Potent Antiretroviral Therapy for Human Immunodeficiency Virus Infection Increases Aortic Stiffness

Background: Highly active antiretroviral therapy for AIDS is known to increase cardiovascular risk, but the effects of potent antiretroviral agents according to gender are unknown. Objective: The present study evaluated the impact of HIV infection treatment on aortic stiffness according to gender. Methods: From university-affiliated hospitals, we recruited 28 AIDS patients undergoing highly active antiretroviral treatment (HAART), 28 treatment-naive HIV-infected patients, 44 patients with type 2 diabetes, and 30 controls. Aortic stiffness was determined by measuring pulse wave velocity (PWV) using a validated and non-invasive automatic device. Results: The crude mean PWV values and 95% confidence intervals (95% CI) for HAART, diabetics, and controls were 9.77 m/s (95% CI 9.17-10.36),, 9.00 m/s (95% CI 8.37-9.63), 9.90 m/s (95% CI 9.32-10.49), and 9.28 m/s (95% CI 8.61-9.95), respectively, for men (P-value for trend = 0.14), and 9.61 m/s (95% CI 8.56-10.66), 8.45 m/s (95% CI 7.51-9.39), 9.83 (95% CI 9.21-10.44), and 7.79 m/s (95% CI 6.99-8.58), respectively, for women (P-value for trend <0.001). Post-hoc analysis revealed a significant difference between the mean PWV values in the HAART group and controls in women (P-value <0.01). After adjusting for other potential covariates, including systolic blood pressure and diabetes, these results did not change. The findings indicate that the impact of HAART treatment on aortic stiffness was amplified in women with hypertension, dyslipidemia, and metabolic syndrome. Conclusion: Potent anti-retroviral agents used in the treatment of HIV infection increases aortic stiffness, mainly among women with higher cardiovascular risk. (Arq Bras Cardiol 2012;99(6):1100-1107)

Antiretrovirals as antimalarial agents

Recent studies have indicated that antiretroviral protease inhibitors may affect outcome in malarial disease. We have investigated the antimalarial activities of 6 commonly used antiretroviral agents. Our data indicate that, in addition to the previously published effects on cytoadherence and phagocytosis, the human immunodeficiency virus (HIV)-1 protease inhibitors saquinavir, ritonavir, and indinavir directly inhibit the growth of Plasmodium falciparum in vitro at clinically relevant concentrations. These findings are particularly important in light of both the high rate of malaria and HIV-1 coinfection in sub-Saharan Africa and the effort to employ highly active antiretroviral therapy in these regions.

Statistical and econometric analysis of the treatment of HIV/AIDS

The epidemic of HIV/AIDS in the United States is constantly changing and evolving, starting from patient zero to now an estimated 650,000 to 900,000 Americans infected. The nature and course of HIV changed dramatically with the introduction of antiretrovirals. This discourse examines many different facets of HIV from the beginning where there wasn't any treatment for HIV until the present era of highly active antiretroviral therapy (HAART). By utilizing statistical analysis of clinical data, this paper examines where we were, where we are and projections as to where treatment of HIV/AIDS is headed.

Chapter Two describes the datasets that were used for the analyses. The primary database utilized was collected by myself from an outpatient HIV clinic. The data included dates from 1984 until the present. The second database was from the Multicenter AIDS Cohort Study (MACS) public dataset. The data from the MACS cover the time between 1984 and October 1992. Comparisons are made between both datasets.

Chapter Three discusses where we were. Before the first anti-HIV drugs (called antiretrovirals) were approved, there was no treatment to slow the progression of HIV. The first generation of antiretrovirals, reverse transcriptase inhibitors such as AZT (zidovudine), DDI (didanosine), DDC (zalcitabine), and D4T (stavudine) provided the first treatment for HIV. The first clinical trials showed that these antiretrovirals had a significant impact on increasing patient survival. The trials also showed that patients on these drugs had increased CD4+ T cell counts. Chapter Three examines the distributions of CD4 T cell counts. The results show that the estimated distributions of CD4 T cell counts are distinctly non-Gaussian. Thus distributional assumptions regarding CD4 T cell counts must be taken, into account when performing analyses with this marker. The results also show the estimated CD4 T cell distributions for each disease stage: asymptomatic, symptomatic and AIDS are non-Gaussian. Interestingly, the distribution of CD4 T cell counts for the asymptomatic period is significantly below that of the CD4 T cell distribution for the uninfected population suggesting that even in patients with no outward symptoms of HIV infection, there exists high levels of immunosuppression.

