The human immunodeficiency virus (HIV) and acquired immune deficiency syndrome (AIDS) that subsequently develops remain major health concerns even after three decades since the first cases were reported. Successful therapeutic measures to address HIV/AIDS consist mostly of combinations of drugs targeting viral enzymes including reverse transcriptase (RT), protease (PR) and integrase (IN) as well as entry steps of the viral life cycle. The remarkable benefits (e.g. improved quality of life) derived from the use of these agents are unfortunately limited by toxicity to the host and the development of drug resistant viral strains. Drug resistance limits the repertoire of drug combinations available. Unfortunately, because latent forms of the virus exists, therapy has to be life-long and with new infections occurring every day, resistant strains tend to spread. To circumvent these problems, new drugs that inhibit resistant strains or work against new viral targets have to be developed. The history of gold compounds as potential inhibitors of HIV prompted this study in which twenty seven compounds consisting of gold(I), gold(III) and precursors from five classes were tested for drug-likeness, anti-HIV and immunomodulatory effects using wet lab and in silico methodologies. Cytotoxicity determination was done using viability dyes and flow cytometry. Cell proliferation profiles were monitored using the carboxyflourescein succinimidyl ester dye dilution technology and a real time cell analyser for confirming viability dye findings. The compounds’ effects on viral enzymes was determined using direct enzyme assays and in silico molecular modelling techniques. H and P nuclear magnetic resonance spectroscopy studies for determining stability revealed that the backbone chemical shifts of the compounds were relatively unchanged after one week (-20 and 37 ºC) when dissolved in dimethylsulfoxide. Eight of the gold compounds had drug-like properties comparable to clinically available drugs when in silico predictions were performed. The 50% cytotoxic dose of the compounds in human cells was between 1 and 20 μM (clinically relevant concentrations for gold compounds). Three gold(I) compounds inhibited viral infectivity at non-toxic concentrations and two gold(III) compounds did so at cytostatic (anti-proliferative mechanism that is also antiviral) concentrations. In the immunomodulatory assay, cytokine levels were altered by five compounds with one gold(I) and a gold(III) compound significantly reducing the frequency of CD4+ cells (an anti-viral function) from HIV+ donors (p= 0.005 and 0.027 respectively) when multi-parametric flow cytometry was performed. Inhibition of RT activity was predicted in in silico studies to be through interactions with the ribonuclease (RNase) H site although with poor stereochemical orientation while favourable binding predictions with the IN cofactor binding site were observed for some gold(III) complexes. Compounds predicted to interact with the RNase H site of RT and the IN cofactor site require structural modification to improve drug-likeness and binding affinity. The drug-like compound(s) which inhibited viral infectivity and lowered CD4+ cell frequency have potential for incorporation into virostatic cocktails (combination of cytostatic and directly anti-viral agent). Cytostatic agents are known to be less prone to drug resistance and because they lower CD4+ cell frequency, such compounds can potentially limit HIV immune activation. / Thesis (PhD)--University of Pretoria, 2011. / Biochemistry / unrestricted

Patients can develop malignancies due to various reasons including genetic factors, chemical carcinogens, radiation, and defects in their immune system. The immune system is postulated to carry out routine surveillance for malignancy. Patients who have defective immune responses may be susceptible to malignancies due to complicated underlying mechanisms. These include defective immune response to cancer-causing bacteria, transforming viruses, and concomitant molecular, cellular and immunoregulatory defects. Common variable immune deficiency (CVID) is characterized by hypogammaglobulinemia, impaired antibody responses and an increased susceptibility to infections. A disorderly immune response, or immune dysregulation, may also lead to autoimmune complications and possibly to malignancy. The treatment of CVID involves infusion of replacement doses of immunoglobulin, either intravenously (IGIV) or subcutaneously (SCIG). However, it is unclear whether adequate replacement of immunoglobulins is sufficient to prevent the increased risk of malignancy seen in this disease. We present two cases of unusual solid tumors complicating CVID treated with adequate doses of intravenous immunoglobulins. In this study we review the occurrence of malignancy in patients with CVID and postulate mechanisms that may be involved indigent to this disease. We will also review the role of replacement immunoglobulin and discuss cancer screening in these high risk individuals.

