Structural characterization and comparative analysis of human and piscine cartilage acidic protein (CRTAC1/CRTAC2)

Dissertação de mestrado, Biotecnologia, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2014

What Determines the Innovativeness of Polish Family Firms? Empirical Results and Theoretical Puzzles

This article will review and synthesize the existing research on the innovativeness of Polish family firms in order to separate universal factors that influence the degree of innovativeness of firms from the factors which distinctively influence the innovativeness of family firms. To better assess the innovation propensity of family firms the author will work out the typology by combining the variety of innovations with particular features of family firms and the industrial context. A more nuanced approach will help to understand why the academic literature is inconclusive with regards to the question of whether family firms are anti-innovative (as some authors claim), pro-innovative or ambivalent with regard to innovations. In particular it will be argued that when assessing family firms’ innovativeness special attention needs to be paid to the impact of the management of intergenerational change on the propensity to innovate, as this process relates to the capacity for investments into innovativeness and the time horizon of the owner’s decisions.

GENETIC VARIATION, LOCAL ADAPTATION AND POPULATION STRUCTURE IN NORTH AMERICAN RED OAK SPECIES, QUERCUS RUBRA L. AND Q. ELLIPSOIDALIS E. J. HILL

Forest trees, like oaks, rely on high levels of genetic variation to adapt to varying environmental conditions. Thus, genetic variation and its distribution are important for the long-term survival and adaptability of oak populations. Climate change is projected to lead to increased drought and fire events as well as a northward migration of tree species, including oaks. Additionally, decline in oak regeneration has become increasingly concerning since it may lead to decreased gene flow and increased inbreeding levels. This will in turn lead to lowered levels of genetic diversity, negatively affecting the growth and survival of populations. At the same time, populations at the species’ distribution edge, like those in this study, could possess important stores of genetic diversity and adaptive potential, while also being vulnerable to climatic or anthropogenic changes. A survey of the level and distribution of genetic variation and identification of potentially adaptive genes is needed since adaptive genetic variation is essential for their long-term survival. Oaks possess a remarkable characteristic in that they maintain their species identity and specific environmental adaptations despite their propensity to hybridize. Thus, in the face of interspecific gene flow, some areas of the genome remain differentiated due to selection. This characteristic allows the study of local environmental adaptation through genetic variation analyses. Furthermore, using genic markers with known putative functions makes it possible to link those differentiated markers to potential adaptive traits (e.g., flowering time, drought stress tolerance). Demographic processes like gene flow and genetic drift also play an important role in how genes (including adaptive genes) are maintained or spread. These processes are influenced by disturbances, both natural and anthropogenic. An examination of how genetic variation is geographically distributed can display how these genetic processes and geographical disturbances influence genetic variation patterns. For example, the spatial clustering of closely related trees could promote inbreeding with associated negative effects (inbreeding depression), if gene flow is limited. In turn this can have negative consequences for a species’ ability to adapt to changing environmental conditions. In contrast, interspecific hybridization may also allow the transfer of genes between species that increase their adaptive potential in a changing environment. I have studied the ecologically divergent, interfertile red oaks, Quercus rubra and Q. ellipsoidalis, to identify genes with potential roles in adaptation to abiotic stress through traits such as drought tolerance and flowering time, and to assess the level and distribution of genetic variation. I found evidence for moderate gene flow between the two species and low interspecific genetic differences at most genetic markers (Lind and Gailing 2013). However, the screening of genic markers with potential roles in phenology and drought tolerance led to the identification of a CONSTANS-like (COL) gene, a candidate gene for flowering time and growth. This marker, located in the coding region of the gene, was highly differentiated between the two species in multiple geographical areas, despite interspecific gene flow, and may play a role in reproductive isolation and adaptive divergence between the two species (Lind-Riehl et al. 2014). Since climate change could result in a northward migration of trees species like oaks, this gene could be important in maintaining species identity despite increased contact zones between species (e.g., increased gene flow). Finally I examined differences in spatial genetic structure (SGS) and genetic variation between species and populations subjected to different management strategies and natural disturbances. Diverse management activities combined with various natural disturbances as well as species specific life history traits influenced SGS patterns and inbreeding levels (Lind-Riehl and Gailing submitted).

