810 research outputs found

    Counter-terrorism in cyber–physical spaces:Best practices and technologies from the state of the art

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    Context: The demand for protection and security of physical spaces and urban areas increased with the escalation of terroristic attacks in recent years. We envision with the proposed cyber–physical systems and spaces, a city that would indeed become a smarter urbanistic object, proactively providing alerts and being protective against any threat. Objectives: This survey intend to provide a systematic multivocal literature survey comprised of an updated, comprehensive and timely overview of state of the art in counter-terrorism cyber–physical systems, hence aimed at the protection of cyber–physical spaces. Hence, provide guidelines to law enforcement agencies and practitioners providing a description of technologies and best practices for the protection of public spaces. Methods: We analyzed 112 papers collected from different online sources, both from the academic field and from websites and blogs ranging from 2004 till mid-2022. Results: (a) There is no one single bullet-proof solution available for the protection of public spaces. (b) From our analysis we found three major active fields for the protection of public spaces: Information Technologies, Architectural approaches, Organizational field. (c) While the academic suggest best practices and methodologies for the protection of urban areas, the market did not provide any type of implementation of such suggested approaches, which shows a lack of fertilization between academia and industry. Conclusion: The overall analysis has led us to state that there is no one single solution available, conversely, multiple methods and techniques can be put in place to guarantee safety and security in public spaces. The techniques range from architectural design to rethink the design of public spaces keeping security into account in continuity, to emerging technologies such as AI and predictive surveillance.</p

    Ovarian hormones shape brain structure, function, and chemistry: A neuropsychiatric framework for female brain health

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    There are robust sex differences in brain anatomy, function, as well as neuropsychiatric and neurodegenerative disease risk (1-6), with women approximately twice as likely to suffer from a depressive illness as well as Alzheimer’s Disease. Disruptions in ovarian hormones likely play a role in such disproportionate disease prevalence, given that ovarian hormones serve as key regulators of brain functional and structural plasticity and undergo major fluctuations across the female lifespan (7-9). From a clinical perspective, there is a wellreported increase in depression susceptibility and initial evidence for cognitive impairment or decline during hormonal transition states, such as the postpartum period and perimenopause (9-14). What remains unknown, however, is the underlying mechanism of how fluctuations in ovarian hormones interact with other biological factors to influence brain structure, function, and chemistry. While this line of research has translational relevance for over half the population, neuroscience is notably guilty of female participant exclusion in research studies, with the male brain implicitly treated as the default model and only a minority of basic and clinical neuroscience studies including a female sample (15-18). Female underrepresentation in neuroscience directly limits opportunities for basic scientific discovery; and without basic knowledge of the biological underpinnings of sex differences, we cannot address critical sexdriven differences in pathology. Thus, my doctoral thesis aims to deliberately investigate the influence of sex and ovarian hormones on brain states in health as well as in vulnerability to depression and cognitive impairment:Table of Contents List of Abbreviations ..................................................................................................................... i List of Figures .............................................................................................................................. ii Acknowledgements .....................................................................................................................iii 1 INTRODUCTION .....................................................................................................................1 1.1 Lifespan approach: Sex, hormones, and metabolic risk factors for cognitive health .......3 1.2 Reproductive years: Healthy models of ovarian hormones, serotonin, and the brain ......4 1.2.1 Ovarian hormones and brain structure across the menstrual cycle ........................4 1.2.2 Serotonergic modulation and brain function in oral contraceptive users .................6 1.3 Neuropsychiatric risk models: Reproductive subtypes of depression ...............................8 1.3.1 Hormonal transition states and brain chemistry measured by PET imaging ...........8 1.3.2 Serotonin transporter binding across the menstrual cycle in PMDD patients .......10 2 PUBLICATIONS ....................................................................................................................12 2.1 Publication 1: Association of estradiol and visceral fat with structural brain networks and memory performance in adults .................................................................................13 2.2 Publication 2: Longitudinal 7T MRI reveals volumetric changes in subregions of human medial temporal lobe to sex hormone fluctuations ..............................................28 2.3 Publication 3: One-week escitalopram intake alters the excitation-inhibition balance in the healthy female brain ...............................................................................................51 2.4 Publication 4: Using positron emission tomography to investigate hormone-mediated neurochemical changes across the female lifespan: implications for depression ..........65 2.5 Publication 5: Increase in serotonin transporter binding across the menstrual cycle in patients with premenstrual dysphoric disorder: a case-control longitudinal neuro- receptor ligand PET imaging study ..................................................................................82 3 SUMMARY ...........................................................................................................................100 References ..............................................................................................................................107 Supplementary Publications ...................................................................................................114 Author Contributions to Publication 1 .....................................................................................184 Author Contributions to Publication 2 .....................................................................................186 Author Contributions to Publication 3 .....................................................................................188 Author Contributions to Publication 4 .....................................................................................190 Author Contributions to Publication 5 .....................................................................................191 Declaration of Authenticity ......................................................................................................193 Curriculum Vitae ......................................................................................................................194 List of Publications ................................................................................................................195 List of Talks and Posters ......................................................................................................19

