MIV/VIGS is 'n mediese toestand wat verreikende gevolge vir die individu en sy omgewing inhou. Dit het 'n direkte fisiologiese impak, asook diverse psigososiale en neurologiese gevolge op die persoon (Lowenthal, 1997). Die mees algemene enkele neurologiese komplikasie van VIGS staan bekend as VIGS-demensiekompleks (VDK) (Cournos&Bakalar, 1999). VDK word gekenmerk deur kognitiewe en motoriese disfunksie sowel as gedragsveranderinge en gemoedsversteurings of affekversteurings in verskillende grade (Price, 1996). Ten spyte van die waardevolle literatuur reeds beskikbaar oor die neurologiese, kognitiewe, motoriese en psigologiese impak van VDK, is daar steeds beperkte data oor die gevolglike invloed daarvan op kommunikasievaardighede en die individu se ervaring daarvan (Zuniga, 1999). Om hierdie rede blyk daar 'n gaping te wees in die effektiewe doelgerigte behandeling van hierdie afwyking (Druck, 2002). Die doel van hierdie studie is om ondersoek in te stel na die individu met VDK se subjektiewe siekte-ervaring met betrekking tot moontlike kommunikasiepatologie, sowel as om die rol van die spraak-taalterapeut met betrekking tot individue met VDK te definieer. Daar is van 'n enkelgevallestudie, nie-eksperimentele, beskrywende opnameontwerp gebruik gemaak. Die navorsingsdeelnemer se narratiewe is ontlok deur middel van natuurlike gesprekvoering en 'n oudiobandopname is daarvan gemaak. Die narratief is getranskribeer en die volgende drie metodes is gebruik om dit volledig en omvattend te analiseer: 1.)Plotanalise, 2.) Strukturele storie-analise volgens Labov (1982) se strukturele benadering, en 3.) Holistiese inhoudsanalise. Uit die plotanalise het dit duidelik geword dat die betrokke narratief regessief van aard is. Tydens die bespreking van die meta-narratiewe het verskeie aspekte van hierdie individu met VDK se persoonlike siekte-ervaring na vore gekom wat betref die invloed daarvan op haar vermoë om steeds haar lewenskwaliteit te handhaaf deur aan haar alledaagse lewensaktiwiteite te kan deelneem. Die ICFkomponente is as raamwerk gebruik om die verkreë inligting op 'n sinvolle wyse voor te stel. Dit wil verder voorkom of die siekte-ervaring nie tot spesifieke aspekte van menslike funksionering, onder andere kommunikasiefunksies, beperk kan word nie. Die ervaring van haar kommunikasieprobleme maak deel uit van haar algehele siekte-ervaring wat binne die konteks van haar alledaagse lewensaktiwiteite en deelname daarin plaasvind. Die identifikasie en analise van die temas het dit verder duidelik gemaak dat narratiewe 'n effektiewe metode is om insig te verkry in die individu met VDK se konteks en ervaring daarvan. 'n Geheelbeeld van die individu se funksionele vaardighede, konteks en emosionele ervarings en behoeftes is verkry, sonder om haar bloot te stel aan formele toeste en evalueringsinstrumente wat haar onbevoeg en minderwaardig kan laat voel. Die verkreë resultate impliseer ook dat die individu met VDK se siekte-ervaring sentraal is tot sy of haar mate van deelname aan alledaagse lewensaktiwiteite asook ervaring van kwaliteit van lewe. Die waarde van narratiewe in die terapeutiese behandelingsproses het ook in hierdie studie duidelik geword. Narratiewe kan suksesvol aangewend word om 'n verbeterde insig in die individu met VDK se lewe en behoeftes te verkry. ENGLISH : HIV/AIDS is a medical condition that has far-reaching consequences for the individual and his or her environment. It has a direct physiological impact, as well as diverse psychological and neurological effects on the individual (Lowenthal, 1997). The most common single neurological complication of AIDS is known as AIDS Dementia Complex (ADC) (Cournos&Bakalar, 1999). ADC is associated with cognitive and motor dysfunction as well as behavioural changes and mood disorders of different degrees (Price, 1996). Despite all the valuable literature already available on the neurological, cognitive, motor and psychological impact of ADC, there is still limited data on the consequential influence thereof on the communication skills and the individual's experience and treatment of this (Zuniga, 1999). Therefore there seems to be a gap in the effective goal-directed treatment of this disorder. (Druck, 2002). The aim of this study is to investigate the subjective illness experience of the individual with ADC in terms of possible communication difficulties, and to define the role of the speech-language therapist in this regard. A single case study, non-experimental, descriptive research design was used. The research participant’s narrative was elicited by means of natural conversation and recorded on audio tape. The narrative was transcribed and analysed, using the following three methods: 1.) Plot analysis, 2.) Structural story analysis according to Labov’s (1982) structural approach, and 3.) Holistic content analysis. According to the plot analysis this was a regressive narrative. During the discussion of the meta-narratives, different aspects of the research participant's personal illness experience were identified as regards the influence thereof on her ability to maintain her quality of life through participating in everyday living activities. The ICF components were used as framework to present the results obtained in a meaningful manner. It also became apparent that the illness experience cannot be isolated to specific human functions, such as communication. The experience of her communication problems is part of her complete illness experience, which takes place in the context of her everyday living activities and participation therein. The identification and analysis of themes also indicated that the narrative is an effective method to gain insight into the individual’s context and experience thereof. A holistic view could be obtained of the individual’s functional skills, context and emotional experience and needs, without subjecting her to formal tests and evaluation instruments which might leave her feeling incompetent and inferior. The results obtained imply that the individual with ADC’s illness experience is central to the extent to which he or she participates in everyday life activities as well as to the quality of life experienced. The value of narratives in the therapeutic treatment process also became apparent. Narratives can successfully be applied to gain better insight into the life and needs of the individual living with ADC. / Dissertation (M (Communication Pathology))--University of Pretoria, 2006. / Speech-Language Pathology and Audiology / unrestricted