Proteases are present in all living organisms and are involved in various biological processes. Inhibition of protease activities in disease-causing agents is one strategy for rational drug design. Characterization of the protease inhibition processes is essential for understanding the inhibition mechanisms and for developing efficient therapeutics. This work represents a major challenge in analytical biochemistry. In this study, a strategy based on mass spectrometry has been developed to characterize irreversible inhibition of proteases. Five proteases representing three of the four protease classes were irreversibly inhibited by various irreversible inhibitors, some of which are potential drug candidates. In all the cases, the stoichiometry of each of the protease/inhibitor complexes was determined by electrospray ionization mass spectrometry through measurement of the complex's molecular weight. The inhibited proteases were then enzymatically cleaved and the resulting peptides isolated for further characterization by high performance tandem mass spectrometry. Attention was focused on the determination of the site(s) of the modification and the reaction mechanisms involved. High energy collision induced dissociation mass spectra of each modified peptide provided information on the exact modification site(s) and the detailed chemical nature of the covalent complex. The serine protease trypsin, the cysteine protease cruzain, and the aspartic proteases, HIV-2 protease and SIV protease, were covalently modified only at one amino acid residue, while the aspartic protease, HIV-1 protease, was found to be modified at three sites by the haloperidol derivative compounds. In addition, mass spectrometry has been applied to characterize the plasma glycoprotein, biotinidase, and to obtain partial peptide sequences of a membrane-bound protein, UDP-GalNac:polypeptide N-acetylgalactosaminyl transferase, using a low picomole quantity of sample.

This study assessed the effectiveness of an HIV/AIDS psychoeducational intervention and skills training package for a high-risk group for HIV/AIDS, the chronic mentally ill. Twenty-seven participants were randomly assigned to either a treatment or no treatment control group. Elements of the intervention were (a) education about HIV transmission and high risk groups, (b) problem-solving skills training, and (c) behavioral skills training based on education needs identified in a previous study of the same population. Pretest and posttest measures were administered to both the treatment and control groups in order to assess differences and improvement. Results showed improvements from pretest to posttest in the treatment group's HIV/AIDS knowledge, behavioral and role-play skills in high-risk situations, self-efficacy in utilizing learned skills, and intention to use these skills. The same improvement did not occur in the control group. Follow-up data and implications of the study are discussed.

No description available.

The population pharmacokinetics and pharmacodynamics (PK/PD) of nevirapine (NVP), zidovudine (ZDV), and didanosine (ddI) were evaluated in 432 pediatric patients with HIV, randomized to receive either a double-therapy of ZDV + ddI or NVP + ddI; or triple-therapy of NVP + ZDV + ddI as a substudy of the AIDS Clinical Trials Group Protocol 245 in 2 phases. In phase 1, nonlinear mixed-effect modeling (NONMEM) analysis was employed for population pharmacokinetics (PPK) study for ZDV, ddI and NVP. One-compartment model with first-order input and first-order elimination was fitted to the NVP, ZDV and ddI data. Final PPK models were as follows: ZDV; CL (1/hr, without nevirapine coadministration) = 52.4 × BSA, CL (1/hr, with nevirapine coadministration) = 65.0 × BSA, Vd/F(1) = 116 × BSA, ddI; CL (1/hr) = 73.4 × BSA + 69.9, Vd/F(1) = 132, and NVP; CL (1/hr) = 2.30 × BSA, Vd/F(1) = 120. In phase 2, the relationship between the predicted serum concentrations of ZDV, ddI, and NVP and pharmacodynamic responses were evaluated via S-Plus ® exploratory data analysis. No apparent relationship between average steady-state serum concentrations and pharmacodynamic variables, such as HIV-1 RNA(RNA) levels, CD4 + count was found. However, the responses of RNA level and CD4 + count to the double therapy (ddI/NVP) versus triple therapy (ddI/NVP/ZDV) were significantly different after 4 weeks of therapy ( P = 0.0014 for RNA level at week 4, P = 0.0454 for CD4 + count at week 12). No significantly different responses were found in weight changes ( P > 0.25 at all weeks). Also, the maximum drop of RNA level throughout the treatment period had a strong relationship to the decline slope of RNA at week 4 as follows: Maximum drop of RNA = 3.1139 × RNA decline slope at week 4 - 0.411. Nevirapine dosing regimens were compared using simulation via Trial Simulator™. Both ACTG regimen (body surface area based) and manufacture's regimen (age and weight based) produced similar concentrations at lower end concentration but manufacture's regimen produce higher concentration at upper end with 1000 simulated patients (ACTG regimen; 2150, 3827, and 4992 ng/ml, manufacturer's regimen; 2066, 4130, and 6568 ng/ml, for 10 th , 50 th , and 96 th percentile, respectively). It is suggested to use body surface area based dosing regimen for NVP.

