Tese (doutorado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2016. / Submitted by Fernanda Percia França (fernandafranca@bce.unb.br) on 2017-03-13T15:40:17Z No. of bitstreams: 1 2016_ÉlidaCleyseGomesdaMata.pdf: 20672845 bytes, checksum: a126098842e2704babe5730ae79a7ae9 (MD5) / Approved for entry into archive by Raquel Viana(raquelviana@bce.unb.br) on 2017-03-28T18:06:09Z (GMT) No. of bitstreams: 1 2016_ÉlidaCleyseGomesdaMata.pdf: 20672845 bytes, checksum: a126098842e2704babe5730ae79a7ae9 (MD5) / Made available in DSpace on 2017-03-28T18:06:09Z (GMT). No. of bitstreams: 1 2016_ÉlidaCleyseGomesdaMata.pdf: 20672845 bytes, checksum: a126098842e2704babe5730ae79a7ae9 (MD5) / O presente trabalho avaliou as atividades antirretroviral e citotóxica de 9 peptídeos sintetizados a partir da análise transcritômica parcial de bibliotecas de cDNA obtidas das glândulas de peçonhas de diferentes espécies de escorpiões. Foram utilizados o Vírus da Imunodeficiência Símia/SIV e o HIV defectivo, em linhagens celulares humanas linfoblásticas, leucocitárias primárias e estabelecidas aderentes para a detecção de atividade antirretroviral, assim como citotóxica dos peptídeos sintéticos. Incluíram-se ensaios de viabilidade celular com azul de Tripan, citotoxicidade quantificando-se LDH e citometria de fluxo com detecção de Anexina/Iodeto de propídio. Os peptídeos escorpiônicos 1, 2, 4 e 6 reduziram a produção de SIV em células HUT-78 e 174X100, dentre os quais, os peptídeos 1 e 6 reduziram em 50% a viabilidade celular, em concentrações de 50 μM e 12,5 μM, respectivamente, tornando-os mais adequados aos ensaios. Em análises finais, o peptídeo 6 a 3,12 μM reduziu em 75% a produção viral em relação ao controle de células unicamente infectadas. Células HUT-78 tratadas com o peptídeo 6 e infectadas com SIV, unicamente produziram IL-8, enquanto que os leucócitos primários apresentaram níveis detectáveis de IFN-γ, IL-4, IL-6 e IL- 10. Em estudos de morfologia ultra-estrutural detectou-se o brotamento de virions de SIV unicamente em leucócitos primários infectados, enquanto que os tratados com o peptídeo 6 e AZT, infectados com SIV, não se detectou o brotamento viral. Observou-se lesões citoplasmáticas, extravasamento de material nuclear e acúmulo de cromatina na membrana perinuclear em leucócitos primários tratados com o peptídeo 6, compatível com os resultados obtidos por citometria de fluxo. Exclui-se a possiblidade de lise celular por destruição de membranas, visto que virions infectivos tratados em altas concentrações do peptídeo 6 não inibiram a infecção viral. Ensaios preliminares com células HeLa P4 previamente tratadas com o peptídeo 6 e transfectadas com plasmídeo pCHIV carreando o genoma defectivo de HIV, apresentaram redução da progênie viral quando comparada à produção de células transfectadas sem tratamento, células transfectadas e tratadas posteriormente com peptídeo 6 e tratadas com AZT. De modo geral, os experimentos de cultivo celular com o peptídeo 6 e posterior infecção viral, demonstraram que, nos intervalos iniciais, ocorre forte redução do número de células nas populações em tratamento, no entanto, após 96 horas as células proliferam, ultrapassando o número de células não tratadas, exercendo forte impacto quando avalia-se a interferência nos mecanismos de replicação viral, resultando na redução de progênie viral. Propõe-se que os mecanismos de proteção celular, em resposta à toxicidade do peptídeo 6, induz a expressão de citocinas, dosadas neste estudo e, muito provavelmente, fatores de restrição viral induzidos principalmente por IFN-γ, e modulados por IL-6, IL-10 e IL-4, ademais de outras citocinas e quimiocinas que não foram quantificadas neste estudo. / The present work evaluated the antiretroviral and cytotoxic activity of 9 synthetized peptides based on the transcriptome of the partial cDNA library of different Scorpions venenom glands. The Simian Immunodeficiency Virus/SIV and the defective HIV were utilized in human cell lineages, lymphoblastic cells, primary leucocytes and adherent cells, to screen for antiretroviral activity as also the peptides cytotoxicity. There were included assays for cell viability with trypan blue, citotoxicity quantifying LDH and flow citometry for detection of annexin/propidium iodide. The scorpionic peptides 1, 2, 4 and 6 reduced the SIV production in HUT-78 and 174XCEM cell lines, and among them, the peptides 1 and 6 reduced by 50 % the cell viability at 50 μM and 12,5 μM concentration, respectively, which was more suitable for the assays. In final analyses, the peptide 6 at 3,12 μM concentration reduced at 75 % the SIV production compared to untreated cells but infected. HUT-78 