The prevalence of HIV/AIDS is increasing with immense influence and pressure on the grandparents as primary caregivers and the main supporters of orphans in families. The grandparents are faced with biophysical, socio–economic and psychosocially challenges and lack of support from the community of Koster in the North–West Province of South Africa. Caring for AIDS orphans who have lost their parents due to AIDS and of whom some are infected is an enormous challenge. There should be a partnership between the grandparents, the community, public sector and private sector to assist the grandparents who care for AIDS orphans. Therefore, there are recommendations made by the researcher as tools for health–care workers in order to support the grandparents as they care for AIDS orphans. The objectives of this research were to explore and describe challenges faced by the grandparents in caring for AIDS orphans in Koster, and how their needs could be met by making recommendations for effective support of the grandparents. A qualitative, explorative and descriptive design was used which enabled the research to understand the challenges faced by grandparents in caring for AIDS orphans and how their needs could be met. Semi–structured interviews were conducted to obtain the data. The population studied in this research consisted of the grandparents caring for AIDS orphans in Koster in the North–West Province, South Africa. Voluntary purposive sampling was used to select participants with the assistance of mediators who are working for the Non– Government Organizations dealing with HIV and AIDS in Koster. The sample size was determined by data saturation, which was reached after 15 interviews. Data analysis was carried out simultaneously with data collection. In consensus discussions, the researcher and the co–coder reached consensus on the main, sub and further sub–categories. From the research findings, four main categories were identified namely; the challenges faced by the grandparents, perceptions of the grandparents on how their need could be met, the impact of the challenge and coping mechanisms. It could be concluded that the grandparents are faced with diverse challenges in caring for AIDS orphans. In order to address these challenges the community and government must be fully involved. The basic, psychosocial, socio–economic and biophysical needs can be addressed through support system. / Thesis (M.Cur.)--North-West University, Potchefstroom Campus, 2011.

The prevalence of HIV/AIDS is increasing with immense influence and pressure on the grandparents as primary caregivers and the main supporters of orphans in families. The grandparents are faced with biophysical, socio–economic and psychosocially challenges and lack of support from the community of Koster in the North–West Province of South Africa. Caring for AIDS orphans who have lost their parents due to AIDS and of whom some are infected is an enormous challenge. There should be a partnership between the grandparents, the community, public sector and private sector to assist the grandparents who care for AIDS orphans. Therefore, there are recommendations made by the researcher as tools for health–care workers in order to support the grandparents as they care for AIDS orphans. The objectives of this research were to explore and describe challenges faced by the grandparents in caring for AIDS orphans in Koster, and how their needs could be met by making recommendations for effective support of the grandparents. A qualitative, explorative and descriptive design was used which enabled the research to understand the challenges faced by grandparents in caring for AIDS orphans and how their needs could be met. Semi–structured interviews were conducted to obtain the data. The population studied in this research consisted of the grandparents caring for AIDS orphans in Koster in the North–West Province, South Africa. Voluntary purposive sampling was used to select participants with the assistance of mediators who are working for the Non– Government Organizations dealing with HIV and AIDS in Koster. The sample size was determined by data saturation, which was reached after 15 interviews. Data analysis was carried out simultaneously with data collection. In consensus discussions, the researcher and the co–coder reached consensus on the main, sub and further sub–categories. From the research findings, four main categories were identified namely; the challenges faced by the grandparents, perceptions of the grandparents on how their need could be met, the impact of the challenge and coping mechanisms. It could be concluded that the grandparents are faced with diverse challenges in caring for AIDS orphans. In order to address these challenges the community and government must be fully involved. The basic, psychosocial, socio–economic and biophysical needs can be addressed through support system. / Thesis (M.Cur.)--North-West University, Potchefstroom Campus, 2011.

