Introdução: A infecção pelo HIV está sendo considerada de caráter crônico e potencialmente controlável desde a instituição da Terapia Antirretroviral (TARV) e disponibilidade de marcadores para acompanhamento da sua evolução. A adesão ao tratamento necessária para garantir a supressão virológica é elevada, necessitando de monitoramento adequado a fim de detectar potenciais pacientes não-aderentes e auxiliar nas intervenções. O uso dos registros de dispensação de medicamentos pode ser uma estratégia simples e factível para identificar potenciais pacientes não-aderentes. Intervenções do profissional farmacêutico podem contribuir para aumentar a adesão e obter desfechos clínicos favoráveis. Objetivos: O objetivo deste estudo é avaliar se os registros de dispensação de medicamentos e intervenções farmacêuticas podem contribuir na avaliação e melhora da adesão ao tratamento antirretroviral e outros desfechos clínicos relevantes, através de duas revisões sistemáticas. Métodos: Foram realizadas duas revisões sistemáticas, com busca nas bases de dados MEDLINE, Registro de ensaios clínicos da Cochrane, EMBASE, IBECS, CINAHL, IPA, SCOPUS, Web Of Science, LILACS, Scielo e Clinical Trials. Para avaliar os registros de dispensação na adesão ao tratamento foram selecionados estudos que estimaram a adesão ao tratamento pelos registros de dispensação de antirretrovirais e compararam com desfechos clínicos (carga viral, contagem de linfócitos CD4, resistência viral ou óbito) ou outro método para estimar a adesão. Para avaliar intervenções farmacêuticas na TARV foram selecionados ensaios clínicos randomizados em que havia a participação de farmacêutico na intervenção profissional, e as principais medidas de desfecho selecionadas foram a adesão ao tratamento, carga viral e CD4. Resultados: Foram encontrados 3551 estudos para a primeira revisão, dos quais 92 foram selecionados. Os estudos selecionados eram heterogêneos, sendo estudos de coorte os mais frequentes, o período mais utilizado para cálculo da adesão de seis meses e o ponto de corte mais utilizado para adesão de 95%. Os resultados dos estudos apontam para associação positiva entre adesão estimada pelos dados de farmácia e carga viral e outros métodos, com melhores resultados desta associação quando avaliada de forma temporal. Para a revisão sobre atenção farmacêutica na TARV foram encontrados 681 estudos, dos quais quatro atenderam aos critérios de inclusão. A adesão ao tratamento foi estimada em todos os estudos, e o odds ratio sumarizado foi de 1,47 (IC95% 0,81 – 2,65). A supressão virológica foi estimada em três estudos, obtendo odds ratio sumarizado de 1,95 (IC95% 0,61 – 6,25). Conclusões: Os resultados dos estudos indicam que os dados da farmácia podem ser úteis na avaliação da adesão ao tratamento, relacionando-se com desfechos clínicos como carga viral e CD4. Os resultados das meta-análises sugerem que a intervenção farmacêutica pode auxiliar na melhora da adesão ao tratamento antirreroviral e carga viral, no entanto, não houve diferença estatisticamente significativa entre os grupos. Em populações com baixa adesão e maior vulnerabilidade, a intervenção farmacêutica pode ser mais eficaz. / Introduction: HIV infection is being considered chronic and potentially manageable since the introduction of Antiretroviral Therapy (ART) and availability of biogical markers for monitoring its evolution. Adherence to treatment necessary to ensure virologic suppression is high, requiring adequate monitoring to detect potential nonadherent patients and plan interventions. The use of medication dispensing records can be a simple and feasible method to identify potential non-adherent patients. Pharmacist interventions can increase adherence and get favorable outcomes. Objectives: The aim of this study is to evaluate whether the dispensing drugs records and pharmaceutical interventions may help in assessing and improving adherence to antiretroviral treatment and other relevant clinical outcomes through two systematic reviews. Methods: Two systematic reviews were performed, with the search strategies performed in MEDLINE, Cochrane registration of clinical trials, EMBASE, IBECS, CINAHL, IPA, SCOPUS, Web of Science, LILACS, SciELO and Clinical Trials database. To evaluate the prescription refill records treatment adherence, were selected studies that had measures treatment adherence by records of antiretrovirals dispensing and compared with clinical outcomes (viral load, CD4 count, viral resistance or death) or alternative method to estimate adherence. To evaluate pharmaceutical interventions on ART, randomized clinical trials in which there was participation of pharmacists in the intervention were selected, and the main outcome measures were treatment adherence, viral load and CD4. Results: 3551 studies were found for the first review, of which 92 were selected. The selected studies were heterogeneous, with cohort studies the most frequent, the period most frequent used to calculate adherence were six months and the cutoff for adherence were 95%. Study results indicate good relationship between adherence estimated by pharmacy data, viral load and other methods, with best results of the association between adherence and viral load as measured temporally. For the review of pharmaceutical care in ART 681 studies were found, of which four met the inclusion criteria. Adherence to treatment was estimated in all studies, and summarized odds ratio was 1.47 (95% CI 0.81 to 2.65). Virological suppression was estimated in three studies, getting summarized odds ratio of 1.95 (95% CI 0.61 to 6.25). Conclusions: The study results indicate that the pharmacy claim data can be useful in assessing adherence to antirretroviral treatment, correlating with clinical outcomes such as viral load and CD4. The results of the meta-analyzes suggest that there was no difference between treatment adherence and virologic suppression in intervention group with pharmacist and the control group, despite overall results being favorable to pharmaceutical intervention. In populations with low compliance and vulnerability, pharmaceutical interventions may be more effective.

