Essays on Innovative Activity and the Protection of Innovations

This dissertation looks at three widely accepted assumptions about how the patent system works: patent documents disclose inventions; this disclosure happens quickly, and patent owners are able to enforce patents. The first chapter estimates the effect of stronger trade secret protection on the number of patented innovations. When firms find it easier to protect business information, there is less need for patent protection, and accordingly less need for the disclosure of technical information that is required by patent law. The novel finding is that when it is easier to keep innovations, there is not only a reduction in the number of patents but also a sizeable reduction in disclosed knowledge per patent. The chapter then shows how this endogeneity of the amount of knowledge per patent can affect the measurement of innovation using patent data. The second chapter develops a game-theoretic model to study how the introduction of fee-shifting in US patent litigation would influence firms’ patenting propensities. When the defeated party to a lawsuit has to bear not only their own cost but also the legal expenditure of the winning party, manufacturing firms in the model unambiguously reduce patenting, with small firms affected the most. For fee-shifting to have the same effect as in Europe, the US legal system would require shifting of a much smaller share of fees. Lessons from European patent litigation may, therefore, have only limited applicability in the US case. The third chapter contains a theoretical analysis of the influence of delayed disclosure of patent applications by the patent office. Such a delay is a feature of most patent systems around the world but has so far not attracted analytical scrutiny. This delay may give firms various kinds of strategic (non-)disclosure incentives when they are competing for more than a single innovation.

Design, production and testing of innovative key components of HEV/BEV powertrains

Through modelling activity, experimental campaigns, test bench and on-field validation, a complete powertrain for a BEV has been designed, assembled and used in a motorsport competition. The activity can be split in three main subjects, representing the three key components of an BEV vehicle. First of all a model of the entire powertrain has been developed in order to understand how the various design choices will influence the race lap-time. The data obtained was then used to design, build and test a first battery pack. After bench tests and track tests, it was understood that by using all the cell charac-teristics, without breaking the rules limitations, higher energy and power densities could have been achieved. An updated battery pack was then designed, produced and raced with at Motostudent 2018 re-sulting in a third place at debut. The second topic of this PhD was the design of novel inverter topologies. Three inverters have been de-signed, two of them using Gallium Nitride devices, a promising semiconductor technology that can achieve high switching speeds while maintaining low switching losses. High switching frequency is crucial to reduce the DC-Bus capacitor and then increase the power density of 3 phase inverters. The third in-verter uses classic Silicon devices but employs a ZVS (Zero Voltage Switching) topology. Despite the in-creased complexity of both the hardware and the control software, it can offer reduced switching losses by using conventional and established silicon mosfet technology. Finally, the mechanical parts of a three phase permanent magnet motor have been designed with the aim to employ it in UniBo Motorsport’s 2020 Formula Student car.

Development and Validation of a Model-Based Combustion Controller for Water Injection Management

This manuscript reports the overall development of a Ph.D. research project during the “Mechanics and advanced engineering sciences” course at the Department of Industrial Engineering of the University of Bologna. The project is focused on the development of a combustion control system for an innovative Spark Ignited engine layout. In details, the controller is oriented to manage a prototypal engine equipped with a Port Water Injection system. The water injection technology allows an increment of combustion efficiency due to the knock mitigation effect that permits to keep the combustion phasing closer to the optimal position with respect to the traditional layout. At the beginning of the project, the effects and the possible benefits achievable by water injection have been investigated by a focused experimental campaign. Then the data obtained by combustion analysis have been processed to design a control-oriented combustion model. The model identifies the correlation between Spark Advance, combustion phasing and injected water mass, and two different strategies are presented, both based on an analytic and semi-empirical approach and therefore compatible with a real-time application. The model has been implemented in a combustion controller that manages water injection to reach the best achievable combustion efficiency while keeping knock levels under a pre-established threshold. Three different versions of the algorithm are described in detail. This controller has been designed and pre-calibrated in a software-in-the-loop environment and later an experimental validation has been performed with a rapid control prototyping approach to highlight the performance of the system on real set-up. To further make the strategy implementable on an onboard application, an estimation algorithm of combustion phasing, necessary for the controller, has been developed during the last phase of the PhD Course, based on accelerometric signals.

