5,424 research outputs found

    An examination of the verbal behaviour of intergroup discrimination

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    This thesis examined relationships between psychological flexibility, psychological inflexibility, prejudicial attitudes, and dehumanization across three cross-sectional studies with an additional proposed experimental study. Psychological flexibility refers to mindful attention to the present moment, willing acceptance of private experiences, and engaging in behaviours congruent with one’s freely chosen values. Inflexibility, on the other hand, indicates a tendency to suppress unwanted thoughts and emotions, entanglement with one’s thoughts, and rigid behavioural patterns. Study 1 found limited correlations between inflexibility and sexism, racism, homonegativity, and dehumanization. Study 2 demonstrated more consistent positive associations between inflexibility and prejudice. And Study 3 controlled for right-wing authoritarianism and social dominance orientation, finding inflexibility predicted hostile sexism and racism beyond these factors. While showing some relationships, particularly with sexism and racism, psychological inflexibility did not consistently correlate with varied prejudices across studies. The proposed randomized controlled trial aims to evaluate an Acceptance and Commitment Therapy intervention to reduce sexism through enhanced psychological flexibility. Overall, findings provide mixed support for the utility of flexibility-based skills in addressing complex societal prejudices. Research should continue examining flexibility integrated with socio-cultural approaches to promote equity

    Some quaternary additive codes outperform linear counterparts

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    The additive codes may have better parameters than linear codes. However, it is still a challenging problem to efficiently construct additive codes that outperform linear codes, especially those with greater distances than linear codes of the same lengths and dimensions. This paper focuses on constructing additive codes that outperform linear codes based on quasi-cyclic codes and combinatorial methods. Firstly, we propose a lower bound on the symplectic distance of 1-generator quasi-cyclic codes of index even. Secondly, we get many binary quasi-cyclic codes with large symplectic distances utilizing computer-supported combination and search methods, all of which correspond to good quaternary additive codes. Notably, some additive codes have greater distances than best-known quaternary linear codes in Grassl's code table (bounds on the minimum distance of quaternary linear codes http://www.codetables.de) for the same lengths and dimensions. Moreover, employing a combinatorial approach, we partially determine the parameters of optimal quaternary additive 3.5-dimensional codes with lengths from 2828 to 254254. Finally, as an extension, we also construct some good additive complementary dual codes with larger distances than the best-known quaternary linear complementary dual codes in the literature

    Seamless Multimodal Biometrics for Continuous Personalised Wellbeing Monitoring

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    Artificially intelligent perception is increasingly present in the lives of every one of us. Vehicles are no exception, (...) In the near future, pattern recognition will have an even stronger role in vehicles, as self-driving cars will require automated ways to understand what is happening around (and within) them and act accordingly. (...) This doctoral work focused on advancing in-vehicle sensing through the research of novel computer vision and pattern recognition methodologies for both biometrics and wellbeing monitoring. The main focus has been on electrocardiogram (ECG) biometrics, a trait well-known for its potential for seamless driver monitoring. Major efforts were devoted to achieving improved performance in identification and identity verification in off-the-person scenarios, well-known for increased noise and variability. Here, end-to-end deep learning ECG biometric solutions were proposed and important topics were addressed such as cross-database and long-term performance, waveform relevance through explainability, and interlead conversion. Face biometrics, a natural complement to the ECG in seamless unconstrained scenarios, was also studied in this work. The open challenges of masked face recognition and interpretability in biometrics were tackled in an effort to evolve towards algorithms that are more transparent, trustworthy, and robust to significant occlusions. Within the topic of wellbeing monitoring, improved solutions to multimodal emotion recognition in groups of people and activity/violence recognition in in-vehicle scenarios were proposed. At last, we also proposed a novel way to learn template security within end-to-end models, dismissing additional separate encryption processes, and a self-supervised learning approach tailored to sequential data, in order to ensure data security and optimal performance. (...)Comment: Doctoral thesis presented and approved on the 21st of December 2022 to the University of Port

