Studio e sviluppo di sistemi RFID in banda UHF

The recent widespread diffusion of radio-frequency identification (RFID) applications operating in the UHF band has been supported by both the request for greater interrogation ranges and greater and faster data exchange. UHF-RFID systems, exploiting a physical interaction based on Electromagnetic propagation, introduce many problems that have not been fully explored for the previous generations of RFID systems (e.g. HF). Therefore, the availability of reliable tools for modeling and evaluating the radio-communication between Reader and Tag within an RFID radio-link are needed. The first part of the thesis discuss the impact of real environment on system performance. In particular an analytical closed form formulation for the back-scattered field from the Tag antenna and the formulation for the lower bound of the BER achievable at the Reader side will be presented, considering different possible electromagnetic impairments. By means of the previous formulations, of the analysis of the RFID link operating in near filed conditions and of some electromagnetic/system-level co-simulations, an in-depth study of the dimensioning parameters and the actual performance of the systems will be discussed and analyzed, showing some relevant properties and trade-offs in transponder and reader design. Moreover a new low cost approach to extend the read range of the RFID UHF passive systems will be discussed. Within the scope to check the reliability of the analysis approaches and of innovative proposals, some reference transponder antennas have been designed and extensive measurement campaign has been carried out with satisfactory results. Finally, some commercial ad-hoc transponder for industrial application have been designed within the cooperation with Datalogic s.p.a., some guidelines and results will be briefly presented.

Integrazione di tecnologie non distruttive per la caratterizzazione di rocce ornamentali

ABSTRACT: The dimension stone qualification through the use of non-destructive tests (NDT) is a relevant research topic for the industrial characterisation of finite products, because the competition of low-costs products can be sustained by an offer of highqualification and a top-guarantee products. The synthesis of potentialities offered by the NDT is the qualification and guarantee similar to the well known agro-industrial PDO, Protected Denomination of Origin. In fact it is possible to guarantee both, the origin and the quality of each stone product element, even through a Factory Production Control on line. A specific disciplinary is needed. A research developed at DICMA-Univ. Bologna in the frame of the “OSMATER” INTERREG project, allowed identifying good correlations between destructive and non-destructive tests for some types of materials from Verbano-Cusio-Ossola region. For example non conventional ultrasonic tests, image analysis parameters, water absorption and other measurements showed to be well correlated with the bending resistance, by relationships varying for each product. In conclusion it has been demonstrated that a nondestructive approach allows reaching several goals, among the most important: 1) the identification of materials; 2) the selection of products; 3) the substitution of DT by NDT. Now it is necessary to move from a research phase to the industrial implementation, as well as to develop new ND technologies focused on specific aims.

Dimensionamento di invasi per il controllo dei deflussi nei bacini urbanizzati

Urbanization is a continuing phenomenon in all the world. Grasslands, forests, etc. are being continually changed to residential, commercial and industrial complexes, roads and streets, and so on. One of the side effects of urbanization with which engineers and planners must deal with, is the increase of peak flows and volumes of runoff from rainfall events. As a result, the urban drainage and flood control systems must be designed to accommodate the peak flows from a variety of storms that may occur. Usually the peak flow, after development, is required not to exceed what would have occurred from the same storm under conditions existing prior to development. In order to do this it is necessary to design detention storage to hold back runoff and to release it downstream at controlled rates. In the first part of the work have been developed various simplified formulations that can be adopted for the design of stormwater detention facilities. In order to obtain a simplified hydrograph were adopted two approaches: the kinematic routing technique and the linear reservoir schematization. For the two approaches have been also obtained other two formulations depending if the IDF (intensity-duration-frequency) curve is described with two or three parameters. Other formulations have been developed taking into account if the outlet have a constant discharge or it depends on the water level in the pond. All these formulations can be easily applied when are known the characteristics of the drainage system and maximum discharge that these is in the outlet and has been defined a Return Period which characterize the IDF curve. In this way the volume of the detention pond can be calculated. In the second part of the work have been analyzed the design of detention ponds adopting continuous simulation models. The drainage systems adopted for the simulations, performed with SWMM5, are fictitious systems characterized by different sizes, and different shapes of the catchments and with a rainfall historical time series of 16 years recorded in Bologna. This approach suffers from the fact that continuous record of rainfall is often not available and when it is, the cost of such modelling can be very expensive, and that the majority of design practitioners are not prepared to use continuous long term modelling in the design of stormwater detention facilities. In the third part of the work have been analyzed statistical and stochastic methodologies in order to define the volume of the detention pond. In particular have been adopted the results of the long term simulation, performed with SWMM, to obtain the data to apply statistic and stochastic formulation. All these methodologies have been compared and correction coefficient have been proposed on the basis of the statistic and stochastic form. In this way engineers which have to design a detention pond can apply a simplified procedure appropriately corrected with the proposed coefficient.

