Ebolavirus (EBOV) est un filovirus responsable de fièvres hémorragiques virales sévères chez l’humain, qui peuvent être létales dans 90% des cas. L’actuelle épidémie en République Démocratique du Congo et l’ampleur démesurée de l'épidémie de 2014-2016 en Afrique de l’Ouest, qui a causé la mort de plus de 11 000 personnes, ont poussé les agences sanitaires internationales à tester plusieurs approches thérapeutiques afin d’essayer d’endiguer rapidement la propagation virale et de limiter la mortalité liée au virus lors de futures épidémies. Parmi toutes les stratégies testées, le virus recombinant réplicatif rVSV-ZEBOV qui exprime la glycoprotéine de surface d’EBOV, semble offrir la meilleur protection, aussi bien en modèle animaliers que sur le terrain. Avant d’être testé chez l’humain, de nombreuses études ont permis de mettre en évidence l’efficacité et l’innocuité de ce vaccin prototype. Pourtant et malgré le fait que de nombreuses études ont démontré l’importance et le rôle de la glycoprotéine GP dans l’efficacité des vaccins contre ce virus, aucune étude n’a encore été réalisé sur la nature des glycoprotéines virales synthétisées par le gène GP d’EBOV inséré dans le génome du virus VSV. Ainsi, les caractérisations moléculaires des protéines virales produites lors de l’infection par le virus rVSV-GP décrites dans ces travaux de thèse offrent de nouvelles perspectives pour comprendre le succès de ce vaccin mais aussi l’origine virales dans les effets secondaires sévères observés lors de la vaccination, et pourront aider à développer un vaccin plus sûr, qui n’est actuellement pas utilisable chez les personnes immunodéprimées / The filovirus Ebolavirus (EBOV) is the causative agent of severe viral hemorrhagic fevers in humans that can be lethal in 90% of cases. The current outbreak in the Democratic Republic of Congo and the extraordinary scale of the 2014-2016 outbreak in West Africa, that caused the death of more than 11 000 disease victims, lead the international public health agencies to test several therapeutic approach to limit viral spreading and mortality. Amongst those, the recombinant replication-competent rVSV-ZEBOV virus, that expressed EBOV GP glycoprotein, appears to offer the best protection in animal models and outbreak settings. While its effectiveness and safety have been widely investigated before human trials and despite numerous studies that showed the importance the nature of the glycoproteins which are produced during the infection from the EBOV GP gene that has been inserted in VSV genome are unknown. In this respect, the molecular characterisations of the viral glycoproteins synthesised during rVSV-GP presented in this thesis, offer new insights with which to understand the success of the rVSV-GP vaccine but also the potential viral origins of the severe adverse side effects observed during vaccination and could help in developing a safer vaccine, which currently cannot be used in an immunocompromised population

