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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
Screening and heterologous expression of chitinases from environmental metagenomic libraries.
No full textMy PhD thesis has been developed in the frame of the EU-funded Metaexplore project. This project, conceived in collaboration with eighteen research groups all over Europe, concerns the development of new metagenomic techniques to detect enzymes involved in the biodegradation of recalcitrant natural molecules such as chitin and lignin. The final goal is the heterologous expression and characterization of the newly discovered chitinases and ligninases to exploit them in industrial bioprocesses. My thesis deals with chitinases that include a group of enzymes acting in a synergic and consecutive way, responsible for the complete hydrolysis of chitin into the constitutive monomers of N-acetylglucosamine. An alternative pathway involves chitin deacetylases for the deacetylation of chitin into chitosan, which is then further hydrolyzed by chitosanases in glucosamine units. Chitin derivatives have a multiplicity of industrial applications, which make chitinases attractive at industrial level. University of Insubria participates at the Metaexplore project in different work-packages. In the first part, my work is devoted to the validation of the functional screening methods to detect chitin degrading activities from metagenomic libraries. The following task regards the preliminary analysis of the most promising clones and the heterologous expression of the genes of interest in conventional and alternative hosts. At the moment, protein expression is the major bottleneck for the biotechnological success of the metagenomic approach to discover new enzymes. The optimization of the heterologous expression of metagenome-sourced genes will allow a complete biochemical characterization of the new enzymes and their production at industrial level.
The following chapters report Material and Methods, Results and Discussion related to the experimental work performed for the Metaexplore project, during the three years of my PhD course.
The first chapter, Screening for chitinase activities in actinomycetes, consist in an exploration of the University of Insubria’s actinomycete culture collection for chitin degrading activities. This culture collection comprises microorganisms belonging to Streptomyces genus, but also to other genera of uncommon actinomycetes named “rare”, since difficult to be isolated, cultivated, and manipulated. All the assayed microorganisms are degrading chitin when it is used as a sole carbon and nitrogen source in plate assay, but only some of them seem to possess chitosan and chitin deacetylase activities. Induction and repression of the chitinolytic system has been investigated in representative strains. Enzymatic assays to detect the extracellular and intracellular levels of chitin depolymerase, chitobiosidase, N-acetylglucosaminidase activities have been developed, validated and transferred to the Metaexplore partners to be applied at the screening of the metagenomic libraries. Interestingly, the rare actinomycete Nonomuraea sp. ATCC 39727, previously never explored for its chitin degrading activity, results positive in the chitin deacetylase plate assay and produces chitosan. To my knowledge, this is the first report on chitin deacetylase enzyme discovered from actinomycetes.
The second paper, Bacterial chitinase with biocontrol capacity from suppressive soil revealed by functional metagenomics, presents the research work performed in collaboration with the School of Life Sciences of Södertörn University (Huddinge, Sweden), on the heterologous expression of the first chitinase isolated from a metagenomic library within the Metaexplore consortium. The metagenomic library originates from a soil considered suppressive for clubroot disease of cabbage. Genetic screening using degenerate primers for the conserved central catalytic domain of family 18 chitinases has selected a putative chitobiosidase gene, called chi18H8. The gene is cloned in BL21(DE3) E. coli cells, in transcriptional fusion with the gluthathione-S-transferase tag. In this host, most of the protein accumulates in the insoluble fraction as inclusion bodies, so expression optimization in E. coli soluble fractions is required for protein purification by affinity chromatography. On the pure enzyme, recovered at very low concentration, preliminary analysis confirms its chitinolytic nature and its antifungal properties against common plant phytopathogens.
In the third chapter, entitled Expression of a metagenome-sourced chitinase in Streptomyces lividans, this actinomycete is presented as alternative candidate to E. coli for the heterologous expression of the chi18H8 chitinase. Streptomyces spp. are typical degraders of biomasses in soil habitats. They possess a developed secretory pathway for the extracellular release of their hydrolytic enzymes. As described in the first paper on screening, they usually produce different chitinases, whose activity can be detected also when cultivated in rich media not containing chitin as carbon source. To use S. lividans as a host for heterologous chitinase expression, the chitinolytic system is metabolically repressed by adding glucose to the culture media, and the chi18H8 gene is cloned in a multicopy plasmid under the control of a heterologus constitutive promoter which is not repressed by the presence of glucose. The transformation of S. lividans TK 24 occurs by E. coli - S. lividans integeneric conjugation after a DNA demethylation step. Enzymatic activity assay and zymogram analysis confirm that the recombinant protein is secreted in the extracellular broth with an estimated volumetric productivity of 66 U/L which represents a good starting point for the further attempts of protein production and purification of Chi18H8 in S. lividans host
Determinants of knowledge flows and their effect on economic growth.