cells treated with peptide 6 and SIV infected solely produced IL-8 while the primary leucocytes displayed detectable levels of IFN-γ, IL-4, IL-6 and IL-10. In morphological ultrastructural studies it could be detected the budding of SIV virions exclusively in infected primary leucocytes, while among leucocytes treated with peptide 6, AZT and SIV infected, no virions budding were observed. Cytoplasmatic lesions, leaking of nuclear material and chromatin condensation in the perinuclear region were evident in leucocytes treated with peptide 6, which was compatible with the results obtained by flow cytometry analysis. It was excluded the possibility of cell lysis by membrane destruction, as infectious virions treated with high concentrations of peptide 6 did not inhibit viral infection. Preliminary assays with HeLa P4 cells previously treated with peptide 6 and transfected with the pCHIV plasmid carrying a defective HIV genome, showed viral progenie reduction when compared with the production of transfected cells without treatment, transfected cells subsequently treated with pepetide 6 and cells treated with AZT. In a general way, the cell culture experiments with peptide 6 and subsequent viral infection showed that, in the initial intervals, a strong cell number reduction occur, but after 96 hours the cells proliferate, outnumbering the untreated cells, exerting significative effect when evaluating the interference in the mechanisms of viral replication, which culminates in the reduction of viral progenie. It is proposed that the mechanisms of cell protection, in response to the peptide 6 toxicity, induce the expression of cytokines, quantified in this work and, very probably, viral restriction factors mainly triggered by IFN-γ, and modulated by IL-6, IL-10 e IL-4, besides other cytokines and chemokines that were not quantified here.

Made available in DSpace on 2014-06-11T19:22:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-12-20Bitstream added on 2014-06-13T19:28:12Z : No. of bitstreams: 1 segatto_afm_me_prud.pdf: 307549 bytes, checksum: b37359a27c088a907d37cb6899f6d330 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Após a introdução da terapia anti-retroviral altamente ativa (TARV), houve significativo aumento da sobrevida e melhora da qualidade de vida de indivíduos portadores de HIV, porém, esses tratamentos têm efeitos colaterais que podem causar transtornos para seus usuários, dentre os quais, a lipodistrofia. Além de causar um novo estigma estético para os portadores do vírus a síndrome pode elevar o risco de doenças cardiovasculares e diabetes. Nesse contexto, torna-se importante dar atenção a estratégias de prevenção e tratamento da síndrome. A atividade física pode ser uma alternativa válida para este fim, no entanto existem poucos estudos que tratam dessa temática. Desse modo, o objetivo do presente estudo foi verificar a possível associação entre o nível de atividade física e a ocorrência de lipodistrofia relacionada ao uso de terapia anti-retroviral em indivíduos portadores de HIV. A casuística foi formada por 42 indivíduos portadores de HIV em uso de TARV, todos pacientes do Centro de Testagem e Aconselhamento da cidade de Presidente Prudente. O nível de atividade física foi obtido pela aplicação do questionário internacional de atividade física (IPAQ), enquanto a lipodistrofia foi diagnosticada pela técnica de auto-relato do paciente... / After the introduction of the Highly Active Antiretroviral Therapy (HAART), there was significant increase in survival and improved quality of life among HIV-infected individuals, however, these treatments have side effects that may cause inconvenience to its users, among which lipodystrophy is. Besides causing a new aesthetic stigma for those with the virus, the syndrome may increase the risk of cardiovascular disease and diabetes. In this context, it becomes important to pay attention to strategies for prevention and treatment of the syndrome. Physical activity may be a valid alternative for this purpose, however there are few studies that address this issue. Thus, the purpose of this study was to investigate the possible association between level of physical activity and lipodystrophy occurence related to the use of antiretroviral therapy in HIV-infected individuals. The sample consisted of 42 HIV patients under antiretroviral therapy, all patients of the Center for Counseling and Testing in the city of Presidente Prudente. The level of physical activity was achieved by using the international physical activity questionnaire (IPAQ), whereas lipodystrophy was diagnosed by the technique of self-report of the patient... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:24:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-07-28Bitstream added on 2014-06-13T20:31:21Z : No. of bitstreams: 1 oliveira_oca_me_botfm.pdf: 592027 bytes, checksum: 64bb656e1e07bb63f132e5aca2ffecd3 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / A aids é uma doença causada pelo HIV, que compromete o sistema imune do organismo. O advento da terapia antirretroviral (TARV) altamente eficaz promoveu melhora substancial do prognóstico dessa doença e da qualidade de vida dos pacientes com HIV/AIDS. Durante seu tratamento prolongado notam-se algumas alterações hematológicas, como anemia e macrocitose, bem como carências de micronutrientes como a vitamina B12 e ácido fólico. O objetivo do presente trabalho é determinar a relação entre macrocitose e anemia e o uso de ARV ou a deficiência de vitamina B12 ou de ácido fólico. Foram avaliados 110 pacientes HIV-positivos, comparando-se aqueles em uso de TARV com AZT (G1), em TARV sem AZT (G2) e sem o uso de TARV (G3). Os pacientes dos três grupos não apresentaram variações significativas quanto aos níveis de hemoglobina (p=0,584). Os pacientes do G1 apresentaram VCM aumentado quando comparados aos do G3 (p<0,05), bem como os do G2 em relação aos do G3 (p<0,001). As dosagens de vitamina B12 do G1 e G3 foram menores do que as encontradas pelo G2 (p=0,008). As dosagens do ácido fólico não apresentaram diferença estatística entre os grupos (p=0,956). Conclui-se que os indivíduos em uso de TARV apresentaram macrocitose, embora esta não pudesse ser relacionada ao tipo de TARV ou à deficiência de vitamina B12, como também a deficiência de ácido fólico não esteve relacionada ao uso de TARV nem à macrocitose / AIDS is a chronic disease characterized by HIV infection and results in immunodeficiency. HAART is an effective approach to this disease, substantially improving quality of life and prognostic factors. It has become frequent the occurrence of hematologic disorders such anemia and macrocytosis, as well as micronutrients deficiency with the outcome of the treatment. The objective of this study is to correlate macrocytosis, anemia and HAART collateral effects with B12 vitamin and folic acid deficiencies. 110 HIV positive patients were included and divided in 3 groups: HAART with AZT (Group 1), HAART without AZT (Group 2) and without any antiretroviral treatment (Group 3). All groups did not have difference related to hemoglobin level (p=0,584). G1 had higher VCM levels than G3 (p<0,05), as well as G2 than G3 (p<0,001). G1 and G3 Vitamin B12 levels were smaller than those from G2 (p=0,008). Folic acid measurements did not differ among groups (p=0,956). We conclude that patients in HAART treatment had macrocytosis, even though this could not be related to an specific drug among HAART treatment or vitamin B12 deficiency. However, folic acid deficiency was not related neither to HAART nor macrocytosis

Ph.D. / Despite nearly three decades of intensive research, the HIV/AIDS pandemic remains a major challenge to modern medicine. The discovery and development of antiretroviral agents acting against various essential viral processes and enzymatic targets have greatly enhanced the quality of life for infected individuals, but no cure or preventative vaccine is available as yet and HIV infection is currently considered irreversible. Furthermore, the emergence of viral resistance to every class and type of antiretroviral treatment agent necessitates the continued discovery of antiretroviral agents with novel mechanisms of action. The first antiretroviral agent targeting the retroviral integrase enzyme (InsentressTM, Raltegravir) received regulatory approval from the United States Food and Drug Administration during 2007, validating HIV-1 integrase as a therapeutic target and providing a much-needed second- or third-line treatment option for treatment experienced patients. This enzyme was selected as a target for the current work. As limited data were available on the primary and secondary structure of the biologically relevant HIV-1 integrase enzyme, a first step in the present work was the construction of monomeric, dimeric and tetrameric models of the enzyme with biologically relevant catalytic centres incorporating both viral and host co-factors and DNA. The models were constructed to identify potential inhibitors of the strandtransfer reaction of HIV-1 integrase and were based on observations and interactions reported in the literature and on crystal structure data of HIV-1 integrase sub-domains and related structures available in the Protein Data Bank. The monomeric model was used as the macromolecular target in docking studies with “drug-like” compound databases, identifying the pyrrolidinone compound class as an in silico hit candidate for further development. Initial activity screening of a number of commercially available pyrrolidinone analogues against recombinant HIV-1 subtype B integrase in direct enzyme assays confirmed the predicted potential for strand transfer inhibition of the compound class, and provided initial support in the further development of this compound class as inhibitors of HIV-1 integrase that target the strand-transfer step. Retrosynthetic analysis of the pyrrolidinone hit candidates provided a facile one-pot, three-component synthetic pathway from readily available starting materials, which generally gave the proposed products cleanly and in acceptable yields. A range of closely related analogues were designed and synthesised. The analogues making up this series generally differed by only one functional group, in order to enable initial structureactivity relationship investigations during later stages of the project. Foreword Page XVI The synthesised pyrrolidinone analogues were screened through a range of direct and cell-based in vitro assays to determine the toxicity and strand-transfer activity of each. In general, the pyrrolidinone compounds proved well-tolerated in PM1 cell culture, with clear potential to further develop the strandtransfer inhibition of the compound family in second- and further-generation optimisation stages. Furthermore, the aqueous solubility and membrane permeability of each compound were determined in vitro, providing initial biological profiles of each test compound. As no in vivo testing was performed with any of the compounds during this first round of drug discovery and optimisation, several computational models were employed to extrapolate the in vitro and structural data to possible in vivo scenarios. Two pyrrolidinone analogues (11.6 and 15.2) were identified as low micro-molar strand-transfer inhibitors of wildtype-equivalent HIV-1 integrase, with low toxicity in cell culture and favourable solubility and permeability profiles. Resistance screening of these two compounds against four mutant HIV-1 integrases (Q148H; Q148H/G140S; N155H and N155H/E92Q) has shown some promise, with compound 15.2 retaining a measure of activity against the Raltegravir-resistant N155-mutants. These hit candidates will form the basis of structure-activity relationship optimisations in second- and further generation design stages.

Magister Public Health - MPH / Background:HIV is a worldwide pandemic with an estimated 2.5 million children under the age of 15 living with HIV in the world in 2009. Children account for approximately 14% of all HIV-related deaths around the world. Several studies have shown that the use of antiretroviral drugs greatly improve the lives of HIV-1 infected individuals, however, most of these studies report on outcomes of ART programmes in developed world and for adult patients. Very few settings have published outcomes of paediatric ART programmes.Objectives This research was aimed at describing the long term (at least one year) treatment outcome of HIV-1 infected children in the HIV/AIDS Prevention Group (HAPG) program in Bela-Bela in the Limpopo province of South Africa.Study design and methods: A quantitative approach involving a retrospective cohort design was used for the study. The study included all children under the age of 15 that were enrolled in the HATG treatment programme in Bela-Bela between February 2004 and December2009.Immunological, virological, clinical outcomes and loss to follow-up were determined for this cohort. Mortality and survival was also determined. Results: The median age of children in this study was 5 years (IQR: 2-7) with 14% (10/71) of them being less than 18 months. Median CD4 count at commencement of ART, viral load and weight were 358 cells/mm3 (IQR 203.5-, 125673 RNA copies/μL (IQR 58094-328424.5) and 14.5Kg (IQR: 11.0-18.35) respectively. CD4 counts and weight showed increase within the study period, and there was also a decline in viral load. Loss to follow-up was 7.04% while mortality was 19% with 21.43% of mortality cases being children who were ≤18months. Mortality occurred within the first year of ART initiation and occurred in cases that had advanced disease.Conclusion: This study shows that the ART program in Bela-Bela has a positive outcome on HIV positive children.The high mortality rate was due to children starting ART at an advanced disease stage. Despite the good outcome, it is recommended that a system be put into place that will aid in identifying children at an early stage of the disease and treatment initiated promptly.