Novel dual D3R/GSK-3β modulators: an innovative multitarget strategy for bipolar disorder

Among the psychiatric diseases, bipolar disorder (BD) is the sixth leading cause of disability with a prevalence up to 4 % worldwide. BD is a complex neuropsychiatric condition which alternates episodes of mania with symptoms of depression. Although the neurobiological pathways are not completely clarified, the dopamine (DA) hypothesis, recognized as the leading theory explaining the pathophysiology of the malady, states that the dramatically compromised homeostatic regulation of dopaminergic circuits leads to alternated changes in DA neurotransmission. Modulation of D2 and D3 receptors (D2/3R) through partial agonists represents the first-line therapeutic strategy for psychiatric diseases. Moreover, a deregulation of the enzyme glycogen synthase kinase-3β (GSK-3β) has been reported as peculiar feature of BD. In this scenario, the concomitant modulation of D3R and GSK-3β, by employing multitarget compounds, could offer promises to achieve an effective cure of this illness. In the light of these findings, we rationally envisaged the pharmacophoric model at the basis of the design of several D3R partial agonists, suitable to be exploited for the dual D3R/GSK-3β ligand design. Thus, synthetic efforts were addressed to develop a first set of hybrid molecules able to concurrently modulate the selected targets. For a chemical structure point of view, we employed different spacers to combine a substituted aryl-piperazine moiety, reported in previously discovered D3R modulators, with a pyrazole-based fragment, already identified in GSK-3β inhibitors. A fluorescent and a cellular functional assays were carried out to assess the activity of all synthetized compounds against GSK-3β and on D3R, respectively. Most of the derivatives proved to effectively modulate both GSK-3β and D3R with potencies in the low-µM and low-nM range, respectively. The consistent biological data allowed us to identify some lead candidates worth to be further modified with the aim to optimize their biological profile and to perform a structure-activity relationship (SAR) study.

Development and testing of innovative methodologies for modelling and control of normal and knocking combustion and implementation of novel rapid prototyping solutions

The thesis work deals with topics that led to the development of innovative control-oriented models and control algorithms for modern gasoline engines. Knock in boosted spark ignition engines is the widest topic discussed in this document because it remains one of the most limiting factors for maximizing combustion efficiency in this kind of engine. First chapter is thus focused on knock and a wide literature review is proposed to summarize the preliminary knowledge that even represents the background and the reference for discussed activities. Most relevant results achieved during PhD course in the field of knock modelling and control are then presented, describing every control-oriented model that led to the development of an adaptive model-based combustion control system. The complete controller has been developed in the context of the collaboration with Ferrari GT and it allowed to completely redefine the knock intensity evaluation as well as the combustion phase control. The second chapter is focused on the activity related to a prototyping Port Water Injection system that has been developed and tested on a turbocharged spark ignition engine, within the collaboration with Magneti Marelli. Such system and the effects of injected water on the combustion process were then modeled in a 1-D simulation environment (GT Power). Third chapter shows the development and validation of a control-oriented model for the real-time calculation of exhaust gas temperature that represents another important limitation to the performance increase in modern boosted engines. Indeed, modelling of exhaust gas temperature and thermocouple behavior are themes that play a key role in the optimization of combustion and catalyst efficiency.

A preclinical study of spinal cord injury focused on cellular and molecular modifications as potential targets for innovative therapies

