Orientador: Sandra Cecilia Botelho Costa / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-07T04:26:19Z (GMT). No. of bitstreams: 1 Bismara_BeatrizAparecidaPassos_M.pdf: 3138714 bytes, checksum: 1bf8ed00aa70e66a625bf970d08a27e9 (MD5) Previous issue date: 2006 / Resumo: Investigamos a presença de mutações em crianças infectadas verticalmente pelo vírus da Imunodeficiência Humana tipo 1 (HIV-1) que conferem resistência aos agentes antiretrovirais. Amostras de sangue periférico foram coletadas de sessenta e seis pacientes em seguimento no do Ambulatório de Pediatria do Hospital das Clínicas da Universidade Estadual de Campinas. A partir de leucócitos destas amostras foi extraído o DNA, após diversas lavagens e precipitações. Foi preparado um Mix para 20 reações contendo 100µl de Buffer (50 mM de cloreto de potássio; 20 mM de Tris-HCl - pH 8,4); 100 µl de cloreto de magnésio (25mM); 12µl da mistura desoxirribonucléica - dNTPs (dATP, dGTP, dCTP; dTTP) a 25mM, 10µl de cada ¿primer¿ (25pmoles/µl); 0,5µl de Taq DNA polimerase e 0,5 µl do DNA a ser estudado, para a realização da PCR. As condições da reação foram: Desnaturação: 95ºC - 3 minutos; Anelamento: 55ºC ¿ 1 minuto; Extensão: 72ºC - 1 minuto (3 ciclos). Desnaturação: 95ºC ¿ 1 minuto; Anelamento: 55ºC ¿ 45 segundos; Extensão: 72ºC ¿ 1 minuto e Extensão final: 72ºC por 10 minutos (35 ciclos). As bandas foram visulizadas em gel de agarose 1%, obtendo-se uma banda de 1008 pares de bases. Os produtos selecionados foram submetidos a seguinte reação de seqüenciamento: 1,0 µl do produto da PCR; 4,0 µl de Premix (Amersham); 1,0 µl de primer 5 µM; Completar com dH2O para 10,0 µl de reação. Após a amplificação, os fragmentos foram analisados pelo Software MegaBACE¿ Sequence Analyzer, em seguida alinhados e comparados com o banco de dados público (GenBank), com o vírus selvagem, e com o programa Phred, Phrap, Consed. As principais mutações encontradas no gene da Transcriptase reversa foram: M184V(42,6%), M41L (39,3%), D67N (27,9%), T215Y (26,2%) e K70R (18%). No gene da Protease as principais encontradas foram: L63P (42,6%), L90M e M36I (32,8%), V77I (29,5%), I93L (24,6%), V82A (16,4%), I54V (14,8%) e K20R (13,1%). Concluímos que este método é muito eficaz para análise das seqüências, auxiliando na observação das mutações. Este exame deveria ser implantado na rotina para os pacientes portadores deste vírus, pois auxiliaria os clínicos na escolha de uma terapia ideal para cada paciente. As crianças que apresentavam estas mutações associadas às drogas antiretrovirais estavam com o quadro clínico afetado e a maioria dos esquemas terapêuticos não reduziam a carga viral em níveis desejáveis / Abstract: In this work our purpose was to determine the subtype, prevalence of drug-resistance mutations, and assess genotypic profiles in HIV-1 infected children under antiretroviral treatment, from Campinas, São Paulo, Brazil. Blood samples from sixty six vertically human immunodeficiency virus type 1 (HIV-1) infected Brazilian children were studied for antiretroviral drug resistance. Combination therapy with protease (PR) and reverse transcriptase (RT) inhibitors can efficiently supress human immunodeficiency virus (HIV) replication, but the emergence of drug resistance variants correlates strongly with therapeutic failure1. DNA was extracted from peripheral blood mononuclear cells (PBMc) samples, and a 1.0 kb fragment containing HIV-1PR and RT-coding sequence were amplified by Nested Polymerase Chain Reaction, sequencing and subtyping. The HIV-1 subtyping based on the polymerase (pol) gene sequences (protease and reverse transcriptase-RT regions) was as follow: subtype B (83,6%), subtype F (9,8%) and B/F viral recombinant forms (6,6%). We found fifty five sequences presented significant mutations and thirty five presented common polymorphism conferring resistance to protease inhibitors (PIs). Forty one presented mutations associated with resistant to nucleoside reverse transcriptase inhibitors (NRTIs). The entire viral protease and codons 1 to 219 of the reverse transcriptase gene from 61 HIV-1 isolates were amplified and sequenced for genotyping. Two major protease inhibitor-resistance associated mutations, M36I and L90M, were most prevalent in our samples (32,8%) and the polymorphism L69P (42,6%). Minor mutation were also found at the protease gene: V77I (29,5%), V82A (16,4%), I93L (24,6%), I54V (14,8%), K20R (13,1%). Many mutations associated with reduced susceptibility to nucleoside or non-nucleoside reverse transcriptase inhibitors were detected: M41L (39,3%), M184V (42,6%), D67N (27,9%), T215Y (26,2%), L210W (21%), K70R (18%) and E44D (11,5%). This study demonstrated that 98,3% of the studied population from the Campinas, showing evidences of therapy failure, presented viral genomic mutations associated with drug resistance. The main antiretroviral to which this population showed resistance were PI nelfinavir (54%), and ritonavir with lopinavir (18%), and the NNRTIs efavirenz (18%) and nevirapine (3,2%), and the NRTIs zidovudine (60,6%), didanosine (47,5%), lamivudine (45,9%). Ninety-eight percent of patients under antiretroviral therapy and with high viral load counts showed resistance to at least one of the antiretroviral analyzed. / Mestrado / Mestre em Farmacologia