This research is aimed at the needs of counsellors working with children infected and affected by HIV/AIDS. A lack of sufficient guidelines for this field has been identified in the relevant literature. This shortage has been confirmed by experienced counsellors working with children. To reach the required goal a number of objectives were set. This included setting up a theoretical framework by way of a literature study as well as consultation with experts in the field; studying the developmental needs of children in the mid childhood phase and how these needs are influenced by HIV/AIDS; the need for guidance of children affected by AIDS as well as the role that counsellors can play in this regard. An empirical study was undertaken during which focus group interviews were held with a group of lay counsellors as well as a group of trained counsellors. Ten participants took part in the focus groups. Applied research was undertaken as the researcher aimed at establishing the true needs of counsellors working in the field. The empirical data showed the following: Ø Counsellors feel that they have a specific role to play during the therapy with AIDS-affected children. Ø Counsellors have definite views on the needs of children affected by AIDS. Ø Counsellors have listed specific skills and training needed by therapists working with these children. Ø Counsellors have listed definite shortcomings and needs that they experience in their work with AIDS-affected children. The basic need for the establishing of a protocol to be used in guidance of children infected and affected by AIDS was one of the key findings of the study. / Dissertation (MSD (Play Therapy))--University of Pretoria, 2005. / Social Work and Criminology / Unrestricted

Los cítricos son el cultivo frutal económicamente más importante tanto en España como en el resto de los países productores. La clave para mantener la competitividad de este sector consiste en obtener material vegetal de alta calidad, para lo cual son indispensables los programas de mejora. La mejora de cítricos por métodos clásicos es muy complicada, por lo que hay que recurrir a las nuevas tecnologías para intentar acelerar y optimizar el procedimiento. La reciente secuenciación del genoma de dos especies de cítricos ha permitido identificar una larga lista de genes candidatos a participar en determinados procesos biológicos. Sin embargo, son necesarios nuevos análisis para asociar cada gen a un fenotipo específico o función biológica. El empleo de vectores virales para determinar la función de genes mediante silenciamiento génico inducido por virus (VIGS) ha demostrado ser una herramienta muy útil para los estudios de genética reversa realizados en plantas. Este sistema presenta ventajas respecto a los métodos tradicionales para estudiar la función de genes como son la mutagénesis o la transformación genética, ya que permite ensayar la función de numerosos genes en un corto periodo de tiempo. Esto es especialmente crítico en el caso de los cítricos, que poseen largos periodos juveniles de entre 6 y 8 años y donde la transformación de plantas adultas es muy difícil. Además, permite estudiar la función de genes que son esenciales para el crecimiento o el desarrollo de la planta y cuyo análisis es inviable con los métodos tradicionales. Al comienzo de la tesis se había desarrollado un vector viral para cítricos basado en el virus de la tristeza de los cítricos (CTV) con el que se pueden expresar proteínas pero que no se ha ensayado para estudiar la función de genes mediante VIGS. En el laboratorio disponíamos de un clon infeccioso de cDNA del genoma completo del virus del manchado foliar de los cítricos (CLBV), un virus que infecta a todas las especies y variedades de cítricos ensayadas y es asintomático en la mayoría de ellas. Este clon infeccioso se ha modificado para obtener vectores virales basados en el genoma de CLBV que pueden servir tanto para expresar proteínas como para silenciar mediante VIGS genes de cítricos para la mejora genética de este cultivo. Para ello, se ha introducido un punto de corte único PmlI en dos zonas del genoma de CLBV: en el extremo 3¿ no traducible (vector clbv3¿) o en la zona intergénica localizada entre los genes de las proteínas de movimiento y cápsida (CP) (vector clbvIN). Para la expresión de secuencias foráneas mediante la formación de un nuevo RNA subgenómico (sgRNA) se delimitó la secuencia mínima promotora del sgRNA CP mediante clonación de fragmentos de distinta longitud en torno al origen de transcripción de dicho sgRNA en el vector clbv3'. El fragmento de 92 bases localizado entre los nt -42 y +50 respecto al inicio de transcripción del sgRNA CP contenía todos los elementos necesarios para la promoción de un nuevo sgRNA in vivo. Esta secuencia mínima promotora se clonó en los 2 vectores virales previamente desarrollados para generar los vectores clbv3¿pr y clbvINpr, respectivamente. Ambos vectores fueron capaces de producir un nuevo sgRNA y de expresar proteínas recombinantes. Para determinar la estabilidad de los vectores obtenidos se clonaron en ellos fragmentos de secuencias lineales de distinto tamaño, o en tándem invertido para la formación de una estructura