Few studies have assessed the optimum second line highly active anti-retroviral therapy (HAART) regimen in patients who had failed on the first-line HAART in resource-limited settings. This study aimed to compare the Protease inhibitor (PI)-based second line HAART regimens used in one clinic in Trinidad by comparing immunological, virological and clinical outcomes of patients on the different second line HAART regimens. The records of 35 treatment-experienced patients, over 21years of age and on PI-based regimens for at least six months, were analysed using SPSS version 20. The regimen containing TDF/FTC/AZT/LPV/r proved to produce superior outcomes compared to the other second line regimens. Due the small number of usable patients’ records, the findings cannot be generalised but indicate directions for future studies attempting to compare the treatment outcomes of different second line HAART regimens / Health Studies / M. A. (Public Health)

El uso de la terapia HAART (Highly-Active-Antiretroviral-Therapy), puede dar lugar a múltiples efectos secundarios. El más frecuente de ellos es el denominado HALS (“HIV-infection, HAARTtreatment- associated-lipodystrophy-syndrome”) que comprende alteraciones como la lipoatrofia periferica; un aumento de tejido adiposo visceral o la lipomatosis del tejido adiposo. Además, estas alteraciones fisiológicas pueden ir acompañadas de alteraciones metabólicas. Para saber la aportación individual de algunos fármacos al desarrollo del síndrome lipodistrófico HALS se realizó un estudio de los efectos de Efavirenz, Nevirapina (NNRTIs) y Kaletra (PI) sobre adipocitos primarios de linaje blanco en cultivo. Efavirenz, que no era considerado un fármaco asociado al síndrome lipodistrófico, es capaz de inhibir la adipogénesis con mayor potencia que Kaletra y Nevirapina. Ninguno de estos fármacos provoca toxicidad mitocondrial por lo que sus efectos ocurren en ausencia de toxicidad mitocondrial. Tanto efavirenz como kaletra reducen la secreción de adipoquinas y aumentan la expresión y secreción de citoquinas relacionadas con la inflamación, pero estos efectos siempre son mayores con el EFV mientras la Nevirapina parece no afectar a este tipo de secreciónes. Para estudiar las características moleculares de los diferentes depósitos de tejido adiposo con comportamiento opuesto como el tejido adiposo subcutáneo lipoatrófico y el tejido adiposo visceral lipohipertrófico o el tejido adiposo lipomatoso de las “buffalo-hump” de los pacientes se han comparado características moleculares de ellas con tejido adiposo de individuos control. En la comparación entre el tejido adiposo subcutáneo lipoatrófico y el visceral se ha observado que en ambos casos el tejido adiposo presenta alteraciones similares en la función mitocondrial. En cambio el descenso de marcadores de adipogénesis observado en el tejido subcutáneo de pacientes no se reproduce en el tejido visceral. Este hecho, acompañado de diferencias en el perfil de expresión de marcadores de inflamación (que parece más leve en el tejido visceral), podría explicar el comportamiento opuesto de ambos depósitos en pacientes. El estudio en el que se compararon el tejido adiposo lipomatosos de las “buffalo-hump” y el tejido adiposo subcutáneo lipoatrófico de pacientes infectados por el HIV- indica que el tejido adiposo de las “buffalo-hump” presenta alteraciones especificas en la expresión génica respecto al tejido lipoatrófico en las que destaca una expresión normal de genes adipogénicos. Así mismo, el tejido lipomatoso es capaz de expresar UCP1, un gen característico del tejido adiposo marrón, y su capacidad proliferativa concuerda más con un fenotipo del tipo marrón, por lo que se puede decir que estos adipocitos tienen un fenotipo intermedio entre blanco y marrón que se mantiene cuando este tejido lipomatoso es utilizado para trasplante autólogo a la zona facial lipoatrófica en la que mantiene la proliferación desarrollándose el síndrome hámster. No se observan diferencias en las alteraciones mitocondriales observadas en ambos tejidos. Por otra parte la ausencia de un estado de inflamación local en BH podría explicar en parte este comportamiento diferente de ambos tejidos. Además se ha observado que los pacientes muestran niveles elevados