A adesão do paciente à terapia antirretroviral é essencial para alcançar os objetivos do tratamento. A taxa de adesão do conjunto de pacientes de um serviço pode ser compreendida como medida proxy da qualidade dos serviços. Realizou-se uma pesquisa avaliativa de caráter qualitativo sobre a aplicabilidade do Questionário Qualiaids de Monitoramento da Adesão ao Tratamento Antirretroviral (WebAd-Q) para uso rotineiro nos serviços. Duas dimensões orientaram o delineamento do estudo: Dimensão de Utilização - que se refere à experiência dos profissionais de utilização do WebAd-Q no ambiente de pesquisa; e Dimensão de Utilidade - que se refere à opinião dos profissionais sobre a importância de conhecer a adesão do conjunto de pacientes e a potencialidade do WebAd-Q para auxiliar a gestão dos serviços. Participaram sete serviços: quatro foram classificados como Serviço de Atenção Especializada (SAE); dois são ambulatórios inseridos em grandes hospitais; e o último é um ambulatório que funciona em Unidade Básica de Saúde (UBS). Foram entrevistados 22 profissionais que atuam na assistência, além dos sete gerentes dos serviços. Foi utilizado um roteiro de entrevista semiestruturado; as entrevistas foram gravadas em áudio, transcritas na íntegra e submetidas à análise temática de conteúdo. Os serviços apresentam diferenças de porte, estrutura e modalidades assistenciais oferecidas, mas são semelhantes na composição das equipes, na organização do trabalho e no modo de gerenciamento. Para os profissionais, a (não) adesão é um problema complexo, multideterminado e de difícil compreensão, cuja solução, muitas vezes, está além das suas possibilidades de intervenção. O WebAd-Q foi avaliado como um questionário simples, simpático, objetivo e de fácil compreensão e manuseio para profissionais e usuários. Além disso, não apresenta dificuldades operacionais para sua implantação. Alguns atributos bastante valorizados são: poder ser respondido anonimamente, combinar linguagens de vídeo e áudio, e ser compreendido por pessoas de todos os níveis de escolaridade. Quanto à utilidade, entendem que sua aplicação pode produzir efeitos importantes: efeito de constatação daquilo que todos sabem e reconhecem como desafio: a insuficiente adesão dos pacientes à terapia antirretroviral; efeito de avaliação da qualidade, pois os profissionais compreendem as medidas de adesão do conjunto de pacientes como um indicador de desempenho do serviço e da importância do monitoramento para melhorar a assistência; efeito demonstrativo, que explicita a necessidade de intervenções mais estruturais que tenham como alvo uma reconceituação dos serviços; e efeito de valorização, referente ao sentimento de \"sentir-se valorizado\" relatado pelos pacientes. Os profissionais se mostraram entusiasmados com as potencialidades do questionário; contudo, não apresentaram proposições tecnológicas e organizacionais concretas para sua utilização, o que pode significar que a plena potencialidade do WebAd-Q como insumo gerencial só será alcançada com o aprimoramento do gerenciamento / Patient adherence to antiretroviral therapy is essential to meet the treatment goals. Adherence rates of a group of patients in a certain service might be considered a measure of quality proxy. A qualitative research study was carried out to assess the feasibility of the Qualiaids Adherence Monitoring Questionnaire in antiretroviral therapy - WebAd-Q as a daily routine. Two dimensions guided the study design: the Application Dimension - about the staff\'s experience in the application of webAD-Q within the research environment; and the and Usefulness Dimension - focusing on the staff\'s opinion on the significance to know the patients\' adherence and the webAD-Q\'s potential to assist them in managing their services. Seven types of service participated in this study. Four of them were ranked as Specialized Care Service (SCS). There were also two clinics located in big hospitals and one clinic which was part of a Basic Health Care Unit (BHU). 22 healh care providers were interviewed, as well as the seven managers of the facilities abovementioned. A semi-structured interview guide was used. The interviews were audio recorded, transcribed and their content was analyzed. The services differ among themselves in size, structure and type of assistance. On the other hand, they are similar in team composition and how they are managed. For the health care providers, (non) adherence is a complex problem, caused by multiple factors, and difficult to understand. The solution is often beyond their means of intervention. The WebAd-Q was assessed as simple, friendly, straight-to-the-point questionnaire, easy to understand and handle, both by providers and users. Also, its application was did not have any operational difficulties. Some of the questionnaire\'s most valued attributes include allowing for anonymous responses, providing video and audio, and being understood by people of all educational levels. The participants also understand that its application eventually results in substantial effects: the effect of finding out something widely known and recognized as a challenge: the insufficient adherence rates of patients to antiretroviral therapy; the effect of assessing quality, because health care providers understand adherence measures of groups of patients as an indicator of service performance and of the importance of monitoring to improve health care procedures; the effect of demonstration, as it shows the need for more structural interventions aiming at a revision of what their services mean; and the effect of valuation, meant as the feeling of \"feeling valued\", as reported by the patients.The healthcare providers showed enthusiasm with the Questionnaire\'s potential. However, they did not proposed concrete technological and organizational plans for its use. That might mean that the WebAd-Q as a management tool will only reach its full potential if management procedures are improved

Orientadores: Marcos Tadeu Nolasco da Silva, Rubens Nelson Amaral de Assis Reimão / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-22T23:19:37Z (GMT). No. of bitstreams: 1 Gazini_CarinaCaires_M.pdf: 1252210 bytes, checksum: 00b55ba57c9260460ffa328a2940bdb8 (MD5) Previous issue date: 2011 / Resumo: INTRODUÇÃO: O sono é fator indispensável para a saúde integral. Sua má qualidade, em adolescentes, pode resultar em alterações cognitivas, psicológicas e clínicas. Crianças e adolescentes infectados pelo Vírus da Imunodeficiência Humana (HIV) são vulneráveis a apresentar distúrbios do sono, como consequência de alterações imunológicas e emocionais. Com base neste contexto, este estudo teve o objetivo de avaliar as características do sono de adolescentes infectados pelo HIV e verificar a associação entre aspectos psicossociais, clínicos, imunológicos e virológicos e alterações no padrão e na qualidade do sono. CASUÍSTICA E MÉTODOS: Estudo observacional, analítico, prospectivo, do tipo corte transversal. Foram avaliados 222 adolescentes de ambos os gêneros, com idade a partir de 10 anos, sendo 102 infectados pelo HIV (59 rapazes) e 120 estudantes (73 garotas) presumidamente saudáveis. Para a coleta de dados aplicaram-se: Escala de distúrbios do sono em crianças (SDSC), Escala de Sonolência de Epworth (ESE) e Questionário genérico sobre Qualidade de Vida Pediátrica (PedsQLTM4.0). Dados clínicos, imunológicos e virológicos foram retirados dos prontuários médicos. Na análise estatística, utilizaram-se os testes de Mann-Whitney, Qui-Quadrado e o Coeficiente de Correlação de Spearman. Resultados foram considerados estatisticamente significativos se P ? 