used to assess the correlation between chronological age and pulp volume. Linear and logistic regression models were developed for age and sex estimation, respectively, and were validated in another sample of 72 teeth. Pearson's correlation coefficients between age and pulp volume were negative and significant (p < 0.0001) for both teeth (r = -0.8782 for central incisors and r = -0.8738 for canines). The age estimation formulas showed good determination coefficients (adjusted R² = 0.7614 to 0.8367). For sex estimation, when the age was known, the coefficients were also good (adjusted R² = 0.649 to 0.812). However, when the age was unknown, the coefficients of the sex estimation formulas were low (adjusted R² = 0.047 to 0.393). Validation showed high accuracy of age estimation in individuals older than 35 years, as well as high accuracy of sex estimation when the age was known. Our formulas provided excellent results and can be applied to the Brazilian population. The best results were observed for age estimation in females and for sex estimation when the age was known.

Methods for age estimation in children involve measurements of: (1) the projections of open apices and tooth heights (T), (2) the total area of the carpal bones and the epiphyses of the ulna and radius (HW), and (3) a combination of these parameters (THW). This study aimed to compare the accuracy of the original formulae for T, HW, and THW in Black South African (BSA) and White South African (WSA) samples of children and adolescents. The sample comprised 556 subjects, 164 BSA (71 males and 93 females) and 392 WSA (171 males and 221 females) aged 6 to 16 years. The main sample comprised 556 subjects and was divided into two parts. The first part was used to develop new formulae specific to the South African population, while the second part, consisting of 183 subjects and referred to as the test sample, was used to evaluate the accuracy of these formulae and to quantify their uncertainty when applied to real cases. Adjusted R2 and standard errors of the estimate (SEEs) were calculated for each regression model. The mean absolute difference or error (MAE) between estimated and chronological age and the percentage of individuals within specific age ranges of ± 0.25, ± 0.5, ± 1, and ± 2 years of chronological age was used to compare the accuracy of formulae. Teeth showed the most significant underestimation in the training sample, -0.49 years in males and - 0.29 years in females, as much as -0.70 years in WSA males, while BSA males were overestimated by + 0.03 years. Combining teeth and hand-wrist, the new formula showed the best performance in age estimation compared to only teeth or hand-wrist, except in WSA males (-0.51 years). New South African-specific formula improved accuracy, especially in males, suggesting their suitability alongside the original Cameriere et al. methods for precise age estimation. The original regression formulae by Cameriere et al., along with the newly developed formulaespecific to South African populations, shows promise for age estimation, considering potential estimation errors and the accuracy rates demonstrated in this study.

Development of a Reference Data Set (RDS) for dental age estimation (DAE) and testing of this with a separate Validation Set (VS) in a southern Chinese population

Aging clocks dissociate biological from chronological age. The estimation of biological age is important for identifying gerontogenes and assessing environmental, nutritional, or therapeutic impacts on the aging process. Recently, methylation markers were shown to allow estimation of biological age based on age‐dependent somatic epigenetic alterations. However, DNA methylation is absent in some species such as Caenorhabditis elegans and it remains unclear whether and how the epigenetic clocks affect gene expression. Aging clocks based on transcriptomes have suffered from considerable variation in the data and relatively low accuracy. Here, we devised an approach that uses temporal scaling and binarization of C. elegans transcriptomes to define a gene set that predicts biological age with an accuracy that is close to the theoretical limit. Our model accurately predicts the longevity effects of diverse strains, treatments, and conditions. The involved genes support a role of specific transcription factors as well as innate immunity and neuronal signaling in the regulation of the aging process. We show that this binarized transcriptomic aging (BiT age) clock can also be applied to human age prediction with high accuracy. The BiT age clock could therefore find wide application in genetic, nutritional, environmental, and therapeutic interventions in the aging process.