de FGF21 y disminuidos de FGF19 (dos agentes homeostaticos) respecto a los controles. Estas diferencias con los controles se acentúan a medida que los pacientes infectados por el virus HIV-1 pasan de ser no tratados a tratados y aun más al desarrollar la lipodistrofia. Los niveles de FGF21 en suero se correlacionaban con indicadores de sensibilidad a insulina o marcadores de síndrome metabólico así como con marcadores de daño hepático que podrían estar relacionados con esteatosis hepática. Los niveles disminuidos en suero de FGF19 se correlacionan negativamente con parámetros indicativos de resistencia a insulina. Además se ha descrito en esta tesis que los receptores de estos agentes endocrinos FGFR1 y β-Klotho aparecen disminuidos en el tejido adiposo de los pacientes infectados por el virus HIV-1. / Molecular basis of adipose tissue alterations and metabolic disturbances associated to HIV-1-infected lipodystrophic patients Disturbances in adipose tissue in HIV-1-infected patients undergoing HAART involve a complex set of alterations known as HAART-associated-lipodystrophy-syndrome (HALS). In most cases, lipoatrophy occur in the face, arms and legs. An enlargement of visceral adipose tissue, reminiscent of visceral obesity, is present often in combination with peripheral lipoatrophy. Lipomatosis is also commonly found in HALS, usually as an enlargement in the dorso-cervical area (buffalo-hump), although the development of lipomas in distinct anatomical sites has also been reported. This adipose tissue redistribution is associated with systemic metabolic alterations such as insulin resistance or dyslipidemia. To determine the individual contribution to HALS of some HAART-prescriptioned drugs, we performed an assessment of the effects of efavirenz, nevirapine and Kaletra on human cultured adipocytes. Our results support the fact that efavirenz and Kaletra impair adipogenesis, reduce the release of adipokines and increase the expression and release of inflammation-related cytokines, while nevirapine does not. Overall, those effects are greater in the case of efavirenz. We compared as well the molecular signature of subcutaneous lipoatrophic, visceral lipohipertrophic and dorso-cervical lipomatous adipose tissues from patients in order to determine the molecular basis causing these fat depots to behave in an opposite way. All fat biopsies from patients exhibited alterations in mitochondrial function marker genes. Visceral and “buffalo hump” fat didn’t show any alterations in the expression of adipogenesis marker genes when compared to healthy controls, while subcutaneous fat showed lower levels. The inflammatory profile was normal in “buffalo hump”, whereas visceral and subcutaneous adipose tissue depots exhibited a distinct, more exacerbated, pro-inflammatory profile. These differences could be part of the explanation of the mentioned different behavior. The serum levels of novel homeostatic agents FGF19 and FGF21 was also assessed in samples from lipodystrophic patients. FGF21 levels were significantly higher in patients and correlated positively with markers of insulin resistance, metabolic syndrome and hepatic damage. On the other hand, FGF19 levels were significantly lower in patients and correlated negatively with markers of insulin resistance. We studied the transcription level of FGF receptors in adipose tissue as well, resulting in a lower expression in biopsies from HALS patients.

This diploma thesis deals with corporate identity of a tractor manufacturer. For this purpose fictive company called Haart was created. The endeavour of this corporate identity is to make an image of a reliable company which specialises in the production of high-quality agricultural machines. This is accomplished by an orange-grey colour scheme where orange stands for living (agricultural production) and grey refers to lifeless (tractor is a machine). The final corporate identity is presented next to company stationery, merchandise, orientation graphics and web page also on the tractor designed within the pre-diploma project. For advertising graphics needs typeface Haart was created. Part of this diploma project is also the creation of a design manual. It describes all elements of Haart‘s corporate identity.

