Climate change and the tourism sector: the clean development mechanism, a market instrument under the Kyoto Protocol to achieve multiple objectives

The main objective of this research is to demonstrate that the Clean Development Mechanism (CDM), an instrument created under a global international treaty, can achieve multiple objectives beyond those for which it has been established. As such, while being already a powerful tool to contribute to the global fight against climate change, the CDM can also be successful if applied to different sectors not contemplated before. In particular, this research aimed at demonstrating that a wider utilization of the CDM in the tourism sector can represent an innovative way to foster sustainable tourism and generate additional benefits. The CDM was created by Article 12 of the Kyoto Protocol of the United Nations Framework Convention on Climate Change (UNFCCC) and represents an innovative tool to reduce greenhouse gases emissions through the implementation of mitigation activities in developing countries which generate certified emission reductions (CERs), each of them equivalent to one ton of CO2 not emitted in the atmosphere. These credits can be used for compliance reasons by industrialized countries in achieving their reduction targets. The logic path of this research begins with an analysis of the scientific evidences of climate change and its impacts on different economic sectors including tourism and it continues with a focus on the linkages between climate and the tourism sector. Then, it analyses the international responses to the issue of climate change and the peculiar activities in the international arena addressing climate change and the tourism sector. The concluding part of the work presents the objectives and achievements of the CDM and its links to the tourism sector by considering case studies of existing projects which demonstrate that the underlying question can be positively answered. New opportunities for the tourism sector are available.

Processing of measured acceleration response for SHM purpose

Structural Health Monitoring (SHM) is the process of characterization for existing civil structures that proposes for damage detection and structural identification. It's based firstly on the collection of data that are inevitably affected by noise. In this work a procedure to denoise the measured acceleration signal is proposed, based on EMD-thresholding techniques. Moreover the velocity and displacement responses are estimated, starting from measured acceleration.

Theoretical and numerical study of the laser-plasma ion acceleration

The laser driven ion acceleration is a burgeoning field of resarch and is attracting a growing number of scientists since the first results reported in 2000 obtained irradiating thin solid foils by high power laser pulses. The growing interest is driven by the peculiar characteristics of the produced bunches, the compactness of the whole accelerating system and the very short accelerating length of this all-optical accelerators. A fervent theoretical and experimental work has been done since then. An important part of the theoretical study is done by means of numerical simulations and the most widely used technique exploits PIC codes (“Particle In Cell'”). In this thesis the PIC code AlaDyn, developed by our research group considering innovative algorithms, is described. My work has been devoted to the developement of the code and the investigation of the laser driven ion acceleration for different target configurations. Two target configurations for the proton acceleration are presented together with the results of the 2D and 3D numerical investigation. One target configuration consists of a solid foil with a low density layer attached on the irradiated side. The nearly critical plasma of the foam layer allows a very high energy absorption by the target and an increase of the proton energy up to a factor 3, when compared to the ``pure'' TNSA configuration. The differences of the regime with respect to the standard TNSA are described The case of nearly critical density targets has been investigated with 3D simulations. In this case the laser travels throughout the plasma and exits on the rear side. During the propagation, the laser drills a channel and induce a magnetic vortex that expanding on the rear side of the targer is source of a very intense electric field. The protons of the plasma are strongly accelerated up to energies of 100 MeV using a 200PW laser.

Dynamic stabilization of Rayleigh-Taylor instability of ablation fronts in inertial confinement fusion

One of the most important problems in inertial confinement fusion is how to find a way to mitigate the onset of the Rayleigh-Taylor instability which arises in the ablation front during the compression. In this thesis it is studied in detail the possibility of using for such a purpose the well-known mechanism of dynamic stabilization, already applied to other dynamical systems such as the inverted pendulum. In this context, a periodic acceleration superposed to the background gravity generates a vertical vibration of the ablation front itself. The effects of different driving modulations (Dirac deltas and square waves) are analyzed from a theoretical point of view, with a focus on stabilization of ion beam driven ablation fronts, and a comparison is made, in order to look for optimization.