Introduction: Recent Ebola epidemic in West Africa have put viral hemorrhagic fever diseases in the spotlight. Uganda has had several outbreaks throughout the years, which have successfully been managed. Nurses’ patient education plays an important role in the work to increase public awareness about viral hemorrhagic fever diseases. Objectives: To assess how nurses at the emergency department educate the patients and relatives about the viral hemorrhagic fever diseases. Methods: An explorative and descriptive qualitative study with qualitative approach have been used. In-depth interviews with 18 open-ended questions have been conducted with nurses in the emergency department. Data was analyzed by qualitative content analysis and analyzed with Peplau’s theory of interpersonal relationship. Results: Through data analysis four categories were developed; Wide variety of educational techniques, Experienced obstacles, How to attain wider audience and Preferable characteristics as an educating nurse. Nurses used many different approaches when educating about viral hemorrhagic fever diseases, often uniquely combined. Lack of time and too unstable patients in the emergency department were seen as the main obstacles to educate. Methods to reach the community and employing a special education-nurse on the ward were suggestions to improve the preventive work against viral hemorrhagic fever diseases. Conclusion: Nurses are aware of the importance of patient education and use the educational methods they believe to be the most effective. However, patients in the emergency department often come in too late and priority should be on preventive measures. Training the nurses in educational techniques and patient education could be a key in decreasing the risk of coming outbreaks. / Bakgrund: Den senaste ebolaepidemin i västafrika har gjort att blödarsjukdomar hamnat i rampljuset. Uganda har genom åren drabbats av flera utbrott som framgångrikt hanterats. Sjuksköterskors patientutbildning spelar en viktig roll i arbetet för att öka allmänhetens medvetenhet om blödarsjudkomar. Syfte: Att undersöka hur sjuksköterskor på akutmottagningen utbildar patienter och anhöriga om blödarsjukdomar. Metod: En explorativoch deskriptiv kvalitativ studie med kvalitativ ansats användes. Sjuksköterskor som arbetar på akutmottagningen har djupintervjuats med 18 öppna frågor. Data har analyserats med en kvalitativ innehållsanalys och analyserats utifrån Peplaus ”interpersonal relationship theory”. Resultat: Genom dataanalysen utvecklades fyra kategorier; Stor variation på utbildningstekniker, Upplevda hinder, Hur man ska nå en bredare publik och Önskvärda egenskaper som utbildande sjuksköterska. Sjuksköterskorna använder många olika metoder för utbilda patienter och anhöriga om blödarsjukdomar, ofta i unika kombinationer. Tidsbrist och alltför instabila patienter på akutmottagningen sågs som de största hindren för att utbilda. Metoder för att nå allmänheten och att anställa en speciell utbildningssjuksköterska på avdelningen var några av förslagen för att förbättra det förebyggande arbetet mot blödarsjukdomar. Slutsats: Sjuksköterskorna är medvetna om vikten av patientutbildning och använder de pedagogiska metoder de anser vara mest effektiva. Då patienterna kommer till akutmottagningen är det dock ofta för sent och preventiva åtgärder borde prioriteras. Att öka sjuksköterskornas kunskap i utbildningstekniker och patientutbildning kan vara en viktig del i det preventiva arbetet för att minska risken för kommande utbrott.

Tübingen, Univ., Diss., 2008.

Les virus appartenant à l’ordre des Mononegavirales possèdent une « large » protéine L, responsable du cycle réplication/transcription et de maturation des ARNs. Six domaines conservés portent les différentes activités de cette protéine dont le site catalytique d’une activité méthyltransférase (MTase) de la coiffe. La coiffe est une structure chimique constituée d’une guanosine méthylée en position N7 reliée à l’extrémité 5’ des ARNm par une liaison 5’-5’ triphosphate. Une seconde méthylation est également présente en position 2’O du ribose du premier nucléotide de l’extrémité 5’ de l’ARNm. Ces méthylations ont un rôle critique chez les virus car elles permettent la traduction efficace des ARNm mais permettent également aux ARNs viraux d’échapper à leur détection par l’immunité innée de l’hôte. Ainsi, la caractérisation de ce domaine chez le virus Ebola serait un point clé pour une meilleure compréhension de la réplication des filovirus et un pas vers l’élaboration d’une nouvelle stratégie thérapeutique. Nous avons donc produit le domaine MTase du virus Soudan (SUDV) afin de caractériser son activité. Il a été démontré que le domaine C-terminal de la protéine L joue un rôle dans le recrutement de l’ARN, crucial pour l’activité MTase. Nous avons pu identifier une activité A-2’O MTase interne originale. Le domaine MTase de SUDV est également capable de méthyler les positions N7 et 2’O de la coiffe mais une caractérisation plus approfondie est nécessaire. Enfin, nous avons identifié des molécules inhibant l’activité MTase des filovirus. Une analyse biochimique plus poussée permettra d’initier le développement d’une nouvelle stratégie antivirale contre le virus Ebola. / In the Mononegavirales order, viruses encode a large protein (L), which is responsible for replication/transcription and RNA modifications. This protein harbours six conserved domains accountable of these different activities. Among these domains, the conserved region VI (CRVI) has been predicted to support cap-methyltransferase (MTase) activity. The cap consists in a N7-methylated guanosine linked to the first nucleotide at the mRNA 5'-end by a 5'-5' triphosphate bond. This structure can also be methylated at the 2'O position of N1 ribose. These methylations play a critical role in virus life cycle as N7 methylation triggers efficient viral RNA translation and 2'O methylation hampers the detection of viral RNA by the host innate immunity. Thus, the characterization of this domain in Ebola virus is a key point to understand replication of mononegaviruses and design new antiviral strategies. We produced the MTase domain of Sudan ebolavirus (SUDV) to characterize its MTase activity. We demonstrated that the protruding C-terminal domain is essential for MTase activity as this domain is a key for the RNA recognition. Using synthetic short RNAs holding different cap structures, we discovered that SUDV MTase harbours an unconventional A-2’O MTase activity. Besides this, the MTase domain is able to methylate the cap structure at N7 and 2'O positions but further characterization would be necessary to fully understand the cap synthesis. Finally, we identified compounds limiting the Ebola virus MTase activity. Further biochemistry and compounds characterization results will thus pave the way towards the development of an innovative antiviral strategy.

