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La cooperazione di polizia e giudiziaria e la circolazione della prova digitale nello spazio giudiziario europeo: un bilanciamento critico tra libertà, sicurezza e giustizia.
No full textThe modern society is the result of a technological progress, as a sign of goodness and richness.
As Mark Mazower observes, it is necessary to consider the “dark side” of the development, in order to avoid the dangers.
The new technologies bring with them a hidden potential of violence, stressed by the relevant role in human life.
Sociologists and psychologists are afraid of the unbalanced relation between man and machine, in which the first one is the slave of the second one.
In fact, many times the computer instruments become a way for the commission (or for a better and fast commission) of a crime.
Everyone would be in condition to know and understand the risks of the progress, in order to avoid them.
This form of perception of the negatives is facilitated by the ways of communication, that allow to know hypothesis of cybercrimes, considered as the crimes committed by or on the computer (or others technological supports).
Remember, for example, the famous case ECHELON, or the case of the website Wikileaks, or the case of the serial killer, Matej Curko and so on.
These are only a little part of many news regarding to computer technologies.
The research is due to this social contemporary panorama, from which are developed the analysis on the transnational crimes, uses of new technologies and the problems of EU criminal justice and procedure.
In this field take importance the police and judicial cooperation (as informative cooperation too) between Member States, especially in the fight against transnational crimes, the circulation of e-evidences and the protection of data and of fundamental rights too.
The Lisbon Treaty traces the state of the project of the European space of freedom, security and justice.
The development of the police and judicial cooperation is one of the elements useful for the realization of this objective.
The European law of cooperation and the interpretation made by the European Court of Justice, provide new powers of the EU institutions and also the recognition of the fundamental rights, written in the Nice Charta and in the ECHR.
An integration of the national judicial orders needs the introduction of communitarian laws.
In cooperation material, from the Convention of 1959 to the Bruxelles Convention of 2000, many things are changed.
Out of these conventional instruments, we can consider also others important elements of development, as the Decision of EAW and the Decision of EEW.
Many communitarian bodies are involved in the cooperation and their powers are increased during the time, as Europol, Eurojust and Olaf.
These organisms are really so important in the fight against transnational and cyber crimes, in particular because they have member of every State and a good sight on the general EU space.
The use of new technologies has increased the number of digital evidence, collected and useful for a decision of a criminal proc eeding.
In the european judicial space there is not a common definition of the concept of digital evidence, but every national order use this.
In the USA were developed the first studies in this material, related to the computer forensics, a method of analysis of the e -evidence.
There is not a list of best practices in computer forensics too, but every police organism that use this method has a internal regulation (not binding).
This condition of no common rules and criteria generates many problems to the judicial authorities, called to use or not use for the decision an e-evidence.
But the digital evidence, in a context of fight against serious transnational crimes, needs to circulate between the States involved, balancing the interest of security and justice and the right to privacy .
In order to realize this objective, all the infrastructure for the collection and circulation of data must be secure and controlled by specialized staff.
In the EU (but also in the member States) there are many databases, useful for the collection of electronic evidences and for the circulation of them, in the procedure of police and judicial cooperation.
The most important are SIS II, EPOC III of Eurojust, TECS of Europol, the database of Olaf, ECRIS, VIS, EURODAC and VIS.
Each of these archives has special rules for access, collection and circulation of data and, in general, operational rules applicable.
In these field we have to consider the protection of fundamental rights and of the procedural rights too. In particular, the right of a fair trial, of translation and interpretation, of defence and of privacy.
In a multilanguage EU space, it is more difficult for the police and judicial authorities to cooperate and understand eachother. The increasing of powers of communitarian bodies (Europol, Eurojust and Olaf) and the development con common rules and principles, can realize a better collaboration between member States, as underlined in the Lisbon Treaty.
The Europen Union, as stressed in the project Europe 2020,intends to put money for growing up the progress, technical and economical.
The innovation needs a clear common response, by a normative way too.
In many situations is better the choice of mutual recognition, as already known in the European context, in many others is better to harmonize (as possible) the principles and the laws between member States.
Regarding the development of the EU judicial space, especially in criminal law and procedure, the communitarian order focus on t he judicial, police and informative cooperation. This is stressed also in the Sthockolm Program and in the Action Plan, in which are provided reforms in these matters.
So, all the European citizen can hope in a EU space of freedom, security and justice and also in a communitarian criminal justice, with guaranties of procedural and fundamental rights, as written in Nice Charta, in ECHR and in the national Constitutions
Methylation profiles of exocrine and neuroendocrine colorectal carcinomas using methylation-specific multiple ligation-dependent probe amplification.