Introdução: A infecção pelo HIV está sendo considerada de caráter crônico e potencialmente controlável desde a instituição da Terapia Antirretroviral (TARV) e disponibilidade de marcadores para acompanhamento da sua evolução. A adesão ao tratamento necessária para garantir a supressão virológica é elevada, necessitando de monitoramento adequado a fim de detectar potenciais pacientes não-aderentes e auxiliar nas intervenções. O uso dos registros de dispensação de medicamentos pode ser uma estratégia simples e factível para identificar potenciais pacientes não-aderentes. Intervenções do profissional farmacêutico podem contribuir para aumentar a adesão e obter desfechos clínicos favoráveis. Objetivos: O objetivo deste estudo é avaliar se os registros de dispensação de medicamentos e intervenções farmacêuticas podem contribuir na avaliação e melhora da adesão ao tratamento antirretroviral e outros desfechos clínicos relevantes, através de duas revisões sistemáticas. Métodos: Foram realizadas duas revisões sistemáticas, com busca nas bases de dados MEDLINE, Registro de ensaios clínicos da Cochrane, EMBASE, IBECS, CINAHL, IPA, SCOPUS, Web Of Science, LILACS, Scielo e Clinical Trials. Para avaliar os registros de dispensação na adesão ao tratamento foram selecionados estudos que estimaram a adesão ao tratamento pelos registros de dispensação de antirretrovirais e compararam com desfechos clínicos (carga viral, contagem de linfócitos CD4, resistência viral ou óbito) ou outro método para estimar a adesão. Para avaliar intervenções farmacêuticas na TARV foram selecionados ensaios clínicos randomizados em que havia a participação de farmacêutico na intervenção profissional, e as principais medidas de desfecho selecionadas foram a adesão ao tratamento, carga viral e CD4. Resultados: Foram encontrados 3551 estudos para a primeira revisão, dos quais 92 foram selecionados. Os estudos selecionados eram heterogêneos, sendo estudos de coorte os mais frequentes, o período mais utilizado para cálculo da adesão de seis meses e o ponto de corte mais utilizado para adesão de 95%. Os resultados dos estudos apontam para associação positiva entre adesão estimada pelos dados de farmácia e carga viral e outros métodos, com melhores resultados desta associação quando avaliada de forma temporal. Para a revisão sobre atenção farmacêutica na TARV foram encontrados 681 estudos, dos quais quatro atenderam aos critérios de inclusão. A adesão ao tratamento foi estimada em todos os estudos, e o odds ratio sumarizado foi de 1,47 (IC95% 0,81 – 2,65). A supressão virológica foi estimada em três estudos, obtendo odds ratio sumarizado de 1,95 (IC95% 0,61 – 6,25). Conclusões: Os resultados dos estudos indicam que os dados da farmácia podem ser úteis na avaliação da adesão ao tratamento, relacionando-se com desfechos clínicos como carga viral e CD4. Os resultados das meta-análises sugerem que a intervenção farmacêutica pode auxiliar na melhora da adesão ao tratamento antirreroviral e carga viral, no entanto, não houve diferença estatisticamente significativa entre os grupos. Em populações com baixa adesão e maior vulnerabilidade, a intervenção farmacêutica pode ser mais eficaz. / Introduction: HIV infection is being considered chronic and potentially manageable since the introduction of Antiretroviral Therapy (ART) and availability of biogical markers for monitoring its evolution. Adherence to treatment necessary to ensure virologic suppression is high, requiring adequate monitoring to detect potential nonadherent patients and plan interventions. The use of medication dispensing records can be a simple and feasible method to identify potential non-adherent patients. Pharmacist interventions can increase adherence and get favorable outcomes. Objectives: The aim of this study is to evaluate whether the dispensing drugs records and pharmaceutical interventions may help in assessing and improving adherence to antiretroviral treatment and other relevant clinical outcomes through two systematic reviews. Methods: Two systematic reviews were performed, with the search strategies performed in MEDLINE, Cochrane registration of clinical trials, EMBASE, IBECS, CINAHL, IPA, SCOPUS, Web of Science, LILACS, SciELO and Clinical Trials database. To evaluate the prescription refill records treatment adherence, were selected studies that had measures treatment adherence by records of antiretrovirals dispensing and compared with clinical outcomes (viral load, CD4 count, viral resistance or death) or alternative method to estimate adherence. To evaluate pharmaceutical interventions on ART, randomized clinical trials in which there was participation of pharmacists in the intervention were selected, and the main outcome measures were treatment adherence, viral load and CD4. Results: 3551 studies were found for the first review, of which 92 were selected. The selected studies were heterogeneous, with cohort studies the most frequent, the period most frequent used to calculate adherence were six months and the cutoff for adherence were 95%. Study results indicate good relationship between adherence estimated by pharmacy data, viral load and other methods, with best results of the association between adherence and viral load as measured temporally. For the review of pharmaceutical care in ART 681 studies were found, of which four met the inclusion criteria. Adherence to treatment was estimated in all studies, and summarized odds ratio was 1.47 (95% CI 0.81 to 2.65). Virological suppression was estimated in three studies, getting summarized odds ratio of 1.95 (95% CI 0.61 to 6.25). Conclusions: The study results indicate that the pharmacy claim data can be useful in assessing adherence to antirretroviral treatment, correlating with clinical outcomes such as viral load and CD4. The results of the meta-analyzes suggest that there was no difference between treatment adherence and virologic suppression in intervention group with pharmacist and the control group, despite overall results being favorable to pharmaceutical intervention. In populations with low compliance and vulnerability, pharmaceutical interventions may be more effective.