No full textThis thesis consists of three essays that analyse the determinants of the diffusion of knowledge and their impact in terms of economic growth. The first essay analyses the patterns of diffusion of knowledge between European regions during the period 1981-2000. Knowledge flows are measured by two variables: patent citations and collaboration among inventors. A modified gravity model is estimated to show that geographical proximity and territorial borders are important barriers to knowledge flows during the period analysed and that their effects are greater for technological collaboration than for patent citation. We show that inventor propensity to collaborate within the same country decreases and inventor propensity to cite inventors residing in the same country increases. We show also that the geographical distance effect decreases only for inventor collaboration. Finally, we show that the EU enlargement processes during the period analysed have favoured knowledge flows between old and new EU members, although with significant differences for the two measures of knowledge flows. The second essay analyses the impact of knowledge capital on the economic growth of Italian regions for the period 1995-2007. To measure regional knowledge capital we use indicators for the processes of knowledge creation, i.e. R&D and patents, and the processes of diffusion of knowledge, i.e. patent citations and inventor mobility. We use a technology gap model to show that the economic growth of Italian regions can be explained by internal R&D activities and by the interregional mobility of inventors. The third essay analyses the impact of knowledge flows on economic performance at firm level. It distinguishes between various sources of knowledge flows, i.e. suppliers, customers, rivals and research institutions, and between two types of innovative activities, i.e. sales realized by products new to the firm but not to the market (imitation) and sales realized by products new to the firm and to the market (innovation).
We show that knowledge flows from rivals lead to more imitation, while knowledge flows from customers and research institutions enhance original innovation
Development and validation of an algorithm to predict the success of the ablation of macroreentrant atrial tachycardia.
No full textAblation of macroreentrant atrial tachycardia (MRAT) is challenging because of complex anatomy and multiple reentrant loops. In order to define an effective ablation strategy, 3-dimensional electroanatomic mapping proved very useful. To identify predictors of ablation procedure failure may be helpful for patients treatment. In the first part of our study we analyzed into details the electroanatomical features of the reentry circuit in MRAT and we compared the characteristics of successfully versus unsuccessfully consecutive treated patients undegoing electroanatomic mapping and ablation of MRAT in order to identify variables predicting the ablation outcome. Ablation was linearly placed at the mid-diastolic isthmus (MDI) to achieve arrhythmia interruption and conduction block. Variables were analyzed for predictors of both procedural failure and cumulative failure. We demonstrated a significant difference as to the electroanatomic mapping characteristics: successfully treated cases showed a narrower target isthmus with a slower conduction velocity across the isthmus itself. In the second part of our research, we analyzed the relation between the strongest predictors of procedure outcome identified in part I (MDI width and conduction velocity across the MDI) and the chance of success of the ablation procedure. In order to analyze this relation and to predict the difficulty of the ablation procedure, we developed an algorithm and we validated prospectively the accuracy of the developed model in a second patient series
Uso di statine e recidiva di fibrillazione atriale dopo ablazione transcatetere o cardioversione elettrica.
No full textmissin
Bio-materials and surgical options for the treatment of lung cancer invading the superior vena cava.
No full textBackground. The oncologic value of superior vena cava (SVC) resection for lung malignancies remains controversial. The aim of this study was to review postoperative outcome and long-term oncologic results in the surgical treatment of locally advanced lung cancer (NSCLC) invading the SVC system.
Methods. The clinical data of 88 consecutive patients who underwent surgery for locally advanced NSCLC invading SVC system from 1998 to november 2011 were retrospectively reviewed. Patients were grouped according to the type of surgical treatment performed. The study endpoints were overall survival, post-operative morbidity, mortality and lenght of postoperative stay.
Results. Forty-seven (72.4%) patient underwent partial SVC resection. Replacement of the SVC system by prosthesis was performed in 18 patients (27.6%). In 20 patients (30.7%) SVC resection was associated with carina resection (SVC-CR). In 23 patients a replacement of the SVC was performed by a biomaterial: polytetrafluoroethylene graft (13 patients), biological custom-made bovine pericardial tube (14 patients) or autologus pericardium patch (5 patients). Twenty-three patients underwent explorative thoracotomy (ET). In the resected group overall postoperative major morbidity and mortality rates were 17% (minor 28%, major 16%) and 9,2% respectively. In multivariate analysis, advanced age (≥60 years) and advanced stage (III-IV) are independent predictors of survival. Mortality was not influenced by any factor. Five-year survival probability was 25% for overall SVC resection, and 20% when resection of SVC was associated with CR. In SVC resection patients, lymph node status was the most important prognostic factor of survival (5-year survival in N0 patients 44%; 22% in N+). In ET group 5-year survival was 12%.
Conclusions. Pursuing extended resection in locally advanced NSCLC invading SVC may achieve permanent cure in selected patients who would otherwise be defined inoperable with very poor prognosis. Satisfactory long-term results in NSCLC infiltrating the SVC system can be achieved only when pathologic N2 disease is excluded
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