Ricerca, mediante tecniche di proteomica, di biomarcatori proteici utili nella valutazione della qualità degli alimenti di origine animale

In the last decades, the increase of industrial activities and of the request for the world food requirement, the intensification of natural resources exploitation, directly connected to pollution, have aroused an increasing interest of the public opinion towards initiatives linked to the regulation of food production, as well to the institution of a modern legislation for the consumer guardianship. This work was planned taking into account some important thematics related to marine environment, collecting and showing the data obtained from the studies made on different marine species of commercial interest (Chamelea gallina, Mytilus edulis, Ostrea edulis, Crassostrea gigas, Salmo salar, Gadus morhua). These studies have evaluated the effects of important physic and chemical parameters variations (temperature, xenobiotics like drugs, hydrocarbons and pesticides) on cells involved in the immune defence (haemocytes) and on some important enzymatic systems involved in xenobiotic biotransformation processes (cytochrome P450 complex) and in the related antioxidant defence processes (Superoxide dismutase, Catalase, Heat Shock Protein), from a biochemical and bimolecular point of view. Oxygen is essential in the biological answer of a living organism. Its consume in the normal cellular breathing physiological processes and foreign substances biotransformation, leads to reactive oxygen species (ROS) formation, potentially toxic and responsible of biological macromolecules damages with consequent pathologies worsening. Such processes can bring to a qualitative alteration of the derived products, but also to a general state of suffering that in the most serious cases can provoke the death of the organism, with important repercussions in economic field, in the output of the breedings, of fishing and of aquaculture. In this study it seemed interesting to apply also alternative methodologies currently in use in the medical field (cytofluorimetry) and in proteomic studies (bidimensional electrophoresis, mass spectrometry) with the aim of identify new biomarkers to place beside the traditional methods for the control of the animal origin food quality. From the results it’s possible to point out some relevant aspects from each experiment: 1. The cytofluorimetric techniques applied to O. edulis and C. gigas could bring to important developments in the search of alternative methods that quickly allows to identify with precision the origin of a specific sample, contributing to oppose possible alimentary frauds, in this case for example related to presence of a different species, also under a qualitative profile, but morpholgically similar. A concrete perspective for the application in the inspective field of this method has to be confirmed by further laboratory tests that take also in account in vivo experiments to evaluate the effect in the whole organism of the factors evaluated only on haemocytes in vitro. These elements suggest therefore the possibility to suit the cytofluorimetric methods for the study of animal organisms of food interest, still before these enter the phase of industrial working processes, giving useful information about the possible presence of contaminants sources that can induce an increase of the immune defence and an alteration of normal cellular parameter values. 2. C. gallina immune system has shown an interesting answer to benzo[a]pyrene (B[a]P) exposure, dose and time dependent, with a significant decrease of the expression and of the activity of one of the most important enzymes involved in the antioxidant defence in haemocytes and haemolymph. The data obtained are confirmed by several measurements of physiological parameters, that together with the decrease of the activity of 7-etossi-resourifine-O-deetilase (EROD linked to xenobiotic biotransformation processes) during exposure, underline the major effects of B[a]P action. The identification of basal levels of EROD supports the possible presence of CYP1A subfamily in the invertebrates, still today controversial, never identified previously in C. gallina and never isolated in the immune cells, as confirmed instead in this study with the identification of CYP1A-immunopositive protein (CYP1A-IPP). This protein could reveal a good biomarker at the base of a simple and quick method that could give clear information about specific pollutants presence, even at low concentrations in the environment where usually these organisms are fished before being commercialized. 3. In this experiment it has been evaluated the effect of the antibiotic chloramphenicol (CA) in an important species of commercial interest, Chamelea gallina. Chloramphenicol is a drug still used in some developing countries, also in veterinary field. Controls to evaluate its presence in the alimentary products of animal origin, can reveal ineffective whereas the concentration results to be below the limit of sensitivity of the instruments usually used in this type of analysis. Negative effects of CA towards the CYP1A- IPP proteins, underlined in this work, seem to be due to the attack of free radicals resultant from the action of the antibiotic. This brings to a meaningful alteration of the biotransformation mechanisms through the free radicals. It seems particularly interesting to pay attention to the narrow relationships in C. gallina, between SOD/CAT and CYP450 system, actively involved in detoxification mechanism, especially if compared with the few similar works today present about mollusc, a group that is composed by numerous species that enter in the food field and on which constant controls are necessary to evaluate in a rapid and effective way the presence of possible contaminations. 4. The investigations on fishes (Gadus morhua, and Salmo salar) and on a bivalve mollusc (Mytilus edulis) have allowed to evaluate different aspects related to the possibility to identify a biomarker for the evaluation of the health of organisms of food interest and consequently for the quality of the final product through 2DE methodologies. In the seafood field these techniques are currently used with a discreet success only for vertebrates (fishes), while in the study of the invertebrates (molluscs) there are a lot of difficulties. The results obtained in this work have underline several problems in the correct identification of the isolated proteins in animal organisms of which doesn’t currently exist a complete genomic sequence. This brings to attribute some identities on the base of the comparison with similar proteins in other animal groups, incurring in the possibility to obtain inaccurate data and above all discordant with those obtained on the same animals by other authors. Nevertheless the data obtained in this work after MALDI-ToF analysis, result however objective and the spectra collected could be again analyzed in the future after the update of genomic database related to the species studied. 4-A. The investigation about the presence of HSP70 isoforms directly induced by different phenomena of stress like B[a]P presence, has used bidimensional electrophoresis methods in C. gallina, that have allowed to isolate numerous protein on 2DE gels, allowing the collection of several spots currently in phase of analysis with MALDI-ToF-MS. The present preliminary work has allowed therefore to acquire and to improve important methodologies in the study of cellular parameters and in the proteomic field, that is not only revealed of great potentiality in the application in medical and veterinary field, but also in the field of the inspection of the foods with connections to the toxicology and the environmental pollution. Such study contributes therefore to the search of rapid and new methodologies, that can increase the inspective strategies, integrating themselves with those existing, but improving at the same time the general background of information related to the state of health of the considered animal organism, with the possibility, still hypothetical, to replace in particular cases the employment of the traditional techniques in the alimentary field.