0,05. O estudo foi aprovado pelo Comitê de Ética em Pesquisa em Seres Humanos da Faculdade de Ciências Médicas da Universidade Estadual de Campinas. RESULTADOS: Verificou-se prevalência de distúrbios do sono, com base nos escores SDSC, de 77,4% nos pacientes e 75% nos controles (P = 0,67). Pacientes infectados pelo HIV apresentaram maior pontuação do escore SDSC nos distúrbios respiratórios do sono com medianas de 3 (3 - 15) vs 3 (3-11) (P = 0,05), e maior pontuação de sonolência diurna excessiva pela ESE, com medianas de 6 (0 - 17) vs 5 (0 - 14) (P = 0,02). Houve correlação entre os escores SDSC e PedsQL em ambos os grupos, com rs = -0,46 no grupo infectado (P < 0,001) e rs = -0,37 no grupo controle (P < 0,001). Não se observaram associações estatisticamente significativas entre os indicadores de qualidade do sono e variáveis clínicas, imunológicas ou virológicas, no grupo infectado. CONCLUSÃO: Os adolescentes infectados pelo HIV apresentaram qualidade do sono semelhante à população saudável, porém com alta prevalência de distúrbios do sono, ocorrendo associação entre qualidade de sono e qualidade de vida em ambos os grupos. Não foram identificados fatores clínicos, imunológicos ou virológicos associados à qualidade do sono, no grupo infectado. Tais resultados decorrem provavelmente da gradual melhora de condições de vida dos adolescentes infectados, resultante do sucesso da terapia antirretroviral, bem como das vulnerabilidades que potencialmente afetam a qualidade do sono em adolescentes brasileiros / Abstract: INTRODUCTION: Sleep is an indispensable feature of integral health. It's bad quality, in adolescents, may result in cognitive, psychological and clinical disturbances. Children and adolescents infected with Human Immunodeficiency Virus (HIV) are vulnerable to present sleep disturbances, due to immunological and emotional abnormalities. Within the above setting, this study aimed to evaluate sleep characteristics of HIV-infected adolescents and to verify the association between psychosocial, clinical, immunological and virological issues and abnormalities in pattern and quality of sleep. PATIENTS AND METHODS: Observational, analytical, prospective, cross-sectional study. Two hundred and twenty-two adolescents from both genders were evaluated, aged 10 or more years-old. One hundred and two (59 boys) were HIV-infected, and 120 (73 girls) were presumedly healthy students. The following tools were used for data collection: Sleep Disturbance Scale for Children (SDSC), Epworth Daytime Sleepiness Scale (EDSS) and Generic Pediatric Quality of Life Questionnaire (PedsQLTM4.0). Clinical, immunological and virological data were retrieved from medical records. For statistical analysis, the Mann-Whitney and Qui-Square tests, and Spearman Correlation Coefficient were used. Results were considered statistically significant if P ? 0.05. The study was approved by the Committee of Ethics in Research in Human Beings from the Faculty of Medical Sciences, State University of Campinas. RESULTS: The prevalence of sleep disturbances, based on SDSC scores, was 77.4% in patients and 75% in controls (P = 0.67). HIV-infected patients showed higher SDSC scores in the respiratory sleep disturbances domain, with medians of 3 (3 - 15) vs 3 (3-11) (P = 0.05), and higher scores in EDSS, with medians of 6 (0 - 17) vs 5 (0 - 14) (P = 0.02). A correlation was observed between SDSC and PedsQL scores in both groups, with rs = -0,46 in the infected group (P < 0.001) and rs = -0.37 in the control group (P < 0.001). No statistically significant associations were observed between quality of sleep markers and clinical, immunological or virological variables, in the infected group. CONCLUSION: HIV-infected adolescents showed similar quality of sleep when compared to the healthy control population, yet with high prevalence of sleep disturbances in both groups. An association between quality of sleep and quality of life was observed in both groups. No clinical, immunological or virological issues were identified in the HIV-infected group in association to sleep quality. Such results are probably due to the gradative improvement of conditions of life in HIV-infected adolescents, as a result of successful antiretroviral therapy, and also to the vulnerabilities which potentially affect the quality of sleep in Brazilian adolescents / Mestrado / Saude da Criança e do Adolescente / Mestre em Saude da Criança e do Adolescente