Chapter Four discusses where we are at present. HIV quickly grew resistant to reverse transcriptase inhibitors which were given sequentially as mono or dual therapy. As resistance grew, the positive effects of the reverse transcriptase inhibitors on CD4 T cell counts and survival dissipated. As the old era faded a new era characterized by a new class of drugs and new technology changed the way that we treat HIV-infected patients. Viral load assays were able to quantify the levels of HIV RNA in the blood. By quantifying the viral load, one now had a faster, more direct way to test antiretroviral regimen efficacy. Protease inhibitors, which attacked a different region of HIV than reverse transcriptase inhibitors, when used in combination with other antiretroviral agents were found to dramatically and significantly reduce the HIV RNA levels in the blood. Patients also experienced significant increases in CD4 T cell counts. For the first time in the epidemic, there was hope. It was hypothesized that with HAART, viral levels could be kept so low that the immune system as measured by CD4 T cell counts would be able to recover. If these viral levels could be kept low enough, it would be possible for the immune system to eradicate the virus. The hypothesis of immune reconstitution, that is bringing CD4 T cell counts up to levels seen in uninfected patients, is tested in Chapter Four. It was found that for these patients, there was not enough of a CD4 T cell increase to be consistent with the hypothesis of immune reconstitution.

In Chapter Five, the effectiveness of long-term HAART is analyzed. Survival analysis was conducted on 213 patients on long-term HAART. The primary endpoint was presence of an AIDS defining illness. A high level of clinical failure, or progression to an endpoint, was found.

Chapter Six yields insights into where we are going. New technology such as viral genotypic testing, that looks at the genetic structure of HIV and determines where mutations have occurred, has shown that HIV is capable of producing resistance mutations that confer multiple drug resistance. This section looks at resistance issues and speculates, ceterus parabis, where the state of HIV is going. This section first addresses viral genotype and the correlates of viral load and disease progression. A second analysis looks at patients who have failed their primary attempts at HAART and subsequent salvage therapy. It was found that salvage regimens, efforts to control viral replication through the administration of different combinations of antiretrovirals, were not effective in 90 percent of the population in controlling viral replication. Thus, primary attempts at therapy offer the best change of viral suppression and delay of disease progression. Documentation of transmission of drug-resistant virus suggests that the public health crisis of HIV is far from over. Drug resistant HIV can sustain the epidemic and hamper our efforts to treat HIV infection. The data presented suggest that the decrease in the morbidity and mortality due to HIV/AIDS is transient. Deaths due to HIV will increase and public health officials must prepare for this eventuality unless new treatments become available. These results also underscore the importance of the vaccine effort.

The final chapter looks at the economic issues related to HIV. The direct and indirect costs of treating HIV/AIDS are very high. For the first time in the epidemic, there exists treatment that can actually slow disease progression. The direct costs for HAART are estimated. It is estimated that the direct lifetime costs for treating each HIV infected patient with HAART is between $353,000 to $598,000 depending on how long HAART prolongs life. If one looks at the incremental cost per year of life saved it is only $101,000. This is comparable with the incremental costs per year of life saved from coronary artery bypass surgery.

Policy makers need to be aware that although HAART can delay disease progression, it is not a cure and HIV is not over. The results presented here suggest that the decreases in the morbidity and mortality due to HIV are transient. Policymakers need to be prepared for the eventual increase in AIDS incidence and mortality. Costs associated with HIV/AIDS are also projected to increase. The cost savings seen recently have been from the dramatic decreases in the incidence of AIDS defining opportunistic infections. As patients who have been on HAART the longest start to progress to AIDS, policymakers and insurance companies will find that the cost of treating HIV/AIDS will increase.