    Rational development of stabilized cyclic disulfide redox probes and bioreductive prodrugs to target dithiol oxidoreductases

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    Countless biological processes allow cells to develop, survive, and proliferate. Among these, tightly balanced regulatory enzymatic pathways that can respond rapidly to external impacts maintain dynamic physiological homeostasis. More specifically, redox homeostasis broadly affects cellular metabolism and proliferation, with major contributions by thiol/disulfide oxidoreductase systems, in particular, the Thioredoxin Reductase Thioredoxin (TrxR/Trx) and the Glutathione Reductase-Glutathione-Glutaredoxin (GR/GSH/Grx) systems. These cascades drive vital cellular functions in many ways through signaling, regulating other proteins' activity by redox switches, and by stoichiometric reductant transfers in metabolism and antioxidant systems. Increasing evidence argues that there is a persistent alteration of the redox environment in certain pathological states, such as cancer, that heavily involve the Trx system: upregulation and/or overactivity of the Trx system may support or drive cancer progression, making both TrxR and Trx promising targets for anti-cancer drug development. Understanding the biochemical mechanisms and connections between certain redox cascades requires research tools that interact with them. The state-of-the-art genetic tools are mostly ratiometric reporters that measure reduced:oxidized ratios of selected redox pairs or the general thiol pool. However, the precise cellular roles of the central oxidoreductase systems, including TrxR and Trx, remain inaccessible due to the lack of probes to selectively measure turnover by either of these proteins. However, such probes would allow measuring their effective reductive activity apart from expression levels in native systems, including in cells, animals, or patient samples. They are also of high interest to identify chemical inhibitors for TrxR/Trx in cells and to validate their potential use as anti-cancer agents (to date, there is no selective cellular Trx inhibitor, and most known TrxR inhibitors were not comprehensively evaluated considering selectivity and potential off-targets). However, small molecule redox imaging tools are underdeveloped: their protein specificity, spectral properties, and applicability remain poorly precedented. This work aimed to address this opportunity gap and develop novel, small molecule diagnostic and therapeutic tools to selectively target the Trx system based on a modular trigger cargo design: artificial cyclic disulfide substrates (trigger) for oxidoreductases are tethered to molecular agents (cargo) such that the cargo’s activity is masked and is re-established only through reduction by a target protein. The rational design of these novel reduction sensors to target the cell's strongest disulfide-reducing enzymes was driven by the following principles: (i) cyclic disulfide triggers with stabilized ring systems were used to gain low reduction potentials that should resist reduction except by the strongest cellular reductases, such as Trx; and (ii) the cyclic topology also offers the potential for kinetic reversibility that should select for dithiol-type redox proteins over the cellular monothiol background. Creating imaging agents based on such two-component designs to selectively measure redox protein activity in native cells required to combine the correct trigger reducibility, probe activation kinetics, and imaging modalities and to consider the overall molecular architecture. The major prior art in this field has applied cyclic 5-membered