    Single-molecule detection and characterisation of alpha-synuclein aggregates

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    Aberrant protein aggregation is a predominant feature of many neurodegenerative disorders. It has long been recognised that aggregates of alpha-synuclein (α-syn) drive pathogenesis in Parkinson’s Disease (PD), and it is widely accepted that small α-syn oligomers are the key cytotoxic species in PD. Notably, however, these oligomeric species are difficult to characterise using traditional biochemical ensemble methods due to their high level of heterogeneity and low abundance. Single-molecule fluorescence microscopy techniques have emerged as a suitable approach to circumventing this problem, enabling the detection of individual aggregates amongst monomeric protein and thus facilitating the identification, quantification, and characterisation of rare oligomeric species. However, cellular mechanisms of α-syn aggregation are poorly understood. Furthermore, there remains some limitations to the singlemolecule techniques currently available. This thesis describes the work completed to address some of these issues. Chapter 1 provides the contextual background for the work presented in this thesis, detailing the biological aspects of α-syn, its aggregation, and its implications in PD, as well as outlining the single-molecule techniques used to investigate aggregate species. Chapter 2 describes the methodologies undertaken in this thesis, and chapters 3 to 5 describe the findings made using the single-molecule techniques which were utilised and developed in this work. One primary approach for studying species in single-molecule experiments involves directly labelling biomolecules of interest with a suitable fluorophore. Early steps in α-syn aggregation have previously been identified using fluorescently tagged α-syn and single-molecule Förster resonance energy transfer (smFRET) in vitro; however, the characterisation of early aggregate formation in cells has thus far been difficult to achieve. Chapter 3 describes the use of duallabelled α-syn to detect and characterise aggregates formed both intracellularly and in vitro via smFRET, using both single-molecule confocal microscopy coupled with microfluidics and iii total internal reflection fluorescence microscopy (TIRFM) to determine both the sizes and structures of the oligomers formed. This work reveals the presence of distinct oligomeric species in vitro and in neurons resulting from structural conversion during early aggregate formation. The approach taken in Chapter 3 is highly suitable for investigating aggregate formation resulting from the addition of exogenous α-syn to samples of interest. However, such an approach is not ideal for the detection and characterisation of endogenous aggregates due to issues with the covalent labelling of cellular protein. Extrinsic amyloid dyes are typically used as an alternative approach to labelled protein; however, such dyes are non-protein-specific and bind to the common amyloid beta-sheet motif. As an alternative, the work presented in Chapter 4 describes a novel single-molecule method to specifically detect and characterise α-syn aggregates with high sensitivity, making use of a high-affinity antibody labelled with orthogonal fluorophores which is combined with fast-flow microfluidics and single-molecule confocal microscopy. This enables the quantification and size approximation of α-syn aggregates at picomolar concentrations, both in vitro and in biological samples. Although the kinetics of α-syn aggregation have been studied extensively, much of our current knowledge stems from ensemble averaging techniques which are associated with high levels of variability and are not conducive to detecting the earliest steps in aggregate formation. In addition, there remains uncertainty surrounding the effect of familial variants and posttranslational modifications (PTM) on aggregation. Chapter 5 encompasses the study of the effects of the ubiquitous N-terminal acetylation PTM, in addition to the familial, rapid-onset G51D mutation, on α-syn aggregation, using the novel detection method developed in Chapter 4. This is used in conjunction with single-molecule detection with thioflavin-T (ThT) to reveal new insights into the aggregation of α-syn variants. Overall, the work presented here provides new insights into the aggregation of α-syn via the use and development of single-molecule techniques. The advancements made have added to the current understanding of the molecular mechanisms of α-syn aggregation, both in vitro and in neurons, and have also been used to develop a novel single-molecule detection method for α-syn aggregates. The work presented in this thesis has resulted in two published papers, ’Pathological structural conversion of alpha-synuclein at the mitochondria induces neuronal toxicity’ in Nature Neuroscience, and ’Single-molecule two-color coincidence detection of unlabeled alpha-synuclein aggregates’ in Angewandte Chemie International Edition. Furthermore, the novel detection method presented here holds promise for measuring α-syn oligomeric load in clinical samples due to its high sensitivity and specificity for α-syn aggregates. This may therefore be used in future studies for identifying, detecting, and studying potential biomarkers in PD, with potential use in disease diagnosis. It is therefore expected that the work from this thesis will be used to aid researchers towards better understanding the mechanisms of α-syn aggregation, both in vitro and in clinical samples

    Aspects of Terahertz Reflection Spectroscopy

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    Deep learning for intracellular particle tracking and motion analysis

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    Deep learning for intracellular particle tracking and motion analysis

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    APPLICATION OF NANOPORE DATA TO THE PIMMS (Pragmatic Insertional Mutation Mapping System) SEQ PIPELINE.