Dolutegravir plus Abacavir-Lamivudine for the treatment of HIV-1 infection

BACKGROUND: Dolutegravir (S/GSK1349572), a once-daily, unboosted integrase inhibitor, was recently approved in the United States for the treatment of human immunodeficiency virus type 1 (HIV-1) infection in combination with other antiretroviral agents. Dolutegravir, in combination with abacavir-lamivudine, may provide a simplified regimen. METHODS: We conducted a randomized, double-blind, phase 3 study involving adult participants who had not received previous therapy for HIV-1 infection and who had an HIV-1 RNA level of 1000 copies per milliliter or more. Participants were randomly assigned to dolutegravir at a dose of 50 mg plus abacavir-lamivudine once daily (DTG-ABC-3TC group) or combination therapy with efavirenz-tenofovir disoproxil fumarate (DF)-emtricitabine once daily (EFV-TDF-FTC group). The primary end point was the proportion of participants with an HIV-1 RNA level of less than 50 copies per milliliter at week 48. Secondary end points included the time to viral suppression, the change from baseline in CD4+ T-cell count, safety, and viral resistance. RESULTS: A total of 833 participants received at least one dose of study drug. At week 48, the proportion of participants with an HIV-1 RNA level of less than 50 copies per milliliter was significantly higher in the DTG-ABC-3TC group than in the EFV-TDF-FTC group (88% vs. 81%, P = 0.003), thus meeting the criterion for superiority. The DTG-ABC-3TC group had a shorter median time to viral suppression than did the EFV-TDF-FTC group (28 vs. 84 days, P<0.001), as well as greater increases in CD4+ T-cell count (267 vs. 208 per cubic millimeter, P<0.001). The proportion of participants who discontinued therapy owing to adverse events was lower in the DTG-ABC-3TC group than in the EFV-TDF-FTC group (2% vs. 10%); rash and neuropsychiatric events (including abnormal dreams, anxiety, dizziness, and somnolence) were significantly more common in the EFV-TDF-FTC group, whereas insomnia was reported more frequently in the DTG-ABC-3TC group. No participants in the DTG-ABC-3TC group had detectable antiviral resistance; one tenofovir DF-associated mutation and four efavirenz-associated mutations were detected in participants with virologic failure in the EFV-TDF-FTC group. CONCLUSIONS: Dolutegravir plus abacavir-lamivudine had a better safety profile and was more effective through 48 weeks than the regimen with efavirenz-tenofovir DF-emtricitabine. Copyright © 2013 Massachusetts Medical Society.

Influence de l'initiation de la traduction sur le changement programmé du cadre de lecture en -1 responsable de la synthèse des enzymes du virus de l’immunodéficience humaine de type 1