Introdução: A infecção pelo HIV está sendo considerada de caráter crônico e potencialmente controlável desde a instituição da Terapia Antirretroviral (TARV) e disponibilidade de marcadores para acompanhamento da sua evolução. A adesão ao tratamento necessária para garantir a supressão virológica é elevada, necessitando de monitoramento adequado a fim de detectar potenciais pacientes não-aderentes e auxiliar nas intervenções. O uso dos registros de dispensação de medicamentos pode ser uma estratégia simples e factível para identificar potenciais pacientes não-aderentes. Intervenções do profissional farmacêutico podem contribuir para aumentar a adesão e obter desfechos clínicos favoráveis. Objetivos: O objetivo deste estudo é avaliar se os registros de dispensação de medicamentos e intervenções farmacêuticas podem contribuir na avaliação e melhora da adesão ao tratamento antirretroviral e outros desfechos clínicos relevantes, através de duas revisões sistemáticas. Métodos: Foram realizadas duas revisões sistemáticas, com busca nas bases de dados MEDLINE, Registro de ensaios clínicos da Cochrane, EMBASE, IBECS, CINAHL, IPA, SCOPUS, Web Of Science, LILACS, Scielo e Clinical Trials. Para avaliar os registros de dispensação na adesão ao tratamento foram selecionados estudos que estimaram a adesão ao tratamento pelos registros de dispensação de antirretrovirais e compararam com desfechos clínicos (carga viral, contagem de linfócitos CD4, resistência viral ou óbito) ou outro método para estimar a adesão. Para avaliar intervenções farmacêuticas na TARV foram selecionados ensaios clínicos randomizados em que havia a participação de farmacêutico na intervenção profissional, e as principais medidas de desfecho selecionadas foram a adesão ao tratamento, carga viral e CD4. Resultados: Foram encontrados 3551 estudos para a primeira revisão, dos quais 92 foram selecionados. Os estudos selecionados eram heterogêneos, sendo estudos de coorte os mais frequentes, o período mais utilizado para cálculo da adesão de seis meses e o ponto de corte mais utilizado para adesão de 95%. Os resultados dos estudos apontam para associação positiva entre adesão estimada pelos dados de farmácia e carga viral e outros métodos, com melhores resultados desta associação quando avaliada de forma temporal. Para a revisão sobre atenção farmacêutica na TARV foram encontrados 681 estudos, dos quais quatro atenderam aos critérios de inclusão. A adesão ao tratamento foi estimada em todos os estudos, e o odds ratio sumarizado foi de 1,47 (IC95% 0,81 – 2,65). A supressão virológica foi estimada em três estudos, obtendo odds ratio sumarizado de 1,95 (IC95% 0,61 – 6,25). Conclusões: Os resultados dos estudos indicam que os dados da farmácia podem ser úteis na avaliação da adesão ao tratamento, relacionando-se com desfechos clínicos como carga viral e CD4. Os resultados das meta-análises sugerem que a intervenção farmacêutica pode auxiliar na melhora da adesão ao tratamento antirreroviral e carga viral, no entanto, não houve diferença estatisticamente significativa entre os grupos. Em populações com baixa adesão e maior vulnerabilidade, a intervenção farmacêutica pode ser mais eficaz. / Introduction: HIV infection is being considered chronic and potentially manageable since the introduction of Antiretroviral Therapy (ART) and availability of biogical markers for monitoring its evolution. Adherence to treatment necessary to ensure virologic suppression is high, requiring adequate monitoring to detect potential nonadherent patients and plan interventions. The use of medication dispensing records can be a simple and feasible method to identify potential non-adherent patients. Pharmacist interventions can increase adherence and get favorable outcomes. Objectives: The aim of this study is to evaluate whether the dispensing drugs records and pharmaceutical interventions may help in assessing and improving adherence to antiretroviral treatment and other relevant clinical outcomes through two systematic reviews. Methods: Two systematic reviews were performed, with the search strategies performed in MEDLINE, Cochrane registration of clinical trials, EMBASE, IBECS, CINAHL, IPA, SCOPUS, Web of Science, LILACS, SciELO and Clinical Trials database. To evaluate the prescription refill records treatment adherence, were selected studies that had measures treatment adherence by records of antiretrovirals dispensing and compared with clinical outcomes (viral load, CD4 count, viral resistance or death) or alternative method to estimate adherence. To evaluate pharmaceutical interventions on ART, randomized clinical trials in which there was participation of pharmacists in the intervention were selected, and the main outcome measures were treatment adherence, viral load and CD4. Results: 3551 studies were found for the first review, of which 92 were selected. The selected studies were heterogeneous, with cohort studies the most frequent, the period most frequent used to calculate adherence were six months and the cutoff for adherence were 95%. Study results indicate good relationship between adherence estimated by pharmacy data, viral load and other methods, with best results of the association between adherence and viral load as measured temporally. For the review of pharmaceutical care in ART 681 studies were found, of which four met the inclusion criteria. Adherence to treatment was estimated in all studies, and summarized odds ratio was 1.47 (95% CI 0.81 to 2.65). Virological suppression was estimated in three studies, getting summarized odds ratio of 1.95 (95% CI 0.61 to 6.25). Conclusions: The study results indicate that the pharmacy claim data can be useful in assessing adherence to antirretroviral treatment, correlating with clinical outcomes such as viral load and CD4. The results of the meta-analyzes suggest that there was no difference between treatment adherence and virologic suppression in intervention group with pharmacist and the control group, despite overall results being favorable to pharmaceutical intervention. In populations with low compliance and vulnerability, pharmaceutical interventions may be more effective.