Orientador: Marcos Tadeu Nolasco da Silva / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-18T22:53:13Z (GMT). No. of bitstreams: 1 Ernesto_AlineSantarem_M.pdf: 2616267 bytes, checksum: 8169b5c577c4b73c8dedf740de1f15b9 (MD5) Previous issue date: 2011 / Resumo: Introdução: A Síndrome de Imunodeficiência Adquirida (Aids) é atualmente uma doença crônica, controlável graças à Terapia Antirretroviral de Alta Atividade (TARV). Em um contexto de acesso universal ao tratamento, a adesão do paciente torna-se um fator limitante e desafiador. Este estudo teve como objetivo avaliar a prevalência da nãoadesão à TARV em uma coorte de crianças e adolescentes com o uso de instrumentos complementares, bem como identificar e compreender os fatores associados a ela. Casuística e Métodos: Estudo analítico, observacional, prospectivo do tipo corte transversal. A população foi composta por 108 pacientes infectados pelo Vírus da Imunodeficiência Humana (HIV) em TARV (60 meninos), com idades entre 8 e 19 anos. A adesão foi avaliada por meio de um questionário padronizado, consulta a registros de dispensação de farmácia (RDF) e uma escala de auto-eficácia. Foram entrevistados os responsáveis pela administração da medicação, cuidadores ou pacientes. Indivíduos que receberam menos de 95% das doses prescritas nas 24 horas ou nos 7 dias anteriores à entrevista, ou que apresentaram um intervalo maior que 37 dias no RDF nos três meses anteriores à entrevista, foram considerados não-aderentes. A escala de auto-eficácia forneceu um escore contínuo, com amplitude de 0 a 100. Foi avaliada a associação de variáveis independentes ligadas a condições demográficas, clínicas, imunológicas, virológicas, e psicossociais aos desfechos de adesão. Na análise estatística univariada foi utilizada a determinação de Odds Ratios (OR) para a comparação entre variáveis categóricas, o teste de Mann-Whitney para a comparação entre variáveis contínuas e categorias, e determinado Coeficiente de Correlação de Spearman (rs) para a comparação entre variáveis contínuas. Resultados foram considerados significativos com valor de p _ 0,05. Para o controle de variáveis de confundimento, foi utilizada a análise multivariada com o uso de regressão logística. O estudo foi aprovado pelo Comitê de Ética em Pesquisa em Seres Humanos da Faculdade de Ciências Médicas da Universidade Estadual de Campinas. Resultados: A prevalência de não-adesão variou entre 11,1% (pacientes não aderentes por 3 instrumentos), 15,8% (auto-relato de 24 horas), 27,8% (auto-relato semanal), 45,4% (RDF) e 56,3% (ao menos um dos 3 desfechos). Os auto-relatos de 24 horas e 7 dias, quando comparados ao RDF, mostraram baixa sensibilidade (29% e 43%, respectivamente) e alta especificidade (95% e 85%, respectivamente). As seguintes variáveis independentes apresentaram associação estatisticamente significativa com a não-adesão na análise univariada, de acordo com o instrumento: Auto-relato de 24 horas: dificuldade de administração pelo cuidador (OR = 9,11; IC95% = 2,87 - 28,98); paciente não-praticante de religião (OR = 2,76; IC95% = 0,92 - 8,32); intolerância à medicação (OR=4,61; IC95% =1,47 - 14,42); renda per capita (medianas de US$ 137,91 vs US$ 208,33; p = 0,016); número de ITRNs com mutações de resistência (medianas de 6 vs 1; p = 0,016); Auto-relato de 7 dias: dificuldade de administração pelo cuidador (OR = 2,91; IC95% = 1,05 - 8,12; administração da TARV pelo paciente (OR = 3,59; IC95% = 1,47 - 8,78); cuidador com 8 ou menos anos de escolaridade (OR = 3,25; IC95% = 1,03 - 10,30); paciente com mais de 8 anos de escolaridade (OR = 3,70; IC95% = 1,41 - 0,70); idade do paciente (medianas de 13,94 vs 12,94; p = 0,03); renda per capita (medianas de US$ 131,67 vs US$ 201,39; p = 0,009); Registro de dispensação de farmácia: dificuldade de administração pelo cuidador (OR = 3,19; IC95% = 1,11 - 9,17); administração da TARV pelo paciente (OR = 2,70; IC95% = 1,15 - 6,33); falha de controle virológico (OR = 3,70; IC95% = 1,67 - 8,33); falta a consulta nos últimos 6 meses (OR = 3,27 IC95% = 1,38 - 7,78); paciente nãopraticante de religião (OR = 2,47 IC95% = 1,10 - 5,57); cuidador não-praticante de religião (OR = 3,19; IC95% = 1,36 - 7,50); cuidador com emprego fora do domicílio (OR = 2,27; IC95% = 1,05 - 4,92); renda per capita (medianas de US$ 166,67 vs US$ 222,22; p = 0,014); As seguintes variáveis independentes apresentaram associação estatisticamente significativa com a não-adesão na análise multivariada, de acordo com o instrumento: Auto-relato de 24 horas: intolerância à medicação (OR = 9,11; IC95% = 2,87 - 28,98); Auto-relato de 7 dias: dificuldade de administração pelo cuidador (OR = 2,91; IC95% = 1,05 - 8,12); administração da TARV pelo paciente (OR = 3,59; IC95% = 1,47 - 8,78); classe socioeconômica C+D (3,54; 0,97 - 2,85); Registro de dispensação de farmácia: falha de controle virológico (OR = 3,73; IC95% = 1,68 - 8,31); falta a consulta nos últimos 6 meses (OR = 3,27 (IC95% = 1,38 - 7,78); cuidador não-praticante de religião (OR = 3,19; IC95% = 1,36 - 7,50); O escore de auto-eficácia teve mediana de 95,20 (11,90 - 100) e mostrou associação significativa com dificuldade de administração da medicação pelo cuidador (mediana de 78,5 vs 95,2; p = 0,001), falha de controle virológico (mediana de 90,4 vs 100; p = 0,001), administração da TARV pelo paciente (mediana de 89,8 vs 95,2; p = 0,05), falta à consulta nos últimos seis meses (mediana de 86,3 vs 100, p < 0,001), categoria clínica N, A ou B (mediana de 90,47 vs 100; p = 0,018), paciente não praticante de religião (mediana de 90,4 vs 95,2, p = 0,037), orfandade (mediana de 95,2 vs 90,4 p = 0,05), relação CD4/CD8 (rs = 0,220; p = 0,025), número de classes de antirretrovirais com resistência viral (rs = 0,583; p < 0,001), número de ITRNs com resistência viral (rs = 0,44; p = 0,009), renda per capita (rs = 0,302; p = 0,001), Escore PedsQL domínio emocional (rs = 0,265; p = 0,007). Conclusão: Na população estudada, observou-se alta prevalência de falha de adesão à TARV, com maior sensibilidade de detecção pela análise da retirada de medicamentos na farmácia. Adicionalmente, observou-se associação entre os escores de auto-eficácia e as categorias de adesão. Os instrumentos utilizados mostraram-se complementares na identificação de fatores de risco para a não-adesão. Com o objetivo de eliminar variáveis de confundimento, sete fatores foram identificados como associados a dificuldade de adesão: intolerância à medicação, dificuldade de administração da medicação pelo cuidador, responsabilidade de administração medicação pelo próprio paciente, classe socioeconômica mais baixa, ausência de controle virológico, cuidador não praticante de religião e faltas às consultas / Abstract: Background: The Acquired Immunodeficiency Syndrome (Aids) is currently a chronic disease, manageable by Highly Active Antiretroviral Therapy (HAART). In a setting of universal access to treatment, patient adherence arises as a limiting and challenging