No full textIn this study, we employed Methylation-Specific Multiple Ligation-dependent Probe Amplification (MS-MLPA) to evaluate the methylation status of 34 genes in a series 104 formalin-fixed and paraffin-embedded specimens, including 83 exocrine adenocarcinomas (CRCs) and 21 neuroendocrine cancers (NECs), previously examined for clinico-pathological and molecular features, as well as in 25 colorectal mucosae from 13 CRC patients and from12 non-neoplastic patients.
We found higher levels of promoter methylation in normal colonic tissue of CRC-patients with respect to the control group. The CpG Island Methylation Phenotype (CIMP) was observed with similar frequency in exocrine CRCs and in NECs, but different methylation profiles were present in the two tumour types. Microsatellite instability was the marker most strongly associated with CIMP. In both CRCs and NECs, CIMP+/MSI+ tumours represented a homogeneous clinicopathological entity, associated with a very good prognosis compared with the other three classes of cancers.
Our results demonstrated that MS-MLPA is a rapid and sensitive method to analyse the methylation status of multiple genes simultaneously and presents innovative aspects that may have important scientific and clinical implications. The use of DNA methylation alterations as a molecular marker system could potentially be a powerful approach to population-based screening for the early detection and for risk assessment of colorectal cancer
Bacterial oxygenases: potential, features and new investigation methods.
No full textBacterial oxygenases, the enzymes involved in the breakdown pathways of aromatic and aliphatic hydrocarbons, have aroused great interest within the scientific community for their potential applications in a number of different fields: environmental (bioremediation and biomonitoring), biotechnological (green chemistry) and biomedical (drug production).
The aims of this study were 1) to clone new bacterial oxygenases using conventional methods and further develop new screening techniques and 2) to assess the ability of the newly-cloned enzyme(s), of toluene/o-xilene monooxygenase (ToMO) and of some of its mutants, to exploit various types of aromatic substrate on which the activity of these enzymes has never previously been assessed.
Using conventional techniques, Pseudomonas sp. N1 genes coding for naphthalene dioxygenase (NDO) and Sphingobium sp. PhS genes, putatively coding for Phenanthrene dioxygenase (PhDO), were cloned; of the two enzymes only NDO appeared to be functional. ToMO is an enzyme complex formed from six different subunits, ABCDEF; subunit A (TouA) is part of the terminal hydroxylase and contains the active site. In this study, in addition to wild type enzymes, the TouA mutants D211A and D211A/E214G, previously created by this laboratory, and TouA E214G, already described in the literature, were also used.
The ability of ToMO and TouA mutants, and of NDO to hydroxylate different aromatic compounds other than their natural substrates (toluene and naphthalene respectively) was evaluated by biotransformation assays using, as substrates, the following molecules: 1,2,3-trimethoxybenzene, anisole, benzophenone, bibenzyl, biphenyl, nitrobenzene, quinoline, and trans-stilbene. These compounds were chosen for the environmental, biotechnological and pharmacological importance of their hydroxylated derivatives. To this end, ToMO, TouA mutants and NDO were expressed in the heterologous host E. coli JM109. For both enzymes, the products of bioconversion were identified by GC-MS and, using the appropriate standard of reference, quantified by HPLC analysis.
ToMO proved active in all substrates tested. The mutant E214G, like the wild type, was capable of oxidising all the compounds tested: in almost all the biotransformations both the mutant form and the wild type behaved in the same way in that they both produced the same isomers in similar proportions. In terms of catalytic efficiency, the mutant E214G was found to be most powerful: the mutants D211A and D211A/E214G were capable of hydroxylating only a few of the substrates used (1,2,3-trimethoxybenzene, anisole, benzophenone, biphenyl) with very low yields. However, considering the fact that strains expressing the mutants D211A e D211A/E214G showed impaired growth and expression problems, probably attributable to unwanted mutations introduced during mutagenesis, these data cannot be considered definitive.
The ability of NDO to perform dihydroxylation processes was confirmed but, in addition, monohydroxylation reactions (quinoline) and dealkylation reactions (anisole and 1,2,3-trimethoxybenzene) were observed. The quantitative analysis of the biotransformation products showed that the enzyme efficiency decreased with the increasing steric footprint of the substrate. The failure of nitrobenzene to be transformed suggests that, in addition to steric footprint, other factors such as the presence of deactivating substituent groups plays a key role in the catalytic process of this enzyme.
Both NDO and ToMO have shown a good potential in the field of biotransformation and further studies could lead to the production of more useful and efficient enzyme variants.