en horquilla, y se inocularon en plantas de N. benthamiana y cítricos. Todas las construcciones derivadas del vector clbv3' se mostraron estables a lo largo de las diferentes brotaciones analizadas durante al menos 3 años, comprobándose la replicación viral e integridad del inserto. Sin embargo, no se detectó multiplicación viral con ninguna de las construcciones derivadas del vector clbvIN. La estabilidad de las construcciones derivadas de los vectores con el promotor duplicado dependía del tamaño del inserto. Con todas ellas se detectó replicación viral pero se observaron eventos de recombinación cuando se clonaban fragmentos superiores a 720 nt en el vector clbvINpr o 408 nt en el vector clbv3'pr. Un factor importante para determinar la eficiencia y funcionalidad de los vectores desarrollados es conocer cómo se mueve y se distribuye el virus en los distintos tejidos de la planta. Para ello se inocularon plantas de N. benthamiana y cítricos con la construcción clbv3¿pr-GFP, que expresa GFP en los tejidos donde se localiza el virus. En N. benthamiana, la observación de GFP permitió detectar la presencia de CLBV en la mayoría de tejidos, acumulándose preferentemente en óvulos y regiones meristemáticas. En cítricos no se pudo visualizar GFP pero el virus se detectó en regiones meristemáticas mediante RT-PCR a tiempo real e hibridación molecular. La acumulación de CLBV en tejidos meristemáticos explicaría la dificultad de eliminar este virus mediante microinjerto. Para evaluar la capacidad de los vectores clbv3'pr y clbvINpr para expresar proteínas se clonó en ellos la secuencia completa del gen gfp y se cuantificó la cantidad de proteína GFP sintetizada en las plantas infectadas. En N. benthamiana la cantidad de GFP estimada para el vector clbv3'pr fue de 16 µg de proteína por gramo de peso fresco, cantidad que resultó entre 5 y 6 veces superior a la estimada para el vector clbvINpr. Sin embargo, en cítricos, debido a la inestabilidad del vector clbv3'pr, sólo se pudo cuantificar la proteína expresada por la construcción del vector clbvINpr, estimándose en 0.6 µg de GFP por gramo de peso fresco. La efectividad de los vectores clbv3', clbv3'pr y clbvINpr para silenciar genes mediante VIGS se ensayó clonando fragmentos de genes tanto endógenos de plantas (pds, actina, sulfur) como el gen gfp introducido experimentalmente en plantas transgénicas. En cítricos todas las construcciones de los tres vectores indujeron fenotipo de silenciamiento del gen ensayado, aunque el vector clbv3' fue el más efectivo para el estudio de VIGS en este huésped. Sin embargo, en N. benthamiana sólo se desencadenó el silenciamiento en las plantas inoculadas con la construcción clbv3¿pr-hp58PDS, que expresa una horquilla de doble cadena de un fragmento de 58 nt del gen pds. En todas las plantas silenciadas se detectó una disminución del correspondiente mRNA del gen ensayado y una acumulación de siRNAs derivados tanto del mRNA del gen insertado como del RNA genómico del virus. Por otro lado, el fenotipo de silenciamiento de los genes ensayados se observó en sucesivas brotaciones, lo que confirma la gran estabilidad de los vectores basados en el genoma de CLBV. Los vectores virales desarrollados en esta tesis constituyen una herramienta eficiente para el estudio de la función de genes mediante genética reversa utilizando la técnica VIGS. También pueden ser útiles para estudio de genética directa mediante expresión de proteínas o para la protección del cultivo frente a enfermedades producidas por virus, bacterias y hongos o frente a plagas de invertebrados. / Agüero González, J. (2013). Desarrollo de vectores virales basados en el virus del manchado foliar de los cítricos (CLBV) [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/34342 / TESIS

Made available in DSpace on 2015-03-26T13:42:06Z (GMT). No. of bitstreams: 1 texto completo.pdf: 737365 bytes, checksum: 3b9e993a4f8a367d7f1b82a2dded22f8 (MD5) Previous issue date: 2009-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Begomoviruses belong to the Geminiviridae family which includes viruses with genomes containing one or two single stranded DNA molecules within icosahedral geminate particles. Begomoviruses are transmitted by the whitefly Bemisia tabaci and may cause diseases of economical importance in several crops mainly in tropical and subtropical regions. In Brazil there are several reports of begomovirus causing serious losses to the common bean and tomato crops, and sporadic reports of infection of soybean. Virus-induced gene silencing (VIGS) is an attractive and fast alternative for studying the expression and function of genes because it does not require plant transformation. Viruses with RNA or DNA genomes have been successfully used as vectors to induce gene silencing in plants. However, there are few examples of efficient vectors for inoculation of tomato and soybean. The main goal of this work was to build a viral vector based on the DNA-A of the begomovirus Tomato rugose mosaic virus (ToRMV) to induce gene silencing in plants of soybean, tomato and Nicotiana benthamiana. The viral vector, designated pToR-A1.4ΔCP, was built from an infectious ToRMV-A clone from which the gene encoding