Magister Public Health - MPH / Aim: To obtain baseline data on adherence levels and the major determinants of adherence among patients on HAART at Rundu Hospital, Namibia. Results: Seventy-eight percent of the 97 participants included in the study were female, resulting in a female to male ratio of 4 :1. The mean age of the participants was 36.7 (SD: 9.00) years with 80% of the participants being in the 20-44 age group. The mean duration on ART treatment was 20 (SD: 10.3) months with 76% of the participants being on ART for 24 or less months. The average adherence rate reported by mean composite of the three measures was 95.1%, while the proportion of patients who achieved adherence levels of 95% and above was 64%. The main barriers to adherence to HAART reported by participants were forgetfulness (28%), lack of food (13%) and being away from the pills (11%): facilitators reported included counselling (19%) and treatment supporters (11%). Having knowledge of the consequences of failing to take HAART as prescribed was significantly associated with adherence (p = 0.03), as was being female (p = 0.04) while living further than 6 km from the hospital was significantly associated with non adherence (p = 0.018).

Magister Public Health - MPH / Background: Namibia is one of the Southern African countries hardest hit by the HIV epidemic, with an estimated one out of every five people infected (MoHSS, 2004). Approximately 80,000 of the infected population currently require antiretroviral treatment (ART). In order to prevent the progression of the HIV infection to AIDS, patients are required to take antiretroviral medicines lifelong. This lifelong use exposes patients to toxicities of these medicines and the only available options of managing the toxicities of ARVs are to treat the toxicity or substitute or switch the offending medicines. Aim: The current study aimed to describe the extent and reasons for substituting and switching HAART at the Katutura Intermediate Hospital in Windhoek, Namibia. Methodology: A descriptive retrospective case series study, in which medical records were reviewed to determine the extent and reasons for substituting and switching HAART was conducted. Random sampling was used to draw a sample of 500 from 3477 adult HAART patients who commenced treatment between 1 January 2002 and 31 December 2006. A prepiloted data collection tool was used to collect the data. The following information was collected: baseline CD4 count, weight, initial ARVs, first and second ARV substitutions, ART switch and the reasons for substituting ARVs or switching ART during the indicated period. Epi Info version 6 was used to analyse frequencies, means and medians of all important variables in the data set. Results: The sample was made up of 500 HAART patients; 60% were females. The median age of the sample was 34 years (Inter-quartile range (IQR) 30 – 40) and the median CD4 cell count was 153 cells/mm3 (IQR 96 – 212) at initiation of therapy. The median time on treatment before first substitution was 28 months (IQR 24 – 34), whereas the median time before second substitution was 10 months (IQR 6 – 15) from the time of the first substitution. The median time before switching was 31 months (IQR 24 - 39). A total of 31% of the study subjects underwent a substitution once, whereas 1.8% underwent a second substitution. Only six (1.2%) patients switched to a second line treatment after the modification of the treatment. The most commonly recorded reason for the first substitution was toxicity (19%). As in other studies, stavudine (D4T), nevirapine (NVP) and efavirenz (EFV) were the ARVs associated with most of the recorded toxicities. High viral load (50%) was the most reported reason for switching. In almost half of the substitution cases the reasons for substitution were not stated, and in a third of the switch cases the reasons for switching were not stated. Conclusion: The rate of substitution at 31% was similar to that found in other resource poor settings, however, the rate of switching (1.2%) was much lower than was found in similar settings. The main reason stated for substituting antiretrovirals was “toxicity”. / South Africa

Thesis (Ph.D. (Physiology and Environmental Health)) -- University of Limpopo, 2021 / Background: Human immunodeficiency virus (HIV)-infection remains a major public health burden where approximately 38 million people are affected globally. Human immunodeficiency virus infection is associated with chronic inflammation which can lead to endothelial dysfunction and thrombosis, which are precursor events for cardiometabolic abnormalities such as dysglycaemia and dyslipidaemia. The degree of chronic inflammation, endothelial dysfunction, and hypercoagulation among HIVpositive adults on highly active antiretroviral therapy are not well understood in Sub- Saharan Africa. The objective of this study was to determine the effect of highly active antiretroviral therapy (HAART) on chronic inflammation, endothelial dysfunction, and hypercoagulation among HAART-exposed adult South African participants in a rural setting. Aim: The study aimed to determine the effects of HAART on early biomarkers