disulfides (1,2 dithiolanes) as substrates for TrxR in a similar way to create such tools. However, this motif was described elsewhere as thermodynamically instable and was due to widely used for dynamic covalent cascade reactions. By comparing a novel 1,2 dithiolane-based probe to the state-of-the-art probes, including commercial TrxR sensors, by screening a conclusive assay panel of cellular TrxR modulations, I clarified that 1,2 dithiolanes are not selective substrates for TrxR in biological settings (Nat Commun 2022). Instead, aiming for more stable ring systems and thus more robust redox probes, during this work, I developed bicyclic 6 membered disulfides (piperidine fused 1,2 dithianes) with remarkably low reduction potentials. I showed that molecular probes using them as reduction sensors can be mostly processed by thioredoxins while being stable against reduction by GSH. The thermodynamically stabilized decalin like topology of the cis-annelated 1,2 dithianes requires particularly strong reductants to be cleaved. They also select for dithiol type redox proteins, like Trx, based on kinetic reversibility and offer fast cyclization due to the preorganization by annelation (JACS 2021). This work further expanded the system’s modularity with structural cores based on piperazine-fused 1,2 dithianes with the two amines allowing independent derivatization. Diagnostic tools using them as reduction sensors proved equally robust but with highly improved activation kinetics and were thus cellularly activated. Cellular studies evolved that they are substrates for both Trxs and their protein cousins Grxs, so measuring the cellular dithiol protein pool rather than solely Trx activity (preprint 2023). Finally, a trigger based on a slightly adapted reduction sensor, a desymmetrized 1,2 thiaselenane, was designed for selective reduction by TrxR’s selenol/thiol active site, then combined with a precipitating large Stokes’ shift fluorophore and a solubilizing group, to evolve the first selective probe RX1 to measure cellular TrxR activity, which even allowed high throughput inhibitor screening (Chem 2022). The central principle of this work was further advanced to therapeutic prodrugs based on the duocarmycin cargo (CBI) with tunable potency (JACS Au 2022) that can be used to create off-to-on therapeutic prodrugs. Such CBI prodrugs employing stabilized 1,2 dichalcogenide triggers proved to be cytotoxins that depend on Trx system activity in cells. They could further be exploited for cell-line dependent reductase activity profiling by screening their redox activation indices, the reduction-dependent part of total prodrug activation, in 177 cell lines. Beyond that, these prodrugs were well-tolerated in animals and showed anti-cancer efficacy in vivo in two distinct mouse tumor models (preprint 2022). Taken together, I introduced unique monothiol-resistant reducible motifs to target the cellular Trx system with chemocompatible units for each for TrxR and Trx/Grx, where the cyclic nature of the dichalcogenides avoids activation by GSH. By using them with distinct molecular cargos, I developed novel selective fluorescent reporter probes; and introduced a new class of bioreductive therapeutic constructs based on a common modular design. These were either applied to selectively measure cellular reductase activity or to deliver cytotoxic anti cancer agents in vivo. Ongoing work aims to differentiate between the two major redox effector proteins Trx and Grx, requiring additional layers of selectivity that may be addressed by tuned molecular recognition. The flexible use of various molecular cargos allows harnessing the same cellular redox machinery