Les propriétés du papier - pompe capillaire, prix et disponibilité - lui permettent de répondre à tout type de contraintes logistiques et économiques d'un diagnostic médical de terrain. En amont de l'application, nous avons étudié les écoulements dans des géométries confinées par des barrières de cire. Une focalisation hydrodynamique associée à l'évaporation, crée un effet de concentrateur atteignant un facteur d'amplification de 1000. La continuité du diagnostic depuis le terrain jusqu'au laboratoire est assurée par des dispositifs hybrides papier-microsystèmes. Dans cette thèse, l'élution d'échantillons séchés dans le papier est quantifiée et sa compatibilité avec différentes fonctions microfluidiques démontrée. Nous nous sommes intéressés à deux applications de diagnostic : la détection d'un biomarqueur cardiaque par immunoessai et celle du virus Ebola par amplification d'acides nucléiques (RT-RPA). Avec un dispositif papier simple et une révélation colorimétrique, la troponine a été détectée jusqu'à une concentration de 1 ng/mL. Le diagnostic précoce de maladies infectieuses est rendu possible par la biologie moléculaire sur papier. Après des développements en laboratoires sur ARN synthétiques, des expériences réalisées en Guinée, sur des échantillons cliniques, à partir de papiers prêts à l'emploi, avec une instrumentation transportable ont atteint une sensibilité à 85.3%. Le multiplexage du diagnostic est obtenu dans des géométries multicouches par la réalisation simultanée de tests et contrôles. Enfin, l'application à d'autres pathogènes comme HIV et la Dengue, a montré les limites du papier et de son environnement biochimique non contrôlé. / Paper properties – such as capillary pump, affordability and availability – made it suitable for medical diagnostics in logistic and economic field constraints. Upstream of application, we studied liquid flows in wax-confined geometries. Hydrodynamic focus coupled to evaporation creates a concentrator effect reaching an amplification factor of 1000. Diagnosis continuity from the point of care to testing laboratory is insured thanks to hybrid paper-microsystem devices. In this thesis, the elution of dried samples in paper is quantified and its compatibility with various microfluidic functions demonstrated. We were interested in two diagnostics application: detection of a cardiac biomarker by immunoassay and of Ebola virus by nucleic acids amplification (RT-RPA). With simple paper devices and a colorimetric signal, Troponin has been detected until a concentration of 1 ng/mL. Early diagnostics of infectious diseases is made possible with molecular biology on paper. After laboratory preliminary developments on synthetic RNA strains, experiments performed in Guinea with clinical samples, from ready to use papers, with carry-on equipments achieved a sensitivity of 85.3%. Multiplexed diagnostics is obtained in multilayered geometries enabling simultaneous tests and controls. Finally, application to other pathogens, like HIV and Dengue, showed paper limits from its uncontrolled bio-chemical environment.

Sierra Leone was heavily affected by the West African Ebola virus disease (EVD) epidemic from 2013 to 2016. Ongoing EVD transmission during the epidemic was connected to several factors including unsafe traditional burial practices. This phenomenological qualitative study addressed Kono members' perceived knowledge, attitudes, and beliefs regarding how burial practices influenced EVD transmission. Rosenstock's health belief model provided the framework for the study. The participants purposefully selected from various religions and professions were interviewed individually and in focus group settings. Similar phrases and comments were identified from the interview responses resulted which resulted in the following 5 main themes: (a) Kono community leaders and public health workers were cognizant of important EVD issues, but there was a knowledge deficit among Konos about EVD and its mode of transmission; (b) although customary burial rituals were temporarily banned from 2014 to 2016, they were practiced among the Konos to promote culture-driven dignity and respect for the dead; (c) many Konos harbored grudges and mistrusted government officials and public health workers; (d) infrastructural deficits were a barrier to health care as private and public sectors lacked training and equipment to mitigate the 2013-2016 EVD outbreak; and (e) participants were willing to adopt safer burial practices if EVD outbreaks were to reemerge. These findings indicated that EVD transmission was connected to unsafe burial practices. Findings may be used to improve community engagement and public health outreach efforts to promote safer burial practices, especially during periods of infectious disease outbreaks.