issue. This study aimed to evaluate the prevalence of nonadherence to HAART in a cohort of children and adolescents, using complementary instruments, and also identify and understand associated factors. Patients and Methods: Observational, analytical, prospective, cross-sectional study. The study population comprised 108 Human Immunodeficiency Virus (HIV) -infected patients on HAART (60 boys), from 8 to 19 years-old. Adherence was evaluated by a standardized questionnaire, pharmacy refill data (PRD) and a self-efficacy scale. Patients or caregivers were interviewed (whoever was in control of medicine administration). Patients who received less than 95% of prescribed doses in the 24 hours of 7 days before the interview, or who had a record of an interval of more than 37 days between refills, were considered nonadherent. The self-efficacy scale provided a continuous score, varying from 0 to 100. The association between adherence outcomes and independent variables related to demographical, clinical, immunological, virological and psychosocial conditions was estimated. Statistical analysis was performed with the use of Odds Ratios (OR) for comparison between categorical variables, Mann-Whitney test for comparison between continuous variables and categories, and Spearman Correlation Coefficient (rs) for comparison between continuous variables. Results were considered statistically significant if p _ 0.05. Confounding variables were controlled by multivariate analysis with logistic regression. The study was approved by the Human Research Ethics Committee of the State University of Campinas Faculty of Medical Sciences. Results: Nonadherence prevalence varied from 11.1% (nonadherent patients in 3 instruments), 15.8% (24-hour self-report), 27.8% (7-day self-report), 45.4% (PRD) and 56.3% (at least one of the outcomes). Self-reports from 24 hours and 7 days, when compared to PRD, showed low sensitivity (29% and 43%, respectively) and high specificity (95% and 85%, respectively). The following independent variables showed statistically significant association with nonadherence on univariate analysis, according to each instrument: Twenty-four hour self-report: difficulty of ministration by caregiver (OR = 9.11 ; 95%CI = 2.87 - 28.98); lack of religious practice by patient (OR = 2.76; 95%CI = 0.92 - 8.32); medication intolerance (OR=4.61; 95%CI =1.47 - 14.42); per capita income (median US$ 137.91 vs US$ 208.33; p = 0.016); number of nucleoside/nucleotide analogues (NRTIs) with resistant mutations (median 6 vs 1; p = 0.016); Seven-day self-report: difficulty of ministration by caregiver (OR = 2.91; 95%CI = 1.05 - 8.12; HAART ministration by the patient (OR = 3.59; 95%CI = 1.47 - 8.78); caregiver with 8 or less years of school attendance (OR = 3.25; 95%CI = 1.03 - 10.30); patient with 8 or more years of school attendance (OR = 3.70; 95%CI = 1.41 - 0.70); patient age (median 13.94 vs 12.94; p = 0.03); per capita income (median US$ 131.67 vs US$ 201.39; p = 0.009); Pharmacy refill data: difficulty of ministration by caregiver (OR = 3.19; 95%CI = 1.11 - 9.17); HAART ministration by the patient (OR = 2.70; 95%CI = 1.15 - 6.33); lack of virological control (OR = 3.70; 95%CI = 1.67 - 8.33); missed consultations in the former 6 months (OR = 3.27 (95%CI = 1.38 - 7.78); lack of religious practice by patient (OR = 2.47 (95%CI = 1.10 - 5.57); lack of religious practice by caregiver (OR = 3.19; 95%CI = 1.36 - 7.50); caregiver working outside the home (OR = 2.27; 95%CI = 1.05 - 4.92); per capita income (median US$ 166.67 vs US$ 222.22 ; p = 0.014); The following independent variables showed statistically significant association with nonadherence on multivariate analysis, according to each instrument: Twenty-four hour self-report: medication intolerance (OR = 9.11; 95%CI = 2.87 - 28.98); Seven-day self-report: difficulty of ministration by caregiver (OR = 2.91; 95%CI = 1.05 - 8.12); HAART ministration by the patient (OR = 3.59; 95%CI = 1.47 - 8.78); socioeconomical classes C+D (3.54; 0.97 - 2.85); Pharmacy refill data: lack of virological control (OR = 3.73; 95%CI = 1.68 - 8.31); missed consultations in the former 6 months (OR = 3.27 (95%CI = 1.38 - 7.78); lack of religious practice by caregiver (OR = 3.19; 95%CI = 1.36 - 7.50); The self-efficacy score had a median of 95.20 (11.90 - 100) and showed significant association with difficulty of ministration by caregiver (median 78.5 vs 95.2; p = 0.001), lack of virological control (median 90.4 vs 100; p = 0.001), HAART ministration by patient (median 89.8 vs 95.2; p = 0.05), missed consultations in the former 6 months (median 86.3 vs 100, p < 0.001), clinical categories N, A or B ( median 90.47 vs 100; p = 0.018), lack of religious practice by patient (median 90.4 vs 95.2, p = 0.037), being orphan (median 95.2 vs 90.4 p = 0.05), CD4/CD8 ratio (rs = 0.220; p = 0.025), number or antiretroviral classes with resistance (rs = 0.583; p < 0.001), number of NRTIs with resistance (rs = 0.44; p = 0.009), per capita income (rs = 0.302; p = 0.001), PedsQL score, emotional domain (rs = 0.265; p = 0.007). Conclusion A high prevalence of HAART nonadherence was observed in the study population, being pharmacy refill data the most sensitive measurement. Additionally, an association was observed between adherence outcomes and self-efficacy scores. The instruments employed showed complementarity in the recognization of nonadherence risk factors. Aiming to eliminate confounding variables, seven factors were identified as associated to lack of adherence: medication intolerance, difficulty of ministration by the caregiver, ministration of medicines by the patient, lower socioeconomic class, lack of virological control, lack of religious practice by the caregiver and missed consultations / Mestrado / Saude da Criança e do Adolescente / Mestre em Ciências