Few studies have assessed the optimum second line highly active anti-retroviral therapy (HAART) regimen in patients who had failed on the first-line HAART in resource-limited settings. This study aimed to compare the Protease inhibitor (PI)-based second line HAART regimens used in one clinic in Trinidad by comparing immunological, virological and clinical outcomes of patients on the different second line HAART regimens. The records of 35 treatment-experienced patients, over 21years of age and on PI-based regimens for at least six months, were analysed using SPSS version 20. The regimen containing TDF/FTC/AZT/LPV/r proved to produce superior outcomes compared to the other second line regimens. Due the small number of usable patients’ records, the findings cannot be generalised but indicate directions for future studies attempting to compare the treatment outcomes of different second line HAART regimens / Health Studies / M. A. (Public Health)

Made available in DSpace on 2016-01-26T12:51:37Z (GMT). No. of bitstreams: 1 arlindoschiesarijunior_dissert.pdf: 1015118 bytes, checksum: a4f2b40a6b49a66d95541b775795efb0 (MD5) Previous issue date: 2010-11-25 / Hereby we present the epidemiological and clinical profile of the HIV-infected group before and during the HAART era from a tertiary care hospital catering to a large population from the Southeastern Brazilian region. A retrospective, cross-sectional and descriptive study was carried out, which involved the analysis of the medical records of patients diagnosed with HIV-1/AIDS admitted to Hospital Escola Emílio Carlos, located in the municipality of Catanduva, State of São Paulo, Brazil. In both pre-HAART and HAART periods, HIV-1 infection was more prevalent in men. Heterosexuality and secondary education were the risk facts for acquisition of the disease in the HAART period. Statistically significant association was only observed for co-infection with HIV-1/Hepatitis C in the pre-HAART era and the number of patients with opportunistic illness (OI) was lower in the HAART period. Among all these OI it is worth mentioning pulmonary pneumocystosis, since despite being frequent in the two periods, its occurrence was considerably greater in the pre-HAART era. Concerning the distribution of OI according to the HIV-1 viral load and serial count of T CD4+ lymphocytes, a significant association was observed. The association between the number of deaths by OI and the survival rate of less than 1 year in the HAART period was significant. The clinical and epidemiological picture of a specialized HIV-1/AIDS Center in a municipality in the southeastern region of Brazil is consistent with the current epidemiology of AIDS in the country. In conclusion, our results indicate that the OI are still important causes of morbi-mortality among HIV-1/AIDS infected patients in the municipality of Catanduva, particularly pulmonary pneumocystosis, tuberculosis and cryptococcal meningoencephalitis. We are aware that retrospective studies such as ours, which involve the review of patients medical records, may present some limitations arising from the scarcity or even absence of information. / Nós apresentamos o perfil clínico e epidemiológico de indivíduos portadores do HIV-1 antes e durante a era da terapia antirretroviral altamente ativa (HAART) de um hospital terciário que atende uma grande população da região Sudeste do Brasil. Estudo retrospectivo, transversal e descritivo que envolveu a análise de prontuários dos pacientes diagnosticados com HIV-1/AIDS atendidos no Hospital Escola Emílio Carlos, localizado no município de Catanduva, Estado de São Paulo, Brasil. Em ambos os períodos pré-HAART e HAART, a infecção por HIV-1 foi mais prevalente em homens. Heterossexualidade e nível de escolaridade do ensino médio foram os fatores de risco para aquisição da doença no período HAART. Associação estatisticamente significante foi observada somente para a coinfecção HIV-1/Hepatite C na era pré-HAART e o número de pacientes com infecções oportunistas (IO) foi menor no período HAART. Entre todas estas IO vale à pena mencionar a pneumocistose pulmonar, pois apesar de ser freqüente nos dois períodos, sua ocorrência foi significativamente maior na era pré-HAART. Quanto à distribuição de IO de acordo com a carga viral do HIV-1 e contagem de linfócitos T CD4 +, uma associação significativa foi observada. A associação entre o número de mortes por IO e a taxa de sobrevivência de menos de um ano na era HAART foi significativa. O quadro clínico e epidemiológico de um centro de atendimento especializado em HIV-1/AIDS em um município na região sudeste do Brasil é compatível com a epidemiologia atual da AIDS no país. Em conclusão, nossos resultados indicam que as IO ainda são importantes causas de morbi-mortalidade entre os pacientes infectados por HIV-1/AIDS no município de Catanduva, particularmente a pneumocistose pulmonar, a tuberculose e a meningoencefalite criptococócica. Estamos cientes de que estudos retrospectivos como o nosso, que envolvem a revisão de prontuários médicos, podem apresentar algumas limitações decorrentes da escassez, ou mesmo da ausência de informações.