Molecular approach to Neurodegenerative Disorders: Role of Nociceptin/Orphanin FQ - NOP System in Parkinson and Epigenetic Mechanism in Alzheimer's Disease

With life expectancies increasing around the world, populations are getting age and neurodegenerative diseases have become a global issue. For this reason we have focused our attention on the two most important neurodegenerative diseases: Parkinson’s and Alzheimer’s. Parkinson’s disease is a chronic progressive neurodegenerative movement disorder of multi-factorial origin. Environmental toxins as well as agricultural chemicals have been associated with PD. Has been observed that N/OFQ contributes to both neurotoxicity and symptoms associated with PD and that pronociceptin gene expression is up-regulated in rat SN of 6-OHDA and MPP induced experimental parkinsonism. First, we investigated the role of N/OFQ-NOP system in the pathogenesis of PD in an animal model developed using PQ and/or MB. Then we studied Alzheimer's disease. This disorder is defined as a progressive neurologic disease of the brain leading to the irreversible loss of neurons and the loss of intellectual abilities, including memory and reasoning, which become severe enough to impede social or occupational functioning. Effective biomarker tests could prevent such devastating damage occurring. We utilized the peripheral blood cells of AD discordant monozygotic twin in the search of peripheral markers which could reflect the pathology within the brain, and also support the hypothesis that PBMC might be a useful model of epigenetic gene regulation in the brain. We investigated the mRNA levels in several genes involve in AD pathogenesis, as well DNA methylation by MSP Real-Time PCR. Finally by Western Blotting we assess the immunoreactivity levels for histone modifications. Our results support the idea that epigenetic changes assessed in PBMCs can also be useful in neurodegenerative disorders, like AD and PD, enabling identification of new biomarkers in order to develop early diagnostic programs.

Evolutionary tendencies in flowers of Marantaceae with special reference to the style movement mechanism

One of the quickest plant movements ever known is made by the ´explosive´ style in Marantaceae in the service of secondary pollen presentation – herewith showing a striking apomorphy to the sister Cannaceae that might be of high evolutionary consequence. Though known already since the beginning of the 19th century the underlying mechanism of the movement has hitherto not been clarified. The present study reports about the biomechanics of the style-staminode complex and the hydraulic principles of the movement. For the first time it is shown by experiment that in Maranta noctiflora through longitudinal growth of the maturing style in the ´straitjacket´ of the hooded staminode both the hold of the style prior to its release and its tensioning for the movement are brought about. The longer the style grows in relation to the enclosing hooded staminode the more does its capacity for curling up for pollen transfer increase. Hereby I distinguish between the ´basic tension´ that a growing style builds up anyway, even when the hooded staminode is removed beforehand, and the ´induced tension´ which comes about only under the pressure of a ´too short´ hooded staminode and which enables the movement. The results of these investigations are discussed in view of previous interpretations ranging from possible biomechanical to electrophysiological mechanisms. To understand furthermore by which means the style gives way to the strong bending movement without suffering outwardly visible damage I examined its anatomical structure in several genera for its mechanical and hydraulic properties and for the determination of the entire curvature after release. The actual bending part contains tubulate cells whose walls are extraordinarily porous and large longitudinal intercellular spaces. SEM indicates the starting points of cell-wall loosening in primary walls and lysis of middle lamellae - probably through an intense pectinase activity in the maturing style. Fluorescence pictures of macerated and living style-tissue confirm cell-wall perforations that do apparently connect neighbouring cells, which leads to an extremely permeable parenchyma. The ´water-body´ can be shifted from central to dorsal cell layers to support the bending. The geometrical form of the curvature is determined by the vascular bundles. I conclude that the style in Marantaceae contains no ´antagonistic´ motile tissues as in Mimosa or Dionaea. Instead, through self-maceration it develops to a ´hydraulic tissue´ which carries out an irreversible movement through a sudden reshaping. To ascertain the evolutionary consequence of this apomorphic pollination mechanism the diversity and systematic value of hooded staminodes are examined. For this hooded staminodes of 24 genera are sorted according to a minimalistic selection of shape characters and eight morphological types are abstracted from the resulting groups. These types are mapped onto an already available maximally parsimonious tree comprising five major clades. An amazing correspondence is found between the morphological types and the clades; several sister-relationships are confirmed and in cases of uncertain position possible evolutionary pathways, such as convergence, dispersal or re-migration, are discussed, as well as the great evolutionary tendencies for the entire family in which – at least as regards the shape of hooded staminodes – there is obviously a tendency from complicated to strongly simplified forms. It suggests itself that such simplifying derivations may very likely have taken place as adaptations to pollinating animals about which at present too little is known. The value of morphological characters in relation to modern phylogenetic analysis is discussed and conditions for the selection of morphological characters valuable for a systematic grouping are proposed. Altogether, in view of the evolutionary success of Marantaceae compared with Cannaceae the movement mechanism of the style-staminode complex can safely be considered a key innovation within the order Zingiberales.