Orientadores: Marcos Tadeu Nolasco da Silva, Maria Marluce dos Santos Vilela / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T12:56:55Z (GMT). No. of bitstreams: 1 Ferreira_JosianeFrancisca_M.pdf: 1674494 bytes, checksum: 65421fb00fd3ba43536a3c494f244ecd (MD5) Previous issue date: 2012 / Resumo: INTRODUÇÃO: A adoção da Terapia Antirretroviral Combinada (TARC), proporcionou uma dramática melhora na sobrevida, na qualidade de vida e no controle clínico da infecção por HIV em pediatria. Em consequência do controle efetivo da replicação viral, observa-se reconstituição do sistema imune, que contribui para a redução da incidência dos quadros infecciosos. Neste contexto, a determinação da reconstituição imune em estudos observacionais pode fornecer marcadores para a avaliação da resposta à TARC. OBJETIVOS: Determinar a prevalência de recuperação imunológica associada à introdução da TARC em uma população pediátrica acompanhada em serviço de referência, bem como fatores associados à obtenção desta meta. MÉTODO: Estudo observacional, analítico, do tipo coorte histórico. Em 127 pacientes em TARC pelo período mínimo de 1 ano, avaliou-se a reconstituição imune, caracterizada por uma porcentagem de linfócitos T CD4+ 'maior ou igual' 25% ou valor absoluto considerado normal para a idade. RESULTADOS: Dos 127 pacientes avaliados, 117 (92,12%) obtiveram reconstituição ou preservação imune. Nos indivíduos com reconstituição ou preservação imune, a idade ao início da TARC foi significativamente inferior (p = 0,007), e os escores-z de peso e IMC foram significativamente superiores (p = 0,028 e 0,032, respectivamente). Em relação às categorias clínicas, observou-se uma proporção significativamente superior de indivíduos nas categorias N, A e B no grupo com reconstituição ou preservação imune (p = 0,05). As porcentagens de linfócitos T CD4+ e a relação CD4 / CD8, bem como a proporção de indivíduos nas categorias imunológicas 1 e 2, foram superiores no grupo com reconstituição ou preservação imune (p = 0,019, 0,018 e 0,005, respectivamente). CONCLUSÃO: Pudemos observar que, em crianças e adolescentes infectados por HIV em TARC, a reconstituição ou preservação imune estiveram associadas a uma idade mais jovem e à maior preservação imunológica e clínica ao início da terapia / Abstract: INTRODUCTION: The adoption of Highly Active Antiretroviral Therapy (HAART) provided a dramatic improvement in survival, quality of life and clinical management of HIV infection in children. As a result the effective control of viral replication, it is observed immune reconstitution, which contributes to reducing the incidence of infectious. In this context, the determination of immune reconstitution in observational studies can provide markers for assessing response to HAART. Objective: To determine the prevalence of immune recovery or preservation associated with the introduction of HAART in a pediatric population followed in a reference service, as well as factors associated with the attainment of this goal. Methods: Observational study, analytic, historical cohort. In 127 patients on HAART for at least one year, we evaluated the immune reconstitution or preservation, characterized by a of CD4+ cell percentage 'more or equal' 25% or absolute values considered normal for their age. Results: Of the 127 patients evaluated, 117 (92.12%) had immune reconstitution or preservation. In individuals with immune reconstitution or preservation, the age at first HAART was significantly lower (p = 0.007), and weight and BMI z-scores for age were significantly higher (p = 0.028 and 0.032, respectively). With respect to clinical categories, there was a significantly higher proportion of individuals in N, A and B categories in the immune reconstitution or preservation group (p = 0.05). The CD4+ cell percentages and CD4/CD8 ratios and the proportion of individuals in immunological categories 1 and 2, were higher in the group with immune reconstitution or preservation (p = 0.019, 0.018 and 0.005, respectively). Conclusion: In our cohort of HIV-infected children and adolescents, on HAART, the preservation of immunocompetence or immune reconstitution were associated to an earlier age and better immune and clinical preservation at the beginning of therapy / Mestrado / Saude da Criança e do Adolescente / Mestra em Saúde da Criança e do Adolescente

Magister Public Health - MPH / Antiretroviral regimens have side effects that can threaten adherence by patients resulting in evolution of viral resistance due to suboptimal drug levels. Studies have shown that drug adherence of at least 80% can result in viral load suppression. There is no literature on the association between the level of adherence to antiretroviral therapy and the degree of virological suppression in Namibia. The aim of the present study was to determine the therapeutic and virological outcomes in HIV/AIDS patients at 6 and 12 months after initiation of highly-active antiretroviral therapy (HAART) in an urban population in Namibia. The distribution of viral load results showed a low uptake (35%) of virological monitoring at 6 month time point and even lower (12%) at 12 months. A conservative viral load threshold for virological response is required in the Namibian setting. The current adherence level of &gt;80% encourage increased ARV therapy rollout. Poor virological outcome was associated with self-reported adherence. / South Africa

Magister Public Health - MPH / Antiretroviral therapy (ART) improves the quality of lives of people living with HIV/AIDS by suppressing HIV replication and improving the patient’s immunity. An improved immunity will help prevent patients from contracting opportunistic infections. Adherence to ART is vital to obtain good clinical outcome for patients. Defaulting ART leads to increase in viral load, decreased host immunity, development of HIV drug resistant strains, exposure to opportunistic infections and ultimately death. HIV positive patients who are on ART face many challenges in adhering to their medications, these challenges act as barriers to their adherence to treatment. This study explores the barriers that adult patients in a hospital in Eastern Cape of South Africa face while on treatment. These barriers include individual factors, socio-economic factors, health service factors, medical regimen factors.

