Development of Imaging Mass Spectrometry Analysis of Lipids in Biological and Clinically Relevant Applications

La spectrométrie de masse mesure la masse des ions selon leur rapport masse sur charge. Cette technique est employée dans plusieurs domaines et peut analyser des mélanges complexes. L’imagerie par spectrométrie de masse (Imaging Mass Spectrometry en anglais, IMS), une branche de la spectrométrie de masse, permet l’analyse des ions sur une surface, tout en conservant l’organisation spatiale des ions détectés. Jusqu’à présent, les échantillons les plus étudiés en IMS sont des sections tissulaires végétales ou animales. Parmi les molécules couramment analysées par l’IMS, les lipides ont suscité beaucoup d'intérêt. Les lipides sont impliqués dans les maladies et le fonctionnement normal des cellules; ils forment la membrane cellulaire et ont plusieurs rôles, comme celui de réguler des événements cellulaires. Considérant l’implication des lipides dans la biologie et la capacité du MALDI IMS à les analyser, nous avons développé des stratégies analytiques pour la manipulation des échantillons et l’analyse de larges ensembles de données lipidiques. La dégradation des lipides est très importante dans l’industrie alimentaire. De la même façon, les lipides des sections tissulaires risquent de se dégrader. Leurs produits de dégradation peuvent donc introduire des artefacts dans l’analyse IMS ainsi que la perte d’espèces lipidiques pouvant nuire à la précision des mesures d’abondance. Puisque les lipides oxydés sont aussi des médiateurs importants dans le développement de plusieurs maladies, leur réelle préservation devient donc critique. Dans les études multi-institutionnelles où les échantillons sont souvent transportés d’un emplacement à l’autre, des protocoles adaptés et validés, et des mesures de dégradation sont nécessaires. Nos principaux résultats sont les suivants : un accroissement en fonction du temps des phospholipides oxydés et des lysophospholipides dans des conditions ambiantes, une diminution de la présence des lipides ayant des acides gras insaturés et un effet inhibitoire sur ses phénomènes de la conservation des sections au froid sous N2. A température et atmosphère ambiantes, les phospholipides sont oxydés sur une échelle de temps typique d’une préparation IMS normale (~30 minutes). Les phospholipides sont aussi décomposés en lysophospholipides sur une échelle de temps de plusieurs jours. La validation d’une méthode de manipulation d’échantillon est d’autant plus importante lorsqu’il s’agit d’analyser un plus grand nombre d’échantillons. L’athérosclérose est une maladie cardiovasculaire induite par l’accumulation de matériel cellulaire sur la paroi artérielle. Puisque l’athérosclérose est un phénomène en trois dimension (3D), l'IMS 3D en série devient donc utile, d'une part, car elle a la capacité à localiser les molécules sur la longueur totale d’une plaque athéromateuse et, d'autre part, car elle peut identifier des mécanismes moléculaires du développement ou de la rupture des plaques. l'IMS 3D en série fait face à certains défis spécifiques, dont beaucoup se rapportent simplement à la reconstruction en 3D et à l’interprétation de la reconstruction moléculaire en temps réel. En tenant compte de ces objectifs et en utilisant l’IMS des lipides pour l’étude des plaques d’athérosclérose d’une carotide humaine et d’un modèle murin d’athérosclérose, nous avons élaboré des méthodes «open-source» pour la reconstruction des données de l’IMS en 3D. Notre méthodologie fournit un moyen d’obtenir des visualisations de haute qualité et démontre une stratégie pour l’interprétation rapide des données de l’IMS 3D par la segmentation multivariée. L’analyse d’aortes d’un modèle murin a été le point de départ pour le développement des méthodes car ce sont des échantillons mieux contrôlés. En corrélant les données acquises en mode d’ionisation positive et négative, l’IMS en 3D a permis de démontrer une accumulation des phospholipides dans les sinus aortiques. De plus, l’IMS par AgLDI a mis en évidence une localisation différentielle des acides gras libres, du cholestérol, des esters du cholestérol et des triglycérides. La segmentation multivariée des signaux lipidiques suite à l’analyse par IMS d’une carotide humaine démontre une histologie moléculaire corrélée avec le degré de sténose de l’artère. Ces recherches aident à mieux comprendre la complexité biologique de l’athérosclérose et peuvent possiblement prédire le développement de certains cas cliniques. La métastase au foie du cancer colorectal (Colorectal cancer liver metastasis en anglais, CRCLM) est la maladie métastatique du cancer colorectal primaire, un des cancers le plus fréquent au monde. L’évaluation et le pronostic des tumeurs CRCLM sont effectués avec l’histopathologie avec une marge d’erreur. Nous avons utilisé l’IMS des lipides pour identifier les compartiments histologiques du CRCLM et extraire leurs signatures lipidiques. En exploitant ces signatures moléculaires, nous avons pu déterminer un score histopathologique quantitatif et objectif et qui corrèle avec le pronostic. De plus, par la dissection des signatures lipidiques, nous avons identifié des espèces lipidiques individuelles qui sont discriminants des différentes histologies du CRCLM et qui peuvent potentiellement être utilisées comme des biomarqueurs pour la détermination de la réponse à la thérapie. Plus spécifiquement, nous avons trouvé une série de plasmalogènes et sphingolipides qui permettent de distinguer deux différents types de nécrose (infarct-like necrosis et usual necrosis en anglais, ILN et UN, respectivement). L’ILN est associé avec la réponse aux traitements chimiothérapiques, alors que l’UN est associé au fonctionnement normal de la tumeur.