A introdução da terapia anti-retroviral de alta potência no tratamento da infecção pelo HIV reduziu significativamente as taxas de morbi-mortalidades relacionadas à imunodeficiência. Entretanto, o tratamento medicamentoso é acompanhado de vários efeitos colaterais, dentre eles, a síndrome da lipodistrofia (SL), caracterizada por alterações morfológicas e metabólicas. Apesar de sua patogenia não estar totalmente esclarecida, é sabido que aumento dos níveis de algumas citocinas inflamatórias estão relacionados com o desenvolvimento da SL. Diversos sítios polimórficos têm sido descritos por influenciarem a transcrição de genes, levando a variações nos níveis de produção de citocinas, como os da região promotora da interleucina-18 (IL-18 -607 C/A e IL-18 -137 C/G) e do gene do interferon gama (IFN- +874 T/A). Diante disso, esse estudo tipificou os polimorfismos da IL-18 e do IFN- em 88 pacientes portadores do HIV com a SL, em 79 portadores do HIV sem a SL, todos sob terapia anti-retroviral e em 133 indivíduos saudáveis, por meio da técnica de reação em cadeia da polimerase com iniciadores de seqüência específica. Este estudo foi aprovado pelo Comitê de Ética em Pesquisa. A presença do alelo -607A e do genótipo -607AA na IL-18 estava significativamente aumentada nos pacientes portadores do HIV com SL quando comparados aos sem a SL, conferindo susceptibilidade ao desenvolvimento da síndrome. De maneira oposta, o alelo -607C e o genótipo -607CC estavam significativamente aumentados em pacientes portadores do HIV sem SL quando comparados aos com a SL, conferindo proteção ao desenvolvimento da síndrome. Os haplótipos -137G/-607A and -137C/-607A, que comportam o alelo -607A, também estavam associados com a susceptibilidade à SL e o haplótipo -137G/-607C estava fortemente associado com proteção contra a SL. Nenhuma diferença significativa na distribuição alélica e genotípica da IL-18 -137 e do IFN- +874 foram observadas entre os grupos de pacientes e o grupo controle. Este é o primeiro estudo que avaliou o polimorfismo da IL-18 e do IFN- na SL e os resultados sugerem que a região promotora da IL-18 está associada com o desenvolvimento da SL em pacientes portadores do HIV. / The introduction of highly active antiretroviral therapy in the treatment of HIV infection significantly reduced the rates of morbidity and mortality related to immunodeficiency. However, the drug treatment is accompanied by various side effects, including the lipodystrophy syndrome (LD), characterized by morphological and metabolic changes. Although its pathogenesis is not totally clear, it is known that increased levels of some inflammatory cytokines are related to the development of LD. Several polymorphic sites have been described by influencing transcription of genes, leading the variations in the levels of cytokine production, such as the promoter region of interleukin-18 (IL-18 -607 C/A and IL-18 -137 C/G) and the interferon gamma gene (IFN- +874 T/A). Thus, this study typifies the polymorphism of the IL-18 and IFN- in 88 HIV-infected patients with LD, in 79 HIV-infected patients without LD, all under antiretroviral therapy and in 133 healthy controls, using the sequence-specific primers-polymerase chain reaction. This study was approved by the Ethics Committee at the place of study. The presence of -607A allele and -607AA genotype in IL-18 gene were significantly increased in HIV patients presenting LD as compared with HIV patients without LD, resulting in susceptibility to the development of LD. Conversely, the -607C allele and -607CC genotype were significantly increased in HIV patients without LD as compared with the HIV patients with LD, offering protection against LD. Haplotypes -137G/-607A and -137C/-607A, carrying the -607A allele, were also associated with susceptibility to LD. The haplotype -137G/-607C was strongly associated with protection against LD. No significant differences in IL-18 -137 and IFN- +874 genotype and allele distribution were observed in patients when compared to a control group. This is the first study evaluating the IL-18 and IFN- polymorphisms in LD and the results suggest that the promoter region of the IL-18 gene is associated with LD development in HIV-infected patients.

