The role of epigenetics in endothelial cell senescence is a cutting‐edge topic in ageing research. However, little is known of the relative contribution to pro‐senescence signal propagation provided by microRNAs shuttled by extracellular vesicles (EVs) released from senescent cells. Analysis of microRNA and DNA methylation profiles in non‐senescent (control) and senescent (SEN) human umbilical vein endothelial cells (HUVECs), and microRNA profiling of their cognate small EVs (sEVs) and large EVs demonstrated that SEN cells released a significantly greater sEV number than control cells. sEVs were enriched in miR‐21‐5p and miR‐217, which target DNMT1 and SIRT1. Treatment of control cells with SEN sEVs induced a miR‐21/miR‐217‐related impairment of DNMT1‐SIRT1 expression, the reduction of proliferation markers, the acquisition of a senescent phenotype and a partial demethylation of the locus encoding for miR‐21. MicroRNA profiling of sEVs from plasma of healthy subjects aged 40–100 years showed an inverse U‐shaped age‐related trend for miR‐21‐5p, consistent with senescence‐associated biomarker profiles. Our findings suggest that miR‐21‐5p/miR‐217 carried by SEN sEVs spread pro‐senescence signals, affecting DNA methylation and cell replication.

Alzheimer's Disease (AD) is the most common neurodegenerative disease and the leading cause of dementia among senile subjects. It has been proposed that AD can be caused by defects in mitochondrial oxidative phosphorylation. Given the fundamental contribution of the mitochondrial genome (mtDNA) for the respiratory chain, there have been a number of studies investigating the association between mtDNA inherited variants and multifactorial diseases, however no general consensus has been reached yet on the correlation between mtDNA haplogroups and AD. We applied for the first time a high resolution analysis (sequencing of displacement loop and restriction analysis of specific markers in the coding region of mtDNA) to investigate the possible association between mtDNA-inherited sequence variation and AD in 936 AD patients and 776 cognitively assessed normal controls from central and northern Italy. Among over 40 mtDNA sub-haplogroups analysed, we found that sub-haplogroup H5 is a risk factor for AD (OR = 1.85, 95% CI:1.04–3.23) in particular for females (OR = 2.19, 95% CI:1.06–4.51) and independently from the APOE genotype. Multivariate logistic regression revealed an interaction between H5 and age. When the whole sample is considered, the H5a subgroup of molecules, harboring the 4336 transition in the tRNA^Gln gene, already associated to AD in early studies, was about threefold more represented in AD patients than in controls (2.0% vs 0.8%; p = 0.031), and it might account for the increased frequency of H5 in AD patients (4.2% vs 2.3%). The complete re-sequencing of the 56 mtDNAs belonging to H5 revealed that AD patients showed a trend towards a higher number (p = 0.052) of sporadic mutations in tRNA and rRNA genes when compared with controls. Our results indicate that high resolution analysis of inherited mtDNA sequence variation can help in identifying both ancient polymorphisms defining sub-haplogroups and the accumulation of sporadic mutations associated with complex traits such as AD.

Summary: Sequence variations in a variety of pro‐ or anti‐inflammatory cytokine genes have been found to influence successful aging and longevity. Because of the role played by the transforming growth factor β1 (TGF‐β1) cytokine in inflammation and regulation of immune responses, the variability of the TGF‐β1 gene may affect longevity by playing a role in inflamm‐aging. Two polymorphisms, G/A₋₈₀₀ and C/T₋₅₀₉, located in the 5′ region, and two missense polymorphisms, T/C₈₆₉ and G/C₉₁₅ which change (Leu > Pro)₁₀ and (Arg > Pro)₂₅, respectively, located in the signal peptide, were analysed in 419 subjects from Northern and Central Italy, including 172 centenarians and 247 younger controls. In addition, the effects of the TGF‐β1 genetic variability on plasma levels of the biologically active form (naturally processed) of this cytokine were studied in 143 randomly selected subjects, including 73 centenarians. Significant differences were found at the +915 site as far as the C allele and GC genotype were concerned, both of them being lower in centenarians than in young controls (P = 0.034 and 0.028, respectively), but none of the other tested genetic variants was significantly different between centenarians and controls. Moreover, a particular haplotype combination (G₋₈₀₀/C₋₅₀₉/C₈₆₉/C₉₁₅) was notably lower in centenarians than in younger individuals (P = 0.007). Finally, active TGF‐β1 plasma levels were significantly increased in the elderly group, but no relationship with TGF‐β1 genotypes was observed. These results suggest that, at least in this population, the variability of the TGF‐β1 gene influences longevity and that the age‐related increase in plasma levels of active TGF‐β1 seems not to be genetically regulated.

N-glycan profiling of the human serum glycoproteins including immunoglobulin fraction on different age groups of healthy persons shows substantial changes with increasing age in three major N-glycan structures. In individuals more than 40–50 years of age, there is an increase in under-galactosylated glycans and a decrease in the core α -1,6-fucosylated bi-galactosylated biantennary structure. These three glycan structures are also substantially changed in a Werner syndrome patient, to a level comparable or even more pronounced than those observed in a healthy Italian centenarian population. These data show that the glycosylation machineries in both liver cells and B-cells are affected in a similar way by the aging process despite their highly different nature. The observed changes in the glycan structures are indicative that biosynthetic processes are at the basis of the changes, possibly together with changes in serum clearing of glycan-altered proteins. Our data suggest that measurement of the N-glycan level changes could provide a noninvasive surrogate marker for general health or for age-related disease progression, and for monitoring the improvement of health after therapy.