Life Cycle Assessment comparativo tra il processo di co-digestione della frazione organica dei rifiuti solidi urbani e dei fanghi di depurazione disidratati e il sistema di gestione attuale. Il caso di Bagnacavallo (RA)

Waste management is becoming, year after year, always more important both for the costs associated with it and for the ever increasing volumes of waste generated. The discussion on the fate of organic fraction of municipal solid waste (OFMSW) leads everyday to new solutions. Many alternatives are proposed, ranging from incineration to composting passing through anaerobic digestion. “For Biogas” is a collaborative effort, between C.I.R.S.A. and R.E.S. cooperative, whose main goal is to generate “green” energy from both biowaste and sludge anaerobic co-digestion. Specifically, the project include a pilot plant receiving dewatered sludge from both urban and agro-industrial sewage (DS) and the organic fraction of MSW (in 2/1 ratio) which is digested in absence of oxygen to produce biogas and digestate. Biogas is piped to a co-generation system producing power and heat reused in the digestion process itself, making it independent from the national grid. Digestate undergoes a process of mechanical separation giving a liquid fraction, introduced in the treatment plant, and a solid fraction disposed in landfill (in future it will be further processed to obtain compost). This work analyzed and estimated the impacts generated by the pilot plant in its operative phase. Once the model was been characterized, on the basis of the CML2001 methodology, a comparison is made with the present scenario assumed for OFMSW and DS. Actual scenario treats separately the two fractions: the organic one is sent to a composting plant, while sludge is sent to landfill. Results show that the most significant difference between the two scenarios is in the GWP category as the project "For Biogas" is able to generate “zero emission” power and heat. It also generates a smaller volume of waste for disposal. In conclusion, the analysis evaluated the performance of two alternative methods of management of OFMSW and DS, highlighting that "For Biogas" project is to be preferred to the actual scenario.

Sistema per l'estrazione e purificazione di sostanze antiossidanti naturali nelle acque di vegetazione delle olive

The olive oil extraction industry is responsible for the production of high quantities of vegetation waters, represented by the constitutive water of the olive fruit and by the water used during the process. This by-product represent an environmental problem in the olive’s cultivation areas because of its high content of organic matter, with high value of BOD5 and COD. For that reason the disposal of the vegetation water is very difficult and needs a previous depollution. The organic matter of vegetation water mainly consists of polysaccharides, sugars, proteins, organic acids, oil and polyphenols. This last compounds are the principal responsible for the pollution problems, due to their antimicrobial activity, but, at the same time they are well known for their antioxidant properties. The most concentrate phenolic compounds in waters and also in virgin olive oils are secoiridoids like oleuropein, demethyloleuropein and ligstroside derivatives (the dialdehydic form of elenolic acid linked to 3,4-DHPEA, or p-HPEA (3,4-DHPEA-EDA or p-HPEA-EDA) and an isomer of the oleuropein aglycon (3,4-DHPEA-EA). The management of the olive oil vegetation water has been extensively investigated and several different valorisation methods have been proposed, such as the direct use as fertilizer or the transformation by physico-chemical or biological treatments. During the last years researchers focused their interest on the recovery of the phenolic fraction from this waste looking for its exploitation as a natural antioxidant source. At the present only few contributes have been aimed to the utilization for a large scale phenols recovery and further investigations are required for the evaluation of feasibility and costs of the proposed processes. The present PhD thesis reports a preliminary description of a new industrial scale process for the recovery of the phenolic fraction from olive oil vegetation water treated with enzymes, by direct membrane filtration (microfiltration/ultrafiltration with a cut-off of 250 KDa, ultrafiltration with a cut-off of 7 KDa/10 KDa and nanofiltration/reverse osmosis), partial purification by the use of a purification system based on SPE analysis and by a liquid-liquid extraction system (LLE) with contemporary reduction of the pollution related problems. The phenolic fractions of all the samples obtained were qualitatively and quantitatively by HPLC analysis. The work efficiency in terms of flows and in terms of phenolic recovery gave good results. The final phenolic recovery is about 60% respect the initial content in the vegetation waters. The final concentrate has shown a high content of phenols that allow to hypothesize a possible use as zootechnic nutritional supplements. The purification of the final concentrate have garanteed an high purity level of the phenolic extract especially in SPE analysis by the use of XAD-16 (73% of the total phenolic content of the concentrate). This purity level could permit a future food industry employment such as food additive, or, thanks to the strong antioxidant activity, it would be also use in pharmaceutical or cosmetic industry. The vegetation water depollutant activity has brought good results, as a matter of fact the final reverse osmosis permeate has a low pollutant rate in terms of COD and BOD5 values (2% of the initial vegetation water), that could determinate a recycling use in the virgin olive oil mechanical extraction system producing a water saving and reducing thus the oil industry disposal costs .