the capsid protein was removed and replaced by the multiple cloning site of the plasmid vector pBluescript KS+ (pKS+). Like other begomoviruses, ToRMV does not require the capsid protein to cause a systemic infection. The construction of the viral vector was confirmed by PCR, enzymatic cleavage and DNA sequencing. To be used on gene silencing experiments, fragments from the following genes were PCR amplified from cDNA clones: phytoene desaturase (PDS) from soybean and tomato, myo-inositol-1-phosphate synthase (MIPS) from soybean and stachyose synthase (STS) from soybean. All fragments were amplified with primers contained restriction sites for cloning. After amplification, the fragments were cloned in pGEM-T Easy vector, sequenced and aligned to the corresponding cDNAs to confirm their identity. To clone these fragments in the viral vector they were cleaved out of the pGEM-T Easy vector, purified and cloned into the pToR-A1.4ΔCP vector previously cleaved at the multiple cloning site with the same enzymes used to cleave the plasmid vector. The ligation reaction used T4 DNA ligase and ultracompetent Escherichia coli cells were transformed by heat shock. Transformants colonies were analyzed by PCR and enzymatic cleaved. Systemic infection of soybean, tomato and Nicotiana benthamiana plants with the empty pToR-A1.4ΔCP vector co-inoculated with ToRMV DNA-B was checked by PCR 21 days after inoculation. In the case of N. benthamiana, the viral vector presented 82% infectivity, indicating the high potential of this vector for VIGS studies in this plant species. In the case of soybean and tomato, the viral vector presented low replication efficiency. New infectivity tests need to be done to confirm these results. It is expected that this viral vector will represent an alternative for functional genomic studies and for the analysis of genes involved in several biological processes. / Os begomovírus pertencem à família Geminiviridae, que inclui vírus com genoma composto por uma ou duas moléculas de DNA circular de fita simples, encapsidado em partículas icosaédricas geminadas. Os begomovírus são transmitidos pela mosca branca Bemisia tabaci e causam doenças de importância econômica em diversas culturas, principalmente em regiões tropicais e subtropicais. No Brasil, há diversos relatos de begomovírus causando sérias perdas nas culturas do feijoeiro e tomateiro e relatos esporádicos de incidência na cultura da soja. O silenciamento gênico induzido por vírus (VIGS) é uma alternativa atraente e rápida para o estudo da expressão e função de genes, pois não há necessidade de transformação genética da planta. Vírus com genoma de RNA e de DNA têm sido utilizados com sucesso como vetores para indução de silenciamento gênico em plantas. Entretanto, existem poucos vetores eficientes para a inoculação em plantas de tomate e de soja. O objetivo deste trabalho foi a construção de um vetor viral, baseado no DNA-A do begomovírus Tomato rugose mosaic virus (ToRMV), para a indução do silenciamento de genes em plantas de soja, tomate e Nicotiana benthamiana. O vetor viral, denominado pToR-A1.4ΔCP, foi construído a partir de um clone infeccioso do ToRMV-A do qual foi removido o gene da proteína capsidial, substituindo pelo sítio múltiplo de clonagem do vetor pBluescript KS+ (pKS+). Assim como outros begomovírus, o ToRMV não requer a proteína capsidial para causar infecção sistêmica. A construção do vetor viral foi confirmada por PCR, clivagem enzimática e sequenciamento. Para serem utilizados em experimentos de silenciamento gênico, fragmentos referentes aos genes fitoeno dessaturase (PDS) de soja e tomate, myo-inositol- 1-fosfato sintase (MIPS) de soja e estaquiose sintase (STS) de soja foram isolados a partir de cDNA dessas plantas, utilizando primers específicos, contendo sítios de restrição adequados para as clonagens. Após a amplificação, todos os fragmentos obtidos foram clonados em pGEM-T Easy, sequenciados e sua identificação foi confirmada por meio de alinhamento utilizando o algoritmo BLASTn. Para a clonagem no vetor viral, os fragmentos foram liberados do vetor pGEM-T Easy com as enzimas de restrição adequadas, purificados e inseridos no vetor pToR-A1.4ΔCP, previamente clivado em seu sítio múltiplo de clonagem com as mesmas enzimas. A reação de ligação foi realizada utilizando T4 DNA ligase e células de Escherichia coli ultracompetentes foram transformadas por meio de choque térmico. Os transformantes foram analisados por PCR e reação de clivagem enzimática. A infecção sistêmica de plantas de soja, tomate e Nicotiana benthamiana pelo vetor pToR-A1.4ΔCP vazio, inoculado conjuntamente com o DNA-B do ToRMV, foi diagnosticada via PCR aos 21 dias após a inoculação. Em N. benthamiana, o vetor viral apresentou 82% de infectividade, indicando um grande potencial de uso em estudos de VIGS nessa planta. Já em plantas de soja e tomate, o vetor viral apresentou baixa taxa de replicação. Portanto, novo teste de infectividade deve ser realizado para confirmar os resultados obtidos. Dessa forma, espera-se que este vetor viral sirva como um complemento e uma alternativa em estudos de genômica funcional e na análise de genes envolvidos em vários processos biológicos.