Multi Processor Systems On Chip with Configurable Hardware Acceleration

The evolution of the electronics embedded applications forces electronics systems designers to match their ever increasing requirements. This evolution pushes the computational power of digital signal processing systems, as well as the energy required to accomplish the computations, due to the increasing mobility of such applications. Current approaches used to match these requirements relies on the adoption of application specific signal processors. Such kind of devices exploits powerful accelerators, which are able to match both performance and energy requirements. On the other hand, the too high specificity of such accelerators often results in a lack of flexibility which affects non-recurrent engineering costs, time to market, and market volumes too. The state of the art mainly proposes two solutions to overcome these issues with the ambition of delivering reasonable performance and energy efficiency: reconfigurable computing and multi-processors computing. All of these solutions benefits from the post-fabrication programmability, that definitively results in an increased flexibility. Nevertheless, the gap between these approaches and dedicated hardware is still too high for many application domains, especially when targeting the mobile world. In this scenario, flexible and energy efficient acceleration can be achieved by merging these two computational paradigms, in order to address all the above introduced constraints. This thesis focuses on the exploration of the design and application spectrum of reconfigurable computing, exploited as application specific accelerators for multi-processors systems on chip. More specifically, it introduces a reconfigurable digital signal processor featuring a heterogeneous set of reconfigurable engines, and a homogeneous multi-core system, exploiting three different flavours of reconfigurable and mask-programmable technologies as implementation platform for applications specific accelerators. In this work, the various trade-offs concerning the utilization multi-core platforms and the different configuration technologies are explored, characterizing the design space of the proposed approach in terms of programmability, performance, energy efficiency and manufacturing costs.

From Folke Medicine to Medicine Chemistry: Study, Using in Vitro and Cellular Assays, of Receptors Mechanism Involved in the Activities of Natural Compounds

My Doctorate Research has been focused on the evaluation of the pharmacological activity of a natural extract of chestnut wood (ENC) towards the cardiovascular and gastrointestinal system and on the identification of the active compounds. The ENC has been shown to contain more than 10% (w/w) of phenolic compounds, of which tannins as Vescalgin and Castalgin are the more representative. ENC cardiovascular effects have been investigated in guinea pig cardiac preparations; furthermore its activity has been evalueted in guinea pig aorta strips. ENC induced transient negative chronotropic effect in isolated spontaneously beating right atria and simultaneously positive inotropic effect in left atria driven at 1 Hz. Cardiac cholinergic receptors are not involved in the negative chronotropic effect and positive inotropic effects are not related to adrenergic receptors. In vascular smooth muscle, natural extract of chestnut did not significantly change the contraction induced by potassium (80 mM) or that induced by noradrenaline (1μM). In guinea pig ileum, ENC reduced the maximum response to carbachol in a concentrationdependent manner and behaved as a reversible non competitive antagonist. In guinea pig ileum, the antispasmodic activity of ENC showed a significant antispasmodic activity against a variety of different spasmogenic agents including histamine, KCl, BaCl2. In guinea pig proximal colon, stomach and jejunum, ENC reduced the maximum response to carbachol in a concentrationdependent manner and behaved as a reversible non competitive antagonist. ENC contracted gallbladder guinea pig in a reversible and concentration-dependent manner. This effect does not involve cholinergic and cholecystokinin receptors and it is reduced by nifedipine. ENC relaxed Oddi sphincter smooth muscle. The cholecystokinetic and Oddi sphincter relaxing activities occurred also in guinea pigs fed a lithogenic diet. The cholecystokinetic occurred also in human gallbladder. The Fractionation of the extract led to the identification of the active fraction.