Magister Public Health - MPH / Namibia is one of the countries in the world most affected by HIV/AIDS with the national prevalence of 18.8% in 2010. In 2010, it was reported that an estimated 180,000 Namibians were living with HIV/AIDS; of which 95,000 adult women, 69,000 adult men and 16,000 children. An estimated 6,700 deaths was recorded in 2009 with an estimated number of 70,000 orphans due to the disease. The introduction of anti-retroviral therapy (ART) in public health facilities in Namibia in 2003 has improved the quality of lives of patients with advanced HIV disease, prolonged their lives and enabled them to be economically productive. By 2010 about 90,000 patients were enrolled on ART program in all 34 district hospitals and 3 intermediate referrer hospitals. Adherence to antiretroviral therapy is a key attribute of clinical HIV care and the overall determining factor in gauging the effectiveness of treatment. Good adherence to ART is vital to sustain low viral loads and prevent the development of drug resistant HIV strains. Although the patient retention rate on ART at the Katima Mulilo Hospital was 98.3%, with increased patient uptake to the program in future, there is a need to be aware of factors that influence adherence to ART as such findings could inform the expanded ART program in Caprivi region. An explorative, qualitative study was conducted where in-depth interviews were conducted with 24 ART patients and key informants interviews with 2 health workers. Data were audiotape recorded and transcribed verbatim. Thematic and content analysis of transcribed data was performed.

Masters of Public Health - see Magister Public Health / Introduction: HIV and AIDS continues to be a major public health challenge for Zambia, which has the highest HIV prevalence rate of 13.1% in sub-Saharan Africa. Although individuals living with HIV/AIDS in Zambia have increased access to antiretroviral treatment (ART), not all patients who are initiated on antiretroviral treatment remain in care; with some patients being lost at different points in the continuum of care. The current study aimed to explore the factors affecting retention in care among patients receiving antiretroviral treatment at three primary health facilities in the Kabwe district in Zambia. Methodology: An exploratory qualitative study design was used to explore the patient, health systems and socio-economic factors that underlie retention on ART in three purposefully selected primary health care facilities in Kabwe district. Data was collected through in-depth interviews with 45 ART patients and three focus group discussions with 20 health care providers. The content of the transcribed interviews was analyzed thematically. Findings: The overall retention rate of the ART sites was found to be 65%. The main patient factors that influenced retention in care were side effects of antiretroviral drugs and weight increase as a sign of good health. The social related factors that influenced patient retention in care were stigma and non-disclosure of HIV status, faith healing, use of herbal remedies and alcohol use. The health system factors that contributed to poor retention of patients in care were long waiting times due to staff shortage, high patient load, travel distance to ART centers and transportation cost. Other health system factors reported by participants included shortage of third line ARV drugs and inadequate space in ART clinic. Finally, food shortage and mobility of patients due to employment were some of the identified economic factors that influenced patient retention in care. Conclusions: A large proportion of adult patients initiating ART in Zambia are poorly retained in care because of patient, health system, social and economic factors. In order to improve retention, more nurses and clinical officers should be trained in ART management to improve skills and address staff shortages. It would also be useful for Zambia to introduce community drug distribution points for delivering ARV refills to reduce the workload on the existing ART sites and reduce on the distances that patients have to travel to ART centers. Additionally, efforts should also be made to improve ART care by extending ART clinic days to include all the days of the week except Sundays.

Class of 2005 Abstract / Objectives: To determine compliance rate of patients enrolled in the AIDS Drug Assistance Program (ADAP) at El Rio Health Center. Methods: This study was a retrospective observational study design that utilized medication refill data obtained from computerized pharmacy records. Lists were created for each anti-retroviral drug that included the following data: medical record number, medication, quantity dispensed, days supply and refill date. Patient age, gender and ethnicity were also obtained. The data was compiled into a database using Microsoft Excel©. The medication possession ratio (MPR) was calculated for each drug as well as for each drug group. The subjects in this study were patients enrolled in the ADAP at El Rio Health Center who obtained prescription refills between December 1, 2003 and November 30, 2004. The mean age was 44.56 (range 25-78); 94.8% were male and 5.2% were female. Ethnic distribution included 52.6% Caucasian, 39.6% Hispanic, 3.2%African American, 1.3% Asian, and 3.2% other. Results: The MPR was calculated for each drug as well as for each drug group. MPRs for individual drugs ranged from 0.586-0.906; MPRs for drug groups ranged from 0.717-0.756. Implications: The results of the study indicated that ADAP patients did not have adequate (>95%) compliance rates. The implications of the results are that patients are not fully benefiting from their medication, while at the same time promoting the development of resistant strains of HIV.