Spinal Cord Injury (SCI) is a devastating condition for human and animal health. In SCI particularly, neurons, oligodendrocytes precursor cells, and mature oligodendrocytes are highly vulnerable to the toxic microenvironment after the lesion and susceptible to the elevated levels of noxious stimuli. Thus the regenerative response of the organism in case of SCI is significantly reduced, and only little spontaneous amelioration is observed in lesioned patients during the early phases. This work mainly focuses on studying and characterizing the modification induced by the SCI in a preclinical animal model. We investigated the ECM composition in the spinal cord segments surrounding the primary lesion site at a gene expression level. We found Timp1 and CD44 as a crucial hub in the secondary cascade of SCI in both spinal cord segments surrounding the lesion site. Interestingly, a temporal and anatomical difference in gene expression, indicating a complex regulation of ECM genes after SCI that could be used as a tool for regenerative medicine. We also investigated the modification in synaptic plasticity-related gene expression in spinal and supraspinal areas involved in motor control. We confirmed the anatomical and temporal difference in gene expression in spinal cord tissue. This analysis suggests that a molecular mapping of the lesion-induced modification could be a useful tool for regenerative medicine. In the last part, we evaluated the efficacy of an implantable biopolymer loaded with an anti-inflammatory drug and a pro-myelinating agent on the acute phase of SCI in our preclinical model. We found a consistent reduction of the inflammatory state in the spinal lesion site and the cord's surrounding segments. Moreover, we found increased preservation of the spinal cord tissue with a related upregulation of neuronal and oligodendroglial markers after lesion. Our treatment showed effective ameliorating functional outcome and reducing the lesion extension in the chronic phase.

Modeling Alzheimer disease with iPSCs and mouse model for the identification of risk factors, new targets, and potential therapeutical strategies

Alzheimer's disease (AD) is the most common neurodegenerative disease in elderly. Donepezil is the first-line drug used for AD. In section one, the experimental activity was oriented to evaluate and characterize molecular and cellular mechanisms that contribute to neurodegeneration induced by the Aβ1-42 oligomers (Aβ1-42O) and potential neuroprotective effects of the hybrids feruloyl-donepezil compound called PQM130. The effects of PQM130 were compared to donepezil in a murine AD model, obtained by intracerebroventricular (i.c.v.) injection of Aβ1-42O. The intraperitoneal administration of PQM130 (0.5-1 mg/kg) after i.c.v. Aβ1-42O injection improved learning and memory, protecting mice against spatial cognition decline. Moreover, it reduced oxidative stress, neuroinflammation and neuronal apoptosis, induced cell survival and protein synthesis in mice hippocampus. PQM130 modulated different pathways than donepezil, and it is more effective in counteracting Aβ1-42O damage. The section two of the experimental activity was focused on studying a loss of function variants of ABCA7. GWA studies identified mutations in the ABCA7 gene as a risk factor for AD. The mechanism through which ABCA7 contributes to AD is not clear. ABCA7 regulates lipid metabolism and critically controls phagocytic function. To investigate ABCA7 functions, CRISPR/Cas9 technology was used to engineer human iPSCs and to carry the genetic variant Y622*, which results in a premature stop codon, causing ABCA7 loss-of-function. From iPSCs, astrocytes were generated. This study revealed the effects of ABCA7 loss in astrocytes. ABCA7 Y622* mutation induced dysfunctional endocytic trafficking, impairing Aβ clearance, lipid dysregulation and cell homeostasis disruption, alterations that could contribute to AD. Though further studies are needed to confirm the PQM130 neuroprotective role and ABCA7 function in AD, the provided results showed a better understanding of AD pathophysiology, a new therapeutic approach to treat AD, and illustrated an innovative methodology for studying the disease.

Development of an innovative strategy to enhance the efficacy of gene therapy for CDKL5 deficiency disorder

CDKL5 (cyclin-dependent kinase-like 5) deficiency disorder (CDD) is a severe X-linked neurodevelopmental disease caused by mutations in the CDKL5 gene, characterized by early-onset epileptic seizures, intellectual disability, motor and visual impairment and respiratory dysregulation. Although pharmacological treatments are used to control seizures, there is currently no cure to ameliorate symptoms for CDD. Albeit delivery of a wild-type copy of the mutated gene to cells represents the most curative approach for a monogenic disease, proof-of-concept studies highlight significant efficacy caveats for brain gene therapy. The major one regards the low efficiency of gene delivery to the CNS by viral vectors. We used a secretable Igk-TATk-CDKL5 protein to enhance the efficiency of a gene therapy for CDD. In view of the properties of the Igk-chain leader sequence, the TATk-CDKL5 protein produced by infected cells is secreted via constitutive secretory pathways. Importantly, due to the transduction property of the TATk peptide, the secreted CDKL5 protein is internalized by cells. We compared the effects of a CDKL5 gene therapy with an IgK-TATk-CDKL5 gene therapy in a Cdkl5 KO mouse model to validate whether the Igk-TATk-CDKL5 approach significantly improve the therapeutic efficacy. We found that, although AAVPHP.B_Igk-TATk-CDKL5 and AAVPHP.B_CDKL5 vectors had similar brain infection efficiency, the AAVPHP.B_Igk-TATk-CDKL5 vector led to a higher CDKL5 protein replacement and Cdkl5 KO mice treated with the AAVPHP.B_Igk-TATk-CDKL5 vector showed a behavioral and neuroanatomical improvement in comparison with Cdkl5 KO mice treated with the AAVPHP.B_CDKL5 vector.