During an internship at the University of Granada in Spain, a new protocol, based on the SIGEX (Substrate Induced Gene Expression) system, was developed to screen bacterial genomic libraries. This method, which is normally used to screen metagenomic libraries, was adapted to analyse libraries created from a single bacterial genome. With this technique it is possible to perform a rapid screening on the basis of a signal emitted from a reporter gene included in the construct used in creating the library. Using a previously constructed library, this approach allowed, in a short time, 60 clones to be isolated that had a high probability of containing naphthalene-dependent regulatory genes. The sequencing of the obtained clones, a necessary step in order to validate the protocol, in currently in progress
zCOSMOS survey galaxy groups: exploring the effect of group environment on galaxy properties
No full textThe evolution of galaxies in groups may have important implications for the global evolution of the galaxy population as a whole. The fraction of galaxies bound in groups at z ∼ 0 is as high as 60% and many processes operating in groups may concur in shaping galaxy evolution.
The rich zCOSMOS spectroscopic data (about 20000 galaxies with IAB ≤ 22.5 up to z ∼ 1) and its excellent group catalog (∼ 200 groups with more that 5 members up to z ∼ 1) coupled with the wide photometric coverage of the COSMOS survey, can shed new light on this topic, enabling us to study in a continuous way, up to z ∼ 1, the complex interplay between environment and galaxy evolution.
In this thesis I present the new results I have obtained on this topic by exploring the group-centric dependence of galaxy colors, masses, morphologies and star formation.
In brief, by building two composite groups at intermediate (0.2 ≤ z ≤ 0.45) and high (0.45 < z ≤ 0.8) redshifts, I was able to study in detail how galaxy stellar masses, colors, morphologies, and spectral features vary as a function of the distance from the group center. My analysis was performed in narrow bin of stellar masses/colors, in order to disentangle the obvious galaxy stellar mass/color dependencies.
To build the composite group I developed an algorithm to incorporate the galaxies brighter than IAB = 22.5 and missing a secure spectroscopic redshift, thus improving the statistics of the sample. To confidently determine all galaxy projected distance and rescale them into the composite group, I defined a new centering technique.
From the color/mass analysis I found that the evolution of most massive galaxies (log(Mgal/M⊙) > 10.6) is mainly driven by internal processes, as no strong group-centric environment dependence is visible.
For galaxies of lower masses (9.8 ≤ log(Mgal/M⊙) ≤ 10.6) there is a radial depen- dence in the changing mix of red and blue galaxies, red galaxies residing preferentially in the group center. Such dependence is most evident in poor groups, whereas richer groups do not display any obvious color trend.
Interestingly mass segregation shows the opposite behavior: it is visible only in rich groups, while poorer groups have a a constant mix of galaxy stellar masses as a function of radius.
The morpho-spectral analysis showed the presence of a mild morphological segregation at fixed galaxy stellar mass, with massive early-type galaxies preferentially located in the core of groups. Galaxies with 9.8 ≤ log(Mgal/M⊙) ≤ 10.6 exhibit the strongest morphological differences between group and field environment. These galaxies also have an excess of red-passive spirals in the group with respect to the field.
Despite the presence of a significant segregation of the spectral properties of group galaxies, such that the typical core galaxy have less intense emission lines, the star forming galaxies share the same level of activity at fixed stellar mass irrespective of the environment they reside in.
This findings can be explained in a simple scenario where color/SFR and mass segregation originates from different physical processes.
Mass segregation is driven by dynamical phenomena within groups, and therefore its presence/absence in rich/poor groups is a possible indication that poorer groups start to assemble later in cosmic time than richer structures.
The parallel absence/presence of color segregation in rich/poor groups hints to the fact that nurture effects are still in action in poorer structures, whereas in richer systems are already largely over, so that all galaxies are red irrespective of their position within the group (at least down to the galaxy stellar masses we explored).
Poorer groups hold the smoking gun of environmental effects in action superimposed to secular galaxy evolution: galaxies display gradually redder colors as a consequence of the still recent accretion history of these groups.
The physical processes causing these environmental effects should act on rather short timescales, 1.5-2 Gyrs, because they are not able to erase the strinking bi-modality of galaxy color distribution and, moreover, we are not able to observed blue active galaxies showing less intense star formation activity in groups than in the field
Environmental exposure to carbon nanotubes results in systemic distribution via the circulatory system: hepatotoxicity and central nervous system involvement.
