Zinco, invecchiamento e sistema immunitario: effetti sull'apoptosi e sulla proliferazione dei linfociti

Immunosenescence is characterized by a complex remodelling of the immune system, mainly driven by lifelong antigenic burden. Cells of the immune system are constantly exposed to a variety of stressors capable of inducing apoptosis, including antigens and reactive oxygen species continuously produced during immune response and metabolic pathways. The overall homeostasis of the immune system is based on the balance between antigenic load, oxidative stress, and apoptotic processes on one side, and the regenerative potential and renewal of the immune system on the other. Zinc is an essential trace element playing a central role on the immune function, being involved in many cellular processes, such as cell death and proliferation, as cofactor of enzymes, nuclear factors and hormones. In this context, the age associated changes in the immune system may be in part due to zinc deficiency, often observed in aged subjects and able to induce impairment of several immune functions. Thus, the aim of this work was to investigate the role of zinc in two essential events for immunity during aging, i.e. apoptosis and cell proliferation. Spontaneous and oxidative stress-induced apoptosis were evaluated by flow cytometry in presence of a physiological concentration of zinc in vitro on peripheral blood mononuclear cells (PBMCs) obtained from healthy subjects of different age: a group of young subjects, a group of old subjects and a group of nonagenarians. In addition, cell cycle phases were analyzed by flow cytometry in PBMCs, obtained from the subjects of the same groups in presence of different concentration of zinc. We also analyzed the influence of zinc in these processes in relation to p53 codon 72 polymorphism, known to affect apoptosis and cell cycle in age-dependent manner. Zinc significantly reduces spontaneous apoptosis in all age-groups; while it significantly increases oxidative stress-induced late apoptosis/necrosis in old and nonagenarians subjects. Some factors involved in the apoptotic pathway were studied and a zinc effect on mitochondrial membrane depolarization, cytochrome C release, caspase-3 activation, PARP cleavage and Bcl-2 expression was found. In conclusion, zinc inhibits spontaneous apoptosis in PBMCs contrasting the harmful effects due to the cellular culture conditions. On the other hand, zinc is able to increase toxicity and induce cell death in PBMCs from aged subjects when cells are exposed to stressing agents that compromise antioxidant cellular systems. Concerning the relationship between the susceptibility to apoptosis and p53 codon 72 genotype, zinc seems to affect apoptosis only in PBMCs from Pro- people suggesting a role of this ion in strengthening the mechanism responsible of the higher propensity of Pro- towards apoptosis. Regarding cell cycle, high doses of zinc could have a role in the progression of cells from G1 to S phase and from S to G2/M phase. These effect seems depend on the age of the donor but seems to be unrelated to p53 codon 72 genotype. In order to investigate the effect of an in vivo zinc supplementation on apoptosis and cell cycle, PBMCs from a group of aged subjects were studied before and after six weeks of oral zinc supplementation. Zinc supplementation reduces spontaneous apoptosis and it strongly reduces oxidative stress-induced apoptosis. On the contrary, no effect of zinc was observed on cell cycle. Therefore, it’s clear that in vitro and in vivo zinc supplementation have different effects on apoptosis and cell cycle in PBMCs from aged subjects. Further experiments and clinical trials are necessary to clarify the real effect of an in vivo zinc supplementation because this preliminary data could encourage the of this element in all that disease with oxidative stress pathogenesis. Moreover, the expression of metallothioneins (MTs), proteins well known for their zinc-binding ability and involved in many cellular processes, i.e. apoptosis, metal ions detoxification, oxidative stress, differentiation, was evaluated in total lymphocytes, in CD4+ and in CD8+ T lymphocytes from young and old healthy subjects in presence of different concentration of zinc in vitro. Literature data reported that during ageing the levels of these proteins increase and concomitantly they lose the ability to release zinc. This fact induce a down-regulation of many biological functions related to zinc, such as metabolism, gene expression and signal transduction. Therefore, these proteins may turn from protective in young-adult age to harmful agents for the immune function in ageing following the concept that several genes/proteins that increase fitness early in life may have negative effects later in life: named “Antagonistic Pleyotropy Theory of Ageing”. Data obtained in this work indicate an higher and faster expression of MTs with lower doses of zinc in total lymphocytes, in CD4+ and in CD8+ T lymphocytes from old subjects supporting the antagonistic pleiotropic role of these proteins.