of cardiovascular disease in the HIV-positive subjects. Methods: The study was cross-sectional, descriptive, and quantitative in design. The research population consisted of 158 participants of males and females within the age range of 18 – 81 years from Mankweng Hospital and surrounding clinics. The study population comprised of three groups, HIV-negative (control group), HIV-positive treatment naïve (HAART-naïve group), and HIV-positive participants on HAART (HAART-exposed group). Weight and height were measured using Omron BF 400 and a portable stadiometer respectively, to calculate the body mass index. Glucose and lipid levels were determined on Cobas® Integra 400 plus auto-analyser. The CD4+ T cell count was determined on the Cytomics FC500 Flow Cytometer Multi-Platform loader. The concentration of fibrinogen, c-reactive protein (CRP), L-selectin, D-dimers, P-selectin, von Willebrand factor (VWF), soluble intercellular adhesion molecule (sICAM-1), and soluble vascular cell adhesion molecule (sVCAM-1) in serum samples were determined on the Luminex 200TM. Data were analysed using SPSS version 25.0. Descriptive statistics were performed on all variables and analysis of covariance was used to determine differences across all groups. Correlation coefficients and multiple regression analyses were used to determine associations. Results: Body mass index (BMI) and glucose metabolism were not significantly affected by HAART exposure. However, the HAART-exposed group had significantly increased LDL-C (F (2, 154) = 7.501, p = 0.001) and TC (F (2, 154) = 9.174, 0.0002) levels. The prevalence of high LDL-C levels was significantly elevated in the HAART-exposed group (29.6%) (p = 0.041). The prevalence of pre-diabetes (11.3%) was the highest among the HAART-exposed group (non-significant), although, no significant difference was observed. While P-selectin was significantly reduced in the HAART-exposed group (F (2, 154 = 7.253, p = 0.001). On the other hand, the HAARTexposed group also significantly increased VWF (F (2, 154 = 4.556, p = 0.011). The HAART-exposed group showed no significant effect on L-selectin, sICAM-1, sVCAM- 1, CRP, fibrinogen and D-dimer levels. However, D-dimer was negatively associated with HAART (r = -0.249, p = 0.011). There were significant independent association between the combined HAART regimens and P-selectin (Std β = 0.219, p = 0.032), first-line regimen with both P-selectin (Std β = 0.434, p = 0.004) and sVCAM-1 Std β = 0.328, p = 0.031), second-line regimens with L-selectin (Std β = 1.032, p = 0.005) and, a positive independent association between first-line regimen and D-dimer (β = 0.741, p = 0.0001). Although BMI and glucose metabolism were not significantly affected in both the HAART-exposed and HAART-naïve groups, dyslipidaemia was present across the three groups (HAART-exposed, HAART-naïve and control). HAART-exposure showed a protective effect by reducing endothelial dysfunction (ED) and hypercoagulation. Yet, ED was still present among this rural South African HAART-exposed population. The HAART-exposed group may be at increased risk for CVD. Therefore, CVD should be regularly monitored in the HAART-exposed population. / National Research Fund, the Health and Welfare Sector Education and Training Authority, and the University of Limpopo (UL)

Thesis (MMedSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Background: Serum free light chains (FLC) are associated with imbalances in heavy and light chain production. Abnormal FLC ratios have been associated with risk of progression in certain diseases. Automated assays are available for their determination and they are used in the followup and management of patients with monoclonal gammopathies. Acceptable imprecision, specificity, accuracy and reproducibility between reagent batches is required to prevent under- or overestimation. Method validation is a standard process in every good laboratory to judge the acceptability of a new method. Reference intervals have been established in an older population, but it was considered important to verify these in our population. HIV is associated with B-cell dysfunction. As B-cell abnormalities are associated with disorders leading to monoclonal gammopathies, we postulated that the FLC levels and FLC ratio would be abnormal in HIV infected individuals. Methods and materials: Controls and pooled patient samples were used for the method validation study which included imprecision studies, linearity, recovery and interference studies, and method comparison studies, the latter compared our method to the same method used in another laboratory. For the reference interval study, blood was obtained from 120 healthy subjects. The following blood tests were performed: total protein, IgG, IgA, IgM, creatinine, protein electrophoresis, kappa FLC and lambda FLC. Using the kappa and lambda FLC results, a FLC ratio was determined. Three hundred and sixty-nine HIV positive subjects were then studied. The same tests were performed, as well as CD4+ counts and viral loads on the majority of them. Results: For the method validation study, precision, linearity and recovery was acceptable. Minimal interference