The Effectiveness of the Clean Development Mechanism – A law and economics analysis

Climate change has been acknowledged as a threat to humanity. Most scholars agree that to avert dangerous climate change and to transform economies into low-carbon societies, deep global emission reductions are required by the year 2050. Under the framework of the Kyoto Protocol, the Clean Development Mechanism (CDM) is the only market-based instrument that encourages industrialised countries to pursue emission reductions in developing countries. The CDM aims to pay the incremental finance necessary to operationalize emission reduction projects which are otherwise not financially viable. According to the objectives of the Kyoto Protocol, the CDM should finance projects that are additional to those which would have happened anyway, contribute to sustainable development in the countries hosting the projects, and be cost-effective. To enable the identification of such projects, an institutional framework has been established by the Kyoto Protocol which lays out responsibilities for public and private actors. This thesis examines whether the CDM has achieved these objectives in practice and can thus be considered an effective tool to reduce emissions. To complete this investigation, the book applies economic theory and analyses the CDM from two perspectives. The first perspective is the supply-dimension which answers the question of how, in practice, the CDM system identified additional, cost-effective, sustainable projects and, generated emission reductions. The main contribution of this book is the second perspective, the compliance-dimension, which answers the question of whether industrialised countries effectively used the CDM for compliance with their Kyoto targets. The application of the CDM in the European Union Emissions Trading Scheme (EU ETS) is used as a case-study. Where the analysis identifies inefficiencies within the supply or the compliance dimension, potential improvements of the legal framework are proposed and discussed.

Gamma-secretase modulators and inhibitors in Alzheimer's disease: influence of genetic background on efficacy and mechanism of action

According to the amyloid hypothesis, Alzheimer’s disease (AD) is caused by aberrant production or clearance of the amyloid-β (Aβ) peptides, and in particular of the longer more aggregation-prone Aβ42. The Aβ peptides are generated through successive proteolytic cleavage of the amyloid precursor protein (APP) by the β-site APP cleaving enzyme (BACE) and γ-secretase. γ-secretase produces Aβ peptides with variable C-termini ranging from Aβ34 to Aβ48, presumably by sequential trimming of longer into shorter peptides. γ-secretase is a multiprotein complex consisting of at least four different proteins and the presenilin proteins (PS1 or PS2) contain the catalytic center of the complex. In 2001 several non-steroidal anti-inflammatory drugs were identified as the founding members of a new class of γ-secretase modulators (GSMs) that can selectively reduce production of Aβ42. Concomitantly, these GSMs increase Aβ38 production indicating closely coordinated generation of Aβ42 and Aβ38 and a potential precursor-product relationship between these peptides. GSMs seem to exert their activity by direct modulation of γ-secretase. Support for this hypothesis is drawn from the finding that some PS mutations associated with early-onset familial AD (FAD) can modulate the cellular response to GSMs and to γ-secretase inhibitors (GSIs), which inhibit production of all Aβ peptides and are known to directly interact with PS. A particularly interesting FAD PS mutation is PS1-ΔExon9, a complex deletion mutant that blocks endoproteolysis of PS1 and renders cells completely non-responsive to GSMs. Studies presented in this thesis show that the diminished response of PS1-ΔExon9 to GSMs is mainly caused by its lack of endoproteolytic cleavage. Furthermore, we were able to demonstrate that a reduced response to GSMs and GSIs is not limited to PS1-ΔExon9 but is a common effect of aggressive FAD-associated PS1 mutations. Surprisingly, we also found that while the Aβ42 response to GSMs is almost completely abolished by these PS1 mutations, the accompanying Aβ38 increase was indistinguishable to wild-type PS1. Finally, the reduced response to GSIs was confirmed in a mouse model with transgenic expression of an aggressive FAD-associated PS1 mutation as a highly potent GSI failed to reduce Aβ42 levels in brain of these mice. Taken together, our findings provide clear evidence for independent generation of Aβ42 and Aβ38 peptides, and argue that the sequential cleavage model might be an oversimplification of the molecular mechanism of γ-secretase. Most importantly, our results highlight the significance of genetic background in drug discovery efforts aimed at γ-secretase, and indicate that the use of cellular models with transgenic expression of FAD-associated PS mutations might confound studies of the potency and efficacy of GSMs and GSIs. Therefore, such models should be strictly avoided in the ongoing preclinical development of these promising and potentially disease-modifying therapeutics for AD.