South Africa, with an estimated 5.7 million people living with HIV, continues to have one of the largest epidemics in the world. The introduction of HAART resulted in prolonged and improved quality of life of many infected patients. However, adverse effects caused by these drugs have become a major concern as they affect the adherence of patients and in some cases even result in the death of patients. Although much research has been and is still being conducted in the area of understanding, preventing and management of ARV adverse effects, there is still a need for patients to be actively involved in self-monitoring for adverse effects. This will assist health care professionals in early identification of serious or potentially serious ARV effects. This study aimed at evaluating the usefulness of strategies developed and employed in the identification, recording and monitoring of adverse effects. The study was conducted with patients receiving HAART from a private HIV and AIDS clinic in Uitenhage, Eastern Cape, South Africa. The research project was approved by the Nelson Mandela Metropolitan University Research and Ethics Committee and the research site. This was an experimental, randomized controlled study carried out over a period of three months (August to October 2009), with a sample size of 160 patients divided into four study groups of 40 patients each. Two monitoring strategies, namely an ARV adverse effect monitoring tool and a patient self-monitoring diary were developed and used for the identification and recording of adverse effects. The four study groups included a Control group, a Tool group, a Diary group and a Tool-Diary group. Willing patients, after signing an informed consent form, were randomly assigned to one of the four groups by participating health care workers at the study site. Data was retrieved from the patient files by the researcher. Descriptive statistical analysis of the findings of the study was conducted using SPSS®. One hundred and forty nine patients were included in the final data analysis. Of the 80 diaries handed out to patients, only 33 were returned and due to errors only 31 were suitable for analysis. Monitoring tools were completed and analysed for 36 patients. The tool was found to be more effective in identifying adverse effects of a physical nature (such as peripheral neuropathy and lipodystrophy) than the usual methods of monitoring employed by the clinic, whilst the diary, used alone, was found to be less effective. Use of the tool and diary combined resulted in the most significant identification and recording of central nervous system related adverse effects and physical adverse effects. However due to the low return rate of the diaries and the majority of the monitoring tool not being completed in many instances the results of this study may not be generalisable. The study results did however suggest that combining the tool and the diary methods of adverse effect identification, yielded the most favourable results when compared to each method alone. This may be attributed to the fact that the tool is useful in identifying objective symptoms and the diaries subjective symptoms, particularly in instances where the patients forget to report their symptoms to healthcare professional whilst at the clinic. The diaries were also reported to improve adherence for more than 90 percentage (n=31) of the patients. More research would be needed in order to verify the exact significance of the tool and the diary in identifying and recording adverse effects and symptoms of adverse effects.

HIV-1 is the causative agent for AIDS and has been a major focus of research for the past two decades. Though there is a combination therapy in place known as the “Highly Active Anti-Retroviral Therapy” (HAART), its usefulness is confounded by the generation of escape mutants, a host of side effects, and its prohibitive cost. The most useful alternative would be the prevention of infection by vaccination. Vaccine research has been focused on the use of recombinant protein sub-units of the virus or combinations thereof to elicit a neutralizing response against the virus. These approaches have mostly resulted in a failure to generate broadly cross reactive neutralizing response against primary strains of the virus. The work reported herein is aimed at designing a rigidified version of gp120/gp120 derivatives and understanding the scope of the various antigenic regions in gp120 in generating a neutralization response. Chapter one discusses some general features of the virus and the immune system. The general nature of AIDS, its spread and its immunological characteristics are also described in this chapter. Chapter two discusses the design and NMR structural analysis of gp120 bridging sheet peptide mimics in methanol and water. The structure of gp120 can be loosely divided into two domains (the outer domain and the inner domain) that are linked together by a discontinuous four stranded antiparallel beta sheet known as the bridging sheet. The bridging sheet is known to overlap with the coreceptor binding site of gp120 and hence is a suitable target for designing virus-entry inhibitors. 17b, a neutralizing antibody isolated from an infected individual, is known to bind to this region of gp120. Our aim in this part of the work was to design a four stranded antiparallel beta sheet, based on the sequence of the bridging sheet, that would contain most of the residues involved in 17b binding. NMR and CD studies confirmed that the peptide was well structured in methanol but the structure was largely lost on addition of aqueous solvent. A small population of the peptide was found to be well-folded in aqueous solution. Chapter three discusses the design and characterization of a gp120-CD4D12 single chain. It is well known that the conformation of gp120 changes upon binding CD4 to expose cryptic epitopes, known as CD4i epitopes. In this work we report the generation of a single chain gp120-CD4 construct that has the cryptic epitopes exposed. The construct bound to 17b, a conformation specific antibody against the bridging sheet of gp120, a cryptic epitope, as well as a non-covalent complex of gp120:CD4D12. There was also very insignificant secondary structural change in gp120 upon complex formation with CD4D12 as observed by CD spectroscopy. Immunological studies with DNA and protein vaccination in guinea-pigs indicated that though 17b like antibodies are generated after immunization, they did not contribute towards the neutralization of primary isolates of the virus. It was also observed that