Effectiveness of three age estimation methods based on dental and skeletal development in a sample of young Brazilians

Background: Response to neoadjuvant chemotherapy (NACT) has been proven to be an established prognostic factor after open radical cystectomy (ORC). We evaluated the impact of NACT on survival outcomes of a single-institution robotic radical cystectomy (RARC) series. Methods: From January 2012 to June 2020, 79 patients were identified. Baseline, demographic, perioperative, and pathologic data were described. Kaplan–Meier with the log-rank test was used to compare overall survival (OS) differences between complete, partial, and no-NACT responders, respectively. Univariable and multivariable regression analyses were performed to identify predictors of OS. Results: Complete, partial, and absent response to NACT were recorded in 43 (54.4%), 21 (19%), and 15 (26.6%) patients, respectively. A complete response to NACT displayed a trend toward significant higher OS (p = 0.03). In univariable analysis, significant predictors of lower OS were hypertension (HR 3.37; CI 95% 1.31–8.62; p = 0.01); advanced nodal involvement (HR 2.41; CI 95% 0.53–10.9; p < 0.001); and incomplete response to NACT (HR 0.41; CI 95% 0.18–0.95; p = 0.039). In multivariable analysis, the only independent predictor of worse OS was advanced pathologic N stages (HR 10.1; CI: 95% CI 2.3–44.3; p = 0.002). Conclusions: Complete response to NACT is associated with increased OS probability, but significant nodal residual disease remains the only independent predictor of OS after RARC.

This study aimed to validate Lee's age estimation method and assess the 18-year threshold in Korean and Japanese populations. We evaluated the maxillary and mandibular second (M2) and third molars (M3) in 2657 orthopantomograms of the Korean and Japanese populations aged 15-23 years (19.47±2.62 years for Koreans, 19.31±2.60 years for Japanese), using Demirjian's criteria. Dental age was estimated, and correlations between chronological and dental ages were analyzed. Classification performance was calculated based on the 18-year threshold. The relationship between developmental stage and chronologic age was analyzed using multiple linear regression. Our results revealed that Lee's method was appropriate for estimation in the Korean population. When the Lee's method was applied to the Japanese population, a lower value of correlation coefficients between estimated and chronological age, and lower specificity were observed. Population differences were observed predominantly in the stages of root development (stages F and G) of M2s and M3s in both jaws and more frequently in females than in males. In the multiple linear regression between developmental stage and chronological age, lower values of adjusted r2 were observed in the Japanese population than in the Koreans. In conclusion, the Lee's method derived from the Korean population data might be unsuitable for Japanese juveniles and adolescents. To support the findings of this study, future studies with samples from multiple institutions should be conducted. Future studies with larger sample sizes are also warranted to improve the accuracy of dental age estimation and confirm the developmental pattern of teeth in the Japanese population.

Simple SummaryMucosal melanoma (MM) is rare and entails a poor prognosis. MM is biologically different from cutaneous melanoma (CM). For advanced CM, overall survival has improved since the introduction of immune and targeted therapy. In contrast, little is known about the effect of their introduction on the survival of MM. This study presents the incidence, clinical characteristics, treatment characteristics, and survival of MM over 30 years (1990–2019) in the Netherlands. We conclude that the incidence of MM remained stable, and survival has slightly improved when comparing the timeframe 2014–2019 with previous years. However, the prognosis of MM remains poor as compared to CM. Future studies addressing the effect of immune and targeted therapy in MM are needed to improve outcomes for patients with MM.Background: Mucosal melanoma (MM) is a rare tumour with a poor prognosis. Over the years, immune and targeted therapy have become available and have improved overall survival (OS) for patients with advanced cutaneous melanoma (CM). This study aimed to assess trends in the incidence and survival of MM in the Netherlands against the background of new effective treatments that became available for advanced melanoma. Methods: We obtained information on patients diagnosed with MM during 1990–2019 from the Netherlands Cancer Registry. The age-standardized incidence rate and estimated annual percentage change (EAPC) were calculated over the total study period. OS was calculated using the Kaplan–Meier method. Independent predictors for OS were assessed by applying multivariable Cox proportional hazards regression models. Results: In total, 1496 patients were diagnosed with MM during 1990–2019, mostly in the female genital tract (43%) and the head and neck region (34%). The majority presented with local or locally advanced disease (66%). The incidence remained stable over time (EAPC 3.0%, p = 0.4). The 5-year OS was 24% (95%CI: 21.6–26.0%) with a median OS of 1.7 years (95%CI: 1.6–1.8). Age ≥ 70 years at diagnosis, higher stage at diagnosis, and respiratory tract location were independent predictors for worse OS. Diagnosis in the period 2014–2019, MM located in the female genital tract, and treatment with immune or targeted therapy were independent predictors for better OS. Conclusion: Since the introduction of immune and targeted therapies, OS has improved for patients with MM. However, the prognosis of MM patients is still lower compared to CM, and the median OS of patients treated with immune and targeted therapies remains fairly short. Further studies are needed to improve outcomes for patients with MM.

