Transient ALT Activation Protects Human Primary Cells From Chromosome Instability Induced by Low Chronic Oxidative Stress

Cells are often subjected to the effect of reactive oxygen species (ROS) as a result of both intracellular metabolism and exposure to exogenous factors. ROS-dependent oxidative stress can induce 8-oxodG within the GGG triplet found in the G-rich human telomeric sequence (TTAGGG), making telomeres highly susceptible to ROS-induced oxidative damage. Telomeres are nucleoprotein complexes that protect the ends of linear chromosomes and their dysfunction is believed to affect a wide range of cellular and/or organismal processes. Acute oxidative stress was shown to affect telomere integrity, but how prolonged low level oxidative stress, which may be more physiologically relevant, affects telomeres is still poorly investigated. Here, we explored this issue by chronically exposing human primary fibroblasts to a low dose of hydrogen peroxide. We observed fluctuating changes in telomere length and fluctuations in the rates of chromosome instability phenotypes, such that when telomeres shortened, chromosome instability increased and when telomeres lengthened, chromosome instability decreased. We found that telomere length fluctuation is associated with transient activation of an alternative lengthening of telomere (ALT) pathway, but found no evidence of cell death, impaired proliferation, or cell cycle arrest, suggesting that ALT activation may prevent oxidative damage from reaching levels that threaten cell survival

Human fibroblasts in vitro exposed to 2.45 GHz continuous and pulsed wave signals: Evaluation of biological effects with a multimethodological approach

The increasing exposure to radiofrequency electromagnetic fields (RF-EMF), especially from wireless communication devices, raises questions about their possible adverse health effects. So far, several in vitro studies evaluating RF-EMF genotoxic and cytotoxic non-thermal effects have reported contradictory results that could be mainly due to inadequate experimental design and lack of well-characterized exposure systems and conditions. Moreover, a topic poorly investigated is related to signal modulation induced by electromagnetic fields. The aim of this study was to perform an analysis of the potential non-thermal biological effects induced by 2.45 GHz exposures through a characterized exposure system and a multimethodological approach. Human fibroblasts were exposed to continuous (CW) and pulsed (PW) signals for 2 h in a wire patch cell-based exposure system at the specific absorption rate (SAR) of 0.7 W/kg. The evaluation of the potential biological effects was carried out through a multimethodological approach, including classical biological markers (genotoxic, cell cycle, and ultrastructural) and the evaluation of gene expression profile through the powerful high-throughput next generation sequencing (NGS) RNA sequencing (RNA-seq) approach. Our results suggest that 2.45 GHz radiofrequency fields did not induce significant biological effects at a cellular or molecular level for the evaluated exposure parameters and conditions

Overview of DISCOVER22 experiment in the framework of INFN-LNGS Cosmic Silence activity: challenges and improvements in underground radiobiology

One of the most intriguing and still pending questions in radiobiology is to understand whether and how natural environmental background radiation has shaped Life over millions of years of evolution on Earth. Deep Underground Laboratories (DULs) represent the ideal below-background exposure facilities where to address such a question. Among the few worldwide DULs, INFN-Laboratorio Nazionale del Gran Sasso (LNGS) is one of the largest in terms of size and infrastructure. Designed and built to host neutrino and dark matter experiments, since the 1990 s the LNGS has been one of the first DULs to systematically host radiobiology experiments. Here we present the DISCOVER22 (DNA Damage and Immune System Cooperation in VEry low Radiation environment 2022) experiment recently started at LNGS. DISCOVER22 aims at investigating how the low radiation background modulates the Immune System (IS) response in in vitro and in vivo models. Underground radiobiology experiments are particularly complex and tricky to design and perform. In these studies, the accurate characterization of exposure scenarios is mandatory, but a challenging aspect is to understand how the very few ionizing tracks in the ultra-Low Radiation Environment (LRE) interact with the living matter in space and time in order to trigger different biological responses. In this Perspective, we describe these challenges and how we address them through a microdosimetric and a radiobiological approaches. We aim at linking physical microdosimetric measurements and the corresponding biological radiation responses by using radiation biophysical models that could shed light on many as yet unresolved questions

The Phylogeny of the Spleen

The spleen can exert different functions: hematopoiesis, immune response, blood filtration, and blood storage. Presence and importance of each function vary among different species and, within a single species, vary during ontogeny. During the course of evolution, loss of hematopoietic functions was accompanied by increased specialization of other functions. Increase in the complexity of immunological features can be seen in the passage from scattered lymphocytes to segregation of the white pulp, organization of T- and B-lymphocyte regions and, finally, appearance of germinal centers. The red pulp remained essentially the same in all nonmammalian vertebrates, with a microarchitecture that concentrates blood and allows its filtration. In our view, this high splenic hematocrit was eventually co-opted in teleosts and mammals to develop a new function: the storage of red blood cells. Finally, mammalian species evolved different features of the red pulp, increasing the specialization of the filtration function (pulp sinuses) and/or storing function (muscularization)

Rates of erythropoiesis in mammals and their relationship with lifespan and hematopoietic stem cells aging

Investigations on possible links between hematological parameters and longevity are nearly absent. We tested the hypothesis that a fast rate of erythropoiesis, causing an earlier aging of the hematopoietic stem cells pool, contributes to a shorter lifespan. With this aim, we employed a new quantity, daily produced red blood cells per gram of body mass, as a measure of mass-specific rate of erythropoiesis. We found that among mammals rate of erythropoiesis and maximum lifespan are significantly correlated, independently from mass residuals. This seems to be confirmed also by intra-species comparisons and, although with limited data, by the significant correlation of rate of erythropoiesis and rate of telomere shortening in leukocytes (a proxy for hematopoietic stem cell telomere shortening). In our view, this may give a link of causality between rate of erythropoiesis and maximum lifespan. Further studies could test a similar hypothesis also for other kinds of stem cells