AFRIKAANS: Die doel van hierdie studie was om te beskryf op welke wyse ek, in samewerking met die deelnemers aan my studie, ‘n MIV&VIGS-skoolplan ontwikkel het. Die behoefte aan ‘n gestruktureerde MIV&VIGS-skoolplan het ontstaan tydens ‘n omvattende navorsingsprojek, waarvan hierdie studie deel uitmaak en waartydens laerskoolonderwysers die behoefte uitgespreek het om MIV&VIGS-geïnfekteerde en -geaffekteerde kinders beter te ondersteun. Ek het my studie vanuit ‘n interpretivistiese benadering onderneem en my navorsingsproses verdeel in drie fases, wat onderlê is deur deelnemende aksienavorsingsbeginsels. ‘n Gevallestudie is as navorsingsontwerp gebruik. Agt laerskoolonderwysers, in ‘n informele nedersettingsgemeenskap binne die Nelson Mandela Metropool, is by wyse van gerieflikheid geselekteer. Deur die verloop van hierdie studie het ek ‘n multi-metodiese benadering geïmplementeer. Gedurende Fase 1 het ek eerstens relevante verwante studies binne die omvattende navorsingsprojek tematies ge-analiseer, om deelnemers se idees en behoeftes betreffende ‘n MIV&VIGS-skoolplan te identifiseer. Tweedens het ek gedurende ‘n eerste veldbesoek twee fokusgroepbesprekings gefasiliteer, om te bepaal wat deelnemers se bestaande kennis met betrekking tot die Departement van Onderwys se Nasionale MIV&VIGS-Beleid was; wat hulle onderliggende rasionaal vir die ontwikkeling van die MIV&VIGS-skoolplan was; wat die moontlike inhoud van ‘n MIV&VIGS-skoolplan sou behels; asook hoe en deur wie die MIV&VIGS-skoolplan aangewend en benut sou kon word. Hierna het ek die rou data getranskribeer en op tematiese wyse ge-analiseer en geïnterpreteer. Ek het voortdurend gebruik gemaak van observasies, wat vasgelê is in die vorm van veldnotas, asook foto’s. Vir Fase 2 het ek tydens ‘n tweede veldbesoek ‘n deelnemende aksienavorsingswerkswinkel gefasiliteer. Gedurende hierdie werkswinkel is die inhoud van die MIV&VIGS-skoolplan verfyn en gefinaliseer. Vervolgens het Fase 3 die dokumentering van die MIV&VIGS-skoolplan behels, in die vorm van ‘n formele dokument en ‘n plakkaat. Tydens my derde veldbesoek, met die bekendstelling van die skool as Resource and Support Centre in die gemeenskap, is die MIV&VIGS-skoolplan in tweeledige vorm (plakkaat en formele dokument) aan die skoolhoof oorhandig. Die gebruik van ‘n navorsingsjoernaal het my in staat gestel om deurgaans my persoonlike opinies, reflektiewe gedagtes en indrukke van my navorsingstudie aan te teken. Tydens data-analise het ek drie temas geïdentifiseer. Eerstens is die onderliggende rasionaal vir ‘n MIV&VIGS-skoolplan bepaal, waar verbandhoudende subtemas aangedui het dat onderwysers hulle kennis en vaardighede wou oordra na ander skole in die omgewing ter ondersteuning van die breër gemeenskap; dat die oordrag van kennis en vaardighede om geïnfekteerde en geaffekteerde kinders in die klaskamer meer effektief te ondersteun ’n behoefte was; en dat die behoefte verder bestaan het om kennis en vaardighede te dokumenteer in die vorm van ‘n MIV&VIGS-skoolplan. Tweedens is die deelnemers se verwagtinge met betrekking tot ‘n MIV&VIGS-skoolplan gepeil, waar moontlike fundamentele beginsels vir ‘n MIV&VIGS-skoolplan en die implementering van ‘n MIV&VIGS-skoolplan, as subtemas geïdentifiseer is. Derdens is die moontlike inhoud van ‘n MIV&VIGS-skoolplan bepaal. Die subtemas wat voorgekom het, was die identifisering en verwysing van geïnfekteerde en geaffekteerde kinders, asook ondersteuning aan hierdie kinders. Op grond van die bevindinge van my studie kan die gevolgtrekking gemaak word dat deelnemende aksienavorsing ‘n moontlike wyse is waarop ‘n MIV&VIGS-skoolplan, in samewerking met onderwysers, ontwikkel kan word. ENGLISH: The purpose of this study was to describe the manner in which I, in collaboration with the participants of the study, developed an HIV&AIDS school plan. The need for a structured HIV&AIDS school plan stemmed from a broad research project of which this study forms part and during which the participating educators expressed the need to support HIV&AIDS infected and affected children more efficiently. I approached the study from an interpretivist perspective, underpinned by action research principles, and divided the research process into three phases. I implemented a case study as research design and selected eight primary school teachers in an informal settlement community in the Nelson Mandela Metropole, by means of convenience sampling. Throughout this study, I followed a multi-methodical approach. During Phase 1, I conducted an analysis of the transcripts of related studies within the broad research project, in order to identify the participants’ needs and ideas, concerning an HIV&AIDS school plan. Secondly, I facilitated two focus group discussions during a first field visit, in an attempt to determine the teachers’ existing knowledge concerning the Department of Education’s National HIV&AIDS Policy; what the underlying rational for the development of an HIV&AIDS school plan could be; what the possible content of such a plan could entail; and how and by whom such a plan would be utilised and used in the classroom. I transcribed the focus group discussions and thematically analysed and interpreted the raw data. Throughout, I made use of observations, captured in the form of field notes and photographs. For Phase 2, I facilitated a participatory action research workshop during a second field visit. During this