Rapporti fra fattori ambientali e proteine di parete in Bifidobacterium

The normal gut microbiota has several important functions in host physiology and metabolism, and plays a key role in health and disease. Bifidobacteria, which are indigenous components of gastrointestinal microbiota, may play an important role in maintaining the well-being of the host although its precise function is very difficult to study. Its physiological and biochemical activities are controlled by many factors, particularly diet and environment. Adherence and colonization capacity are considered as contributing factors for immune modulation, pathogen exclusion, and enhanced contact with the mucosa. In this way, bifidobacteria would fortify the microbiota that forms an integral part of the mucosal barrier and colonization resistance against pathogens. Bifidobacteria are not only subjected to stressful conditions in industrial processes, but also in nature, where the ability to respond quickly to stress is essential for survival. Bifidobacteria, like other microorganisms, have evolved sensing systems for/and defences against stress that allow them to withstand harsh conditions and sudden environmental changes. Bacterial stress responses rely on the coordinated expression of genes that alter various cellular processes and structures (e.g. DNA metabolism, housekeeping genes, cell-wall proteins, membrane composition) and act in concert to improve bacterial stress tolerance. The integration of these stress responses is accomplished by regulatory networks that allow the cell to react rapidly to various and sometimes complex environmental changes. This work examined the effect of important stressful conditions, such as changing pH and osmolarity, on the biosynthesis of cell wall proteins in B. pseudolongum subsp. globosum. These environmental factors all influence heavily the expression of BIFOP (BIFidobacterial Outer Proteins) in the cell-wall and can have an impact in the interaction with host. Also evidence has been collected linking the low concentration of sugar in the culture medium with the presence or absence of extracromosomal DNA.

Sviluppo di metodi innovativi per la sintesi di nanoparticelle di interesse industriale

Among various nanoparticles, noble metal nanoparticles have attracted considerable attention due to their optical, catalytic and conducting properties. This work has been focused on the development of an innovative method of synthesis for the preparation of metal nanosuspensions of Au, Ag, Cu, in order to achieve stable sols, showing suitable features to allow an industrial scale up of the processes. The research was developed in collaboration with a company interested in the large scale production of the studied nanosuspensions. In order to develop a commercial process, high solid concentration, long time colloidal stability and particle size control, are required. Two synthesis routes, differing by the used solvents, have been implemented: polyol based and water based synthesis. In order to achieve a process intensification the microwave heating has been applied. As a result, colloidal nanosuspensions with suitable dimensions, good optical properties, very high solid content and good stability, have been synthesized by simple and environmental friendly methods. Particularly, due to some interesting results an optimized synthesis process has been patented. Both water and polyol based synthesis, developed in the presence of a reducing agent and of a chelating polymer, allowed to obtain particle size-control and colloidal stability by tuning the different parameters. Furthermore, it has been verified that microwave device, due to its rapid and homogeneous heating, provides some advantages over conventional method. In order to optimize the final suspensions properties, for each synthesis it has been studied the effect of different parameters (temperature, time, precursors concentrations, etc) and throughout a specific optimization action a right control on nucleation and growth processes has been achieved. The achieved nanoparticles were confirmed by XRD analysis to be the desired metal phases, even at the lowest synthesis temperatures. The particles showed a diameter, measured by STEM and dynamic light scattering technique (DLS), ranging from 10 to 60 nm. Surface plasmon resonance (SPR) was monitored by UV-VIS spectroscopy confirming its dependence by nanoparticles size and shape. Moreover the reaction yield has been assessed by ICP analysis performed on the unreacted metal cations. Finally, thermal conductivity and antibacterial activity characterizations of copper and silver sols respectively are now ongoing in order to check their application as nanofluid in heat transfer processes and as antibacterial agent.