Le virus de l’immunodéficience humaine de type 1 (VIH-1) est responsable du syndrome de l’immunodéficience acquise (SIDA). Il faut identifier de nouvelles cibles pour le développement d’agents anti-VIH-1, car ce virus développe une résistance aux agents présentement utilisés. Notre but est d’approfondir la caractérisation de l’étape du changement de cadre de lecture ribosomique en -1 (déphasage -1) nécessaire à la production du précurseur des enzymes du VIH-1. Ce déphasage est programmé et effectué par une minorité de ribosomes lorsqu’ils traduisent la séquence dite glissante à un endroit spécifique de l’ARN messager (ARNm) pleine-longueur du VIH-1. L’efficacité de déphasage est contrôlée par le signal stimulateur de déphasage (SSF), une tige-boucle irrégulière située en aval de la séquence glissante. La structure du SSF est déroulée lors du passage d’un ribosome, mais elle peut se reformer ensuite. Nous avons montré que des variations de l’initiation de la traduction affectent l’efficacité de déphasage. Nous avons utilisé, dans des cellules Jurkat-T et HEK 293T, un rapporteur bicistronique où les gènes codant pour les luciférases de la Renilla (Rluc) et de la luciole (Fluc) sont séparés par la région de déphasage du VIH-1. La Rluc est produite par tous les ribosomes traduisant l’ARNm rapporteur alors que la Fluc est produite uniquement par les ribosomes effectuant un déphasage. L’initiation de ce rapporteur est coiffe-dépendante, comme pour la majorité des ARNm cellulaires. Nous avons examiné l’effet de trois inhibiteurs de l’initiation et montré que leur présence augmente l’efficacité de déphasage. Nous avons ensuite étudié l’effet de la tige-boucle TAR, qui est présente à l’extrémité 5’ de tous les ARNm du VIH-1. TAR empêche la liaison de la petite sous-unité du ribosome (40S) à l’ARNm et module aussi l’activité de la protéine kinase dépendante de l’ARN double-brin (PKR). L’activation de PKR inhibe l’initiation en phosphorylant le facteur d’initiation eucaryote 2 (eIF2) alors que l’inhibition de PKR a l’effet inverse. Nous avons étudié l’effet de TAR sur la traduction et le déphasage via son effet sur PKR en utilisant TAR en trans ou en cis, mais à une certaine distance de l’extrémité 5’ afin d’éviter l’interférence avec la liaison de la 40S. Nous avons observé qu’une faible concentration de TAR, qui active PKR, augmente l’efficacité de déphasage alors qu’une concentration élevée de TAR, qui inhibe PKR, diminue cette efficacité. Nous avons proposé un modèle où des variations de l’initiation affectent l’efficacité de déphasage en modifiant la distance entre les ribosomes parcourant l’ARNm et, donc, la probabilité qu’ils rencontrent un SSF structuré. Par la suite, nous avons déterminé l’effet de la région 5’ non traduite (UTR) de l’ARNm pleine-longueur du VIH-1 sur l’efficacité de déphasage. Cette 5’UTR contient plusieurs régions structurées, dont TAR à l’extrémité 5’, qui peut interférer avec l’initiation. Cet ARNm a une coiffe permettant une initiation coiffe-dépendante ainsi qu’un site d’entrée interne des ribosomes (IRES), permettant une initiation IRES-dépendante. Nous avons introduit cette 5’UTR, complète ou en partie, comme 5’UTR de notre ARNm rapporteur bicistronique. Nos résultats démontrent que cette 5’UTR complète inhibe l’initiation coiffe dépendante et augmente l’efficacité de déphasage et que ces effets sont dus à la présence de TAR suivie de la tige-boucle Poly(A). Nous avons aussi construit un rapporteur tricistronique où les ribosomes exprimant les luciférases utilisent obligatoirement l’IRES. Nous avons observé que cette initiation par l’IRES est faible et que l’efficacité de déphasage correspondante est également faible. Nous avons formulé une hypothèse pour expliquer cette situation. Nous avons également observé que lorsque les deux modes d’initiation sont disponibles, l’initiation coiffe dépendante est prédominante. Finalement, nous avons étudié l’effet de la protéine virale Tat sur l’initiation de la traduction et sur l’efficacité de déphasage. Nous avons montré qu’elle augmente l’initiation de la traduction et que son effet est plus prononcé lorsque TAR est située à l’extrémité 5’ des ARNm. Nous proposons un modèle expliquant les effets de Tat sur l’initiation de la traduction par l’inhibition de PKR ainsi que par des changements de l’expression de protéines cellulaires déroulant TAR. Ces résultats permettent de mieux comprendre les mécanismes régissant le déphasage du VIH-1, ce qui est essentiel pour le développement d’agents anti-déphasage.

