Manipolazione del metabolismo degli xenobiotici da frutta convenzionale ed attività chemiopreventiva

A reduced cancer risk associated with fruit and vegetable phytochemicals initially dictated chemopreventive approaches focused on specific green variety consumption or even single nutrient supplementations. However, these strategies not only failed to provide any health benefits but gave rise to detrimental effects. In parallel, public-health chemoprevention programmes were developed in the USA and Europe to increase whole vegetable consumption. Among these, the National Cancer Institute (NCI) sponsored plan “5 to 9 a day for a better health” was one of the most popular. This campaign promoted wide food choice through the consumption of at least 5 to 9 servings a day of colourful fruits and vegetables. In this study the effects of the diet suggested by NCI on transcription, translation and catalytic activity of both xenobiotic metabolizing (XME) and antioxidant enzymes were studied in the animal model. In fact, the boost of both antioxidant defences and “good” phase-II together with down-regulation of “bad” phase-I XMEs is still considered one of the most widely-used strategies of cancer control. Six male Sprague Dawley rats for each treatment group were used. According to the Italian Society of Human Nutrition, a serving of fruit, vegetables and leafy greens corresponds to 150, 250 and 50 g, respectively, in a 70 kg man. Proportionally, rats received one or five servings of lyophilized onion, tomato, peach, black grape or lettuce – for white, red, yellow, violet or green diet, respectively - or five servings of each green (“5 a day” diet) by oral gavage daily for 10 consecutive days. Liver subcellular fractions were tested for various cytochrome P450 (CYP) linked-monooxygenases, phase-II supported XMEs such as glutathione S-transferase (GST) and UDP-glucuronosyl transferase (UDPGT) as well as for some antioxidant enzymes. Hepatic transcriptional and translational effects were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. dROMs test was used to measure plasmatic oxidative stress. Routine haematochemical parameters were also monitored. While the five servings administration didn’t significantly vary XME catalytic activity, the lower dose caused a complex pattern of CYP inactivation with lettuce exerting particularly strong effects (a loss of up to 43% and 45% for CYP content and CYP2B1/2-linked XME, respectively; P<0.01). “5 a day” supplementation produced the most pronounced modulations (a loss of up to 60% for CYP2E1-linked XME and a reduction of CYP content of 54%; P<0.01). Testosterone hydroxylase activity confirmed these results. RT-PCR and Western blot analysis revealed that the “5 a day” diet XMEs inactivations were a result of both a transcriptional and a translational effect while lettuce didn’t exert such effects. All administrations brought out none or fewer modulation of phase-II supported XMEs. Apart from “5 a day” supplementation and the single serving of lettuce, which strongly induced DT- diaphorase (an increase of up to 141 and 171%, respectively; P<0.01), antioxidant enzymes were not significantly changed. RT-PCR analysis confirmed DT-diaphorase induction brought about by the administration of both “5 a day” diet and a single serving of lettuce. Furthermore, it unmasked a similar result for heme-oxygenase. dROMs test provided insight into a condition of high systemic oxidative stress as a consequence of animal diet supplementation with “5 a day” diet and a single serving of lettuce (an increase of up to 600% and 900%, respectively; P<0.01). Haematochemical parameters were mildly affected by such dietary manipulations. According to the classical chemopreventive theory, these results could be of particular relevance. In fact, even if antioxidant enzymes were only mildly affected, the phase-I inactivating ability of these vegetables would be a worthy strategy to cancer control. However, the recorded systemic considerable amount of reactive oxygen species and the complexity of these enzymes and their functions suggest caution in the widespread use of vegan/vegetarian diets as human chemopreventive strategies. In fact, recent literature rather suggests that only diets rich in fruits and vegetables and poor in certain types of fat, together with moderate caloric intake, could be associated with reduced cancer risk.