Erosione e sollevamento nell'arco calabro-peloritano

The Calabrian-Peloritani arc represents key site to unravel evolution of surface processes on top of subducting lithosphere. During the Pleistocene, in fact the arc uplifted at rate of the order of about 1mm/yr, forming high-standing low-relief upland (figure 2). Our study is focused on the relationship between tectonic and land evolution in the Sila Massif, Messina strait and Peloritani Mts. Landforms reflect a competition between tectonic, climatic, and surficial processes. Many landscape evolution models that explore feedbacks between these competing processes, given steady forcing, predict a state of erosional equilibrium, where the rates of river incision and hillslope erosion balance rock uplift. It has been suggested that this may be the final constructive stage of orogenic systems. Assumptions of steady erosion and incision are used in the interpretation of exhumation and uplift rates from different geologic data, and in the formulation of fluvial incision and hillslope evolution models. In the Sila massif we carried out cosmogenic isotopes analysis on 24 samples of modern fluvial sediments to constrain long-term (~103 yr) erosion rate averaged on the catchment area. 35 longitudinal rivers profiles have been analyzed to study the tectonic signal on the landscape evolution. The rivers analyzed exhibit a wide variety of profile forms, diverging from equilibrium state form. Generally the river profiles show at least 2 and often 3 distinct concave-up knickpoint-bounded segments, characterized by different value of concavity and steepness indices. River profiles suggest three main stages of incision. The values of ks and θ in the lower segments evidence a decrease in river incision, due probably to increasing uplift rate. The cosmogenic erosion rates pointed out that old landscape upland is eroding slowly at ~0.1 mm/yr. In the contrary, the flanks of the massif is eroding faster with value from 0.4 to 0.5 mm/yr due to river incision and hillslope processes. Cosmogenic erosion rates mach linearly with steepness indices and with average hillslope gradient. In the Messina area the long term erosion rate from low-T thermochronometry are of the same order than millennium scale cosmogenic erosion rate (1-2 mm/yr). In this part of the chain the fast erosion is active since several million years, probably controlled by extensional tectonic regime. In the Peloritani Mts apatite fission-track and (U-Th)/He thermochronometry are applied to constraint the thermal history of the basement rock. Apatite fission-track ages range between 29.0±5.5 and 5.5±0.9 Ma while apatite (U-Th)/He ages vary from 19.4 to 1.0 Ma. Most of the AFT ages are younger than the overlying terrigenous sequence that in turn postdates the main orogenic phase. Through the coupling of the thermal modelling with the stratigraphic record, a Middle Miocene thermal event due to tectonic burial is unravel. This event affected a inner-intermediate portion of the Peloritani belt confined by young AFT data (<15 Ma) distribution. We interpret this thermal event as due to an out-of–sequence thrusting occurring in the inner portion of the belt. Young (U-Th)/He ages (c. 5 Ma) record a final exhumation stage with increasing rates of denudation since the Pliocene times due to postorogenic extensional tectonics and regional uplift. In the final chapter we change the spatial scale to insert digital topography analysis and field data within a geodynamic model that can explain surface evidence produced by subduction process.

Fracture Mechanics Investigation of Structures with Defects

Fracture mechanics plays an important role in the material science, structure design and industrial production due to the failure of materials and structures are paid high attention in human activities. This dissertation, concentrates on some of the fractural aspects of shaft and composite which have being increasingly used in modern structures, consists four chapters within two parts. Chapters 1 to 4 are included in part 1. In the first chapter, the basic knowledge about the stress and displacement fields in the vicinity of a crack tip is introduced. A review involves the general methods of calculating stress intensity factors are presented. In Chapter 2, two simple engineering methods for a fast and close approximation of stress intensity factors of cracked or notched beams under tension, bending moment, shear force, as well as torque are presented. New formulae for calculating the stress intensity factors are proposed. One of the methods named Section Method is improved and applied to the three dimensional analysis of cracked circular section for calculating stress intensity factors. The comparisons between the present results and the solutions calculated by ABAQUS for single mode and mixed mode are studied. In chapter 3, fracture criteria for a crack subjected to mixed mode loading of two-dimension and three-dimension are reviewed. The crack extension angle for single mode and mixed mode, and the critical loading domain obtained by SEDF and MTS are compared. The effects of the crack depth and the applied force ratio on the crack propagation angle and the critical loading are investigated. Three different methods calculating the crack initiation angle for three-dimension analysis of various crack depth and crack position are compared. It should be noted that the stress intensity factors used in the criteria are calculated in section 2.1.