was observed with haemolysis, lipaemia, bilirubin and rheumatoid factor. Our method showed comparable performance with the established method. For the reference interval study, all the creatinine values were normal, as were serum protein values. The serum protein electrophoreses were independently reviewed by 3 pathologists. Most were normal, with a few polyclonal increases seen, but no definite monoclonal bands. The 95% reference intervals for FLC’s as well as the FLC ratio were not statistically significantly different to the manufacturer’s recommendations. When examining the HIV positive study population, we found that FLC and FLC ratio were influenced by markers of HIV disease severity, such as CD4+ count, IgG, viral load, use of antiretroviral treatment and abnormal serum protein electrophoreses. Conclusion: The validation study of FLC showed excellent precision, acceptable bias, good linearity, good recovery and minimal interference, allowing routine introduction of the test. The 95% reference intervals obtained for our population were slightly higher than those recommended by the manufacturer. However, as most of the values fell within the manufacturer’s limits, we could accept the manufacturer’s recommended cut-offs. We found that FLC levels were definitely influenced by markers of HIV disease severity in our population and we postulate that they may be of use for follow-up of patients with HIV. / AFRIKAANSE OPSOMMING: Agtergrond: Serum vry ligte kettings (VLK) word geassosieer met ‘n wanbalans van ligte en swaar ketting produksie. Abnormale VLK ratios is geassosieer met ‘n risiko van verloop in sekere siektes. Geoutomatiseerde laboratorium toetse vir VLK is beskikbaar vir hul bepaling en word gebruik om pasiënte met monoklonale gammopatieë op te volg en te behandel. Aanvaarbare impresisie, spesifisiteit, akkuraatheid en herhaalbaarheid tussen reagens besendings is belangrik om onder- of oorbepaling te verhoed. Metode validasie is ’n standaard proses in elke goeie laboratorium om die aanvaarbaarheid van ’n nuwe metode te bepaal. Verwysingswaardes is al bepaal in ’n ouer populasie. Ons het besluit om die verwysingswaardes in ons populasie te bepaal. Mens-immuungebrekvirus (MIV) word geassosieer met B-sel disfunksie. Omdat B-sel abnormaliteite geassosieer word met afwykings wat tot monoklonale gammopatieë lei, het ons gepostuleer dat die VLK vlakke en VLK ratio abnormaal sal wees in MIV geïnfekteerde persone. Metodes en Materiale: Kontroles en pasiënt monsters is gebruik vir die metode validasie studie wat impresisie studies, lineariteit, herwinning, inmenging en metode korrelasie studies ingesluit het. In laasgenoemde geval is ons metode met dieselfde metode van ’n ander laboratorium vergelyk. Vir die verwysingswaardes studie is 120 gesonde persone se bloed gebruik. Die volgende toetse is bepaal: totale proteïen, IgG, IgA, IgM, kreatinien, proteïen elektroferese, kappa en lambda VLK. Die VLK ratio is bepaal deur die kappa en lambda resultate te gebruik. Driehonderd nege en sestig MIV-positiewe pasiente is gebruik vir die studie. Dieselfde toetse was gedoen, asook CD4+ tellings en virale ladings op die meerderheid van pasiente. Resultate: Vir die metode validasie studie, was presisie, lineariteit en herwinning aanvaarbaar. Minimale inmenging van hemolise, lipemie, bilirubien en rumatoïede factor is waargeneem. Ons metode het goed gekorreleer met die bepaalde metode. Die serum kreatinien en serum totale proteïen waardes was normaal tydens die verwysingswaardes studie. Die serum proteïen elektroferese was onafhanklik beoordeel deur 3 patoloë. Die meeste was normaal met enkele poliklonale verhogings, maar geen definitiewe monoklonale bande nie. Die 95% verwysings intervalle vir VLK en VLK ratio het nie statisties betekenisvol verskil van die vervaardiger se aanbevelings nie. In die studie van die MIV-positiewe studie populasie, het ons gevind dat VLK en VLK ratio beïnvloed word deur merkers van ernstige MIV siekte, soos CD4+ telling, IgG, virale lading, die gebruik van antiretrovale medikasie en abnormale serum proteïen elektroferese. Gevolgtrekking: Die validasie studie van VLK het uitstekende presisie, aanvaarbare partydigheid, goeie lineariteit, goeie herwinning en minimale inmenging gewys, wat die roetine instelling van die toets toegelaat het. Die 95% verwysingsintervalle wat vir ons populasie bepaal is, was effens hoër as die vervaardiger se aanbeveling. Die meeste van die waardes het egter binne die vervaardiger se limiete geval, dus kon ons die vervaardiger se afsnypunte aanvaar. Ons het gevind dat VLK vlakke definitief beïnvloed word deur merkers van die ernstigheidsgraad van MIV siekte in ons populasie en ons postuleer dat VLK van waarde kan wees met die opvolg van MIV pasiente. / NHLS / Harry Crossley for funding obtained

