Enabling a direct path from end-user specifications to executable protocols in a biology laboratory environment

Biomedical analyses are becoming increasingly complex, with respect to both the type of the data to be produced and the procedures to be executed. This trend is expected to continue in the future. The development of information and protocol management systems that can sustain this challenge is therefore becoming an essential enabling factor for all actors in the field. The use of custom-built solutions that require the biology domain expert to acquire or procure software engineering expertise in the development of the laboratory infrastructure is not fully satisfactory because it incurs undesirable mutual knowledge dependencies between the two camps. We propose instead an infrastructure concept that enables the domain experts to express laboratory protocols using proper domain knowledge, free from the incidence and mediation of the software implementation artefacts. In the system that we propose this is made possible by basing the modelling language on an authoritative domain specific ontology and then using modern model-driven architecture technology to transform the user models in software artefacts ready for execution in a multi-agent based execution platform specialized for biomedical laboratories.

Architectures for Context Aware Services in Smart Environments

This thesis deals with Context Aware Services, Smart Environments, Context Management and solutions for Devices and Service Interoperability. Multi-vendor devices offer an increasing number of services and end-user applications that base their value on the ability to exploit the information originating from the surrounding environment by means of an increasing number of embedded sensors, e.g. GPS, compass, RFID readers, cameras and so on. However, usually such devices are not able to exchange information because of the lack of a shared data storage and common information exchange methods. A large number of standards and domain specific building blocks are available and are heavily used in today's products. However, the use of these solutions based on ready-to-use modules is not without problems. The integration and cooperation of different kinds of modules can be daunting because of growing complexity and dependency. In this scenarios it might be interesting to have an infrastructure that makes the coexistence of multi-vendor devices easy, while enabling low cost development and smooth access to services. This sort of technologies glue should reduce both software and hardware integration costs by removing the trouble of interoperability. The result should also lead to faster and simplified design, development and, deployment of cross-domain applications. This thesis is mainly focused on SW architectures supporting context aware service providers especially on the following subjects: - user preferences service adaptation - context management - content management - information interoperability - multivendor device interoperability - communication and connectivity interoperability Experimental activities were carried out in several domains including Cultural Heritage, indoor and personal smart spaces – all of which are considered significant test-beds in Context Aware Computing. The work evolved within european and national projects: on the europen side, I carried out my research activity within EPOCH, the FP6 Network of Excellence on “Processing Open Cultural Heritage” and within SOFIA, a project of the ARTEMIS JU on embedded systems. I worked in cooperation with several international establishments, including the University of Kent, VTT (the Technical Reserarch Center of Finland) and Eurotech. On the national side I contributed to a one-to-one research contract between ARCES and Telecom Italia. The first part of the thesis is focused on problem statement and related work and addresses interoperability issues and related architecture components. The second part is focused on specific architectures and frameworks: - MobiComp: a context management framework that I used in cultural heritage applications - CAB: a context, preference and profile based application broker which I designed within EPOCH Network of Excellence - M3: "Semantic Web based" information sharing infrastructure for smart spaces designed by Nokia within the European project SOFIA - NoTa: a service and transport independent connectivity framework - OSGi: the well known Java based service support framework The final section is dedicated to the middleware, the tools and, the SW agents developed during my Doctorate time to support context-aware services in smart environments.

Flexible Computing Systems For AI Acceleration At The Extreme Edge Of The IoT

Embedding intelligence in extreme edge devices allows distilling raw data acquired from sensors into actionable information, directly on IoT end-nodes. This computing paradigm, in which end-nodes no longer depend entirely on the Cloud, offers undeniable benefits, driving a large research area (TinyML) to deploy leading Machine Learning (ML) algorithms on micro-controller class of devices. To fit the limited memory storage capability of these tiny platforms, full-precision Deep Neural Networks (DNNs) are compressed by representing their data down to byte and sub-byte formats, in the integer domain. However, the current generation of micro-controller systems can barely cope with the computing requirements of QNNs. This thesis tackles the challenge from many perspectives, presenting solutions both at software and hardware levels, exploiting parallelism, heterogeneity and software programmability to guarantee high flexibility and high energy-performance proportionality. The first contribution, PULP-NN, is an optimized software computing library for QNN inference on parallel ultra-low-power (PULP) clusters of RISC-V processors, showing one order of magnitude improvements in performance and energy efficiency, compared to current State-of-the-Art (SoA) STM32 micro-controller systems (MCUs) based on ARM Cortex-M cores. The second contribution is XpulpNN, a set of RISC-V domain specific instruction set architecture (ISA) extensions to deal with sub-byte integer arithmetic computation. The solution, including the ISA extensions and the micro-architecture to support them, achieves energy efficiency comparable with dedicated DNN accelerators and surpasses the efficiency of SoA ARM Cortex-M based MCUs, such as the low-end STM32M4 and the high-end STM32H7 devices, by up to three orders of magnitude. To overcome the Von Neumann bottleneck while guaranteeing the highest flexibility, the final contribution integrates an Analog In-Memory Computing accelerator into the PULP cluster, creating a fully programmable heterogeneous fabric that demonstrates end-to-end inference capabilities of SoA MobileNetV2 models, showing two orders of magnitude performance improvements over current SoA analog/digital solutions.