by either probes or prodrugs. This allows predictive conclusions from diagnostics to be directly translated into therapy and offers great potential for future adaptation to other enzyme classes and therapeutic venues.Die zellulĂ€re Redox-Homöostase hĂ€ngt von Thiol/Disulfid-Oxidoreduktasen ab, die den Stoffwechsel, die Proliferation und die antioxidative Antwort von Zellen beeinflussen. Die wichtigsten Netzwerke sind die Thioredoxin Reduktase-Thioredoxin (TrxR/Trx) und Glutathion Reduktase-Glutathion-Glutaredoxin (GR/GSH/Grx) Systeme, die ĂŒber Redox-Schalter in Substratproteinen lebenswichtige zellulĂ€re Funktionen steuern und so an der Redox-Regulation und -SignalĂŒbertragung beteiligt sind. Persistente VerĂ€nderungen des Redoxmilieus in pathologischen ZustĂ€nden, wie z. B. bei Krebs, sind in hohem Maße mit dem Trx-System verbunden. Eine Hochregulierung und/oder ÜberaktivitĂ€t des Trx-Systems, die bei vielen Krebsarten auftreten, unterstĂŒtzt zudem das Fortschreiten des Krebswachstums, was TrxR/Trx zu vielversprechenden Zielproteinen fĂŒr die Entwicklung neuer Krebsmedikamente macht. Um die biochemischen Prozesse dahinter zu erforschen, sind spezielle Techniken zur Visualisierung und Messung enzymatischer AktivitĂ€t nötig. Die hierzu geeigneten, meist genetischen Sensoren messen ratiometrisch das VerhĂ€ltnis reduzierter/oxidierter Spezies in zellulĂ€rem Umfeld oder spezifisch ausgewĂ€hlte Redoxpaare. Die weitere Erforschung der exakten Funktion von TrxR/Trx und deren Substrate ist jedoch durch mangelnde Nachweismethoden limitiert. Diese sind außerdem zur Validierung chemischer Hemmstoffe fĂŒr TrxR/Trx in Zellen und deren potenziellen Verwendung als Krebsmittel von großem Interesse. Bislang gibt es keinen selektiven zellulĂ€ren Trx-Inhibitor und potenzielle Off-Target-Effekte der bekannten TrxR-Inhibitoren wurden nicht abschließend bewertet. Ziel dieser Arbeit ist die Entwicklung niedermolekularer, diagnostischer und therapeutischer Werkzeuge, die selektiv auf das Trx-System abzielen und auf einem modularen Trigger-Cargo Design basieren. Hierzu werden zyklische Disulfid-Substrate (Trigger) fĂŒr Oxidoreduktasen so mit molekularen Wirkstoffen (Cargo) verknĂŒpft, dass dabei die WirkstoffaktivitĂ€t maskiert, und erst nach Reduktion durch ein Zielprotein wiederhergestellt wird. Diese neuartigen, synthetischen Reduktionssensoren basieren auf den folgenden Grundprinzipien: (i) Zyklische Disulfide sind thermodynamisch stabilisiert und können nur durch die stĂ€rksten Reduktasen gespalten werden; und (ii) die zyklische Topologie ermöglicht die kinetische ReversibilitĂ€t der zwei Thiol-Disulfid-Austauschreaktionen, die eine erste Reaktion mit Monothiolen, wie z. B. GSH, sofort umkehrt und so eine vollstĂ€ndige Reduktion verhindert. Die meisten frĂŒheren Arbeiten auf diesem Gebiet verwendeten ein zyklisches, fĂŒnfgliedriges Disulfid (1,2 Dithiolan) als Substrat fĂŒr TrxR. Das gleiche Strukturmotiv wurde jedoch an anderer Stelle als thermodynamisch instabil beschrieben und aufgrund dieser Eigenschaft explizit fĂŒr dynamische Kaskadenreaktionen verwendet. Deshalb vergleicht diese Arbeit zu Beginn einen neuen 1,2 Dithiolan basierten fluorogenen Indikator mit bestehenden, z. T. kommerziellen, Redox Sonden fĂŒr TrxR in einer Reihe von Zellkultur-Experimenten unter Modulation der zellulĂ€ren TrxR AktivitĂ€t und stellt so einen Widerspruch in der Literatur klar: 1,2 Dithiolane eignen sich nicht als selektive Substrate fĂŒr TrxR, da sie labil sowohl gegen die Reduktion durch andere Redoxproteine, als auch gegen den Monothiol Hintergrund in Zellen sind (Nat. Commun. 