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    Significant developments in next generation sequencing technologies have over the past 15 years allowed novel and exciting opportunities to further our understanding of genomic science. Transposons are mobile regions of DNA that occur frequently throughout the genome, the insertion of these transposable elements play an essential role in both gene regulation and evolution. Locating the positions of transposon insertion into a genome can help our understanding of the role they play in such areas as gene expression and structural variation. Random transposon mediated mutagenesis has proven to be a useful tool in the identification of essential and conditionally essential genes within various bacterial species. Various methods have been generated to identify the regions within a given genome where transposon insertion has occurred. Some of these methods involve complex experimental design in the laboratory, and a high degree of competency in bioinformatics post sequencing. PIMMS (Pragmatic Insertion Mutation Mapping System) Seq was introduced in 2016 to speed up and simplify the bioinformatic pipeline when mapping transposon insertion sites using random mutagenesis sequencing data. The bacterial samples in question undergoing random mutagenesis, followed by inverse PCR, library preparation and sequencing (using Illumina technology). The use of Nanopore sequencing technologies has steadily increased since its introduction in 2014. Although the data quality is not yet as accurate as Illumina, Nanopore sequencing does offer some unique advantages, including the ability to sequence native DNA, generate long sequencing reads, and facilitate real time data analysis. Oxford Nanopore Technologies (ONT), the company that developed nanopore sequencing also offer some unique sample enrichment options such as Cas-9 targeted sequencing which, for PIMMS could potentially be used as an alternative to inverse PCR, targeting sequences that contain known transposon motifs that flank a genomic region of interest. Nanopore sequencing data was generated using S. agalactiae insertion mutant libraries, and uploaded to the PIMMS Seq bioinformatic pipeline to generate a database of transposon insertion sites. When comparing PIMMS output data generated using Nanopore sequencing to the established Illumina short read sequencing method, although the genomic distribution of insertions where consistent, differences where observed with Nanopore sequencing identifying 405 more unique insertion events, reducing the list of genes that were considered to be essential using short read sequencing. However, any advantages of this long read sequencing method must be off set against Nanopore’s sequencing base calling accuracy, which was significantly lower than that generated using Illumina technologies. The Cas-9 Targeted library preparation from ONT has been proven to enrich for transposon sequences that flank genomic regions of interest, with on average 68.1% of all reads mapping to the insert sequence. This establishes its potential as an attractive alternative to amplification based methods. This method may significantly reduce the total amount of sequencing required to run an experiment, which in turn will reduce time and costs. Any potential wet lab sample loses are minimised due to a far smaller amount of sample processing steps. Moving forward, nanopore long read sequencing is a technology that offers some unique advantages over short read technologies, and its application to the PIMMS pipeline has been demonstrated to work well. The added option of Cas- 9 Targeted nanopore sequencing, saves not only lab time and costs, but more importantly removes any potential amplification bias

    New insights along the gut-liver axis in cardiometabolic disease

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    In this thesis we targeted the human gut microbiome for the development of therapeutic strategies in metabolic disorders. In chapter 3 we performed a randomized placebo-controlled cross-over study in individuals with the metabolic syndrome in which we showed that a single duodenal infusion of A. soehngenii improved peripheral glycemic control. In chapter 4 we studied the effect of a 2 weeks oral A. soehngenii treatment in individuals with T2D treated with metformin on their glycemic control.The second part of the thesis focused on MASLD, currently the most common cause of chronic liver dysfunction worldwide. In chapter 5 we reviewed the gut microbial and gut microbial-derived metabolite signatures associated with the development and disease progression of MASLD. To dissect causality of intestinal microbiota in MASLD, in chapter 6 we performed a single-center, double-blind, randomized controlled proof-of-principle pilot study comparing the effect of three 8-weekly lean vegan donor FMT versus autologous FMT on the severity of MASLD, using liver biopsies in individuals with hepatic steatosis on ultrasound. Moreover, we aimed to identify and validate noninvasive diagnostic methods in disease progression in MASLD. Hence, in chapter 7 we examined the diagnostic performance of multiparametric MRI for the assessment of disease severity along the MASLD disease spectrum with comparison to histological scores

    Gurus and Media: Sound, image, machine, text and the digital

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    Gurus and Media is the first book dedicated to media and mediation in domains of public guruship and devotion. Illuminating the mediatisation of guruship and the guru-isation of media, it bridges the gap between scholarship on gurus and the disciplines of media and visual culture studies. It investigates guru iconographies in and across various time periods and also the distinctive ways in which diverse gurus engage with and inhabit different forms of media: statuary, games, print publications, photographs, portraiture, films, machines, social media, bodies, words, graffiti, dolls, sound, verse, tombs and more. The book’s interdisciplinary chapters advance, both conceptually and ethnographically, our understanding of the function of media in the dramatic production of guruship, and reflect on the corporate branding of gurus and on mediated guruship as a series of aesthetic traps for the captivation of devotees and others. They show how different media can further enliven the complex plurality of guruship, for instance in instantiating notions of ‘absent-present’ guruship and demonstrating the mutual mediation of gurus, caste and Hindutva. Throughout, the book foregrounds contested visions of the guru in the development of devotional publics and pluriform guruship across time and space. Thinking through the guru’s many media entanglements in a single place, the book contributes new insights to the study of South Asian religions and to the study of mediation more broadly
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