Étude de la traduction IRES-dépendante du VIH-1

Le virus de l’immunodéficience humaine de type 1 (VIH-1) est responsable de la pandémie du SIDA (syndrome de l’immunodéficience acquise). Des souches virales résistantes aux antirétroviraux actuellement utilisés apparaissent rapidement. Il est donc important d’identifier de nouvelles cibles dans le cycle de réplication du VIH-1 pour développer de nouveaux agents contre ce virus. La traduction des protéines de structure et des enzymes du VIH-1 est une étape essentielle du cycle de réplication virale. Ces protéines sont exprimées à partir de l’ARN messager (ARNm) pleine-longueur (ARNmPL) à la fin du cycle de réplication. L’ARNmPL du VIH-1 peut utiliser un mode d’initiation de la traduction coiffe-dépendant, comme la majorité des ARNm cellulaires, mais peut aussi utiliser un mode d’initiation alternatif, car sa région 5’ non-traduite (5’UTR) contient un site interne d’entrée du ribosome (IRES), ce qui lui permet d’initier la traduction suivant un mode IRES-dépendant. L’initiation IRES-dépendante permet à l’ARNmPL d’être traduit quand l’initiation coiffe-dépendante est inhibée. L’activité de l’IRES de la région 5’UTR de l’ARNmPL du VIH-1 (IRES5’UTR) est faible dans des conditions physiologiques, mais est stimulée lorsque la cellule est arrêtée à la transition G2/M du cycle cellulaire, un arrêt qu’induit l’infection par le VIH-1. Une grande portion de l’IRES5’UTR, que nous nommons IRES5’UTRc, est présente dans tous les ARNm viraux et a une activité semblable à celle de l’ IRES5’UTR, ce qui indique que le mode IRES-dépendant peut être utilisé par tous les messagers du VIH-1. Lors de mes études doctorales, j’ai caractérisé le fonctionnement de l’IRES5’UTR du VIH-1. J’ai transfecté des cellules lymphocytaires Jurkat T, dérivées des cibles naturelles du VIH-1, avec un vecteur dual-luciférase contenant les séquences codantes des luciférases de la Renilla (Rluc) et de la luciole (Fluc) séparées par la région 5’UTR de l’ARNmPL du VIH-1. La traduction de la Rluc est coiffe-dépendante alors que celle de la Fluc dépend de l’IRES5’UTR. J’ai d’abord effectué une analyse mutationnelle et j’ai identifié trois régions qui stimulent l’activité de l’IRES5’UTR et une tige-boucle qui réprime l’activité de cet IRES, que j’ai nommée IRENE (IRES negative element). J’ai montré que l’effet répresseur d’IRENE est aboli lorsque les cellules sont soumises à un stress oxydatif, un type de stress induit lors d’une infection par le VIH-1. Nous proposons que IRENE maintiendrait l’IRES5’UTR dans une conformation peu active dans des conditions physiologiques. On sait que les IRES sont activés par divers facteurs cellulaires, appelés ITAF (IRES trans-acting factors). Nous proposons que l’IRES5’UTR adopterait une conformation active suite à la liaison d’un ITAF exprimé ou relocalisé lors d’un stress oxydatif. Ces travaux ont fait l’objet d’une publication (Gendron et al., 2011, Nucleic Acids Research, 39, 902-912). J’ai ensuite étudié l’effet de la protéine virale Tat sur l’activité de l’IRES5’UTR. En plus de son rôle essentiel dans la transactivation de la transcription des ARNm viraux, Tat stimule leur traduction coiffe-dépendante, en empêchant l’inhibition d’un facteur d’initiation canonique, eIF2, induite par la protéine kinase modulée par l’ARN double-brin (PKR) et en déroulant la structure TAR présente à l’extrémité 5’ de tous les ARNm du VIH-1. Elle affecte aussi l’expression de plusieurs gènes cellulaires. J’ai montré que les isoformes Tat86 et Tat72, mais non Tat101, stimulent l’activité de l’IRES5’UTR. Cet effet est indépendant de PKR et de TAR, mais dépendrait de la conformation de Tat. Nous proposons que Tat activerait un facteur de transcription cellulaire qui déclenche l’expression d’un ITAF de l’IRES5’UTR ou encore qu’elle activerait directement un tel ITAF. J’ai de plus montré que PKR stimule l’activité de l’IRES5’UTR, ce qui est surprenant puisque PKR est une protéine antivirale. Cet effet est indépendant de l’inhibition d’eIF2 par PKR et pourrait résulter de l’activation d’un ITAF. Sachant qu’une portion active de l’IRES5’UTR, IRES5’UTRc, est présente dans tous les ARNm viraux, notre hypothèse est que la stimulation de cet IRES par PKR permettait de traduire l’ARNm de Tat au début du cycle de réplication, ce qui permettrait ensuite la traduction coiffe-dépendante des ARNm du VIH-1, qui est stimulée par Tat. Ces travaux font l’objet d’un manuscrit (Gendron et al., soumis à RNA). Mes résultats, couplés aux données de la littérature, me conduisent à la conclusion que, à la fin du cycle de réplication du VIH-1, l’activité de l’IRES5’UTR est stimulée par le stress oxydatif, l’arrêt en G2/M et la présence de quantités élevées de Tat, alors que la traduction coiffe-dépendante est compromise. L’initiation IRES-dépendante serait alors indispensable pour que le VIH-1 traduise l’ARNmPL. L’IRES5’UTR constituerait donc une cible très intéressante pour développer des agents anti-VIH.