workshop we finalised the content of the school’s HIV&AIDS plan. The third phase entailed the documentation of the HIV&AIDS school plan, in the form of a formal document and a poster. During my third field visit, at the launch of the school as a Resource and Support Centre in the community, I presented the schools’ principal with the HIV&AIDS school plan, in the form of the formal document and poster. I continuously relied on a research journal to document my personal opinions, reflective thoughts and impressions of the study. Subsequent to the data analysis I completed, three themes emerged. Firstly, the underlying rational for an HIV&AIDS school plan were determined, with sub-themes indicating that educators wanted to transfer their knowledge and skills to other schools in the area to support the wider community; that teachers wanted to transfer their knowledge and skills to support infected and affected children more effectively, and that the need existed for knowledge and skills to be documented in the form of an HIV&AIDS school plan. Secondly, the participants’ expectations of an HIV&AIDS school plan were determined, with the fundamental principles of an HIV&AIDS school plan and the implementation of the plan as related sub-themes. The third main theme indicated the possible content of an HIV&AIDS school plan. This theme comprises sub-themes relating to the identification, referral and support of infected and affected children. Based on the findings of my study, I can conclude that participatory action research might be used to develop an HIV&AIDS school plan, in collaboration with teachers. / Dissertation (MEd)--University of Pretoria, 2010. / Educational Psychology / unrestricted

During the past decade, South Africa has become the country with the highest number of HIV/AIDS infections in the world. In the wake of the widely spread HIV/AIDS epidemic in South Africa, hundreds of thousands of children are left parentless. Since limited research exists on the psycho-educational needs of children orphaned by AIDS in South Africa, the focus of this study was on their psycho-educational development. A literature study was conducted to investigate the HIV/AIDS epidemic in South Africa and its consequences for the psycho-educational development of AIDS orphans. A qualitative investigation was undertaken and interviews were conducted with caregivers of AIDS orphans at two care centres in rural KwaZulu-Natal. The aim of the investigation was to identify the psycho-educational needs of AIDS orphans as perceived by their caregivers. Moreover, the investigation also aimed at identifying factors that impede the psycho-educational development of AIDS orphans, and to determine the extent to which these day care centres meet the psycho-educational needs of these orphans. The following psycho-educational needs of AIDS orphans emerged from the investigation: - the need to socialise and communicate - the need to cope with parental death - the need for acceptance - the need for security and care - the need for love and belonging - the need to deal with negative emotions Physical and material needs were added to the above needs, since these were frequently mentioned during the course of the interviews. Factors that seriously impeded the psycho-educational development of AIDS orphans were abuse and neglect. The investigation also revealed that the care centres were to a large extent able to meet a broad spectrum of these orphans’ psycho-educational needs, especially at the care centre where the AIDS orphans were able to sleep over and received the full-time attention caregivers. Since the extended family system can no longer fully carry the burden of hundreds of thousands of AIDS orphans in South Africa, care centres situated in the affected communities can offer a viable alternative to traditional models of care. The study emphasised the need for further research on the needs of AIDS orphans as well as increased support of AIDS orphans on local, provincial and national levels. / MEd (Learner support), North-West University, Potchefstroom Campus, 2012

During the past decade, South Africa has become the country with the highest number of HIV/AIDS infections in the world. In the wake of the widely spread HIV/AIDS epidemic in South Africa, hundreds of thousands of children are left parentless. Since limited research exists on the psycho-educational needs of children orphaned by AIDS in South Africa, the focus of this study was on their psycho-educational development. A literature study was conducted to investigate the HIV/AIDS epidemic in South Africa and its consequences for the psycho-educational development of AIDS orphans. A qualitative investigation was undertaken and interviews were conducted with caregivers of AIDS orphans at two care centres in rural KwaZulu-Natal. The aim of the investigation was to identify the psycho-educational needs of AIDS orphans as perceived by their caregivers. Moreover, the investigation also aimed at identifying factors that impede the psycho-educational development of AIDS orphans, and to determine the extent to which these day care centres meet the psycho-educational needs of these orphans. The following psycho-educational needs of AIDS orphans emerged from the investigation: - the need to socialise and communicate - the need to cope with parental death - the need for acceptance - the need for security and care - the need