Secondary Microseisms Characterization and Green’s Function Extraction at the Larderello-Travale Geothermal Field (Italy)

Over the past ten years, the cross-correlation of long-time series of ambient seismic noise (ASN) has been widely adopted to extract the surface-wave part of the Green’s Functions (GF). This stochastic procedure relies on the assumption that ASN wave-field is diffuse and stationary. At frequencies <1Hz, the ASN is mainly composed by surface-waves, whose origin is attributed to the sea-wave climate. Consequently, marked directional properties may be observed, which call for accurate investigation about location and temporal evolution of the ASN-sources before attempting any GF retrieval. Within this general context, this thesis is aimed at a thorough investigation about feasibility and robustness of the noise-based methods toward the imaging of complex geological structures at the local (∼10-50km) scale. The study focused on the analysis of an extended (11 months) seismological data set collected at the Larderello-Travale geothermal field (Italy), an area for which the underground geological structures are well-constrained thanks to decades of geothermal exploration. Focusing on the secondary microseism band (SM;f>0.1Hz), I first investigate the spectral features and the kinematic properties of the noise wavefield using beamforming analysis, highlighting a marked variability with time and frequency. For the 0.1-0.3Hz frequency band and during Spring- Summer-time, the SMs waves propagate with high apparent velocities and from well-defined directions, likely associated with ocean-storms in the south- ern hemisphere. Conversely, at frequencies >0.3Hz the distribution of back- azimuths is more scattered, thus indicating that this frequency-band is the most appropriate for the application of stochastic techniques. For this latter frequency interval, I tested two correlation-based methods, acting in the time (NCF) and frequency (modified-SPAC) domains, respectively yielding esti- mates of the group- and phase-velocity dispersions. Velocity data provided by the two methods are markedly discordant; comparison with independent geological and geophysical constraints suggests that NCF results are more robust and reliable.

Kinematic description of the rupture from strong motion data: strategies for a robust inversion

We present a non linear technique to invert strong motion records with the aim of obtaining the final slip and rupture velocity distributions on the fault plane. In this thesis, the ground motion simulation is obtained evaluating the representation integral in the frequency. The Green’s tractions are computed using the discrete wave-number integration technique that provides the full wave-field in a 1D layered propagation medium. The representation integral is computed through a finite elements technique, based on a Delaunay’s triangulation on the fault plane. The rupture velocity is defined on a coarser regular grid and rupture times are computed by integration of the eikonal equation. For the inversion, the slip distribution is parameterized by 2D overlapping Gaussian functions, which can easily relate the spectrum of the possible solutions with the minimum resolvable wavelength, related to source-station distribution and data processing. The inverse problem is solved by a two-step procedure aimed at separating the computation of the rupture velocity from the evaluation of the slip distribution, the latter being a linear problem, when the rupture velocity is fixed. The non-linear step is solved by optimization of an L2 misfit function between synthetic and real seismograms, and solution is searched by the use of the Neighbourhood Algorithm. The conjugate gradient method is used to solve the linear step instead. The developed methodology has been applied to the M7.2, Iwate Nairiku Miyagi, Japan, earthquake. The estimated magnitude seismic moment is 2.6326 dyne∙cm that corresponds to a moment magnitude MW 6.9 while the mean the rupture velocity is 2.0 km/s. A large slip patch extends from the hypocenter to the southern shallow part of the fault plane. A second relatively large slip patch is found in the northern shallow part. Finally, we gave a quantitative estimation of errors associates with the parameters.

Innovative techniques for sport turf managing: Agronomic and phyisiological implications

The growing substrate of the putting greens is considered a key factor for a healthy turf ecosystem. Actually detailed study on the effects of growth promoting bacteria and biostimulants on a professional sport turf are very limited. This thesis aimed to study the effectiveness of different microorganisms and biostimulants in order to improve the knowledge relative to the relationship between the beneficial microflora and root apparatus of sport turfs. The research project was divided in three principal steps: Initially, commercial products based on biostimulants and microorganisms were tested on a Lolium perenne L. essence grown in a controlled-environment. The principal evaluations were the study of the habitus of plants, biomass production and length of leaves and roots. Were studied the capacity of colonization of microorganisms within root tissues and rhizosphere. In the second step were developed two different biostimulant solutions based on effective microorganisms, mycorrhizae and humic acids. This test was conducted both on an Agrostis stolonifera putting green (Modena Golf & Country Club) in a semi-field condition and within a growth chamber on a Lolium perenne L. essence. Fungicide and chemicals applications were suspended in order to assess the effectiveness of the inoculants for nutrition and control of pests. In the last step, different microorganism mixes and biostimulants were tested on an experimental putting green in the Turf Research Center (TRC) (Virginia Tech, United States) in a real managing situation. The effects of different treatments were studied maintaining all chemicals and mechanicals managements scheduled during a sport season. Both growth-chamber and field results confirmed the capacity of microorganisms based biostimulants to promote the physiologic conditions of the plants, improve the growth of the roots and enhance the aesthetic performance of the turf. Molecular analysis confirmed the capacity of microorganisms to colonize the root tissues.