Studio della gestione integrata dei rifiuti urbani nell'ambito della provincia di Bologna

This work is an analysis of integrated urban waste management in the province of Bologna. It consists of five chapters and one enclosure. Primarily, it focuses on the legislative framework at european, national and local level. Then the study analyses the situation of urban waste system adopted in the reference territory from 2003 to 2007 to show its evolution process. Chapter 3 is based on considerations about the percentage of effective recover of materials derived from separate collection that has been reached in the province of Bologna in 2006. The following chapter describes the urban waste management at national level using dates of 2005 and 2006 by APAT (National Agency for environmental protection). Then, it has been made a comparison with Emilia-Romagna and district of Bologna. Chapter 5 focuses on the description of innovative strategies introduced in the district of Bologna to increase separate collection level and optimize waste management. In particular, it analyses two sperimental projects: one based on door to door collection and the other founded on an integrated collection system which provides the application of two collection models (door to door collection in industrial areas and collection by containers in urban ones). Finally, in the enclosure, it is also descrided best practices of waste management sector about collection models, treatment plants and innovative strategies available at that moment in Europe.

Dealogenazione riduttiva dei policlorobifenili (PCB) in sedimenti anaerobici marini della laguna di Venezia: arricchimento e identificazione dei microrganismi dealogenanti