2022). Als alternatives Strukturmotiv wird in dieser Arbeit ein bizyklisches sechsgliedriges Disulfid (anneliertes 1,2 Dithian) etabliert. Durch sein niedriges Reduktionspotenzial, also seine hohe Resistenz gegen Reduktion, werden molekulare Sonden basierend auf diesem 1,2 Dithian als Reduktionssensor fast ausschließlich von Trx aktiviert, nicht aber von TrxR oder GSH (JACS 2021). Dieses Kernmotiv bestimmt dabei die Reduzierbarkeit, und damit die EnzymspezifitĂ€t, durch seine zyklische Natur und die Annelierung, auch unter Verwendung unterschiedlicher Farb-/Wirkstoffe. Auf dieser Grundlage konnte die molekulare Struktur durch einen weiteren Modifikationspunkt fĂŒr die flexible Verwendung weiterer funktioneller Einheiten ergĂ€nzt werden. Obwohl zellulĂ€re Studien ergaben, dass diese neuartigen 1,2 Dithian Einheiten in Zellen sowohl Trx als auch das strukturell verwandte Grx adressieren, sind die daraus resultierenden diagnostischen MolekĂŒle wertvoll, um den katalytischen Umsatz zellulĂ€rer Dithiol-Reduktasen, der sogenannten Trx Superfamilie, selektiv anzuzeigen (Preprint 2023). BegĂŒnstigt durch das modulare MolekĂŒldesign stellt diese Arbeit zudem das erste Reportersystem RX1 zum selektiven Nachweis der TrxR-AktivitĂ€t in Zellen vor. Es basiert auf der Verwendung eines zyklischen, unsymmetrischen Selenenylsulfid-Sensors (1,2 Thiaselenan), der selektiv von dem einzigartigen Selenolat der TrxR angegriffen wird, und dadurch letztlich nur von TrxR reduziert werden kann. RX1 eignete sich zudem fĂŒr eine Hochdurchsatz-Validierung bestehender TrxR Inhibitoren und unterstreicht dadurch den kommerziellen Nutzen derartiger Diagnostika (Chem 2022). Das zentrale Trigger-Cargo Konzept dieser Arbeit wurde fĂŒr therapeutische Zwecke weiterentwickelt und nutzt dabei den einzigartigen Wirkmechanismus der Duocarmycin-Naturstoffklasse (CBI) (JACS Au 2022) zur Entwicklung reduktiv aktivierbarer Therapeutika. CBI Prodrugs basierend auf stabilisierten Redox-Schaltern (1,2 Dithiane fĂŒr Trx; 1,2 Thiaselenan fĂŒr TrxR) reagierten signifikant auf TrxR-Modulation in Zellen. Sie wurden darĂŒber hinaus durch das Referenzieren ihrer AktivitĂ€t gegenĂŒber nicht-reduzierbaren KontrollmolekĂŒle fĂŒr die Erstellung zelllinienabhĂ€ngiger Profile der ReduktaseaktivitĂ€t in 177 Zelllinien genutzt. Schließlich waren diese neuen Krebsmittel im Tiermodell gut vertrĂ€glich und zeigten in zwei verschiedenen Mausmodellen eine krebshemmende Wirkung (Preprint 2022b). Zusammenfassend prĂ€sentiert diese Dissertation monothiol-resistente reduzierbare Trigger-Einheiten fĂŒr das zellulĂ€re Trx-System zur Entwicklung neuartiger, selektiver Reporter-Sonden, sowie eine neue Klasse reduktiv aktivierbarer Krebsmittel auf Basis eines adaptierbaren Trigger-Cargo Designs. Diese fanden entweder zur selektiven Messung zellulĂ€rer ProteinaktivitĂ€t oder zum Einsatz als Antikrebsmittel Verwendung. Es wurden chemokompatible Motive sowohl fĂŒr TrxR als auch fĂŒr Trx/Grx identifiziert, wobei deren zyklische Natur eine Aktivierung durch GSH verhindert. Eine weitere Differenzierung zwischen den beiden Redox-Proteinen Trx und Grx und anderen Proteinen der Trx-Superfamilie erfordert eine zusĂ€tzliche Ebene der Selektierung, z. B. durch molekulare Erkennung, und ist Gegenstand laufender Arbeiten. Die flexible Verwendung verschiedener molekularer Wirkstoffe ermöglicht dabei die „Pipeline-Entwicklung“ von Diagnostika und Therapeutika, die von der zellulĂ€ren Redox-Maschinerie analog umgesetzt werden, und dadurch Schlussfolgerungen aus der Diagnostik direkt auf eine Therapie ĂŒbertragbar machen. Dies birgt großes Potenzial fĂŒr kĂŒnftige Entwicklungen bei einer potenziellen Übertragung des modularen Konzepts auf andere Enzymklassen und therapeutische Einsatzgebiete