Network Construction and Graph Theoretical Analysis of Functional Language Networks in Pediatric Epilepsy

This dissertation introduces a new approach for assessing the effects of pediatric epilepsy on the language connectome. Two novel data-driven network construction approaches are presented. These methods rely on connecting different brain regions using either extent or intensity of language related activations as identified by independent component analysis of fMRI data. An auditory description decision task (ADDT) paradigm was used to activate the language network for 29 patients and 30 controls recruited from three major pediatric hospitals. Empirical evaluations illustrated that pediatric epilepsy can cause, or is associated with, a network efficiency reduction. Patients showed a propensity to inefficiently employ the whole brain network to perform the ADDT language task; on the contrary, controls seemed to efficiently use smaller segregated network components to achieve the same task. To explain the causes of the decreased efficiency, graph theoretical analysis was carried out. The analysis revealed no substantial global network feature differences between the patient and control groups. It also showed that for both subject groups the language network exhibited small-world characteristics; however, the patient’s extent of activation network showed a tendency towards more random networks. It was also shown that the intensity of activation network displayed ipsilateral hub reorganization on the local level. The left hemispheric hubs displayed greater centrality values for patients, whereas the right hemispheric hubs displayed greater centrality values for controls. This hub hemispheric disparity was not correlated with a right atypical language laterality found in six patients. Finally it was shown that a multi-level unsupervised clustering scheme based on self-organizing maps, a type of artificial neural network, and k-means was able to fairly and blindly separate the subjects into their respective patient or control groups. The clustering was initiated using the local nodal centrality measurements only. Compared to the extent of activation network, the intensity of activation network clustering demonstrated better precision. This outcome supports the assertion that the local centrality differences presented by the intensity of activation network can be associated with focal epilepsy.

Expression and purification of tau protein and its frontotemporal dementia variants using a cleavable histidine tag

Recombinant tau protein is widely used to study the biochemical, cellular and pathological aspects of tauopathies, including Alzheimer's disease and frontotemporal dementia with Parkinsonism linked to chromosome 17 (FTPD-17). Pure tau in high yield is a requirement for in vitro evaluation of the protein's physiological and toxic functions. However, the preparation of recombinant tau is complicated by the protein's propensity to aggregate and form truncation products, necessitating the use of multiple, time-consuming purification methods. In this study, we investigated parameters that influence the expression of wild type and FTPD-17 pathogenic tau, in an attempt to identify ways to maximise expression yield. Here, we report on the influence of the choice of host strain, induction temperature, duration of induction, and media supplementation with glucose on tau expression in Escherichia coli. We also describe a straightforward process to purify the expressed tau proteins using immobilised metal affinity chromatography, with favourable yields over previous reports. An advantage of the described method is that it enables high yield production of functional oligomeric and monomeric tau, both of which can be used to study the biochemical, physiological and toxic properties of the protein.

Regional diversity and economic performance

This thesis consists of an introductory chapter and four individual papers. In each paper the relationship between some form of spatial diversity and economic performance is analyzed. Diversity is treated as a potential source of externality effects, mainly in the form of knowledge spillovers. The first paper studies the impact of a broad range of spatial externalities on the productivity of manufacturing plants. While finding positive effects of specialization and competition, there is no support for positive spillovers of either related or unrelated industry diversity. The second paper argues that relatedness should be framed at the level of individuals and consequently should be measured in terms of, for example, education and occupation rather than industry belonging. The results show that educational- and occupational related diversity matter for regional productivity growth, while related industry diversity is positively related to employment growth. The third paper analyzes the importance of neighborhood related diversity, in terms of both industries and education, and internal human capital for firms’ propensity to innovate. The findings support that education and skills are strongly related to firm innovation. Additionally, firms in metropolitan regions are more innovative in neighborhoods with more related diversity in industries, while firms in rural regions seem to benefit more from related diversity in education. In the fourth paper, the location factor of interest is segregation, which may be regarded as inverse diversity. The results show that neighborhood segregation has a negative effect on individual employment. However, it is not the spatial separation of individuals with different backgrounds that causes lower employment but rather the distress of segregated neighborhoods.

Essays in Applied Insurance Economics

The papers included in this thesis deal with a few aspects of insurance economics that have seldom been dealt with in the applied literature. In the first paper I apply for the first time the tools of the economics of crime to study the determinants of frauds, using data on Italian provinces. The contributions to the literature are manifold: -The price of insuring has a positive correlation with the propensity to defraud -Social norms constraint fraudulent behavior, but their strength is curtailed in economic downturns -I apply a simple extension of the Random Coefficient model, which allows for the presence of time invariant covariates and asymmetries in the impact of the regressors. The second paper assesses how the evolution of macro prudential regulation of insurance companies has been reflected in their equity price. I employ a standard event study methodology, deriving the definition of the “control” and “treatment” groups from what is implied by the regulatory framework. The main results are: -Markets care about the evolution of the legislation. Their perception has shifted from a first positive assessment of a possible implicit “too big to fail” subsidy to a more negative one related to its cost in terms of stricter capital requirement -The size of this phenomenon is positively related to leverage, size and on the geographical location of the insurance companies The third paper introduces a novel methodology to forecast non-life insurance premiums and profitability as function of macroeconomic variables, using the simultaneous equation framework traditionally employed macroeconometric models and a simple theoretical model of insurance pricing to derive a long term relationship between premiums, claims expenses and short term rates. The model is shown to provide a better forecast of premiums and profitability compared with the single equation specifications commonly used in applied analysis.