BACKGROUND: Highly active antiretroviral therapy(HAART) is used in pregnancy to suppress viral load(pVL) before delivery, reducing risk of vertical HIV-transmission. Nelfinavir(NFV) containing HAART has been highly used in pregnancy, but dosages may be inadequate due to the physiologic changes that occur. Given concerns regarding optimal viral suppression in pregnancy, drug toxicity and resistance development, NFV levels need to be evaluated in this population to guide dosing recommendations. METHODS: As part of a prospective cohort study maternal blood was collected at 18-28wks, 32-37wks and at delivery. Times of last medication dose and blood sampling were recorded and drug levels were measured using HPLC MS-MS. NFV concentration-ratios(NFV-CRs) were calculated by dividing individual levels by a time-adjusted population value. Plasma NFV concentrations and NFV-CRs were compared across gestational age and correlated to variables of interest. Rate and maintenance of viral suppression were analyzed in relation to NFV concentrations and CRs. Statistical tests included ANOVA, χ2, linear regression, and Kaplan Meier estimates. RESULTS: 113 samples were collected from 32 subjects. Samples were eliminated if not in steady state (n=20); 93 samples from 32 subjects were analyzed. Mean NFV-CR at 18-28wks (1.1±0.73) and 32-37wks (0.86±0.73) were not significantly different but were both significantly higher by ANOVA (p=0.049) than the mean NFV-CR at delivery (0.44±0.50). CRs were highly variable. Of 49 antepartum samples, 49%(24) had a CR<0.90 (clinically relevant threshold). Four women reached a pVL <50 copies/mL by 34wks but had a detectable pVL at delivery. One woman never reached an undetectable pVL in pregnancy. Minimum and mean NFV-CRs in these 5 women were not significantly different than those who achieved and maintained virologic suppression. Vertical HIV transmission rate was 0%. CONCLUSIONS: There were no HIV transmissions but 16% (5/32) of women were inadequately suppressed at delivery, which is of concern. Factors associated with inadequate suppression and NFV-CRs need to be explored in conjunction with patient/physician reported adherence and viral resistance profiles. Extreme variability in CRs may limit the potential usefulness of random timed drug levels in all pregnant women. / Medicine, Faculty of / Obstetrics and Gynaecology, Department of / Graduate

A dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in partial fulfilment of the requirements for the Degree of Master of Science in Epidemiology and Biostatistics. Johannesburg, November 2017. / Introduction Viral load is the most reliable indicator of poor adherence to anti-retroviral therapy (ART). However, this assay is difficult to implement in resource-limited settings due to financial and technical constraints. Laboratory markers, combined with the patient’s demographic and clinical details, have been described as better proxies of adherence than the current self-reported adherence measures. However, the real diagnostic value of these biomarkers remains unknown. Therefore, the aim of this study was to assess the usefulness of a composite marker to identify poor adherence to ART defined as a detectable plasma viral load in HIV-positive patients on first-line regimen at Themba Lethu Clinic (TLC) in Johannesburg, South Africa Materials and Methods: This study was retrospective cohort analysis of data collected on HIV-positive ART naïve adults initiating first line antiretroviral regimen at TLC following the 2010 South African antiretroviral treatment guidelines. The data collection was carried out as part of the low-cost monitoring (LCM) study at Themba Lethu Clinic in Johannesburg from February 2012 to 2014. The LCM cohort which aims to look at low cost monitoring of HIV treatment in resource limited settings was initiated in 2009 in Johannesburg, South Africa. The study or treatment outcome was failure to suppress viral load (VL ≥ 400 copies/ml) at 6 and at 12 months. Adherence to antiretroviral treatment was assessed using four (4) self-reported adherence (SRA) measures namely: a self-reporting questionnaire, a Visual Analogue Scale (VAS), a pill identification test (PIT) and the Simplified Medication Adherence Questionnaire (SMAQ). The result of each self-reported measure was classified as either positive or negative given a conventional threshold. In our study three (3) self-reported adherence (SRA) measures were combined into a multi-method approach tool which included self-reports combined