for love and belonging - the need to deal with negative emotions Physical and material needs were added to the above needs, since these were frequently mentioned during the course of the interviews. Factors that seriously impeded the psycho-educational development of AIDS orphans were abuse and neglect. The investigation also revealed that the care centres were to a large extent able to meet a broad spectrum of these orphans’ psycho-educational needs, especially at the care centre where the AIDS orphans were able to sleep over and received the full-time attention caregivers. Since the extended family system can no longer fully carry the burden of hundreds of thousands of AIDS orphans in South Africa, care centres situated in the affected communities can offer a viable alternative to traditional models of care. The study emphasised the need for further research on the needs of AIDS orphans as well as increased support of AIDS orphans on local, provincial and national levels. / MEd (Learner support), North-West University, Potchefstroom Campus, 2012

Résumé : Pour se propager dans les cellules de son hôte et évader les réponses immunitaires, les virus végétaux ont développé plusieurs stratégies de défense. Ici, nous avons investigué les structures génétiques du Apple stem pitting virus (ASPV). Nous avons aussi étudié la diversité moléculaire des isolats d’ASPV provenant des poires en regardant les séquences des gènes CP et TGB afin de mieux comprendre les mécanismes évolutionnaires utilisés par ASPV. Nos études ont démontré que les mutations, incluant les insertions et les délétions, la sélection purificatrice et la recombinaison furent des facteurs importants dans l’évolution du l’ASPV en Chine et possiblement mondialement. Comme tous les virus végétaux, l’ASPV se défend contre le RNA silencing de l’hôte grâce à un suppresseur de RNA silencing (VSR) et nous avons montré que le VSR de l’ASPV est la protéine de capside (CP) du virus. Nous avons aussi établi que la diversité moléculaire cause non seulement une variété de symptômes chez son hôte, Nicotiana occidentalis. Cependant elle cause aussi de la variabilité antigénique chez différents isolats, ce qui mène à des écarts de réactivité sérologique entre isolats. Les plantes ont développé plusieurs stratégies pour se défendre contre les virus. Ici, nous avons étudié comment la plante Arabidopsis se défend contre le Tobacco rattle virus (TRV) via le RNA silencing. Nous avons constaté que les phénomènes de susceptibilité, récupération et virus induced gene silencing (VIGS) sont des mécanismes séparables. Nous avons démontré que les protéines AGO2 et AGO4 sont nécessaires à la susceptibilité initiale au TRV, tandis qu’AGO1 est importante pour les VIGS, tandis que la récupération est médiée par d’autres acteurs qui n’ont pas encore été identifiés. Nos résultats suggèrent l’existence de complexes distincts ciblant différentes populations d’ARN viral et cellulaire. De plus, nous avons montré que la répression de la traduction est un mécanisme important durant la récupération de la plante suite à une infection virale, et que les complexes de décoiffage et de RNA processing jouent des rôles importants dans la dégradation des ARNs viraux. Finalement, nous avons montré que les plantes ayant une mutation dans le gène DCP2 présentent un niveaux de VIGS accrue, ainsi qu’une augmentation des niveaux d’ARN viral. Puisque DCP2 fait partie des complexes de décoiffage qui se trouvent dans des granules spécialisés nommés processing bodies (PBs), cela suggère que les PBs jouent un rôle important dans l’élimination les virus. / Abstract : To live in host cells or to escape from host immunity, plant viruses involved a series of defense strategies. Here we investigated Apple stem pitting virus (ASPV) population structures and molecular diversity of ASPV pear isolates based on its function important gene CP and TGB in China, so as to infer the evolution mechanisms of ASPV. Our study showed that mutations (including insertions or deletions), purifying selection, and recombination were important factors driving ASPV evolutions in China or maybe even in the world. And also ASPV defends against it hosts by encoding a VSR. We also showed that ASPV molecular diversity not only induced different biological properties on its herbaceous host N. occidentails but also resulted in antigenic variation of different ASPV CP isolates, which leaded to differences in serological reactivity among rCPs of different ASPV isolates. Plants have developed a series of mechanisms to defend themselves against viruses. Here we how Arabidopsis defend against. We show that virus susceptibility, recovery, and virus induced gene silencing (VIGS) appear to be separable phenomena, with AGO2 and AGO4 playing important roles in the initial susceptibility to TRV, AGO1 playing an important role in VIGS, and as yet unidentifid players mediating recovery. These results suggest the existence of distinct RNA-induced silencing complexes that target different RNA populations within the cell and over time. Furthermore, we showed that translational repression of viral RNA is likely to play an important role in virus recovery and that decapping function plays an important role in clearing viral RNA from the cell. We also showed that a decapping mutant (DCP2) displayed an increased VIGS and virus RNA accumulation, an important role for PBs in eliminating viral RNA.