Investigation of innovative combustion systems for reduced fuel consumption and increased specific power production

This work resumes a wide variety of research activities carried out with the main objective of increasing the efficiency and reducing the fuel consumption of Gasoline Direct Injection engines, especially under high loads. For this purpose, two main innovative technologies have been studied, Water Injection and Low-Pressure Exhaust Gas Recirculation, which help to reduce the temperature of the gases inside the combustion chamber and thus mitigate knock, being this one of the main limiting factors for the efficiency of modern downsized engines that operate at high specific power. A prototypal Port Water Injection system was developed and extensive experimental work has been carried out, initially to identify the benefits and limitations of this technology. This led to the subsequent development and testing of a combustion controller, which has been implemented on a Rapid Control Prototyping environment, capable of managing water injection to achieve knock mitigation and a more efficient combustion phase. Regarding Low-Pressure Exhaust Gas Recirculation, a commercial engine that was already equipped with this technology was used to carry out experimental work in a similar fashion to that of water injection. Another prototypal water injection system has been mounted to this second engine, to be able to test both technologies, at first separately to compare them on equal conditions, and secondly together in the search of a possible synergy. Additionally, based on experimental data from several engines that have been tested during this study, including both GDI and GCI engines, a real-time model (or virtual sensor) for the estimation of the maximum in-cylinder pressure has been developed and validated. This parameter is of vital importance to determine the speed at which damage occurs on the engine components, and therefore to extract the maximum performance without inducing permanent damages.

Development and Detailed Characterization of Innovative, High-Performance Membrane Materials for CO2 Capture Processes

The scope of this dissertation is to study the transport phenomena of small molecules in polymers and membranes for gas separation applications, with particular attention to energy efficiency and environmental sustainability. This work seeks to contribute to the development of new competitive selective materials through the characterization of novel organic polymers such as CANALs and ROMPs, as well as through the combination of selective materials obtaining mixed matrix membranes (MMMs), to make membrane technologies competitive with the traditional ones. Kinetic and thermodynamic aspects of the transport properties were investigated in ideal and non-ideal scenarios, such as mixed-gas experiments. The information we gathered contributed to the development of the fundamental understanding related to phenomenon like CO2-induced plasticization and physical aging. Among the most significant results, ZIF-8/PPO MMMs provided materials whose permeability and selectivity were higher than those of the pure materials for He/CO2 separation. The CANALs featured norbornyl benzocyclobutene backbone and thereby introduced a third typology of ladder polymers in the gas separation field, expanding the structural diversity of microporous materials. CANALs have a completely hydrocarbon-based and non-polar rigid backbone, which makes them an ideal model system to investigate structure-property correlations. ROMPs were synthesized by means of the ring opening metathesis living polymerization, which allowed the formation of bottlebrush polymers. CF3-ROMP reveled to be ultrapermeable to CO2, with unprecedented plasticization resistance properties. Mixed-gas experiments in glassy polymer showed that solubility-selectivity controls the separation efficiency of materials in multicomponent conditions. Finally, it was determined that plasticization pressure in not an intrinsic property of a material and does not represent a state of the system, but rather comes from the contribution of solubility coefficient and diffusivity coefficient in the framework of the solution-diffusion model.