with VAS and the pill identification test (PIT). Continuous variables were summarized by median with interquartile range. Categorical variables were summarized by giving their frequencies. To compare continuous variables, we used an unpaired t-test if the variable was normally distributed. When continuous variables were compared from baseline to the previous 6 months, a paired t-test was done. In the case of skewed distribution, we used a non-parametric variant of the t-test such as the Mann-Whitney U-test. To compare categorical variables, we used cross-tables with corresponding chi-square test or Fisher exact test. A Modified Poisson Generalized Linear Model (GLM) with robust variance was used to estimate adjusted relative risks (aRR) of failing to suppress viral load at 6 and at 12 months adjusting for age age, gender, self-reported adherence measures, changes in laboratory markers and missed appointments at 6 and 12 months after ART initiation. As there was missing values in the covariatess and the outcome, we performed a multiple imputation technique under missing at random (MAR) assumption in order to compare the robustness of the estimations between the complete case analysis and the imputation model under MAR after imputing missing values. with the imputed dataset. Additionally, we calculated the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for each self-reported adherence measure using viral load as the reference standard. Thus, we derived two diagnostic risk scores from rounding and adding together the adjusted regression coefficients used to estimate adjusted relative risk and following the Spiegelhalter and Knill-Jones approach, at 6 and at 12 months. The Receiver Operating characteristic (ROC) curves were computed to see the overall discriminative value of each continuous risk score. To assess the clinical usefulness of the continuous riskscores we dichotomized them from 2 ≥ vs < 1 to 5 ≥ vs < 5 and calculated the sensitivity (Se), specificity (Sp), positive predictive value (PPV) and negative predictive value (NPV) at each cut-off, taking detectable viral load as a gold standard. Results: There were 353 HIV-positive patients initiated on first line ART at TLC for the LCM cohort study. Of these, 80.7% did not suppress viral load after 6 months while 30.1% did not suppress viral load at 12 months. The proportion of patients classified as being highly adherent was 86.7% but this proportion decreased to 60% at 12 months. By 6 months, after adjusting for gender and age, the variables that were significantly associated with detectable viral load included: having missed at least two ARV visits by ≥ 7 days (aRR: 2.35 95% CI: 1.08 -5.11); platelet count < 150 cells/mm3 (aRR: 2.73 95% CI: 1.04 -7.18) and VAS ≤ 95% (aRR: 1.65. 95% CI: 1.01-2.71). At 12 months, the estimates showed a positive relationship only with age group and unemployment. There were no similarities in the results found using complete case analysis and analysis with imputed datasets. However, the largest standard errors were obtained from the complete case analysis. At 6 months, the AUC ROC curve was calculated as 0.63 (95% CI, 0.53 - 0.72) while, for the visual analogue scale, the AUC decreased to 0.55 (95% CI, 0.49 - 0.62); for the Simplified Medication Adherence Questionnaire (SMAQ), the AUC decreased to 0.52 (95%CI, 0.45 - 0.60), while for the multi-method approach, it decreased to 0.53 (95% CI, 0.46 - 0.58). The optimal diagnostic accuracy was obtained with the score 5 (≥5 vs <5 Se: 64% and a Sp: 50.0%) followed by a risk score of 4 (Se of 76.0%, Sp of 34.7%). At 12 months, the AUC of the diagnostic risk score was calculated as 0.44 (95%CI, 0.40 - 0.60) while for the three self-reported adherence methods, it decreased to 0.48 (95% CI, 0.40 - 0.60), 0.51 (95%CI, 0.40 - 0.60) and 0.50 (95%CI, 0.41 - 0.59) respectively for the visual analogue scale, the SMAQ and the multi-method approach method respectively. Conclusion. This study shows that after ART initiation, the 6-month’s adherence can be better diagnosed using laboratory markers combined with patient’s information and traditional self-reported adherence measures at Themba Lethu Clinic. The advantage of this proposed method is that it is based on routine and accessible informations collected during HIV-positive patient visits, thus incurring no additional cost for its implementation. An external validation of this diagnostic risk score is needed for its translation into clinical practice in resource-limited settings. / LG2018