Made available in DSpace on 2014-12-17T14:44:19Z (GMT). No. of bitstreams: 1 JasonAM_DISSERT.pdf: 1206570 bytes, checksum: 6ff0e9d810dfe142512fa532ed9d6df2 (MD5) Previous issue date: 2014-02-21 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Introduction: The emergence of High Active Antiretroviral Therapy (HAART) increase the life expectancy of the persons living with HIV/AIDS (PLHIV), therefore the prolonged use cause metabolic implications and influences on body fat distribution and increase the cardiovascular diseases prevalence. Aims: Evaluate the effect of resistance training on heart rate variability, biochemical parameters and somatotype on PLHIV. Methods: Participated this study seven sedentary men, with age above 25 years old, living with HIV/AIDS, under HAART use. Were submitted a 16 week intervention with resistance training. Evaluated the heart rate variability, biochemical parameters and somatotype, before, after 8 weeks and 16 weeks, all in paired form. It was found the data normality by Shapiro-Wilk test and conducted the Anova one way combined with Tukey post hoc to samples in each evaluate moment, adopting significance level p<0,05. Also were calculated percentage change deltas. For somatotype was used the somatotype spatial distance (DES), obeying the significance value DES&#8805;1. Results: Was found significance differences only in variable final heart rate delta 60s (p=0,01), however, is not showed changes on heart rate variability, biochemical parameters and somatotype components. Conclusion: 16 weeks of resistance training showed improvement on heart rate recovery after submaximal effort and, despite is not enough to produce significance differences on biochemical parameters and somatotype components, could be realize improvement on average value of fasting glucose and lipid profile, as well as reducing the endomorphic component / Introdu??o: O surgimento da terapia antirretroviral altamente ativa (HAART) aumentou a expectativa de vida das pessoas vivendo com HIV/AIDS (PVHIV), por?m o seu uso prolongado ocasiona implica??es metab?licas e influencia na redistribui??o de gordura corporal e aumento da preval?ncia de doen?as cardiovasculares. Objetivo: Avaliar o efeito do treinamento resistido na variabilidade da frequ?ncia card?aca, par?metros bioqu?micos e somatotipo de PVHIV. M?todos: Participaram do estudo 7 homens sedent?rios, com idade acima de 25 anos, vivendo com HIV/AIDS, sob uso de HAART. Foram submetidos a uma interven??o de 16 semanas com treinamento resistido. Avaliou-se a variabilidade da frequ?ncia card?aca, par?metros bioqu?micos e o somatotipo, inicialmente e ap?s 8 e 16 semanas de interven??o. Todos eles de forma pareada. Constatou-se a normalidade dos dados pelo teste de Shapiro-Wilk e realizou-se uma Anova one way combinado ao post hoc de Tukey para as amostras em cada momento de avalia??o adotando um valor de p<0,05. Tamb?m foram calculados deltas de mudan?as percentual. Para o somatotipo utilizou-se a dist?ncia espacial dos somatotipos (DES), obedecendo um valor de signific?ncia de DES&#8805;1. Resultados: Encontrou-se diferen?a significativa, somente para a vari?vel delta da frequ?ncia card?aca final de 60s (p=0,01), no entanto, n?o foram observadas modifica??es na variabilidade de frequ?ncia card?aca, vari?veis bioqu?micas e componentes do somatotipo. Conclus?o: 16 semanas de treinamento resistido demonstrou melhoria na recupera??o da frequ?ncia card?aca ap?s um esfor?o subm?ximo e, apesar de n?o ser suficiente para produzir diferen?as significativas nos par?metros bioqu?micos e nos componentes do somatotipo, p?de-se perceber melhoria nos valores m?dios de glicemia e perfil lip?dico, assim como redu??o do componente de endomorfia