    Systemic Circular Economy Solutions for Fiber Reinforced Composites

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    This open access book provides an overview of the work undertaken within the FiberEUse project, which developed solutions enhancing the profitability of composite recycling and reuse in value-added products, with a cross-sectorial approach. Glass and carbon fiber reinforced polymers, or composites, are increasingly used as structural materials in many manufacturing sectors like transport, constructions and energy due to their better lightweight and corrosion resistance compared to metals. However, composite recycling is still a challenge since no significant added value in the recycling and reprocessing of composites is demonstrated. FiberEUse developed innovative solutions and business models towards sustainable Circular Economy solutions for post-use composite-made products. Three strategies are presented, namely mechanical recycling of short fibers, thermal recycling of long fibers and modular car parts design for sustainable disassembly and remanufacturing. The validation of the FiberEUse approach within eight industrial demonstrators shows the potentials towards new Circular Economy value-chains for composite materials

    ‘Inner qualities versus inequalities’: A case study of student change learning about Aboriginal health using sequential, explanatory mixed methods

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    Racism and lack of self-determination in health care perpetuate injury and injustice to Aboriginal people. To instil cultural safety at individual, organisational, community and systems levels, a key site of action has been health professional education that seeks to elicit reflexivity, cultural humility and a working understanding of Aboriginal health concepts. Studies in Aboriginal community settings show Family Well Being (FWB) empowerment education is effective in supporting personal and collective reflexivity and transformation through empowering life skills development. Implementation of FWB within educational settings shows early signs of effectiveness among students. Yet knowledge of the steps and processes of student change is lacking. This mixed methods explanatory case study sought to measure and understand change in postgraduate students of a leading Australian university learning about Aboriginal health and wellbeing through blended delivery, including through face-to-face immersion in FWB in an urban classroom. Three interrelated studies investigated fidelity and acceptability of the program, measured and analysed growth and empowerment in students, and explained processes of change observed, through thematic analysis of asynchronous online discussions using lenses based on transformative learning and empowerment. Researcher reflexivity was promoted by Aboriginal supervision. Over six years, 194 students enrolled in two different Aboriginal public health courses, 85 of them in the FWB course. As well as achieving program fidelity and acceptability, pre/post-course change in students across a range of emotional empowerment, personal growth and life-long learning processes was measured in the FWB group. Thematic analysis revealed students’ fluid and recursive processes of transformative learning in their professional selves and capacities to act in domains important to Aboriginal health. This case study contributes new knowledge critical to strengthening health professional capabilities for ever more complex, uncertain and emotionally demanding sites of practice, and to work in empowering ways—with, not for, Aboriginal people and communities

    Ciguatoxins

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    Ciguatoxins (CTXs), which are responsible for Ciguatera fish poisoning (CFP), are liposoluble toxins produced by microalgae of the genera Gambierdiscus and Fukuyoa. This book presents 18 scientific papers that offer new information and scientific evidence on: (i) CTX occurrence in aquatic environments, with an emphasis on edible aquatic organisms; (ii) analysis methods for the determination of CTXs; (iii) advances in research on CTX-producing organisms; (iv) environmental factors involved in the presence of CTXs; and (v) the assessment of public health risks related to the presence of CTXs, as well as risk management and mitigation strategies

    On Efficient Zero-Knowledge Arguments

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    Endogenous measures for contextualising large-scale social phenomena: a corpus-based method for mediated public discourse