Les inhibiteurs de la protéase virale (IPs) sont utilisés avec succès dans le cadre d'une thérapie anti-VIH-1. L'efficacité de ce traitement est incontestable notamment en terme de réduction de la charge virale et de maintient du taux de lymphocytes CD4 circulants. Depuis l'incorporation des IPs dans la prise en charge thérapeutique, on note une réduction significative de la morbidité et de la mortalité ainsi qu'un allongement de la durée de vie des patients. Cependant, la prise de ces molécules anti-rétrovirales s'accompagne de nombreux effets secondaires. Les plus préoccupants d'entre eux sont : une lipodystrophie partielle, une hyperlipidémie, une insulino-résistance, une athérosclérose prématurée, ainsi que des infarctus du myocarde. Les patients sont également confrontés à l'apparition de maladies généralement associées à l'âge telles que la neurodégenérescence, l'ostéopénie et le développement de tumeurs malignes. Les mécanismes cellulaires et moléculaires impliqués dans ces altérations métaboliques n'ont pas encore été élucidés. L'objectif de cette étude était d'étudier les effets cellulaires de quatre IPs (Atazanavir, Lopinavir, Ritonavir et Saquinavir) sur la cellule musculaire striée squelettique humaine. Ces travaux ont permis d'identifier une augmentation de la production d'espèces oxygénées réactives (ERO), une altération morphologique du réticulum sarco/endoplasmique (RS/RE) ainsi qu'une augmentation de l'expression de CHOP, un marqueur du stress de ce compartiment et une diminution de l'expression et de la localisation dans les microdomaines membranaires (lipid rafts) de la cavéoline 3 et de la flotilline 1. Enfin, l'utilisation d'un antioxydant, le Resvératrol protége le myotube primaire humain de ces différentes altérations engendrées par les IPs. Ces données suggèrent un rôle central de la surproduction d'ERO dans le développement du stress du RE/RS et de la perte de localisation des protéines résidantes des microdomaines membranaires. De plus , en l'absence de persceptive vaccinale concrète, le Resvératrol, au travers de ces effets protecteurs, pourrait se révéler un atout de choix dans l'atténuation des effets secondaires des IPs en contribuant ainsi à l'amélioration de la prise en charge du patient séropositif. / HIV protease inhibitors (PI) have been successfully used in highly active antiretroviral therapy (HAART) of HIV-1 infection, the most effective treatment currently available. Incorporation of protease inhibitors in HAART has significantly reduced the morbidity and mortality and prolonged the lifespan of patients with HIV infection. Protease inhibitor benefits are unfortunately compromised by a number of clinically important adverse side-effects. Most patients on HAART develop a metabolic syndrome associated with partial lipodystrophy, hyperlipidemia, insulin resistance, premature atherosclerosis and myocardial infarction. Moreover, these patients face a growing number of other age-related comorbidities, such as neurodegeneration, osteopenia and malignancies. The cellular and molecular mechanisms underlying protease inhibitor-associated metabolic abnormalities remain elusive, but they seem to be related to overproduction of reactive oxygen speci es (ROS), induction of endoplasmic reticulum stress and activation of the unfolded protein response (UPR). The objective of this thesis was to determine the cell effects of four PIs (Atazanavir, Lopinavir, Ritonavir and Saquinavir) in cultures of primary human skeletal myotubes. This study showed that PIs increased ROS production, altered sarco/endoplasmic reticulum (SR/ER) morphology, increased expression of C/EBP homologous protein, a SR/ER stress marker, and decreased expression and localization at lipid rafts of Caveolin 3 and Flotillin 1. In addition, we showed that the antioxidant Resveratrol protected the human primary myotube of these iatrogenic effects. These data suggest a central role of the overproduction of ERO in the development of SR/ER stress and mislocalization of lipid raft proteins induced by PIs. Besides, in absence of vaccine, Resveratrol may be used by a potentiel therapeutic agent to attenuate PI-induced side effects

L’histoire naturelle de l’infection anale par le virus du papillome de type 16 (VPH-16) est mal définie pour les hommes ayant des relations sexuelles avec d’autres hommes (HARSAHs) VIH-séropositifs. Le but de cette étude était d’évaluer l’association entre la charge épisomale et intégrée du VPH-16 et la progression de la néoplasie intraépithéliale anale (AIN). Les charges épisomales et intégrées du VPH-16 furent mesurées par PCR quantitatif en temps réel sur 665 spécimens anaux obtenus de 135 hommes VPH-16-positifs participant à l’étude prospective HIPVIRG (Human Immunodeficiency and Papilloma VIrus Research Group). Le grade de l’AIN fut déterminé sur des biopsies obtenues lors des anuscopies à haute résolution périodiques. L’intégration du VPH-16 fut confirmée par DIPS-PCR pour démontrer la présence de jonctions virales-cellulaires. La charge épisomale du VPH-16 [ratio de cote (OR) 1.5, intervalle de confiance (IC) à 95%=1.1–2.1], le nombre de types de VPH [OR 1.4 (IC 95%=1.1–1.8)] et le tabagisme actuel [OR 4.8 (IC 95%=1.3–18.6)], mais non la charge intégrée, furent associés aux lésions de haut-grade (AIN-2,3) après ajustement pour l’âge et le décompte des lymphocytes CD4. La charge épisomale du VPH-16 était le seul facteur prédictif de progression de l’AIN de bas-grade (AIN-1) vers l’AIN-2,3 [OR 8.0 (IC 95%=1.2–55.4)]. Les spécimens avec une charge épisomale