The research questioned the efficacy of standard biomedical information sharing and communication processes in ensuring rapid and reliable behavioural changes in the control of epidemics, especially in high-context cultures. Information processing arousals and behaviour change motivations are subject to the level of interactions in the extrinsic and intrinsic elements of an information. Following, epidemic control can only be successful if relevant elements of a system’s values, norms, beliefs and practices for information processing are superimposed on scientific communication to create shared meanings. An empirical research approach in grounded theory underscore the data collection of this research with the data analogy utilising the MAXQDA Analytics Pro software. Ebola behavioural changes were identified to be enabled by the functional properties of community mobilisation as a structure and process for meaning making and behavioural motivation. A contextual health communication model dubbed the ecological collegial communication model has been modelled for epidemiological control as the output of the research. Specific to the methodology, a systematic qualitative and data analysis process in grounded theory was adopted for conducting the research and the dissertation writing. Commencing the process was the identification and analysis of the problem from the perspectives of the challenges to the Ebola communication management. This was comprehensively identified from the fundamentals of the process of communication to the communication itself and was assessed from the motivational factors underlying the behaviours within which the rationality of the behaviours could be understood for their inflexibility to change or their insensitivity to the Ebola messages. The mediations of the behavioural motivators in the cognitive processes to information processing were considered for their intrinsic and extrinsic values to arouse information processing and persuade change. To explore the interface between communication and culture in cognitive processes of information processing and decision making, literatures on behavioural theories, including anthropological theories from which the processes and determinants of behavioural enactment are predicted were reviewed in chapters two to four. Intention (also used interchangeably in this dissertation as motivation) was unanimously construed as proximal in determining behaviours in the literatures. However, intention was also construed to have linkages with other factors in the determination of behaviours.:Dedication ii Declaration iii Acknowledgements iv Table of contents v List of figures vi List of photos vii List of matrices vii List of tables vii List of appendices viii Abbreviations ix 1 Communication and culture of the 2014/2015 West Africa Ebola outbreak 1 1.1 Introduction 1 1.2 Conceptualisation of the research problem – the key factors of the Ebola outbreak 4 1.2.1 Structural violence 7 1.2.2 Communication deficiency 10 1.2.3 Cultural models (values and practices 20 1.2.3a Death and funerals 21 1.2.3b Caregiving 26 1.2.3c Reliance on traditional healers 31 1.3 Research objectives 37 1.4 Definitions 38 1.5 Questions formulation and research questions 42 1.6 Justification 52 1.7 Conclusion 58 2 Theoretical frameworks consistent with the 2014/2015 Ebola outbreak health communication approaches – A discourse 59 2.1 Introduction 59 2.2 Psychological/behaviour science models 60 2.2.1 Health belief model 61 2.2.2 Protection motivation theory 64 2.2.3 Theory of planned behavior /reasoned action 71 2.2.4 Social cognitive theory / social learning theory 76 2.3 Summary 79 3 Information processing/communication theories 81 3.1Introduction 81 3.2 Elaboration likelihood model 81 3.3 Activation model 86 3.4 Narrative theory and entertainment education 88 3.5 Summary 95 4 Ecological theories / framework 97 4.1 Introduction 97 4.2 The PEN-3 Model 98 4.2.1 Health education (cultural identity)100 4.2.2 Cultural appropriateness of health behavior (cultural empowerment) 101 4.3 Bioecological theory 103 4.4 Developmental process of Bronfenbrenner’s model in the framework of the 2014/2015 Ebola outbreak 108 4.5 Theoretical framework of this dissertation 119 5 Research process and methodologies 125 5.1 Introduction 125 5.2 Justification of the research methodology 128 5.3 Overview of Monteserrado County 134 5.4 Techniques/procedures 137 5.4.1 Archival materials/documents 138 5.4.2 Ethnographic/observations 139 5.4.3 Key informants/in-depth interviews 142 5.4.4 Focus group discussions 143 5.5 Data analysis 146 5.5.1 Codes 147 5.5.2 Qualitative analysis employed in the research 152 5.6 Role of the researcher 153 5.6.1 Origins of the project 153 5.6.2 The discourse - philosophical worldview 156 5.6.3 Concluding thoughts 157 6 Data analysis: cultural practices, health and communication in the Liberian context 160 6.1 Introduction 160 6.2 Ethnicity and religion 162 6.3 Social organization 171 6.4 Aspects of death and burial practices 179 6.5 Concept of health and health care 186 6.6 Communication and information sharing approach in Liberia 193 6.6.1Traditional communication and the town crier in Liberia 195 6.6.2 Contribution of Crusaders for Peace 201 6.6.3 Development of overarching Ebola communication messages 206 6.7 Conclusion 210 7 Data analysis: Socio-cultural patterns in Ebola perceptions, content of messages and behavioural outcomes 212 7.1 Introduction 212 7.2 Parent codes – summative description and discussions 214 7.3 Understanding the socio-cultural patterns in Ebola knowledge and behaviours: Perceptions of Ebola transmissions 226 7.4 Content and nature of Ebola messages in perceptions and behaviours 237 7.5 Conclusion 276 8 Data analysis: Understanding the motivators of Ebola behaviours – an analytical interrelationships model perspective 278 8.1 Introduction 278 8.2 Patterns of Ebola behaviours 279 8.3 Conclusion 317 9 Decoding: the interface between culture and communication in the Ebola communication management 319 9.1 Introduction 319 9.2 Contextual elements of effective communication – the interface 321 9.3 Cognitive heuristics to “…protect yourself…” 336 9.4 Processes of moderations of “protect yourself” in cognitions 339 9.5 Conclusion 343 10 Theoretical and conceptual inferences from empirical data and framework for a culturally appropriate communication 344 10.1 Introduction 344 10.2 Research questions 344 10.3 Epidemic control: The cultural model framework to persuasive communication for epidemic management 359 10.3.1 The composite conceptual analytical elements of the model 364 10.3.1a Model definition and assumptions 365 10.3.1b The ECCM – the interactive elements of a system 367 10.3.1c Pattern of communication in the ECCM 371 10.3.2 Summary 374 10.4 Processes of how to apply the ECCM 375 10.5 Limitations of the model 382 10.6 Conclusion 383 11 Conclusions and recommendations 385 11.1 Introduction 385 11.2 Key conclusions 385 11.3 Implications 387 11.3.1 Policy framework implications 387 11.3.2 Theoretical implications 390 11.4 Further research 393 11.4.1 Approach to communication 393 11.4.2 Cultural dynamics 396 11.4.3 Health perceptions 398 11.4.4 Ebola orphans and victims 398 11. 5 Research limitations 399 References 401