Regional Workshop for designers monitors responsible for training programs for staff in public and school libraries

In Bogota, from 22 to 26 September 1980 a Seminar Workshop on Education and Training of Personnel in the organization and management of school libraries, sponsored by UNESCO, CERLAL, SECAB and the National Ministry of Colombia. The meeting reiterated that one of the weaknesses of library services is the lack of qualified personnel. As an immediate action is decided to set up a working group composed of experts from those agencies that have had experiences in the field of personnel training, such as Colombia, Venezuela and Costa Rica.

The effect of pre- and postharvest calcium applications on 'Hayward' kiwifruit storage ability

The benefits of calcium applications pre and postharvest on fruit storage ability have been mentioned in the bibliography. It was objective of this work to study the effect of calcium preharvest application in two different forms and calcium chloride application postharvest on 'Hayward' kiwifruit storage ability. Kiwifruit vines were sprayed with 0.03% CaCl2 or 0.03% CaO at one, three and four months before harvest. The control did not have any treatment. After harvest, half fruits were dipped for 2 min in a solution of 1% CaCl2, left to dry and stored at 0 degrees C. The other half was stored at the same temperature without any treatment. The commercial yield was not affected by treatments. During storage, fruits dipped in 1% CaCl2 softened slower and than fruits not treated. Weight loss was higher in fruits treated with CaO preharvest. SSC showed a significant decrease in fruits sprayed with CaO from 4 to 6 months storage. This work suggests that immersion of kiwifruit in 1% CaCl2 postharvest benefits storage life capacity; preharvest spraying with CaCl2 seems to be better than with CaO. However, we have to try higher calcium concentrations in order to get better results in storage ability but, without causing toxicity on the vines.

The effect of pre- and postharvest calcium applications on 'Hayward' kiwifruit storage ability

The benefits of calcium applications pre and postharvest on fruit storage ability have been mentioned in the bibliography. It was objective of this work to study the effect of calcium preharvest application in two different forms and calcium chloride application postharvest on 'Hayward' kiwifruit storage ability. Kiwifruit vines were sprayed with 0.03% CaCl2 or 0.03% CaO at one, three and four months before harvest. The control did not have any treatment. After harvest, half fruits were dipped for 2 min in a solution of 1% CaCl2, left to dry and stored at 0 degrees C. The other half was stored at the same temperature without any treatment. The commercial yield was not affected by treatments. During storage, fruits dipped in 1% CaCl2 softened slower and than fruits not treated. Weight loss was higher in fruits treated with CaO preharvest. SSC showed a significant decrease in fruits sprayed with CaO from 4 to 6 months storage. This work suggests that immersion of kiwifruit in 1% CaCl2 postharvest benefits storage life capacity; preharvest spraying with CaCl2 seems to be better than with CaO. However, we have to try higher calcium concentrations in order to get better results in storage ability but, without causing toxicity on the vines.

An empirical study of the recreationalexpressive and referential roles of the cell phone among young students and their potential applications to advertising

The cell phone has become a means of finding persuasive content, making friends and accessing information. For this reason it is of great interest in analyzing is use by young people during their formative years. Regarding this point, the objectives of the research are: to obtain percentage data on the distraction call phones cause among students; to define their habits re fun activities and their study responsibility in their personal and academic lives; and to identify leisure-expressive and referential trends. It was decided to analyze the Spanish discourse found on Twitter. To do so, a quantitative and qualitative methodology was used, with a linguistic pattern extraction tool which allows us to obtain categories of meaning. The sample was 10,000 tweets in Spanish, with key words such as “cell” and “study” and all possible derivatives. The data were gathered between 30 May and the 6 June 2014, which coincides with the start of the exam period in Spain. Finally, the potential applications of the research to the specific field of publicity and advertising is discussed as a possible solution to the current needs of the brands, which have to find participative formats taking into account the students’ leisure trends.