Influencable autonomy and predictable freedom in the IoE

This thesis investigates how individuals can develop, exercise, and maintain autonomy and freedom in the presence of information technology. It is particularly interested in how information technology can impose autonomy constraints. The first part identifies a problem with current autonomy discourse: There is no agreed upon object of reference when bemoaning loss of or risk to an individual’s autonomy. Here, thesis introduces a pragmatic conceptual framework to classify autonomy constraints. In essence, the proposed framework divides autonomy in three categories: intrinsic autonomy, relational autonomy and informational autonomy. The second part of the thesis investigates the role of information technology in enabling and facilitating autonomy constraints. The analysis identifies eleven characteristics of information technology, as it is embedded in society, so-called vectors of influence, that constitute risk to an individual’s autonomy in a substantial way. These vectors are assigned to three sets that correspond to the general sphere of the information transfer process to which they can be attributed to, namely domain-specific vectors, agent-specific vectors and information recipient-specific vectors. The third part of the thesis investigates selected ethical and legal implications of autonomy constraints imposed by information technology. It shows the utility of the theoretical frameworks introduced earlier in the thesis when conducting an ethical analysis of autonomy-constraining technology. It also traces the concept of autonomy in the European Data Lawsand investigates the impact of cultural embeddings of individuals on efforts to safeguard autonomy, showing intercultural flashpoints of autonomy differences. In view of this, the thesis approaches the exercise and constraint of autonomy in presence of information technology systems holistically. It contributes to establish a common understanding of (intuitive) terminology and concepts, connects this to current phenomena arising out of ever-increasing interconnectivity and computational power and helps operationalize the protection of autonomy through application of the proposed frameworks.

B-type cytochromes of the DOMON domain superfamily (Novel redox elements of plant plasma membrane)

The aim of the present study is understanding the properties of a new group of redox proteins having in common a DOMON-type domain with characteristics of cytochromes b. The superfamily of proteins containing a DOMON of this type includes a few protein families. With the aim of better characterizing this new protein family, the present work addresses both a CyDOM protein (a cytochrome b561) and a protein only comprised of DOMON(AIR12), both of plant origin. Apoplastic ascorbate can be regenerated from monodehydroascorbate by a trans-plasma membrane redox system which uses cytosolic ascorbate as a reductant and comprises a high potential cytochrome b. We identified the major plasma membrane (PM) ascorbate-reducible b-type cytochrome of bean (Phaseolus vulgaris) and soybean (Glycine max) hypocotyls as orthologs of Arabidopsis auxin-responsive gene air12. The protein, which is glycosylated and glycosylphosphatidylinositol-anchored to the external side of the PM in vivo, was expressed in Pichia pastoris in a recombinant form, lacking the glycosylphosphatidylinositol-modification signal, and purified from the culture medium. Recombinant AIR12 is a soluble protein predicted to fold into a β-sandwich domain and belonging to the DOMON superfamily. It is shown to be a b-type cytochrome with a symmetrical α-band at 561 nm, to be fully reduced by ascorbate and fully oxidized by monodehydroascorbate. Redox potentiometry suggests that AIR12 binds two high-potential hemes (Em,7 +135 and +236 mV). Phylogenetic analyses reveal that the auxin-responsive genes AIR12 constitute a new family of plasma membrane b-type cytochromes specific to flowering plants. Although AIR12 is one of the few redox proteins of the PM characterized to date, the role of AIR12 in trans-PM electron transfer would imply interaction with other partners which are still to be identified. Another part of the present project was aimed at understanding of a soybean protein comprised of a DOMON fused with a well-defined b561 cytochrome domain (CyDOM). Various bioinformatic approaches show this protein to be composed of an N-terminal DOMON followed by b561 domain. The latter contains five transmembrane helices featuring highly conserved histidines, which might bind haem groups. The CyDOM has been cloned and expressed in the yeast Pichia pastoris, and spectroscopic analyses have been accomplished on solubilized yeast membranes. CyDOM clearly reveal the properties of b-type cytochrome. The results highlight the fact that CyDOM is clearly able to lead an electron flux through the plasmamembrane. Voltage clamp experiments demonstrate that Xenopus laevis oocytes transformed with CyDOM of soybean exhibit negative electrical currents in presence of an external electron acceptor. Analogous investigations were carried out with SDR2, a CyDOM of Drosophila melanogaster which shows an electron transport capacity even higher than plant CyDOM. As quoted above, these data reinforce those obtained in plant CyDOM on the one hand, and on the other hand allow to attribute to SDR2-like proteins the properties assigned to CyDOM. Was expressed in Regenerated tobacco roots, transiently transformed with infected a with chimeral construct GFP: CyDOM (by A. rhizogenes infection) reveals a plasmamembrane localization of CyDOM both in epidermal cells of the elongation zone of roots and in root hairs. In conclusion. Although the data presented here await to be expanded and in part clarified, it is safe to say they open a new perspective about the role of this group of proteins. The biological relevance of the functional and physiological implications of DOMON redox domains seems noteworthy, and it can but increase with future advances in research. Beyond the very finding, however interesting in itself, of DOMON domains as extracellular cytochromes, the present study testifies to the fact that cytochrome proteins containing DOMON domains of the type of “CyDOM” can transfer electrons through membranes and may represent the most important redox component of the plasmamembrane as yet discovered.