Anti-Idiotype Antibody Response after Vaccination Correlates with Better Overall Survival in Follicular Lymphoma.

To explore expression and clinical relevance of phosphohistidine phosphatase 1 (PHPT1) in clear-cell renal cell carcinoma. Patients with clear-cell renal cell carcinoma who underwent radical or nephron-sparing nephrectomy were enrolled. Correlations between PHPT1 expression and demographic and clinical characteristics were analysed prospectively. In total, 122 patients (78 male/44 female) were included. In normal kidney tissue, PHPT1 expression was observed only in the proximal tubule. High PHPT1 expression levels were associated with larger tumour size, higher Fuhrman nuclear grade and advanced pathological tumour-node-metastasis (pTNM) stage compared with low PHPT1 expression levels. Patients with low PHPT1 expression showed better overall survival and progression-free survival compared with those with high PHPT1 expression. In addition, multivariate analysis showed that nuclear grade and pTNM stage were independent predictors of progression-free survival and overall survival in patients with clear-cell renal cell carcinoma. PHPT1 expression was also an independent predictor of overall survival but not progression-free survival. PHPT1 was expressed in the epithelium of proximal tubuli and nuclei of clear-cell renal cell carcinoma tissue samples. High levels of 14 kDa phosphohistidine phosphatase protein were negatively associated with overall survival and progression-free survival in patients with clear-cell renal cell carcinoma.

In several malignancies extracapsular lymph node involvement (ECLNI) identifies a subgroup of patients with worse prognosis but no data are available on its significance in pancreatic ductal adenocarcinoma (PDAC). The aim of our study was to assess the prognostic relevance of ECLNI in resectable PDAC. A retrospective analysis was performed of 137 consecutive pancreatic resections for PDAC. Two investigators blinded for survival data systematically reviewed all pathological data. Survival curves were estimated using the Kaplan-Meier method. Multivariable Cox regression models were used to identify predictors of disease-free survival (DFS) and overall survival (OS). The median follow-up after surgery was 19 months. ECLNI was identified in 59 of 99 node-positive patients. The median DFS in patients with ECLNI vs. intracapsular LNI (ICLNI) was 6.8 vs. 12.0 months, respectively (P=.027). The median OS in patients with ECLNI vs. ICLNI was 16.1 and 21.8 months, respectively (P=.098). On multivariable analysis extracapsular lymph node ratio (ECLNR) was identified as an independent predictor of OS (P=.003). In patients with ECLNI, both OS and DFS were improved after adjuvant chemoradiation compared with those who did not receive adjuvant treatment (P=.01). Extracapsular lymph node ratio is an independent predictor of survival in patients with PDAC. Patients with ECLNI from pancreatic cancer seem to benefit from adjuvant chemoradiation but not from chemotherapy alone.

Real-world Outcomes and Factors Predicting Survival and Completion of Radium 223 in Metastatic Castrate-resistant Prostate Cancer

The conditions which must be satisfied by an index of biological age are discussed, and a high correlation with chronological age is shown to be neither a necessary nor a sufficient condition to be satisfied by any such index.