Master of Science / Department of Biological & Agricultural Engineering / Caterina M. Scoglio / Lisa R. Wilken / Between the years 2014 and 2015, the world experienced a catastrophic outbreak of Ebola virus, which killed over 26,000 people. Several authorities and organizations actively participated in fighting the epidemic. Infectious disease modelers proved to be invaluable towards this goal. This report provides a background on the Ebola epidemic in West Africa and reviews the biological features of the Ebola virus. Moreover, this report applies a new model for Ebola propagation using data collected by the World Health Organization during the span of the outbreak. The model estimates the reproduction number and assesses the role of mitigation strategies in slowing down the progress of the disease. The report also concludes a review of recent advancements in vaccine production against Ebola.

Ebolavirus is responsible for a deadly hemorrhagic fever that has claimed thousands of lives in Africa and could become a global health threat. Because of the danger of infection, novel Ebola research is restricted to BSL-4 laboratories; this slows progress due to both the cost and expertise required to operate these laboratories. The development of a safe surrogate would speed research and reduce risk to researchers. Two highly conserved Ebola gene segments—from the glycoprotein and nucleoprotein genes—were designed with modifications preventing expression while maintaining sequence integrity, spliced into high copy number plasmids, cloned into E.coli, and tested for stability, safety, and potential research applications. The surrogates were stable over 2-3 months, had a negligible mutation rate (<0.165% over the experiment), and were detectable in human blood down to 5.8E3-1.17E4 surrogates/mL. These protocols could be used to safely simulate other pathogens and promote infectious disease treatment and detection research. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2018. / FAU Electronic Theses and Dissertations Collection

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