Essays on employment / education, investment in education and student achievement

Méthodologie: Modèle de régression quantile de variable instrumentale pour données de Panel utilisant la fonction de production partielle

Attention to Diversity from an Interdisciplinary Perspective: Results from the Final Graduation Project from a Group of Students Majoring in Preschool Education and Special Education

Nowadays, the topic of diversity is being studied, particularly in the field of the formation of future educators, where it is clearly evident in each one of the students. In order to understand this concept and meet the challenges it demands, this investigation, through the experience of action research, looks for a real picture of how this diversity is served in Guanacaste’s rural contexts. This is accomplished by identifying those ways to guide a better teachers’ work, and by taking into account the educational planning and the participation of the different sectors involved in the process of teaching and learning.

Rotational and ro-vibrational spectroscopy of small nitrogen-containing species and their astrophysical applications

This thesis presents the study of small nitrogen-bearing molecules, from diatomic radicals to complex organic molecules, by means of rotational and ro-vibrational spectroscopy. Besides their theoretical relevance, which spans from anharmonic force field analyses to energetic and structural properties, I have chosen this family of species because of their astrochemical importance. After some basic knowledge of molecular spectroscopy and astrochemistry is introduced, the instrumentation used during the course of my PhD school is described. Then, the most relevant studies I conducted during the last three years are presented. Generally speaking, a number of molecules of astrophysical relevance have been characterized by means of rotational and ro-vibrational spectroscopy. The sample of studied species is constituted by small radicals (imidogen, amidogen, and titanium nitride), cyanopolyynes (cyanoacetylene) and pre-biotic molecules (aminoacetonitrile): these studies are presented in great detail. Among the results, the first astronomical detection of two deuterated radicals (NHD and ND2) is presented in this thesis.Thanks to our studies, it was possible to clearly identify molecular absorptions of these species towards the pre-stellar core IRAS16293-2422, as recorded by the Herschel Space Observatory mission. These observations confirm the strong deuterium enhancement generally observed in this cloud but they reveal that models underestimate the abundances of NHD and ND2. I also report the detection of vibrationally excited aminoacetonitrile (NH2CH2CN) in Sagittarius B2, as observed in the ReMoCa survey. This is the second detection of aminoacetonitrile in the interstellar medium and the first astronomical observation of its vibrationally hot lines. This represents a small step toward the comprehension on how complex organic molecules are formed and which processes can lead to the formation of glycine. Finally, few general remarks are discussed and the importance of future laboratory studies is pointed out, along with possible perspectives.

Nanofibrous-structured polymers and their engineering applications

Monolithic materials cannot always satisfy the demands of today’s advanced requirements. Only by combining several materials at different length-scales, as nature does, the requested performances can be met. Polymer nanocomposites are intended to overcome the common drawbacks of pristine polymers, with a multidisciplinary collaboration of material science with chemistry, engineering, and nanotechnology. These materials are an active combination of polymers and nanomaterials, where at least one phase lies in the nanometer range. By mimicking nature’s materials is possible to develop new nanocomposites for structural applications demanding combinations of strength and toughness. In this perspective, nanofibers obtained by electrospinning have been increasingly adopted in the last decade to improve the fracture toughness of Fiber Reinforced Plastic (FRP) laminates. Although nanofibers have already found applications in various fields, their widespread introduction in the industrial context is still a long way to go. This thesis aims to develop methodologies and models able to predict the behaviour of nanofibrous-reinforced polymers, paving the way for their practical engineering applications. It consists of two main parts. The first one investigates the mechanisms that act at the nanoscale, systematically evaluating the mechanical properties of both the nanofibrous reinforcement phase (Chapter 1) and hosting polymeric matrix (Chapter 2). The second part deals with the implementation of different types of nanofibers for novel pioneering applications, trying to combine the well-known fracture toughness enhancement in composite laminates with improving other mechanical properties or including novel functionalities. Chapter 3 reports the development of novel adhesive carriers made of nylon 6,6 nanofibrous mats to increase the fracture toughness of epoxy-bonded joints. In Chapter 4, recently developed rubbery nanofibers are used to enhance the damping properties of unidirectional carbon fiber laminates. Lastly, in Chapter 5, a novel self-sensing composite laminate capable of detecting impacts on its surface using PVDF-TrFE piezoelectric nanofibers is presented.