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    This work presents an interdisciplinary methodology for developing endogenous measures of group membership through analysis of pervasive linguistic patterns in public discourse. Focusing on political discourse, this work critiques the conventional approach to the study of political participation, which is premised on decontextualised, exogenous measures to characterise groups. Considering the theoretical and empirical weaknesses of decontextualised approaches to large-scale social phenomena, this work suggests that contextualisation using endogenous measures might provide a complementary perspective to mitigate such weaknesses. This work develops a sociomaterial perspective on political participation in mediated discourse as affiliatory action performed through language. While the affiliatory function of language is often performed consciously (such as statements of identity), this work is concerned with unconscious features (such as patterns in lexis and grammar). This work argues that pervasive patterns in such features that emerge through socialisation are resistant to change and manipulation, and thus might serve as endogenous measures of sociopolitical contexts, and thus of groups. In terms of method, the work takes a corpus-based approach to the analysis of data from the Twitter messaging service whereby patterns in users’ speech are examined statistically in order to trace potential community membership. The method is applied in the US state of Michigan during the second half of 2018—6 November having been the date of midterm (i.e. non-Presidential) elections in the United States. The corpus is assembled from the original posts of 5,889 users, who are nominally geolocalised to 417 municipalities. These users are clustered according to pervasive language features. Comparing the linguistic clusters according to the municipalities they represent finds that there are regular sociodemographic differentials across clusters. This is understood as an indication of social structure, suggesting that endogenous measures derived from pervasive patterns in language may indeed offer a complementary, contextualised perspective on large-scale social phenomena

    Development and characterisation of phage display-derived antibodies for synovial specific drug delivery in Rheumatoid Arthritis

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    OBJECTIVE: Rheumatoid Arthritis (RA) is a systemic autoimmune disease with a prevalence of 0.5-1% worldwide. In the last two decades we observed the rapid evolution of RA therapeutics, and currently, a wide range of anti-rheumatic drugs is available. In particular, the discovery of anti-Tumor Necrosis Factor-α (TNF-α) antibodies has significantly improved disease management. However, the lack of therapeutic efficacy in a significant proportion of patients and the potential systemic implications (e.g. risk of serious infections) still represent an unmet need in RA therapy. Developing novel agents with synovial targeting specificity might increase the therapeutic index while reducing systemic side effects of RA therapeutics. This project aims to design, construct and evaluate in vitro and ex vivo the targeting and anti-inflammatory capability of a novel bispecific tandem single-chain variable fragment (scFv)-Fc antibody combining synovium specificity with anti-TNFα activity. The potential advantage of this construct, called F7-TNF scFv-Fc, is a reduced systemic TNF-binding activity and increased delivery and activation of TNF-neutralizing capacity at the inflamed joints. METHODS: The first part of this work focuses on the optimisation of synovium specific scFv domains previously identified by in vivo phage display using a SCID mouse model transplanted with human synovium, A7 scFv and F7 scFv. The second part of this project moves to the generation and evaluation of a tandem bispecific scFv-Fc antibody comprising F7 scFv external arms with synovium specific targeting ability linked through a metalloproteinase (MMP) cleavable linker to the anti-TNFα variable regions of Adalimumab fused to Fc domain of human IgG1. RESULTS: F7 scFv-Fc and the tandem bispecific F7/TNF scFv-Fc antibodies proved to satisfy the essential properties as joint-targeted agents showing specific reactivity to human arthritic synovium and no reactivity to normal tissues and other chronic diseases. In addition, F7/TNF scFv-Fc demonstrated ex vivo specific synovium targeting ability and intended reduced anti-TNFα activity in its intact form prior to reaching the joint tested by ELISA and NF-Kb reporter cell assay. Human synovial fluid and recombinant human MMP-1 showed efficient cleavage of the external arms of the antibody, releasing anti-TNF scFv-Fc with same TNF inhibitory capacity/potency as Adalimumab. CONCLUSION: Overall, F7-TNF scFv-Fc has the potential of decreasing the anti-TNFα off-site activity and consequently, reduce systemic toxicity while maintaining high on-site activity. Also, the presence of a synovium targeting domain has the advantage of increasing the delivery and retention within the inflamed synovium and possibly increase the therapeutic index of anti-TNF therapeutics
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