Low CD4(+) T-Cell Levels and B-Cell Apoptosis in Vertically HIV-exposed Noninfected Children and Adolescents

Lymphocyte subsets, activation markers and apoptosis were assessed in 20 HIV-exposed noninfected (ENI) children born to HIV-infected women who were or not exposed to antiretroviral (ARV) drugs during pregnancy and early infancy. ENI children and adolescents were aged 6-18 years and they were compared to 25 age-matched healthy non-HIV-exposed children and adolescents (Control). ENI individuals presented lower CD4(+) T cells/mm(3) than Control group (control: 1120.3 vs. ENI: 876.3; t-test, p=0.030). ENI individuals had higher B-cell apoptosis than Control group (Control: 36.6%, ARV exposed: 82.3%, ARV nonexposed: 68.5%; Kruskal-Wallis, p < 0.05), but no statistical difference was noticed between those exposed and not exposed to ARV. Immune activation in CD4(+) T, CD8(+) T and in B cells was comparable in ENI and in Control children and adolescents. Subtle long-term immune alterations might persist among ENI individuals, but the clinical consequences if any are unknown, and these children require continued monitoring.

Microbiota intestinal de indivíduos que sofreram acidente ocupacional com materiais biológicos e que realizaram profilaxia anti-retroviral

Avaliar a microbiota intestinal de indivíduos que sofreram acidente ocupacional com materiais biológicos e receberam anti-retrovirais foi o objetivo deste estudo. O grupo de estudo constou de 23 indivíduos com idade entre 18-45 anos, sendo 13 doadores de sangue e 10 que sofreram acidente ocupacional. Foram avaliados a microbiota intestinal, antropometria e exames laboratoriais pré, pós e 30 dias após o término da medicação. Zidovudina mais lamivudina foi utilizada em 70% dos indivíduos associado ao nelfinavir, 20% ao efavirenz e 10% ao ritonavir. As alterações nutricionais e dietéticas-laboratoriais e de microbiota intestinal foram analisadas em três momentos. M1: até dois dias do início da profilaxia; M2: no último dia da profilaxia e M3: 30 dias após o término da profilaxia. Náuseas, vômitos e diarréia estiveram presentes em 50% no segundo momento do estudo. Sobrepeso em 70%, desnutrição e eutrofia em 10%, dos indivíduos, não se modificaram durante o estudo. Transaminases, triglicérides, LDL-colesterol se elevaram no segundo momento e normalizaram 30 dias após término da medicação. Houve redução significativa dos Lactobacillus, Bifidobacterium e Bacteróides nos três momentos. Uso de anti-retrovirais provocou impacto significativo na microbiota intestinal dos indivíduos, sem recuperação em 30 dias.

Avaliação da resistência primária aos inibidores de integrase em pacientes soropositivos para o HIV-1

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Desenvolvimento e caracterização de dispersão sólida com propriedade mucoadesiva para liberação de zidovudina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da microbiota intestinal de indivíduos que sofreram acidente com materiais biológicos que realizaram profilaxia anti-retroviral

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desenvolvimento de nanopartículas de quitosana para adminstração nasal de zidovudina

Zidovudine (AZT) is the drug most commonly used in AIDS treatment, isolated or in combination with other antiretroviral agents, but it has certain limitations due to its therapeutic dose-dependent haematological toxicity. In addition, it has low oral bioavailability, since it undergoes pre-systemic metabolism. The nasal route has been used as an alternative route for drug administration, because it can promote its direct absorption to blood circulation, avoiding hepatic metabolism. However, this route presents as a factor limiting the mucociliary clearance mechanisms that remove quickly the formulation of the nasal cavity. To prolong the residence time of formulations, in this direction, has been proposed the development of mucoadhesive systems. Among the various existing systems, the use of chitosan (QS), as mucoadhesive polymer, has been widely exploited in the preparation of nanoparticles (NPs). The objective of this study was to develop and characterize QS’s NPs for intranasal administration of AZT. For both NPs have been developed by ionic crosslinking of QS with sodium tripolyphosphate (TPP). These NPs were characterized by studies of particle size distribution, zeta potential, morphology, mucoadhesion tests, assessing the ability of encapsulation of the drug and permeation profile of AZT. The evaluation of AZT in the NPs was determined by UV-Vis spectroscopy. Mucoadhesion measures were made using a texture analyzer, using a mucin disk and porcine mucous membrane , and permeation assay were conducted using porcine nasal mucous membrane adapted to the Franz cell. These results suggest that the systems in hand have great potential for nasal AZT administration