Practical assessment on the run – iPads as an effective mobile and paperless tool in physical education and teaching

This paper investigates the use of iPads in the assessment of predominantly second year Bachelor of Education (Primary/Early Childhood) pre-service teachers undertaking a physical education and health unit. Within this unit, practical assessment tasks are graded by tutors in a variety of indoor and outdoor settings. The main barriers for the lecturer or tutor for effective assessment in these contexts include limited time to assess and the provision of explicit feedback for large numbers of students, complex assessment procedures, overwhelming record-keeping and assessing students without distracting from the performance being presented. The purpose of this pilot study was to investigate whether incorporating mobile technologies such as iPads to access online rubrics within the Blackboard environment would enhance and simplify the assessment process. Results from the findings indicate that using iPads to access online rubrics was successful in streamlining the assessment process because it provided pre-service teachers with immediate and explicit feedback. In addition, tutors experienced a reduction in the amount of time required for the same workload by allowing quicker forms of feedback via the iPad dictation function. These outcomes have future implications and potential for mobile paperless assessment in other disciplines such as health, environmental science and engineering.

Collective actions for environmental monitoring and conservation: citizen science, informal environmental education, and participatory approach

This PhD was driven by an interest for inclusive and participatory approaches. The methodology that bridges science and society is known as 'citizen science' and is experiencing a huge upsurge worldwide, in the scientific and humanities fields. In this thesis, I have focused on three topics: i) assessing the reliability of data collected by volunteers; ii) evaluating the impact of environmental education activities in tourist facilities; and iii) monitoring marine biodiversity through citizen science. In addition to these topics, during my research stay abroad, I developed a questionnaire to investigate people's perceptions of natural areas to promote the implementation of co-management. The results showed that volunteers are not only able to collect sufficiently reliable data, but that during their participation in this type of project, they can also increase their knowledge of marine biology and ecology and their awareness of the impact of human behaviour on the environment. The short-term analysis has shown that volunteers are able to retain what they have learned. In the long term, knowledge is usually forgotten, but awareness is retained. Increased awareness could lead to a change in behaviour and in this case a more environmentally friendly attitude. This aspect could be of interest for the development of environmental education projects in tourism facilities to reduce the impact of tourism on the environment while adding a valuable service to the tourism offer. We also found that nature experiences in childhood are important to connect to nature in adulthood. The results also suggest that membership or volunteering in an environmental education association could be a predictor of people's interest in more participatory approaches to nature management. In most cases, the COVID -19 pandemic had not changed participants' perceptions of the natural environment.

Enhanced financial literacy through financial education and its impact on financial behaviour

This dissertation aims to contribute to the ongoing discourse on the effect an enhanced financial literacy, through financial education, has on financial behaviour. We posit that financial literacy is enhanced through financial education courses, but it also significantly impacts the financial behaviour of individuals. Moreover, we argue that improved financial literacy plays a significant role in mitigating behavioural biases and an asset price bubble. Chapter 1 analyzes the impact of a financial education course in enhancing financial literacy in a high- school context. Students at specific schools in Tirana, Albania, are delivered a financial education course, which lasts one academic year. To understand the impact of this financial education course in enhancing financial literacy, PISA (2012) questionnaire on financial literacy is delivered to the students before and after the course is delivered. Chapter 2 analysis the impact of financial literacy in mitigating behavioural biases. We focus on the impact that enhanced financial literacy through the financial education course and financial education plays in reducing the propensity to mental accounting bias. Chapter 3 investigates how financial literacy affects the propensity to an asset price bubble occurrence. We posit that enhanced financial literacy through financial education reduces the probability of an asset price bubble occurrence. We find that financial literacy enhanced through financial education has a significant impact in the financial behaviour of the individuals.