Disturbi dell'informazione, nuovi media e libertà di manifestazione del pensiero

L’elaborato affronta il tema della massiccia diffusione di disinformazione tramite internet sotto il profilo del diritto costituzionale, nell’ottica di indagare se le dimensioni del fenomeno richiedano l’elaborazione di principi e soluzioni nuove ovvero se i paradigmi costituzionali enucleati dalla tradizione costituzionalistica con riferimento alla libertà d’espressione forniscano gli strumenti sufficienti per un intervento efficace. In tale ottica, il lavoro premette una ricostruzione del fenomeno della disinformazione nel tentativo di individuare il perimetro socio-cognitivo e tecnologico entro cui la medesima prolifera, descrivendo gli elementi che distinguono l’informazione diffusa dai nuovi media rispetto a quella veicolata dai media mainstream. Ciò premesso, lo studio passa a delineare lo “statuto costituzionale” del falso, indagando la rilevanza che la menzogna assume sotto il profilo costituzionale nel suo rapporto con la libertà di manifestazione del pensiero negli ordinamenti interno, europeo e statunitense. L’analisi prosegue poi con l’esame delle politiche di contrasto alla disinformazione introdotte da singoli Stati (prevalentemente Germania, Francia e Italia), Unione europea, e piattaforme, con l’obiettivo di mettere in luce vantaggi e limiti dei modelli di eteroregolamentazione, coregolamentazione e autoregolamentazione. Da ultimo, l’elaborato scompone alcune delle azioni e misure passate in rassegna e le analizza con la lente d’ingrandimento della libertà di manifestazione del pensiero, ordinandone i contenuti secondo la specifica propensione a comprimere la libertà d’espressione. L’indagine si conclude con alcuni brevi spunti conclusivi che evidenziano l’esigenza, in base ai principi costituzionali analizzati, che eventuali interventi normativi siano se del caso volti alla regolazione delle piattaforme, ovvero dei “contenitori”, lasciano i contenuti al libero scambio delle idee.

Structural and biophysical characterization of RAD52 as novel pharmacological target for synthetic lethality

RAD52 is a protein involved in various DNA reparation mechanisms. In the last few years, RAD52 has been proposed as a novel pharmacological target for cancer synthetic lethality strategies. Hence, this work has the purpose to investigate RAD52 protein, with biophysical and structural tools to shed light on proteins features and mechanistic details that are, up to now poorly described, and to design novel strategies for its inhibition. My PhD work had two goals: the structural and functional characterization of RAD52 and the identification of novel RAD52 inhibitors. For the first part, RAD52 was characterized both for its DNA interaction and oligomerization state together with its propensity to form high molecular weight superstructures. Moreover, using EM and Cryo-EM techniques, additional RAD52 structural hallmarks were obtained, valuable both for understanding protein mechanism of action and for drug discovery purpose. The second part of my PhD project focused on the design and characterization of novel RAD52 inhibitors to be potentially used in combination therapies with PARPi to achieve cancer cells synthetic lethality, avoiding resistance occurrence and side effects. With this aim we selected and characterized promising RAD52 inhibitors through three different approaches: 19F NMR fragment-based screening; virtual screening campaign; aptamers computational design. Selected hits (fragments, molecules and aptamers) were investigated for their binding to RAD52 and for their mechanism of inhibition. Collected data highlighted the identification of hits worthy to be developed into more potent and selective RAD52 inhibitors. Finally, a side project carried out during my PhD is reported. GSK-3β protein, an already validated pharmacological target was investigated using biophysical and structural biology tools. Here, an innovative and adaptable drug discovery screening pipeline able to directly identify selective compounds with binding affinities not higher than a reference binder was developed.

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.