No full textCarbon nanotubes (CNTs) are among the most promising nanomaterials and their interesting properties are now exploited in many fields of technology and biomedical applications. However, the rapid growth of CNT employment raises concerns about the potential risks and toxicities to public health, environment and workers associated with the manufacture and use of these new materials. Here we investigate the main routes of entry following environmental exposure to CNTs, their localization and possible role in inducing inflammation and specific pathologies, using a novel rodent model. Following environmental exposure, we observed that CNTs rapidly accumulate in the lungs and brain, later reaching other organs including the liver, where steatosis was found after chronic exposure to CNTs. Since SWCNTs are considered for potential medical applications ad their toxicity is similar to that of MWCNTs, we further examined the effects of chronic intravenous SWCNT administration, which resulted in a chronic inflammatory condition particularly evident in the liver that was associated with a choleostasis-like syndrome. Our data suggest that after environmental exposure, CNTs can rapidly enter and diffuse in the organism via the blood stream, in addition to the lungs, which appear to have a capacity to clear CNTs, the liver may be the major site of CNT induced damage, where long term accumulation results in inflammation, steatosis and a choleostasis-like condition
Functional analysis of the human RNASET2 gene: non-cell autonomous tumor suppression and putative stress-induced cell autonomous boles.
No full textRNASET2 is an atypical tumor suppressor gene, which behaves as a member of the "tumor antagonizing/malignancy suppressor genes" class. Indeed, in our recent works we have demonstrated that RNASET2 overexpression in ovarian cancer cells is associated with a strong suppression of their tumorigenicity in vivo, without affecting any in vitro cancer-related growth parameter. Noteworthy, the control of tumor growth apparently relies on a non-cell autonomous mechanism which involves the establishment of a cross-talk between RNASET2-overexpressing cancer cells and the tumor microenvironment, where the monocyte/macrophage lineage turned out to represent the most likely target of RNASET2 activity.
Within this framework, the principal aim of my Ph.D. project was to investigate the putative occurrence of a functional cross-talk between the RNASET2-overexpressing tumor cell and the innate immune target cells.
I demonstrate here that RNASET2 protein binds the monocyte/macrophage cell surface via a receptor-mediated interaction and exerts a chemotactic activity on this cell lineage. Thus, I propose that the recruitment and the functional activation of such cells within the tumor microenvironment could be responsible for the RNASET2-mediated tumor suppression that occurs in vivo.
Moreover, I present here data demonstrating both the overexpression and sub-cellular re-localization of RNASET2 protein in response to different stress-inducing treatments. Thus, as for other members of the T2 ribonucleases family, I propose that human RNASET2 could play a cell autonomous role in orchestrating defense mechanisms against stress conditions.
Collectively, these data suggest that at the tumor microenvironment level human RNASET2 could act as an "alarmin", both overexpressed and secreted during the early stages of tumor progression in order to activate innate immune defense mechanisms against the tumor
Caratterizzazione del trasportatore vescicolare delle monoamine di tipo 2 (VMAT2) nei linfociti umani.
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A new isoform of peptide transporter PepT1 (sbPepT1) and ion channels in touch sensation.
No full textPepT1 is an electrogenic transporter expressed at the small intestinal brush-border membrane. It transports di- and tripeptides derived from diet using a H+-electrochemical gradient as driving force. The characterization of the oligopeptide transporters is important not only in nutrition science but also in drug delivery because these proteins are also able to transport a large number of peptido-mimetic drugs and therefore they are responsible for the high levels of oral bioavailability of many pharmaceutical compounds. Examples of PepT1 substrates include angiotensin-converting enzyme (ACE) inhibitors, anticancer drugs, penicillin and cephalosporin antibiotics and the antiviral agents Acyclovir.
The research of my PhD was focused on an sea bass isoform PepT1 cloned at the University of Insubria from the intestine of sea bass (Dicentrarcus labbrax).
The injection of cRNA into Xenopus laevis oocytes allows to obtain a high functional membrane expression of the protein that can be investigated with the use of electrophysiological techniques.
Biophysical and kinetics features of the sea bass oligopeptides transporter 1 were characterized with the two electrode voltage clamp. Exposure of the oocytes to external solutions at different pHs, to substrate concentrations (between 0.03 and 10 mM) and at different membrane voltages allowed to investigate kinetic values as the maximal current (Imax) and the substrate apparent affinity (K0.5).
sbPepT1, as other peptide transporters, exhibits transport-associated currents when different di- and tripeptides are applied to the external solution and presteady-state currents in the absence of substrate.
To better understand the structure-function relation of sbPepT1, the research was extended to the other oligopeptide transporters classes (zebrafish and mammalian). The comparison of the PepT1 transport cycles among the three considered species showed a similar effect of external pH. In fact the presteady-state currents analysis presented a positive shift in the Q/V and τ/V curves and a slowing down of the decay time constants induced by acidity in all the three tested isoforms, especially in the rabbit transporter. These data were used to propose a new kinetic model.
The simulation curves of the presteady-state and the transport currents calculated with the new model that includes the effect of the acidification of external environment were in good agreement with the experimental data