Aureobasidium pullulans as biological control agent: modes of action

The postharvest phase has been considered an environment very suitable for successful application of biological control agents (BCAs). However, the tri-interaction between fungal pathogen, host (fruit) and antagonist is influenced by several parameters such as temperature, oxidative stresses, oxygen composition, water activity, etc. that could be determining for the success of biocontrol. Knowledge of the modes of action of BCAs is essential in order to enhance their viability and increase their potentialities in disease control. The thesis focused on the possibility to explain the modes of action of a biological control agent (BCA): Aureobasidium pullulans, in particular the strains L1 and L8, control effective against fruit postharvest fungal pathogen. In particular in this work were studied the different modes of action of BCA, such as: i) the ability to produce volatile organic compounds (VOCs), identified by SPME- gas chromatography-mass spectrometry (GC-MS) and tested by in vitro and in vivo assays against Penicillium spp., Botrytis cinerea, Colletotrichum acutatum; ii) the ability to produce lytic enzymes (exo and endo chitinase and β-1,3-glucanase) tested against Monilinia laxa, causal agent of brown rot of stone fruits. L1 and L8 lytic enzymes were also evaluated through their relative genes by molecular tools; iii) the competition for space and nutrients, such as sugars (sucrose, glucose and fructose) and iron; the latter induced the production of siderophores, molecules with high affinity for iron chelation. A molecular investigation was carried out to better understand the gene regulation strictly correlated to the production of these chelating molucules. The competition for space against M. laxa was verified by electron microscopy techniques; iv) a depth bibliographical analysis on BCAs mechanisms of action and their possible combination with physical and chemical treatments was conducted.

Spinal cord injury: assessment of autonomic state-dependent control of cardiovascular system and body core temperature

Spinal cord injury (SCI) results not only in paralysis; but it is also associated with a range of autonomic dysregulation that can interfere with cardiovascular, bladder, bowel, temperature, and sexual function. The entity of the autonomic dysfunction is related to the level and severity of injury to descending autonomic (sympathetic) pathways. For many years there was limited awareness of these issues and the attention given to them by the scientific and medical community was scarce. Yet, even if a new system to document the impact of SCI on autonomic function has recently been proposed, the current standard of assessment of SCI (American Spinal Injury Association (ASIA) examination) evaluates motor and sensory pathways, but not severity of injury to autonomic pathways. Beside the severe impact on quality of life, autonomic dysfunction in persons with SCI is associated with increased risk of cardiovascular disease and mortality. Therefore, obtaining information regarding autonomic function in persons with SCI is pivotal and clinical examinations and laboratory evaluations to detect the presence of autonomic dysfunction and quantitate its severity are mandatory. Furthermore, previous studies demonstrated that there is an intimate relationship between the autonomic nervous system and sleep from anatomical, physiological, and neurochemical points of view. Although, even if previous epidemiological studies demonstrated that sleep problems are common in spinal cord injury (SCI), so far only limited polysomnographic (PSG) data are available. Finally, until now, circadian and state dependent autonomic regulation of blood pressure (BP), heart rate (HR) and body core temperature (BcT) were never assessed in SCI patients. Aim of the current study was to establish the association between the autonomic control of the cardiovascular function and thermoregulation, sleep parameters and increased cardiovascular risk in SCI patients.

How teachers think about the role of digital technologies in student assessment in mathematics

This study concerns teachers’ use of digital technologies in student assessment, and how the learning that is developed through the use of technology in mathematics can be evaluated. Nowadays math teachers use digital technologies in their teaching, but not in student assessment. The activities carried out with technology are seen as ‘extra-curricular’ (by both teachers and students), thus students do not learn what they can do in mathematics with digital technologies. I was interested in knowing the reasons teachers do not use digital technology to assess students’ competencies, and what they would need to be able to design innovative and appropriate tasks to assess students’ learning through digital technology. This dissertation is built on two main components: teachers and task design. I analyze teachers’ practices involving digital technologies with Ruthven’s Structuring Features of Classroom Practice, and what relation these practices have to the types of assessment they use. I study the kinds of assessment tasks teachers design with a DGE (Dynamic Geometry Environment), using Laborde’s categorization of DGE tasks. I consider the competencies teachers aim to assess with these tasks, and how their goals relate to the learning outcomes of the curriculum. This study also develops new directions in finding how to design suitable tasks for student mathematical assessment in a DGE, and it is driven by the desire to know what kinds of questions teachers might be more interested in using. I investigate the kinds of technology-based assessment tasks teachers value, and the type of feedback they give to students. Finally, I point out that the curriculum should include a range of mathematical and technological competencies that involve the use of digital technologies in mathematics, and I evaluate the possibility to take advantage of technology feedback to allow students to continue learning while they are taking a test.