Nietzsche's will-to-power ontology : an interpretation of Beyond Good and Evil

Abstract: Nietzsche's Will-to-Power Ontology: An Interpretation of Beyond Good and Evil § 36 By: Mark Minuk Will-to-power is the central component of Nietzsche's philosophy, and passage 36 of Beyond Good and Evil is essential to coming to an understanding of it. 1 argue for and defend the thesis that will-to-power constitutes Nietzsche's ontology, and offer a new understanding of what that means. Nietzsche's ontology can be talked about as though it were a traditional substance ontology (i.e., a world made up of forces; a duality of conflicting forces described as 'towards which' and 'away from which'). However, 1 argue that what defines this ontology is an understanding of valuation as ontologically fundamental—^the basis of interpretation, and from which a substance ontology emerges. In the second chapter, I explain Nietzsche's ontology, as reflected in this passage, through a discussion of Heidegger's two ontological categories in Being and Time (readiness-to-hand, and present-at-hand). In a nutshell, it means that the world of our desires and passions (the most basic of which is for power) is ontologically more fundamental than the material world, or any other interpretation, which is to say, the material world emerges out of a world of our desires and passions. In the first chapter, I address the problematic form of the passage reflected in the first sentence. The passage is in a hypothetical style makes no claim to positive knowledge or truth, and, superficially, looks like Schopenhaurian position for the metaphysics of the will, which Nietzsche rejects. 1 argue that the hypothetical form of the passage is a matter of style, namely, the style of a free-spirit for whom the question of truth is reframed as a question of values. In the third and final chapter, 1 address the charge that Nietzsche's interpretation is a conscious anthropomorphic projection. 1 suggest that the charge rests on a distinction (between nature and man) that Nietzsche rejects. I also address the problem of the causality of the will for Nietzsche, by suggesting that an alternative, perspectival form of causality is possible.

Application of hydrogen-bond propensity calculations to an indomethacin–nicotinamide (1:1) co-crystal

The crystal structure of an indomethacin–nicotinamide (1 : 1) cocrystal produced by milling has been determined from laboratory powder X-ray diffraction (PXRD) data. The hydrogen bonding motifs observed in the structure represent one of the most probable of all the possible combinations of donors and acceptors in the constituent molecules.

Development of novel methods to determine crystalline glucose content of honey based on DSC, HPLC, and viscosity measurements and their use to examine the setting propensity of honey

Crystallization must occur in honey in order to produce set or creamed honey; however, the process must occur in a controlled manner in order to obtain an acceptable product. As a consequence, reliable methods are needed to measure the crystal content of honey (φ expressed as kg crystal per kg honey), which can also be implemented with relative ease in industrial production facilities. Unfortunately, suitable methods do not currently exist. This article reports on the development of 2 independent offline methods to measure the crystal content in honey based on differential scanning calorimetry and high-performance liquid chromatography. The 2 methods gave highly consistent results on the basis of paired t-test involving 143 experimental points (P > 0.05, r**2 = 0.99). The crystal content also correlated with the relative viscosity, defined as the ratio of the viscosity of crystal containing honey to that of the same honey when all crystals are dissolved, giving the following correlation: μr = 1 + 1398.8∅**2.318. This correlation can be used to estimate the crystal content of honey in industrial production facilities. The crystal growth rate at a temperature of 14 ◦C—the normal crystallization temperature used in practice—was linear, and the growth rate also increased with the total glucose content in the honey.

Automated peritoneal dialysis is associated with better survival rates compared to continuous ambulatory peritoneal dialysis: a propensity score matching analysis

The impact of peritoneal dialysis modality on patient survival and peritonitis rates is not fully understood, and no large-scale randomized clinical trial (RCT) is available. In the absence of a RCT, the use of an advanced matching procedure to reduce selection bias in large cohort studies may be the best approach. The aim of this study is to compare automated peritoneal dialysis (APD) and continuous ambulatory peritoneal dialysis (CAPD) according to peritonitis risk, technique failure and patient survival in a large nation-wide PD cohort. This is a prospective cohort study that included all incident PD patients with at least 90 days of PD recruited in the BRAZPD study. All patients who were treated exclusively with either APD or CAPD were matched for 15 different covariates using a propensity score calculated with the nearest neighbor method. Clinical outcomes analyzed were overall mortality, technique failure and time to first peritonitis. For all analysis we also adjusted the curves for the presence of competing risks with the Fine and Gray analysis. After the matching procedure, 2,890 patients were included in the analysis (1,445 in each group). Baseline characteristics were similar for all covariates including: age, diabetes, BMI, Center-experience, coronary artery disease, cancer, literacy, hypertension, race, previous HD, gender, pre-dialysis care, family income, peripheral artery disease and year of starting PD. Mortality rate was higher in CAPD patients (SHR1.44 CI95%1.21-1.71) compared to APD, but no difference was observed for technique failure (SHR0.83 CI95%0.69-1.02) nor for time till the first peritonitis episode (SHR0.96 CI95%0.93-1.11). In the first large PD cohort study with groups balanced for several covariates using propensity score matching, PD modality was not associated with differences in neither time to first peritonitis nor in technique failure. Nevertheless, patient survival was significantly better in APD patients.