Waste prevention: policy instruments and business model innovation

Waste prevention (WP) is a strategy which helps societies and individuals to strive for sufficiency in resource consumption within planetary boundaries alongside sustainable and equitable well-being and to decouple the concepts of well-being and life satisfaction from materialism. Within this dissertation, some instruments to promote WP are analysed, by adopting two perspectives: firstly, the one of policymakers, at different governance levels, and secondly, the one of business in the electrical and electronic equipment (EEE) sector. At a national level, the role of WP programmes and market-based instruments (extended producer responsibility, pay-as-you-throw schemes, deposit-refund systems, environmental taxes) in boosting prevention of municipal solid waste is investigated. Then, focusing on the Emilia-Romagna Region (Italy), the performances of the waste management system are assessed over a long period, including some years before and after an institutional reform of the waste management governance regime. The impact of a centralisation (at a regional level) of both planning and economic regulation of the waste services on waste generation and WP is analysed. Finally, to support the regional decision-makers in the prioritisation of publicly funded projects for WP, a framework for the sustainability assessment, the evaluation of success, and the prioritisation of WP measures was applied to some projects implemented by Municipalities in the Region. Trying to close the research gap between engineering and business, WP strategies are discussed as drivers for business model (BM) innovation in EEE sector. Firstly, an innovative approach to a digital tracking solution for professional EEE management is analysed. New BMs which facilitate repair, reuse, remanufacturing, and recycling are created and discussed. Secondly, the impact of BMs based on servitisation and on producer ownership on the extension of equipment lifetime is analysed, by performing a review of real cases of organizations in the EEE sector applying result- and use-oriented BMs.

Discharge characteristics and discharge simulation model of long air gaps containing a floating conductor

Long air gaps containing a floating conductor are common insulation types in power grids. During the transmission line live-line work, the process of lineman entering the transmission line air gap constitutes a live-line work combined air gap, which is a typical long air gap containing a floating conductor. This thesis investigates the discharge characteristics, the discharge mechanism and a discharge simulation model of long air gaps containing a floating conductor in order to address the engineering issues in live-line work. The innovative achievements of the thesis are as follows: (1) The effect of the gap distance, the floating electrode structure, the switching impulse wavefront time, the altitude, and the deviation of the floating conductor from the axis on the breakdown voltage was determined. (2) The physical process of the discharges in long air gaps containing a floating conductor was determined. The reason why the discharge characteristics of long air gaps containing a floating electrode with complex geometrics and sharp protrusions and long air gaps with a rod-shaped floating electrode are similar has been studied. The formation mechanism of the lowest breakdown voltage area of a long air gap containing a floating conductor is explained. (3) A simulation discharge model of long air gaps containing a floating conductor was established, which can describe the physical process and predict the breakdown voltage. The model can realize the accurate prediction of the breakdown voltage of typical long air gaps containing a floating conductor and live-line work combined air gaps in transmission lines. The findings of the study can provide theoretical reference and technical support for improving the safety of live-line work.

Development of a porcine lactation model for the evaluation of mammary transfer of exogenous molecules - A contribution from the conception project

Around 5 million women give birth each year in Europe and, while breastfeeding, the majority of them may need to take medications, either occasionally or continuously. Unfortunately, there is often scarce evidence of trustworthy information about how a specific molecule might affect the physiology of lactation. This is the reason that brought a European public-private partnership to fund the development of a reliable platform to provide women and health-care professionals a helpful instrument to reduce uncertainty about the effects of medication used during breastfeeding. On April 1st 2019, the ConcePTION project (Grant Agreement n°821520) started to develop such envisaged platform. The 3rd Work Package was in charge of the validation of in vitro, in vivo and in silico lactation models. Between the numerous species currently used in preclinical studies, pigs’ similarities with humans’ anatomy, physiology and genomics make them extremely useful as translational models, when proper veterinary expertise is applied. The ASA team from the University of Bologna, went first to characterize the translational lactation model using the swine species, chosen upon literature review. The aim of this work was to lay the foundations of a porcine lactation model that could be suitable for application within pharmaceutical tests, to study drug transfer through milk prior approval and commercialization. The obtained results highlighted both strengths and critical points of the study design, allowing a significant improvement in the knowledge of pharmacokinetic physiology in lactating mammals. Lastly, this project allowed the assessment of microbial changes in gut resident bacteria of newborns through an innovative in vitro colonic model. Indeed, even if there were no evident adverse effects determined by drug residues in milk, possible alterations in the delicate microbial ecology of newborns’ gastrointestinal tract was considered pivotal, giving its possible impact on the individual health and growth.