du VPH-16 élevée étaient moins susceptibles de contenir de l’intégration [OR 0.5 (IC 95%=0.3–0.8)]. L’intégration du VPH-16 fut détectée en absence d’AIN, dans l’AIN-1 et dans l’AIN-2,3. L’analyse des jonctions virales-cellulaires ne permit pas d’identifier un site d’intégration spécifique. / The natural history of human papillomavirus type 16 (HPV-16) anal infection is undefined among HIV-seropositive men having sex with men (MSM). The aim of this study was to assess the association between HPV-16 episomal and integrated viral loads and the progression of anal intraepithelial neoplasia (AIN). HPV-16 episomal and integrated loads were measured on 665 specimens from 135 HPV-16-positive men participating in the prospective HIPVIRG (Human Immunodeficiency and Papilloma VIrus Research Group) study. AIN grade was evaluated on biopsies obtained during periodical high-resolution anoscopies. HPV-16 integration was confirmed by DIPS-PCR to demonstrate the presence of viral-cellular junctions. HPV-16 episomal loads [odds ratio (OR) 1.5, 95% confidence interval (CI)=1.1–2.1], burden of HPV infection [OR 1.4 (95% CI=1.1–1.8)] and current smoking [4.8 (95% CI=1.3–18.6)], but not integrated loads, were associated with high-grade lesions (AIN-2,3) after age and CD4 counts adjustment. A high HPV-16 episomal load was the only predictive factor of progression from low-grade AIN to high-grade AIN [OR 8.0 (95% CI=1.2–55.4)]. Specimens with higher HPV-16 episomal loads were less likely to contain integration [OR 0.5 (95% CI=0.3–0.8)]. HPV-16 integration was detected in the absence of AIN, in AIN-1 and in AIN-2,3. The analysis of the viral-cellular junctions did not allow identifying a specific site of integration.

Background: Highly Active Antiretroviral Treatment (HAART) has reduced the frequency of weight loss/wasting associated with HIV infection. However, weight loss remains a problem, even in the HAART era. Objectives: This study was carried out to assess weight change in a cohort of HIV-infected women on HAART in Rwanda, from 2005 to 2008, and to identify factors that predict weight loss in this cohort. Methods: Data from a cohort of 449 HIV-positive women on HAART enrolled in the Rwanda Women’s Inter-association Study and Assessment (RWISA), starting in May 2005, and followed at six monthly intervals until December 2008, were analysed. The outcome assessed in this study was change in weight, measured in kilograms at 6, 12 and 24 months after HAART initiation. Nutritional status was recorded and laboratory measurements (weight, height and CD4 cell count) were taken prior and after HAART initiation. All covariates were time dependent, except for the history of weight loss which was recorded at baseline only. Generalized Estimating Equation (GEE) using the linear link (Gaussian [normal]), exchangeable covariance structure and robust standard error was used to assess the factors associated with changes in weight (weight loss or weight gain) and to control for potential confounders. Results: Prior to HAART initiation, the mean weight of the study participants was 53.1 kg (SD 9.5). The mean BMI was 21.3 kg/m2 (SD 3.6) and the mean CD4 cell count was 222.9 cells/μL (SD 120.6) [47.6% had CD4 cell counts <200 cells/μL, 52.2% had CD4 cell counts ≥200 cells/μL]. Overall, the participants gained weight from baseline to 12 months after HAART initiation. The mean weight change was 1.9 kg (SD 7.8) (p<0.001) 6 months after HAART initiation, 2.9 kg (SD 5.9) (p <0.001) 12 months after HAART initiation, and 2.4 kg (SD 6.5) (p <0.001) 24 months after HAART initiation. Six months after HAART initiation, 48.3% of participants had gained weight, and 21.0% had lost weight. Twelve months after HAART initiation, 56.9% had gained weight, and 18.3% had lost weight, Twenty-four months after HAART initiation, 56.6% had gained weight, and 22.6% had lost weight. Participants with CD4 cell counts ≤ 200 cells/μL at baseline gained more weight than those with CD4 cell counts > 200 cells/μL at 6, 12 and 24 months after HAART initiation. Participants who were underweight (BMI <18.5 kg/m2) at baseline gained more weight than other participants three months after HAART initiation. Time-dependent diarrhoea for more than two weeks and a CD4 cell count of 200 - 350 cells/μL were significantly associated with weight loss (p≤ 0.05). Others factors, such as time-dependent education level (completion of secondary school), marital status (married legally and status other than married legally or widowed), and increases in CD4 cell counts, were associated with weight gain (p≤ 0.05). Conclusion: Although the majority of participants gained weight during the first 12 months of being on HAART, a significant proportion of participants lost weight while on HAART. The findings on the predictors of weight change in HIV-positive women on HAART can be used to promote weight gain in women who start HAART. Clinicians who take care of HIV-infected patients on HAART should pay attention to those who lose weight, and those who present with diarrhoea or with CD4 cell counts of <350 cells/μL at follow-up visits, since these factors are associated with weight loss in the HAART era.