Desenvolvimento de dispersões sólidas e nanopartículas poliméricas mucoadesivas de Zidovudina e avaliação da interação biológica com a mucosa intestinal

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Quality of life among Brazilian women living with HIV/AIDS

The purpose of the present study was to assess quality of life (QoL) in Brazilian women living with HIV/AIDS, according to the World Health Organization Quality of Life HIV-BREF (WHOQoL-HIV-BREF) domains. A quantitative-based, cross-sectional, analytical study was carried out in healthcare centers specialized in assisting people living with HIV/AIDS, located in a municipality of the state of Sao Paulo, Brazil. One hundred and six women of age 18 years or more, users of the public healthcare system, participated in the study. Socio-demographic and clinical variables were collected using a specific questionnaire. Quality of life related variables were collected by means of the WHOQoL-HIV-BREF instrument. As per the QoL domains, study results show that the Spirituality domain reached a standardized mean score of 65.7, followed by the Physical (64.7), Psychological (60.6), Social Relationships (59.5), Independence (58.6), and Environment (54.5) domains. Results of the multiple regression analysis indicate that the women's employment or retirement, income greater than the minimum wage, and higher educational level were associated with a higher standardized mean score of QoL. However, recent HIV/AIDS diagnosis and exposure to antiretroviral agents for a period shorter than two years were negatively associated with QoL. It is critical that public policies favor an all-embracing social inclusion of these women, thus promoting better social conditions. Counseling, clinical follow-up immediately after the infection diagnosis, and initiation of antiretroviral treatment are crucial moments in the lives of these individuals.

A validated assay by liquid chromatography-tandem mass spectrometry for the simultaneous quantification of elvitegravir and rilpivirine in HIV positive patients

Because of the large variability in the pharmacokinetics of anti-HIV drugs, therapeutic drug monitoring in patients may contribute to optimize the overall efficacy and safety of antiretroviral therapy. An LC–MS/MS method for the simultaneous assay in plasma of the novel antiretroviral agents rilpivirine (RPV) and elvitegravir (EVG) has been developed to that endeavor. Plasma samples (100 μL) extraction is performed by protein precipitation with acetonitrile, and the supernatant is subsequently diluted 1:1 with 20-mM ammonium acetate/MeOH 50:50. After reverse-phase chromatography, quantification of RPV and EVG, using matrix-matched calibration samples, is performed by electrospray ionization–triple quadrupole mass spectrometry by selected reaction monitoring detection using the positive mode. The stable isotopic-labeled compounds RPV-13C6 and EVG-D6 were used as internal standards. The method was validated according to FDA recommendations, including assessment of extraction yield, matrix effects variability (<6.4%), as well as EVG and RPV short and long-term stability in plasma. Calibration curves were validated over the clinically relevant concentrations ranging from 5 to 2500 ng/ml for RPV and from 50 to 5000 ng/ml for EVG. The method is precise (inter-day CV%: 3–6.3%) and accurate (3.8–7.2%). Plasma samples were found to be stable (<15%) in all considered conditions (RT/48 h, +4°C/48 h, −20°C/3 months and 60°C/1 h). Selected metabolite profiles analysis in patients' samples revealed the presence of EVG glucuronide, that was well separated from parent EVG, allowing to exclude potential interferences through the in-source dissociation of glucuronide to parent drug. This new, rapid and robust LCMS/MS assay for the simultaneous quantification of plasma concentrations of these two major new anti-HIV drugs EVG and RPV offers an efficient analytical tool for clinical pharmacokinetics studies and routine therapeutic drug monitoring service.