Taste receptors in the gut: a chemosensitive mechanism from fish to human

The ingestion of a meal evokes a series of digestive processes, which consist of the essential functions of the digestive system: food transport, secretory activity, absorption of nutrients and the expulsion of undigested residues do not absorbed. The gastrointestinal chemosensitivity is characterized by cellular elements of the endocrine gastrointestinal mucosa and nerve fibers, in particular of vagal nature. A wide range of mediators endocrine and/or paracrine can be released from various endocrine cells in response to nutrients in the diet. These hormones, in addition to their direct activity, act through specific receptors activating some of the most important functions in the control of energy intake and energy homeostasis in the body. For integration of this complex system of control of gastrointestinal chemosensitivity, recent evidence demonstrates the presence of taste receptors (TR) belonging to the family of G proteins coupled receptor expressed in the mucosa of the gastrointestinal tract of different mammals and human. This thesis is divided into several research projects that have been conceived in order to clarify the relationship between TR and nutrients. To define this relationship I have used various scientific approaches, which have gone on to evaluate changes in signal molecules of TR, in particular of the α-transducin in the fasting state and after refeeding with standard diet in the gastrointestinal tract of the pig, the mapping of the same molecule signal in the gastrointestinal tract of fish (Dicentrarchus labrax), the signaling pathway of bitter TR in the STC-1 endocrine cell line and finally the involvement of bitter TR in particular of T2R38 in patients with an excessive caloric intake. The results showed how there is a close correlation between nutrients, TR and hormonal release and how they are useful both in taste perception but also likely to be involved in chronic diseases such as obesity.

On the inheritance and mechanism of baculovirus resistance of the codling moth, Cydia pomonella (L.)