Molecular architecture of fur binding to iron-induced and - repressed genes in Helicobacter pylori

The ferric uptake regulator protein Fur regulates iron-dependent gene expression in bacteria. In the human pathogen Helicobacter pylori, Fur has been shown to regulate iron-induced and iron-repressed genes. Herein we investigate the molecular mechanisms that control this differential iron-responsive Fur regulation. Hydroxyl radical footprinting showed that Fur has different binding architectures, which characterize distinct operator typologies. On operators recognized with higher affinity by holo-Fur, the protein binds to a continuous AT-rich stretch of about 20 bp, displaying an extended protection pattern. This is indicative of protein wrapping around the DNA helix. DNA binding interference assays with the minor groove binding drug distamycin A, point out that the recognition of the holo-operators occurs through the minor groove of the DNA. By contrast, on the apo-operators, Fur binds primarily to thymine dimers within a newly identified TCATTn10TT consensus element, indicative of Fur binding to one side of the DNA, in the major groove of the double helix. Reconstitution of the TCATTn10TT motif within a holo-operator results in a feature binding swap from an holo-Fur- to an apo-Fur-recognized operator, affecting both affinity and binding architecture of Fur, and conferring apo-Fur repression features in vivo. Size exclusion chromatography indicated that Fur is a dimer in solution. However, in the presence of divalent metal ions the protein is able to multimerize. Accordingly, apo-Fur binds DNA as a dimer in gel shift assays, while in presence of iron, higher order complexes are formed. Stoichiometric Ferguson analysis indicates that these complexes correspond to one or two Fur tetramers, each bound to an operator element. Together these data suggest that the apo- and holo-Fur repression mechanisms apparently rely on two distinctive modes of operator-recognition, involving respectively the readout of a specific nucleotide consensus motif in the major groove for apo-operators, and the recognition of AT-rich stretches in the minor groove for holo-operators, whereas the iron-responsive binding affinity is controlled through metal-dependent shaping of the protein structure in order to match preferentially the major or the minor groove.

The Perception of American Adolescent Culture Through the Dubbing and Fansubbing of a Selection of US Teen Series from 1990 to 2013

The present dissertation aims at analyzing the construction of American adolescent culture through teen-targeted television series and the shift in perception that occurs as a consequence of the translation process. In light of the recent changes in television production and consumption modes, largely caused by new technologies, this project explores the evolution of Italian audiences, focusing on fansubbing (freely distributed amateur subtitles made by fans for fan consumption) and social viewing (the re-aggregation of television consumption based on social networks and dedicated platforms, rather than on physical presence). These phenomena are symptoms of a sort of ‘viewership 2.0’ and of a new type of active viewing, which calls for a revision of traditional AVT strategies. Using a framework that combines television studies, new media studies, and fandom studies with an approach to AVT based on Descriptive Translation Studies (Toury 1995), this dissertation analyzes the non-Anglophone audience’s growing need to participation in the global dialogue and appropriation process based on US scheduling and informed by the new paradigm of convergence culture, transmedia storytelling, and affective economics (Jenkins 2006 and 2007), as well as the constraints intrinsic to multimodal translation and the different types of linguistic and cultural adaptation performed through dubbing (which tends to be more domesticating; Venuti 1995) and fansubbing (typically more foreignizing). The study analyzes a selection of episodes from six of the most popular teen television series between 1990 and 2013, which has been divided into three ages based on the different modes of television consumption: top-down, pre-Internet consumption (Beverly Hills, 90210, 1990 – 2000), emergence of audience participation (Buffy the Vampire Slayer, 1997 – 2003; Dawson’s Creek, 1998 – 2003), age of convergence and Viewership 2.0 (Gossip Girl, 2007 – 2012; Glee, 2009 – present; The Big Bang Theory, 2007 - present).

Innovative asymmetric organocatalytic processess: en route to the synthesis of biologically relevant compounds

This doctoral thesis deals with the development of novel organocatalytic strategies for asymmetric transformation. The intrinsic versatility of organocatalysis and the use of different activation modes have been exploited to achieve new catalytic enantioselective processes, towards the synthesis of biologically relevant scaffolds. The most investigated organocatalytic system have been those based on H-bond interaction (such as chiral thioureas or phosphoric acids) as well as the ones based on aminocatalysis. Despite conceptually distinct, the transformations detailed in this Thesis are linked together by simple and recurring modes of activation, induction and reactivity, promoted by the catalysts employed. The chemical diversity of the challenges encountered allows to get a precious overall view on organocatalysis, highlighting that enormous chemical diversity can be created by judicious choice of select catalyst.