A introdução da terapia anti-retroviral de alta potência no tratamento da infecção pelo HIV reduziu significativamente as taxas de morbi-mortalidades relacionadas à imunodeficiência. Entretanto, o tratamento medicamentoso é acompanhado de vários efeitos colaterais, dentre eles, a síndrome da lipodistrofia (SL), caracterizada por alterações morfológicas e metabólicas. Apesar de sua patogenia não estar totalmente esclarecida, é sabido que aumento dos níveis de algumas citocinas inflamatórias estão relacionados com o desenvolvimento da SL. Diversos sítios polimórficos têm sido descritos por influenciarem a transcrição de genes, levando a variações nos níveis de produção de citocinas, como os da região promotora da interleucina-18 (IL-18 -607 C/A e IL-18 -137 C/G) e do gene do interferon gama (IFN- +874 T/A). Diante disso, esse estudo tipificou os polimorfismos da IL-18 e do IFN- em 88 pacientes portadores do HIV com a SL, em 79 portadores do HIV sem a SL, todos sob terapia anti-retroviral e em 133 indivíduos saudáveis, por meio da técnica de reação em cadeia da polimerase com iniciadores de seqüência específica. Este estudo foi aprovado pelo Comitê de Ética em Pesquisa. A presença do alelo -607A e do genótipo -607AA na IL-18 estava significativamente aumentada nos pacientes portadores do HIV com SL quando comparados aos sem a SL, conferindo susceptibilidade ao desenvolvimento da síndrome. De maneira oposta, o alelo -607C e o genótipo -607CC estavam significativamente aumentados em pacientes portadores do HIV sem SL quando comparados aos com a SL, conferindo proteção ao desenvolvimento da síndrome. Os haplótipos -137G/-607A and -137C/-607A, que comportam o alelo -607A, também estavam associados com a susceptibilidade à SL e o haplótipo -137G/-607C estava fortemente associado com proteção contra a SL. Nenhuma diferença significativa na distribuição alélica e genotípica da IL-18 -137 e do IFN- +874 foram observadas entre os grupos de pacientes e o grupo controle. Este é o primeiro estudo que avaliou o polimorfismo da IL-18 e do IFN- na SL e os resultados sugerem que a região promotora da IL-18 está associada com o desenvolvimento da SL em pacientes portadores do HIV. / The introduction of highly active antiretroviral therapy in the treatment of HIV infection significantly reduced the rates of morbidity and mortality related to immunodeficiency. However, the drug treatment is accompanied by various side effects, including the lipodystrophy syndrome (LD), characterized by morphological and metabolic changes. Although its pathogenesis is not totally clear, it is known that increased levels of some inflammatory cytokines are related to the development of LD. Several polymorphic sites have been described by influencing transcription of genes, leading the variations in the levels of cytokine production, such as the promoter region of interleukin-18 (IL-18 -607 C/A and IL-18 -137 C/G) and the interferon gamma gene (IFN- +874 T/A). Thus, this study typifies the polymorphism of the IL-18 and IFN- in 88 HIV-infected patients with LD, in 79 HIV-infected patients without LD, all under antiretroviral therapy and in 133 healthy controls, using the sequence-specific primers-polymerase chain reaction. This study was approved by the Ethics Committee at the place of study. The presence of -607A allele and -607AA genotype in IL-18 gene were significantly increased in HIV patients presenting LD as compared with HIV patients without LD, resulting in susceptibility to the development of LD. Conversely, the -607C allele and -607CC genotype were significantly increased in HIV patients without LD as compared with the HIV patients with LD, offering protection against LD. Haplotypes -137G/-607A and -137C/-607A, carrying the -607A allele, were also associated with susceptibility to LD. The haplotype -137G/-607C was strongly associated with protection against LD. No significant differences in IL-18 -137 and IFN- +874 genotype and allele distribution were observed in patients when compared to a control group. This is the first study evaluating the IL-18 and IFN- polymorphisms in LD and the results suggest that the promoter region of the IL-18 gene is associated with LD development in HIV-infected patients.