Electrochemiluminescence: insights into the mechanisms and nanomaterial-based applications

Biomarkers are biological indicators of human health conditions. Their ultra-sensitive quantification is of paramount importance in clinical monitoring and early disease diagnosis. Biosensors are simple and easy-to-use analytical devices and, in their world, electrochemiluminescence (ECL) is one of the most promising analytical techniques that needs an ever-increasing sensitivity for improving its clinical effectiveness. Scope of this project was the investigation of the ECL generation mechanisms for enhancing the ECL intensity also through the identification of suitable nanostructures. The combination of nanotechnologies, microscopy and ECL has proved to be a very successful strategy to improve the analytical efficiency of ECL in one of its most promising bioanalytical approaches, the bead-based immunoassay. Nanosystems, such as [Ru(bpy)3]2+-dye-doped nanoparticles (DDSNPs) and Bodipy Carbon Nanodots, have been used to improve the sensitivity of ECL techniques thanks to their advantageous and tuneable properties, reaching a signal increase of 750% in DDSNPs-bead-based immunoassay system. In this thesis, an investigation of size and distance effects on the ECL mechanisms was carried out through the innovative combination of ECL microscopy and electrochemical mapping of radicals. It allowed the discovery of an unexpected and highly efficient mechanistic path for ECL generation at small distances from the electrode surface. It was exploited and enhanced through the addition of a branched amine DPIBA to the usual coreactant TPrA solution for enhancing the ECL efficiency until a maximum of 128%. Finally, a beads-based immunoassay and an immunosensor specific for cardiac Troponin I were built exploiting previous results and carbon nanotubes features. They created a conductive layer around beads enhancing the signal by 70% and activating an ECL mechanism unobserved before in such systems. In conclusion, the combination of ECL microscopy and nanotechnology and the deep understanding of the mechanisms responsible for the ECL emission led to a great enhancement in the signal.

Effective field theories and their phenomenological applications

Effective field theories (EFTs) are ubiquitous in theoretical physics and in particular in field theory descriptions of quantum systems probed at energies much lower than one or few characterizing scales. More recently, EFTs have gained a prominent role in the study of fundamental interactions and in particular in the parametriasation of new physics beyond the Standard Model, which would occur at scales Λ, much larger than the electroweak scale. In this thesis, EFTs are employed to study three different physics cases. First, we consider light-by-light scattering as a possible probe of new physics. At low energies it can be described by dimension-8 operators, leading to the well-known Euler-Heisenberg Lagrangian. We consider the explicit dependence of matching coefficients on type of particle running in the loop, confirming the sensitiveness to the spin, mass, and interactions of possibly new particles. Second, we consider EFTs to describe Dark Matter (DM) interactions with SM particles. We consider a phenomenologically motivated case, i.e., a new fermion state that couples to the Hypercharge through a form factor and has no interactions with photons and the Z boson. Results from direct, indirect and collider searches for DM are used to constrain the parameter space of the model. Third, we consider EFTs that describe axion-like particles (ALPs), whose phenomenology is inspired by the Peccei-Quinn solution to strong CP problem. ALPs generically couple to ordinary matter through dimension-5 operators. In our case study, we investigate the rather unique phenomenological implications of ALPs with enhanced couplings to the top quark.

3D Printing and bioprinting of osteomimetic materials for 3D in vitro models and tissue repair applications

Bone disorders have severe impact on body functions and quality life, and no satisfying therapies exist yet. The current models for bone disease study are scarcely predictive and the options existing for therapy fail for complex systems. To mimic and/or restore bone, 3D printing/bioprinting allows the creation of 3D structures with different materials compositions, properties, and designs. In this study, 3D printing/bioprinting has been explored for (i) 3D in vitro tumor models and (ii) regenerative medicine. Tumor models have been developed by investigating different bioinks (i.e., alginate, modified gelatin) enriched by hydroxyapatite nanoparticles to increase printing fidelity and increase biomimicry level, thus mimicking the organic and inorganic phase of bone. High Saos-2 cell viability was obtained, and the promotion of spheroids clusters as occurring in vivo was observed. To develop new syntethic bone grafts, two approaches have been explored. In the first, novel magnesium-phosphate scaffolds have been investigated by extrusion-based 3D printing for spinal fusion. 3D printing process and parameters have been optimized to obtain custom-shaped structures, with competent mechanical properties. The 3D printed structures have been combined to alginate porous structures created by a novel ice-templating technique, to be loaded by antibiotic drug to address infection prevention. Promising results in terms of planktonic growth inhibition was obtained. In the second strategy, marine waste precursors have been considered for the conversion in biogenic HA by using a mild-wet conversion method with different parameters. The HA/carbonate ratio conversion efficacy was analysed for each precursor (by FTIR and SEM), and the best conditions were combined to alginate to develop a composite structure. The composite paste was successfully employed in custom-modified 3D printer for the obtainment of 3D printed stable scaffolds. In conclusion, the osteomimetic materials developed in this study for bone models and synthetic grafts are promising in bone field.