I policlorobifenili (PCB) sono inquinanti tossici e fortemente recalcitranti che contaminano suoli e sedimenti di acqua dolce e marini. Le tecnologie attualmente impiegate per la loro rimozione (dragaggio e trattamento chimoco-fisico o conferimento in discarica) sono molto costose, poco efficaci o ad alto impatto ambientale. L’individuazione di strategie alternative, di natura biologica, consentirebbe lo sviluppo di un processo alternativo più sostenibile. Nel processo di declorurazione riduttiva i congeneri di PCB a più alto grado di clorurazione, che sono i più tossici, recalcitranti e maggiormente tendenti al bioaccumulo, vengono utilizzati da alcuni microrganismi anaerobici come accettori finali di elettroni nella catena respiratoria e bioconvertiti in congeneri a minor grado di clorurazione, meno pericolosi, che possono essere mineralizzati da parte di batteri aerobi. La declorurazione riduttiva dei PCB è stata spesso studiata in colture anaerobiche di arricchimento in terreno minerale ottenute a partire da sedimenti di acqua dolce; questi studi hanno permesso di dimostrare che batteri del phylum dei Chloroflexi e appartenenti al genere Dehalococcoides o filogeneticamente facenti parte del gruppo dei Dehalococcoides-like sono i decloruranti. Sono tuttavia scarse le informazioni riguardanti l'occorrenza della declorurazione dei PCB in ambienti marini, nei quali l'alta salinità e concentrazione di solfati influenzano diversamente l'evoluzione delle popolazioni microbiche. In sedimenti contaminati della laguna di Venezia è stata osservata declorurazione sia dei PCB preesistenti che di congeneri esogeni; questi studi hanno permesso l'ottenimento di colture di arricchimento fortemente attive nei confronti di 5 congeneri di PCB coplanari. In questa tesi, a partire dalle colture capaci di declorurare i PCB coplanari, sono stati allestiti nuovi passaggi di arricchimento su Aroclor®1254, una miscela di PCB più complessa e che meglio rappresenta la contaminazione ambientale. Le colture sono state allestite come microcosmi anaerobici in fase slurry, preparati risospendendo il sedimento nell'acqua superficiale, ricreando in tal modo in laboratorio le stesse condizioni biogeochimiche presenti in situ; gli slurry sterili sono stati inoculati per avviare le colture. Per favorire la crescita dei microrganismi decloruranti e stimolare così la decloruraazione dei PCB sono stati aggiunti inibitori selettivi di metanogeni (Bromoetansulfonato o BES) e solfato-riduttori (molibdato), sono state fornite fonti di carbonio ed energia (eD), quali acidi grassi a corta catena e idrogeno, utilizzate di batteri decloruranti noti, e per semplificare la comunità microbica sono stati aggiunti antibiotici cui batteri decloruranti del genere Dehalococcoides sono resistenti. Con questo approccio sono stati allestiti passaggi di arricchimento successivi e le popolazioni microbiche delle colture sono state caratterizzate con analisi molecolari di fingerprinting (DGGE). Fin dal primo passaggio di arricchimento nei microcosmi non ammendati ha avuto luogo un'estesa declorurazione dell'Aroclor®1254; nei successivi passaggi si è notato un incremento della velocità del processo e la scomparsa della fase di latenza, mentre la stessa stereoselettività è stata mantenuta a riprova dell’arricchimento degli stessi microrganismi decloruranti. Le velocità di declorurazione ottenute sono molto alte se confrontate con quelle osservate in colture anaerobiche addizionate della stessa miscela descritte in letteratura. L'aggiunta di BES o molibdato ha bloccato la declorurazione dei PCB ma in presenza di BES è stata riscontrata attività dealogenante nei confronti di questa molecola. La supplementazione di fonti di energia e di carbonio ha stimolato la metanogenesi e i processi fermentativi ma non ha avuto effetti sulla declorurazione. Ampicillina e vancomicina hanno incrementato la velocità di declorurazione quando aggiunte singolarmente, insieme o in combinazione con eD. E' stato però anche dimostrato che la declorurazione dei PCB è indipendente sia dalla metanogenesi che dalla solfato-riduzione. Queste attività respiratorie hanno avuto velocità ed estensioni diverse in presenza della medesima attività declorurante; in particolare la metanogenesi è stata rilevata solo in dipendenza dall’aggiunta di eD alle colture e la solfato-riduzione è stata inibita dall’ampicillina in microcosmi nei quali un’estesa declorurazione dei PCB è stata osservata. La caratterizzazione delle popolazioni microbiche, condotte mediante analisi molecolari di fingerprinting (DGGE) hanno permesso di descrivere le popolazioni batteriche delle diverse colture come complesse comunità microbiche e di rilevare in tutte le colture decloruranti la presenza di una banda che l’analisi filogenetica ha ascritto al batterio m-1, un noto batterio declorurante in grado di dealogenare un congenere di PCB in colture di arricchimento ottenute da sedimenti marini appartenente al gruppo dei Dehalococcoides-like. Per verificare se la crescita di questo microrganismo sia legata alla presenza dei PCB, l'ultimo passaggio di arricchimento ha previsto l’allestimento di microcosmi addizionati di Aroclor®1254 e altri analoghi privi di PCB. Il batterio m-1 è stato rilevato in tutti i microcosmi addizionati di PCB ma non è mai stato rilevato in quelli in cui i PCB non erano presenti; la presenza di nessun altro batterio né alcun archebatterio è subordinata all’aggiunta dei PCB. E in questo modo stato dimostrato che la presenza di m-1 è dipendente dai PCB e si ritiene quindi che m-1 sia il declorurante in grado di crescere utilizzando i PCB come accettori di elettroni nella catena respiratoria anche in condizioni biogeochimiche tipiche degli habitat marini. In tutte le colture dell'ultimo passaggio di arricchimento è stata anche condotta una reazione di PCR mirata alla rilevazione di geni per dealogenasi riduttive, l’enzima chiave coinvolto nei processi di dealogenazione. E’ stato ottenuto un amplicone di lughezza analoga a quelle di tutte le dealogenasi note in tutte le colture decloruranti ma un tale amplificato non è mai stato ottenuto da colture non addizionate di PCB. La dealogenasi ha lo stesso comportamento di m-1, essendo stata trovata come questo sempre e solo in presenza di PCB e di declorurazione riduttiva. La sequenza di questa dealogenasi è diversa da tutte quelle note sia in termini di sequenza nucleotidica che aminoacidica, pur presentando due ORF con le stesse caratteristiche e domini presenti nelle dealogenasi note. Poiché la presenza della dealogenasi rilevata nelle colture dipende esclusivamente dall’aggiunta di PCB e dall’osservazione della declorurazione riduttiva e considerato che gran parte delle differenze genetiche è concentrata nella parte di sequenza che si pensa determini la specificità di substrato, si ritiene che la dealogenasi identificata sia specifica per i PCB. La ricerca è stata condotta in microcosmi che hanno ricreato fedelmente le condizioni biogeochimiche presenti in situ e ha quindi permesso di rendere conto del reale potenziale declorurante della microflora indigena dei sedimenti della laguna di Venezia. Le analisi molecolari condotte hanno permesso di identificare per la prima volta un batterio responsabile della declorurazione dei PCB in sedimenti marini (il batterio m-1) e una nuova dealogenasi specifica per PCB. L'identificazione del microrganismo declorurante permette di aprire la strada allo sviluppo di tecnologie di bioremediation mirata e il gene della dealogenasi potrà essere utilizzato come marker molecolare per determinare il reale potenziale di declorurazione di miscele complesse di PCB in sedimenti marini.