Natural compounds Camptothecin and Triptolide: highly specific enzyme inhibitors and tools to dissect transcriptional functions

With this work I elucidated new and unexpected mechanisms of two strong and highly specific transcription inhibitors: Triptolide and Campthotecin. Triptolide (TPL) is a diterpene epoxide derived from the Chinese plant Trypterigium Wilfoordii Hook F. TPL inhibits the ATPase activity of XPB, a subunit of the general transcription factor TFIIH. In this thesis I found that degradation of Rbp1 (the largest subunit of RNA Polymerase II) caused by TPL treatments, is preceded by an hyperphosphorylation event at serine 5 of the carboxy-terminal domain (CTD) of Rbp1. This event is concomitant with a block of RNA Polymerase II at promoters of active genes. The enzyme responsible for Ser5 hyperphosphorylation event is CDK7. Notably, CDK7 downregulation rescued both Ser5 hyperphosphorylation and Rbp1 degradation triggered by TPL. Camptothecin (CPT), derived from the plant Camptotheca acuminata, specifically inhibits topoisomerase 1 (Top1). We first found that CPT induced antisense transcription at divergent CpG islands promoter. Interestingly, by immunofluorescence experiments, CPT was found to induce a burst of R loop structures (DNA/RNA hybrids) at nucleoli and mitochondria. We then decided to investigate the role of Top1 in R loop homeostasis through a short interfering RNA approach (RNAi). Using DNA/RNA immunoprecipitation techniques coupled to NGS I found that Top1 depletion induces an increase of R loops at a genome-wide level. We found that such increase occurs on the entire gene body. At a subset of loci R loops resulted particularly stressed after Top1 depletion: some of these genes showed the formation of new R loops structures, whereas other loci showed a reduction of R loops. Interestingly we found that new peaks usually appear at tandem or divergent genes in the entire gene body, while losses of R loop peaks seems to be a feature specific of 3’ end regions of convergent genes.

Science of retracted science: a citation analysis of the arts and humanities domain

In the scholarly publishing domain, a retraction is raised when a specific publication is considered erroneous by the venue in which it appeared after it was published. The aim of this work is uncovering new insights and learn new important information to help us understand the retraction phenomenon in the arts and humanities domain. Our investigation is based on a methodology defined using quantitative and qualitative measures derived from previous studies in the transdisciplinary research field of “science of science” (SciSci). The designed methodology takes into account a general case of retraction and applies a citation analysis based on five phases. Citations to retracted publications (before and after their retraction) are gathered and characterized with a set of attributes, including general metadata and information extracted from citing entities’ full text. The annotated characteristics are further considered for a statistical and a textual analysis (i.e., a topic modeling analysis). The contribution of this thesis is grounded by addressing the following research questions: (RQ1) How did scholarly research cite retracted humanities publications before and after their retraction? (RQ2) Did all the humanities areas behave similarly concerning the retraction phenomenon? (RQ3) What are the main differences and similarities in the retraction dynamics between the humanities domain and the STEM disciplines? RQ1 and RQ2 are addressed by tuning and applying the methodology on the analysis of the retracted publications in the humanities domain. RQ3 is addressed on two levels, i.e., considering and comparing: (L1) the outcomes of the past studies on the retraction in STEM, and (L2) the results obtained from an analysis of a retraction case in STEM using the defined methodology.