Advanced studies on two animal models, murine and fish. RadKI21A626T mouse model: new insights into molecular mechanisms of enteric neuro-epithelial pathology. European sea bass: study of the effects of essential oils in different diets on the gastric system.

My PhD research period was focused on the anatomical, physiological and functional study of the gastrointestinal system on two different animal models. In two different contexts, the purpose of these two lines of research was contribute to understand how a specific genetic mutation or the adoption of a particular dietary supplement can affect gastrointestinal function. Functional gastrointestinal disorders are chronic conditions characterized by symptoms for which no organic cause can be found. Although symptoms are generally mild, a small subset of cases shows severe manifestations. This subset of patients may also have recurrent intestinal sub-occlusive episodes, but in absence of mechanical causes. This condition is referred to as chronic intestinal pseudo-obstruction, a rare, intractable chronic disease. Some mutations have been associated with CIPO. A novel causative RAD21 missense mutation was identified in a large consanguineous family, segregating a recessive form of CIPO. The present thesis was aimed to elucidate the mechanisms leading to neuropathy underlying CIPO via a recently developed conditional KI mouse carrying the RAD21 mutation. The experimental studies are based on the characterization and functional analysis of the conditional KI Rad21A626T mouse model. On the other hand aquaculture is increasing the global supply of foods. The species selected and feeds used affects the nutrients available from aquaculture, with a need to improve feed efficiency, both for economic and environmental reasons, but this will require novel innovative approaches. Nutritional strategies focused on the use of botanicals have attracted interest in animal production. Previous research indicates the positive results of using essential oils (EOs) as natural feed additives for several farmed animals. Therefore, the present study was designed to compare the effects of feed EO supplementation in two different forms (natural and composed of active ingredients obtained by synthesis) on the gastric mucosa in European sea bass.

TiMMing: developing an innovative suite of bioinformatic tools to harmonize and track the origin of copy number alterations in the evolutive history of multiple myeloma

Multiple Myeloma (MM) is a hematologic cancer with heterogeneous and complex genomic landscape, where Copy Number Alterations (CNAs) play a key role in the disease's pathogenesis and prognosis. It is of biological and clinical interest to study the temporal occurrence of early alterations, as they play a disease "driver" function by deregulating key tumor pathways. This study presents an innovative bioinformatic tools suite created for harmonizing and tracing the origin of CNAs throughout the evolutionary history of MM. To this aim, large cohorts of newly-diagnosed MM (NDMM, N=1582) and Smoldering-MM (SMM, N=282) were aggregated. The tools developed in this study enable the harmonization of CNAs as obtained from different genomic platforms in such a way that a high statistical power can be obtained. By doing so, the high numerosity of those cohorts was harnessed for the identification of novel genes characterized as "driver" (NFKB2, NOTCH2, MAX, EVI5 and MYC-ME2-enhancer), and the generation of an innovative timing model, implemented with a statistical method to introduce confidence intervals in the CNAs-calls. By applying this model on both NDMM and SMM cohorts, it was possible to identify specific CNAs (1q(CKS1B)amp, 13q(RB1)del, 11q(CCND1)amp and 14q(MAX)del) and categorize them as "early"/ "driver" events. A high level of precision was guaranteed by the narrow confidence intervals in the timing estimates. These CNAs were proposed as critical MM alterations, which play a foundational role in the evolutionary history of both SMM and NDMM. Finally, a multivariate survival model was able to identify the independent genomic alterations with the greatest effect on patients’ survival, including RB1-del, CKS1B-amp, MYC-amp, NOTCH2-amp and TRAF3-del/mut. In conclusion, the alterations that were identified as both "early-drivers” and correlated with patients’ survival were proposed as biomarkers that, if included in wider survival models, could provide a better disease stratification and an improved prognosis definition.