Das Cydia pomonella Granulovirus (CpGV, Baculoviridae) wird seit Ende der 1980er Jahre als hoch-selektives und effizientes biologisches Bekämpfungsmittel zur Kontrolle des Apfelwicklers im Obstanbau eingesetzt. Seit 2004 wurden in Europa verschiedene Apfelwicklerpopulationen beobachtet die resistent gegenüber dem hauptsächlich angewendeten Isolat CpGV-M aufweisen. Die vorliegende Arbeit befasst sich mit der Untersuchung der Vererbung und des Mechanismus der CpGV Resistenz. Einzelpaarkreuzungen zwischen einem empfindlichen Laborstamm (CpS) und einem homogen resistenten Stamm (CpRR1) zeigten, dass die Resistenz durch ein einziges dominantes Gen, das auf dem Z-Chromosom lokalisiert ist, vererbt wird. Massernkreuzungen zwischen CpS und einer heterogen resistenten Feldpopulation (CpR) deuteten zunächst auf einen unvollständig dominanten autosomalen Erbgang hin. Einzelpaarkreuzungen zwischen CpS und CpR bewiesen jedoch, dass die Resistenz in CpR ebenfalls monogen dominant und geschlechtsgebunden auf dem Z-Chromosom vererbt wird. Diese Arbeit diskutiert zudem die Vor- und Nachteile von Einzelpaarkreuzungen gegenüber Massernkreuzungen bei der Untersuchung von Vererbungsmechanismen. Die Wirksamkeit eines neuen CpGV Isolates aus dem Iran (CpGV-I12) gegenüber CpRR1 Larven, wurde in Bioassays getestet. Die Ergebnisse zeigen, dass CpGV-I12 die Resistenz in allen Larvenstadien von CpRR1 brechen kann und fast so gut wirkt wie CpGV-M gegenüber CpS Larven. Daher ist CpGV-I12 für die Kontrolle des Apfelwicklers in Anlagen wo die Resistenz aufgetreten ist geeignet. Um den der CpGV Resistenz zugrunde liegenden Mechanismus zu untersuchen, wurden vier verschiedene Experimente durchgeführt: 1) die peritrophische Membran degradiert indem ein optischer Aufheller dem virus-enthaltenden Futtermedium beigefügt wurde. Das Entfernen dieser mechanischen Schutzbarriere, die den Mitteldarm auskleidet, führte allerdings nicht zu einer Reduzierung der Resistenz in CpR Larven. Demnach ist die peritrophische Membran nicht am Resistenzmechanismus beteiligt. 2) Die Injektion von Budded Virus in das Hämocoel führte nicht zur Brechung der Resistenz. Folglich die die Resistenz nicht auf den Mitteldarm beschränkt, sondern auch in der Sekundärinfektion wirksam. 3) Die Replikation von CpGV in verschiedenen Geweben (Mitteldarm, Hämolymphe und Fettkörper) von CpS und CpRR1 wurde mittels quantitativer PCR verfolgt. In CpS Larven konnte in allen drei Gewebetypen sowohl nach oraler als auch nach intra-hämocoelarer Infektion eine Zunahme der CpGV Genome in Abhängigkeit der Zeit festgestellt werden. Dagegen konnte in den Geweben aus CpRR1 nach oraler sowie intra-hämocoelarer Infektion keine Virusreplikation detektiert werden. Dies deutet darauf hin, dass die CpGV Resistenz in allen Zelltypen präsent ist. 4) Um zu untersuchen ob ein humoraler Faktor in der Hämolymphe ursächlich an der Resistenz beteiligt ist, wurde Hämolymphe aus CpRR1 Larven in CpS Larven injiziert und diese anschließend oral mit CpGV infiziert. Es konnte jedoch keine Immunreaktion beobachtet und kein Faktor in der Hämolymphe identifiziert werden, der Resistenz induzieren könnte. Auf Grundlage dieser Ergebnisse kann festgestellt werden, dass in resistenten Apfelwicklerlarven die virale Replikation in allen Zelltypen verhindert wird, was auf eine Virus-Zell Inkompatibilität hinweist. Da in CpRR1 keine DNA Replikation beobachtet wurde, wird die CpGV Resistenz wahrscheinlich durch eine frühe Unterbindung der Virusreplikation verursacht.Das früh exprimierte Gen pe38 codiert für ein Protein, das wahrscheinlich für die Resistenzbrechung durch CpGV-I12 verantwortlich ist. Interaktionen zwischen dem Protein PE38 und Proteinen in CpRR1 wurden mit Hilfe des Yeast Two-Hybrid (Y2H) Systems untersucht. Die detektierten Interaktionen sind noch nicht durch andere Methoden bestätigt, jedoch wurden zwei mögliche Gene auf dem Z-Chromosom und eines auf Chromosom 15 gefunden, wie möglicherweise an der CpGV Resistenz beteiligt sind.

Approccio agli spostamenti per la valutazione della vulnerabilità sismica di telai in conglomerato cementizio armato.

Il presente lavoro ha lo scopo di presentare gli studi e i risultati ottenuti durante l’attività di ricerca svolta sul Displacement-based Assessment (DBA) dei telai in cemento armato. Dopo alcune considerazioni iniziali sul tema della vulnerabilità sismica e sui metodi di analisi e verifica, si procede alla descrizione teorica del metodo. Sono stati analizzati tre casi studio di telai piani, progettati per soli carichi verticali e secondo normative non più in vigore che non prevedevano l’applicazione della gerarchia delle resistenze. I telai considerati, destinati ad abitazione civile, hanno diversa altezza e numero di piani, e diverso numero di campate. Si è proceduto all’applicazione del metodo, alla valutazione della vulnerabilità sismica in base alla domanda in termini di spostamento costituita da uno spettro elastico previsto dall’EC8 e alla validazione dei risultati ottenuti mediante analisi non lineari statiche e dinamiche e mediante l’applicazione dei teoremi dell’Analisi limite dei telai, proposta come procedura alternativa per la determinazione del meccanismo anelastico e della capacità in termini di taglio alla base. In ultimo si è applicata la procedura DBA per la valutazione della vulnerabilità sismica di un edificio scolastico, realizzato tra il 1969 e il 1975 in un sito caratterizzato da una accelerazione di picco orizzontale pari a 0,24g e una probabilità di superamento del 10% in 75 anni.