it was the anti-CD4D12 antibodies that were responsible for the neutralization by the sera. These studies indicated towards the inability of the bridging sheet to generate effective neutralization response in case of vaccination with gp120/CD4 complexes. Chapter four discusses the design of a mimic of the gp120/CD4 complex. Since it was seen from our previous work that gp120/CD4 complexes generate a large fraction of antiCD4 antibodies and hence are unsuitable for vaccination purposes, we generated a construct with the minimal binding region of CD4. The small fragment of CD4 spanning from 21st residue to 64th residue was inserted in the V1/V2 loop of gp120. The insertion site was designed based on the region of gp120 closest to this fragment and capable of tolerating insertions. This protein did not bind to 17b as well as gp120/CD4 complex but showed a higher binding compared to full length gp120. Further immunological characterization with this protein revealed that it was not capable of generating neutralizing antibodies against the virus. Chapter five discusses the design and execution of a SPR based solution phase competition experiment to find the solution phase binding constant of CD4 and CD4 analogs to gp120. A major problem during the analysis of binding data obtained by SPR is the accurate determination of Rmax, a parameter needed to obtain an accurate equilibrium dissociation constant. In this chapter we have developed a binary as well as a ternary solution phase SPR based assay to accurately determine a solution phase equilibrium binding constant. The binding constants were determined for gp120 binding to CD4D12 and other CD4 analogs. To confirm the validity of the assay, a control antigen:antibody interaction whose equilibrium dissociation constant has been determined by other methods has been used as a test case. Chapter six discusses the design and characterization of V3 peptides inserted in the loop regions of E. coli Thioredoxin (Trx). Trx has earlier been used to display random peptide libraries between the 33rd and the 34th residue. We have constructed three constructs where the peptide has been inserted between the 33rd and 34th residue, between the 74th and 75th residue and between the 84th and 85th residue. The insertion between 74th and 75th position (74V3Trx) was found to be superior to the other two and would be a suitable alternative for display of a random peptide library. The binding of these constructs to 447-52D, a V3 peptide specific antibody was characterized. These were also characterized immunologically, and 74V3Trx was found to generate weakly neutralizing activity against the MN strain of HIV-1. Competition experiments with 447-52D with these sera indicated that there were antibodies generated that could compete out 447-52D binding to gp120 but not in sufficient concentration to provide broad neutralization. Appendix 1 discusses the rational design of disulfides to stabilize proteins based on the analysis of naturally occurring disulfides. In our attempts to design a rigidified version of gp120 we had designed disulfides in gp120 based on its crystal structure. Many of these were disulfides that would span antiparallel adjacent strands. In order to improve the design principles, we analyzed naturally occurring disulfides that span antiparallel adjacent strands and characterized them in terms of their positional preference in a beta sheet. It was found that these disulfides mostly occur on edge strands and are found exclusively between non-hydrogen bonded registered pairs of adjacent antiparallel strands. Mutagenesis on Thioredoxin was performed to verify our results. It was found that disulfides designed between the non-hydrogen bonded pairs of antiparallel strands could significantly stabilize the protein whereas the ones between hydrogen bonded pairs destabilized the protein.

L’épidémie du VIH/SIDA a particulièrement touché l’Afrique Sub-Saharienne. En 1998, le gouvernement sénégalais a mis en place une cohorte observationnelle d’adultes infectés par le VIH-1 afin d’évaluer l’impact des antirétroviraux donnés au sein de l’Initiative Sénégalaise d’Accès aux Antirétroviraux (ISAARV). Plus spécifiquement, cette évaluation portait sur l’efficacité clinique et biologique des traitements, leur tolérance clinique et biologique, l’observance et l’émergence de résistances virales. La cohorte ANRS 1215 a été mise en place entre 1998 et 2002 à partir des 404 premiers patients inclus dans l’ISAARV. Cette thèse a utilisé les données recueillies au sein de cette cohorte pour analyser certains aspects de la réponse bio-cliniques aux traitements antirétroviraux en Afrique. Dans la première partie de ce travail, différents aspects de l’infection par le VIH en Afrique Sub-Saharienne et les questions relatives à la réponse bio-cliniques aux traitements antirétroviraux ont été passés en revus. Dans la seconde partie, les aspects méthodologiques de l’analyse des données longitudinales de cohorte ainsi que les problématiques associées ont été étudiés. Enfin, différents aspects de la réponse bio-cliniques après initiation des antirétroviraux ont été analysés au travers de 5 articles portant sur la mortalité précoce, la reconstitution immunologique, la survenue d’évènements classant, la réponse et la tolérance à deux régimes thérapeutiques différents. / Sub-Saharan Africa has been heavily affected by the HIV/AIDS epidemic. In 1998, the Sengalese government launched an observational cohort of HIV-1 infected adults that aimed to assess the impact of antiretroviral therapy (ART) provided by the “l’Initiative Sénégalaise d’Accès aux Antirétroviraux” (ISAARV). This assessment focused more specifically on the clinical and biological effectiveness of ART, on the clinical and biological tolerance and on the emergence of virological resistances. The cohort was made of the first 404 patients included in the ISAARV between 1998 and 2002. The data collected through this cohort have been used to analyse in this work various aspects of biological and clinical answer to ART in Africa. In the first section, different aspects of the HIV/AIDS infection in Sub-Saharan countries have been reviewed with an emphasis put on the remaining bio-clinical questions. In the second, methodological aspects of the analysis of longitudinal data and related questions have been studied. Then, various aspects of the biological and clinical answer to ART have been analysed through five articles about early mortality, immune reconstitution, AIDS-defining illness occurrence, and the effectiveness and tolerance of two regimen.