Studio dei profili di espressione dei microRNA nell'epatocarcinoma umano

Il carcinoma epatocellulare (HCC) è il più frequente tumore maligno del fegato e rappresenta il sesto tipo di tumore più comune nel mondo. Spesso i pazienti con HCC vengono diagnosticati a stadi piuttosto avanzati, quando le uniche opzioni terapeutiche in grado di migliorarne la sopravvivenza sono la chemoembolizzazione dell'arteria epatica ed il trattamento con l'inibitore multi-cinasico, Sorafenib. In questo contesto, la scoperta del ruolo centrale dei microRNA (miRNA) nella tumorigenesi umana risulta di fondamentale importanza per lo sviluppo di nuovi marcatori diagnostici e bersagli terapeutici. I microRNA (miRNA) sono delle piccole molecole di RNA non codificante, della lunghezza di 19-22 nucleotidi, filogeneticamente molto conservati, ed esercitano un ruolo cruciale nella regolazione di importanti processi fisiologici, quali sviluppo, proliferazione, differenziamento, apoptosi e risposta a numerosi segnali extracellulari e di stress. I miRNA sono inoltre responsabile della fine regolazione dell'espressione di centinaia di geni bersaglio attraverso il blocco della traduzione o la degradazione dell'mRNA target. Studi di profiling hanno evidenziato l'espressione aberrante di specifici miRNA in numerosi tipi di tumore umano. Lo scopo del presente lavoro è stato quello di individuare un pannello di miRNA deregolati nell'epatocarcinoma umano e di caratterizzare il ruolo biologico di tre miRNA deregolati nell'HCC, al fine di individuare alcuni dei meccanismi molecolari alla base della trasformazione maligna miRNA-associata. La nostra ricerca è stata inoltre focalizzata nell'individuazione di nuovi bersagli e strumenti terapeutici, quali i microRNA, per il trattamento combinato di HCC in stadio intermedio-avanzato.

Rapporto tra struttura e funzione della cistatina B e dei suoi mutanti in relazione all'epilessia mioclonica progressiva di tipo 1 (EPM1)

This work shows for the first time that native CSTB polymerizes on addition of Cu2+ and DnaK (Hsp70). Cysteines are involved in the polymerization process and in particular at least one cysteine is necessary. We propose that Cu2+ interacts with the thiol group of cysteine and oxidize it. The oxidized cysteine modifies the CSTB structure allowing interaction with DnaK/Hsp70 to occur. Thus, Cu2+ binding to CSTB exposes a site for DnaK and such interaction allows the polymerization of CSTB. The polymers generated from native CSTB monomers, are DTT sensitive and they may represent physiological polymers. Denatured CSTB does not require Cu2+ and polymerizes simply on addition of DnaK. The polymers generated from denatured CSTB do not respond to DTT. They have characteristics similar to those of the CSTB toxic aggregates described in vivo in eukaryotic cells following CSTB over-expression. Interaction between CSTB and Hsp70 is shown by IP experiments. The interaction occurs with WT CSTB and not with the cys mutant. This suggests that disulphur bonds are involved. Methal-cathalyzed oxidation of proteins involves reduction of the metal ion(s) bound to the protein itself and oxidation of neighboring ammino acid residues resulting in structural modification and de-stabilization of the molecule. In this work we propose that the cysteine thyol residue of CSTB in the presence of Cu2+ is oxidized, and cathalyzes the formation of disulphide bonds with Hsp70, that, once bound to CSTB, mediates its polymerization. In vivo this molecular mechanism of CSTB polymerization could be regulated by redox environment through the cysteine residue. This may imply that CSTB physiological polymers have a specific cellular function, different from that of the protease inhibitor known for the CSTB monomer. This hypothesis is interesting in relation to Progressive Myoclonus Epilepsy of type 1 (EPM1). This pathology is usually caused by mutations in the CSTB gene. CSTB is a ubiquitous protein, but EPM1 patients have problems only in the central nervous system. Maybe physiological CSTB polymers have a specific function altered in people affected by EPM1.

Architectures and design patterns for functional design of logic control and diagnostics in industrial automation