Étant donnée la séquence hautement conservée de la NC et son rôle crucial dans le cycle viral de VIH-1, les molécules inhibant la NC sont susceptibles d'agir comme complément aux thérapies anti-rétrovirales à haute activité (HAART) basées sur des médicaments ciblant les enzymes virales, Des médicaments anti-NC sont ainsi susceptibles d'entraîner un maintien de l'inhibition de la réplication d'un large panel d'isolats VIH-1 incluant des lignées virales résistantes aux médicaments ciblant les enzymes virales. Récemment, dans le cadre du consortium Européen TRIoH, de nouvelles stratégies visant à cibler spécifiquement les propriétés chaperonnes de la NC sur les acides nucléiques ont été développées. Selon une stratégie protégée par un brevet soumis, une série de peptides a été conçue afin d'agir comme compétiteurs de la NC et pouyant ainsi inhiber la réplication du virus. Au sein de cette série, plusieurs peptides ont montré une inhibition efficace des propriétés de déstabilisation des acides nucléiques par la NC. Quatre de ces peptides ont été testés en milieu cellulaire et trois d'entre eux ont montré qu'ils pouvaient inhiber efficacement la réplication du HIV-1 dans les lymphocytes. Dans ce contexte, un premier objectif de cette thèse fût de caractériser avec précision les propriétés de ces peptides. En outre, un objectif supplémentaire fût de caractériser le mécanisme moléculaire vis-à-vis de la NC de petites molécules anti-virales développées par les groupes de D. Daelemans (Leuven) et M. Botta (Sienne). / Due to the highly conserved §equence ofNC and its crucial function during HIV-I life cycle, molecules directedagainst NC are believed to be able to complement the highly active anti-retroviral therapies (HAART) based on drugstargeting the viral enzymes. Anti-NC drugs are thought to provide a sustained replication inhibition of a large panel ofHIV-I isolates including virus strains resistant to drugs targeting viral enąrmes. Recently, within the Europeanconsortium TRIoH, new strategies to specifically target the nucleic acid chaperone properties ofNC were developed.According to a strategy protected by a submitted patent, a series of peptides have been desigrred to act as competitorsforNC and thus, inhibit virus replication. Among this series, several peptides were found to efficiently ińibit therrrrcleic acid destabilization properties ofNC. Four of them have been tested in the cellular context and t}ree out ofthemwere found to efficientĘ ińibit the replication of HIV-I in lymphocytes. In this context, the objective of the thesis wasto characterize in depth the properties ofthese peptides. Moreover, an additional objective was to characterize themolecular mechanism in respect to NC of small antiviral drugs developed by the group of D. Daelemans (Leuven) andM. Botta (Siena).

PhD (Molecular Medicine and Haematology), Faculty of Health Sciences, University of the Witwatersrand / Background: The availability of highly active antiretroviral (ARV) treatment in the South African government sector has reduced the morbidity and mortality associated with HIV-1 infection. However, ARV drug resistance and toxicity are major obstacles to achieving and maintaining virus suppression, but there is no provision for ARV drug resistance testing in the public sector. To date, most studies of ARV drug resistance in HIV-1 reverse transcriptase (RT) and protease (PR), are based on sequence data from HIV-1 subtype B, whereas subtype C is the predominant circulating subtype in South Africa. Moreover, host genetic polymorphisms associated with ARV drug toxicity have not been investigated in South African populations. This study evaluated viral and host genetic factors associated with ARV treatment outcome in 812 ARV drug-naive South African AIDS participants enrolled on the CIPRA-SA study from Johannesburg and Cape Town. Methodology: An affordable in-house genotyping protocol (subtype C specific) was established and validated to monitor the emergence of ARV drug resistance. This assay was used to genotype all CIPRA-SA participants failing the first- and second-line ARV drug regimens. Allellic discrimination assays to identify the G1344A, A6986G, G516T and C3435T SNPs in CYP3A4, 3A5, 2B6 and MDR-1, respectively, associated with ARV metabolism and absorption were performed. Results: The in-house ARV drug resistance assay successfully genotyped 95% of patient samples, including non-C subtypes from 8 African sites. Treatment failure was experienced in 371 participants, mainly due to toxicity (n=134) or virological failure (n=83). Overall, CIPRA-SA participants with a lower CD4+ T-cell count at study onset were more likely to experience viral failure. Genotyping using the in-house assay revealed that 6 participants had ARV drug resistance mutations at study entry. Treatment failure of 58 participants was a result of ARV drug resistance mutations, whereas 19 had no known ARV drug resistance mutations. The most frequent mutations were M184V (67%) and K103N (50%). K65R was present (3%) and one participant harboured TAMs. Longitudinal genotypic analysis showed that NNRTI mutations accumulated at a rate of one per three months left on failing therapy. No PR mutations were detected amongst participants experiencing second-line failure. The four SNPs analysed occured in similar frequencies between a background and the CIPRA-SA cohort. Furthermore, no statistically significant association could be found between these four SNPs and viral failure and/or toxicity. Conclusion: Overall, HIV-1 subtype C-infected individuals receiving ARV therapy develop many of the known subtype B drug resistance mutations. However, the ARV drug resistance patterns in the closely monitored CIPRA-SA cohort were less complex compared to published data from the region, confirming that more frequent viral load monitoring, genotyping, and a virological failure cut-off value of 1000 RNA copies/ml ensure a better prognosis, and preserve future ARV treatment options.