Novel isolates of Cydia pomonella granulovirus (CpGV): deciphering the molecular mechanism for overcoming CpGV resistance in codling moth (Cydia pomonella)

During the last twenty years, Cydia pomonolla granulovirus (CpGV, Baculoviridae) has become the most important biological control agent for the codling moth (CM) in organic and integrated apple production. All registered products in Europe are based on the isolate CpGV-M, which was discovered 1964 in Mexico. A serious threat to future application of CpGV is the occurrence of CM field populations resistant to CpGV. Since 2003, populations with up to 10,000-fold reduced susceptibility were reported from orchards in Germany, France, Italy, Switzerland, Austria and the Netherlands. A putative alternative to CpGV-M are novel CpGV isolates which are able to overcome CM resistance. This thesis focuses on the identification and characterisation of resistance overcoming CpGV isolates and the analysis of their molecular difference to CpGV-M.rnSixteen CpGV isolates were tested against CM lab strains in bioassays. Hereby, five isolates were identified which were able to completely overcome resistance. The genomes of these isolates were compared to CpGV-M by restriction fragment length polymorphism (RFLP) analysis. To identify the molecular factor responsible for improved virulence of some CpGV isolates, major genomic differences were sequenced and analysed. A 0.7 kb insertion was found in CpGV-I01, -I12 and -E2, but not in other resistance overcoming isolates. Analysis of the insertions sequence revealed that it might be due to a transposition event, but not involved in overcoming resistance. rnFor unequivocal identification of CpGV isolates, a new method based on molecular analysis was established. Partial sequencing of the conserved polyhedrin/granulin (polh/gran), late expression factor-8 (lef-8) and late expression factor-9 (lef-9) genes revealed single nucleotide polymorphisms (SNPs). SNP analysis correlated with the grouping obtained by RFLP analysis. A phylogenetic classification due to different genome types A-E is proposed. Phylogenetic analysis suggested that CpGV-M was the phylogenetically youngest of the tested CpGV isolates.rnWhole genome sequencing of two resistance overcoming isolates CpGV-I12 (type D genome) and -S (type E genome) and CpGV-M (type A genome) was performed. Comparison of the three genomes revealed a high sequence identity. Several insertions and deletions ranging from 1-700 nucleotides (nt) were found. Comparison on open reading frame (ORF) level revealed that CpGV-I12 and -S shared only one protein alteration when compared to CpGV-M: a stretch of 24 nt present in ORF cp24 was not found in any of the resistance overcoming isolates. Cp24 codes for the early gene pe38. Combined with the results of phylogenetic analysis, it is proposed that these 24 nt are a recent insertion into the CpGV-M genome. The role of pe38 in overcoming resistance was investigated by knocking out pe38 of a CpGV-M based bacmid and swapping of CpGV-I12 pe38 of into the k.o. bacmid. When pe38 of CpGV-I12 was inserted into the k.o. bacmid, the infectivity could not be rescued, suggesting that the genomic portion of pe38 might play a role in its function.rnIt can be concluded that the recently observed CpGV resistance in CM is only related to type A genomes. RFLP and SNP analysis provide tools for identifying and characterising different CpGV isolates reliably, a pre-condition for a future registration of CpGV products based on novel CpGV isolates.rnrnrn

Laser driven proton acceleration and beam shaping

In the race to obtain protons with higher energies, using more compact systems at the same time, laser-driven plasma accelerators are becoming an interesting possibility. But for now, only beams with extremely broad energy spectra and high divergence have been produced. The driving line of this PhD thesis was the study and design of a compact system to extract a high quality beam out of the initial bunch of protons produced by the interaction of a laser pulse with a thin solid target, using experimentally reliable technologies in order to be able to test such a system as soon as possible. In this thesis, different transport lines are analyzed. The first is based on a high field pulsed solenoid, some collimators and, for perfect filtering and post-acceleration, a high field high frequency compact linear accelerator, originally designed to accelerate a 30 MeV beam extracted from a cyclotron. The second one is based on a quadruplet of permanent magnetic quadrupoles: thanks to its greater simplicity and reliability, it has great interest for experiments, but the effectiveness is lower than the one based on the solenoid; in fact, the final beam intensity drops by an order of magnitude. An additional sensible decrease in intensity is verified in the third case, where the energy selection is achieved using a chicane, because of its very low efficiency for off-axis protons. The proposed schemes have all been analyzed with 3D simulations and all the significant results are presented. Future experimental work based on the outcome of this thesis can be planned and is being discussed now.