Variations in epiphyseal fusion and persistence of the epiphyseal line in the appendicular skeleton of two identified modern (19th-20th c.) adult Portuguese and Italian samples.

OBJECTIVES: The aim of this work is to study age, sex, and population variations in epiphyseal fusion and persistence of the epiphyseal line in the appendicular skeleton of two identified modern (19th-20th c.) adult skeletal samples, using a specifically designed macroscopic scoring method. The use of epiphyseal closure and persistence of the epiphyseal line as an adult-age marker is also discussed. MATERIALS AND METHODS: This study examined 981 adult skeletons of both sexes from two identified modern (20th c.) skeletal samples from the Sassari Collection (Museum of Anthropology, University of Bologna, Italy) and the Colecção de Esqueletos Identificados (Museum of Anthropology, University of Coimbra, Portugal). Our scoring method considers a five-degree scale, from nonfusion (Degree 0) to complete fusion (Degree 4). In addition, the persistence of the epiphyseal line, a feature that is not commonly collected during routine anthropological analyses, is taken into account here as Degree 3. RESULTS: Intra- and interobserver errors of 1.2% and 5.2%, respectively, were found, suggesting a good reproducibility of this scoring method. Some sites show variable degrees of epiphyseal fusion still in adult skeletons (e.g., secondary center of ossification of the clavicle, iliac crest, ischial tuberosity, distal epiphysis of the radius and ulna). CONCLUSIONS: Population differences have been observed, showing a delay in the complete epiphyseal closure for the Sassari sample compared to the Coimbra sample. Degree 3 seems to be a good adult-age indicator for individuals less than 35-year-old.

Profiling age and body fluid DNA methylation markers using nanopore adaptive sampling.

DNA methylation plays essential roles in regulating physiological processes, from tissue and organ development to gene expression and aging processes and has emerged as a widely used biomarker for the identification of body fluids and age prediction. Currently, methylation markers are targeted independently at specific CpG sites as part of a multiplexed assay rather than through a unified assay. Methylation detection is also dependent on divergent methodologies, ranging from enzyme digestion and affinity enrichment to bisulfite treatment, alongside various technologies for high-throughput profiling, including microarray and sequencing. In this pilot study, we test the simultaneous identification of age-associated and body fluid-specific methylation markers using a single technology, nanopore adaptive sampling. This innovative approach enables the profiling of multiple CpG marker sites across entire gene regions from a single sample without the need for specialized DNA preparation or additional biochemical treatments. Our study demonstrates that adaptive sampling achieves sufficient coverage in regions of interest to accurately determine the methylation status, shows a robust consistency with whole-genome bisulfite sequencing data, and corroborates known CpG markers of age and body fluids. Our work also resulted in the identification of new sites strongly correlated with age, suggesting new possible age methylation markers. This study lays the groundwork for the systematic development of nanopore-based methodologies in both age prediction and body fluid identification, highlighting the feasibility and potential of nanopore adaptive sampling while acknowledging the need for further validation and expansion in future research.

Redox Parameters as Markers of the Rate of Aging and Predictors of Life Span.

Oxidative stress has been reported to increase with aging, and although several age-related changes in redox parameters have been described, none of them have been verified as markers of the rate of aging and life span. Therefore, antioxidant (catalase, glutathione peroxidase, reductase activities, and reduced glutathione) and oxidant (oxidized glutathione, basal superoxide anion, and malondialdehyde concentrations) parameters were studied in whole blood cells from humans divided into different age groups (adult, mature, older adult, nonagenarian, and centenarian) in a cross-sectional study. Moreover, the same parameters were investigated in peritoneal leukocytes of mice at the analogous human ages (adult, mature, old, very old, and long-lived) in a longitudinal study as well as in adult prematurely aging mice. The results reveal that the age-related alterations of these markers are similar in humans and mice, with decreased antioxidants and increased oxidants in old participants, whereas long-lived individuals show similar values to those in adults. In addition, adult prematurely aging mice showed similar values to those in chronologically old mice and had a shorter life span than nonprematurely aging mice. Thus, these parameters could be proposed as markers of the rate of aging and used to ascertain biological age in humans.

Aging markers in human urine: A comprehensive, non-targeted LC-MS study.

Metabolites in human biofluids document the physiological status of individuals. We conducted comprehensive, non-targeted, non-invasive metabolomic analysis of urine from 27 healthy human subjects, comprising 13 young adults (30 ± 3 years) and 14 seniors (76 ± 4 years). Quantitative analysis of 99 metabolites revealed 55 that displayed significant differences in abundance between the two groups. Forty-four did not show a statistically significant relationship with age. These include 13 standard amino acids, 5 methylated, 4 acetylated, and 9 other amino acids, 6 nucleosides, nucleobases, and derivatives, 4 sugar derivatives, 5 sugar phosphates, 4 carnitines, 2 hydroxybutyrates, 1 choline, and 1 ethanolamine derivative, and glutathione disulfide. Abundances of 53 compounds decreased, while 2 (glutathione disulfide, myo-inositol) increased in elderly people. The great majority of age-linked markers were highly correlated with creatinine. In contrast, 44 other urinary metabolites, including urate, carnitine, hippurate, and betaine, were not age-linked, neither declining nor increasing in elderly subjects. As metabolite profiles of urine and blood are quite different, age-related information in urine offers additional valuable insights into aging mechanisms of endocrine system. Correlation analysis of urinary metabolites revealed distinctly inter-related groups of compounds.