Recently in most of the industrial automation process an ever increasing degree of automation has been observed. This increasing is motivated by the higher requirement of systems with great performance in terms of quality of products/services generated, productivity, efficiency and low costs in the design, realization and maintenance. This trend in the growth of complex automation systems is rapidly spreading over automated manufacturing systems (AMS), where the integration of the mechanical and electronic technology, typical of the Mechatronics, is merging with other technologies such as Informatics and the communication networks. An AMS is a very complex system that can be thought constituted by a set of flexible working stations, one or more transportation systems. To understand how this machine are important in our society let considerate that every day most of us use bottles of water or soda, buy product in box like food or cigarets and so on. Another important consideration from its complexity derive from the fact that the the consortium of machine producers has estimated around 350 types of manufacturing machine. A large number of manufacturing machine industry are presented in Italy and notably packaging machine industry,in particular a great concentration of this kind of industry is located in Bologna area; for this reason the Bologna area is called “packaging valley”. Usually, the various parts of the AMS interact among them in a concurrent and asynchronous way, and coordinate the parts of the machine to obtain a desiderated overall behaviour is an hard task. Often, this is the case in large scale systems, organized in a modular and distributed manner. Even if the success of a modern AMS from a functional and behavioural point of view is still to attribute to the design choices operated in the definition of the mechanical structure and electrical electronic architecture, the system that governs the control of the plant is becoming crucial, because of the large number of duties associated to it. Apart from the activity inherent to the automation of themachine cycles, the supervisory system is called to perform other main functions such as: emulating the behaviour of traditional mechanical members thus allowing a drastic constructive simplification of the machine and a crucial functional flexibility; dynamically adapting the control strategies according to the different productive needs and to the different operational scenarios; obtaining a high quality of the final product through the verification of the correctness of the processing; addressing the operator devoted to themachine to promptly and carefully take the actions devoted to establish or restore the optimal operating conditions; managing in real time information on diagnostics, as a support of the maintenance operations of the machine. The kind of facilities that designers can directly find on themarket, in terms of software component libraries provides in fact an adequate support as regard the implementation of either top-level or bottom-level functionalities, typically pertaining to the domains of user-friendly HMIs, closed-loop regulation and motion control, fieldbus-based interconnection of remote smart devices. What is still lacking is a reference framework comprising a comprehensive set of highly reusable logic control components that, focussing on the cross-cutting functionalities characterizing the automation domain, may help the designers in the process of modelling and structuring their applications according to the specific needs. Historically, the design and verification process for complex automated industrial systems is performed in empirical way, without a clear distinction between functional and technological-implementation concepts and without a systematic method to organically deal with the complete system. Traditionally, in the field of analog and digital control design and verification through formal and simulation tools have been adopted since a long time ago, at least for multivariable and/or nonlinear controllers for complex time-driven dynamics as in the fields of vehicles, aircrafts, robots, electric drives and complex power electronics equipments. Moving to the field of logic control, typical for industrial manufacturing automation, the design and verification process is approached in a completely different way, usually very “unstructured”. No clear distinction between functions and implementations, between functional architectures and technological architectures and platforms is considered. Probably this difference is due to the different “dynamical framework”of logic control with respect to analog/digital control. As a matter of facts, in logic control discrete-events dynamics replace time-driven dynamics; hence most of the formal and mathematical tools of analog/digital control cannot be directly migrated to logic control to enlighten the distinction between functions and implementations. In addition, in the common view of application technicians, logic control design is strictly connected to the adopted implementation technology (relays in the past, software nowadays), leading again to a deep confusion among functional view and technological view. In Industrial automation software engineering, concepts as modularity, encapsulation, composability and reusability are strongly emphasized and profitably realized in the so-calledobject-oriented methodologies. Industrial automation is receiving lately this approach, as testified by some IEC standards IEC 611313, IEC 61499 which have been considered in commercial products only recently. On the other hand, in the scientific and technical literature many contributions have been already proposed to establish a suitable modelling framework for industrial automation. During last years it was possible to note a considerable growth in the exploitation of innovative concepts and technologies from ICT world in industrial automation systems. For what concerns the logic control design, Model Based Design (MBD) is being imported in industrial automation from software engineering field. Another key-point in industrial automated systems is the growth of requirements in terms of availability, reliability and safety for technological systems. In other words, the control system should not only deal with the nominal behaviour, but should also deal with other important duties, such as diagnosis and faults isolations, recovery and safety management. Indeed, together with high performance, in complex systems fault occurrences increase. This is a consequence of the fact that, as it typically occurs in reliable mechatronic systems, in complex systems such as AMS, together with reliable mechanical elements, an increasing number of electronic devices are also present, that are more vulnerable by their own nature. The diagnosis problem and the faults isolation in a generic dynamical system consists in the design of an elaboration unit that, appropriately processing the inputs and outputs of the dynamical system, is also capable of detecting incipient faults on the plant devices, reconfiguring the control system so as to guarantee satisfactory performance. The designer should be able to formally verify the product, certifying that, in its final implementation, it will perform itsrequired function guarantying the desired level of reliability and safety; the next step is that of preventing faults and eventually reconfiguring the control system so that faults are tolerated. On this topic an important improvement to formal verification of logic control, fault diagnosis and fault tolerant control results derive from Discrete Event Systems theory. The aimof this work is to define a design pattern and a control architecture to help the designer of control logic in industrial automated systems. The work starts with a brief discussion on main characteristics and description of industrial automated systems on Chapter 1. In Chapter 2 a survey on the state of the software engineering paradigm applied to industrial automation is discussed. Chapter 3 presentes a architecture for industrial automated systems based on the new concept of Generalized Actuator showing its benefits, while in Chapter 4 this architecture is refined using a novel entity, the Generalized Device in order to have a better reusability and modularity of the control logic. In Chapter 5 a new approach will be present based on Discrete Event Systems for the problemof software formal verification and an active fault tolerant control architecture using online diagnostic. Finally conclusive remarks and some ideas on new directions to explore are given. In Appendix A are briefly reported some concepts and results about Discrete Event Systems which should help the reader in understanding some crucial points in chapter 5; while in Appendix B an overview on the experimental testbed of the Laboratory of Automation of University of Bologna, is reported to validated the approach presented in chapter 3, chapter 4 and chapter 5. In Appendix C some components model used in chapter 5 for formal verification are reported.