This study examined the effects of publicly modeling the HIV testing procedure on testing rates at a university health center. Thirteen models, demographically similar to the university student population, modeled the HIV counseling and testing procedure at a campus student center. The intervention was employed twice during the fall semester. HIV testing rates for that semester were compared to the two previous fall semesters. Results showed that HIV testing rates during the semester of intervention were not significantly higher than previous semesters. Results did not demonstrate a marked increase in the number of tests administered within 2 weeks following each modeling intervention. Modeling was not an effective means to increasing utilization of HIV testing centers on this university campus.

Pas de résumé / Chronic mucocutaneous candidiasis (CMC) is seen in human patients with a variety of conditions and refers to recurrent or persistent infection of the skin, nails and/or mucosae by commensal Candida species. Its pathogenesis had long remained elusive, until human genetic studies of rare patients with inherited forms of idiopathic CMC (whether isolated or syndromic), incriminated impaired interleukin (IL)-17A/F immunity. The first genetic etiologies of idiopathic isolated CMC, autosomal dominant (AD) IL-17F and autosomal recessive (AR) IL-17 receptor A (IL-17RA) deficiencies, were reported in 2011 in a multiplex and in a sporadic case, respectively. Using Whole Exome Sequencing (WES), we identified 26 novel patients bearing 15 different homozygous mutations in the IL17RA gene. The mutations identified are either nonsense; missense; frameshift deletions; frameshift insertions; or non-coding essential splice site mutations. Interestingly, 2 alleles encode for surface expressed receptors, whereas all the other tested alleles are not detected at the surface of the patient’s cells (fibroblasts or leucocytes). IL-17RA deficiency is a fully penetrant AR disease, with early onset symptoms, usually within the first year of life. CMC is always present. In addition, 17 patients present with staphylococcal skin infections, and some patients with pyogenic infections of the respiratory tract, including pneumonia. Interestingly, tuberculosis occurred in two unrelated BCG-vaccinated patients. The response to IL-17A and IL-17F homo- and heterodimers is abrogated in fibroblasts, as well as the response to IL-17E/IL-25 in T cells. Human IL-17RA is thus essential for mucocutaneous immunity against Candida and Staphylococcus, but otherwise largely redundant. AR IL-17RA deficiency should be considered in children or adults with CMC, cutaneous staphylococcal disease, or both. In a separate project, I investigated a female child patient born to consanguineous parents who suffered from CMC, recurrent viral infections, disseminated BCG disease and biliary cryptosporidiosis, suggestive of combined immunodeficiency, and who is homozygous for a mutation in REL, encoding the NF-kB protein c-REL. Sanger sequencing confirmed that the patient is homozygous and that both parents are heterozygous for the mutation, consistent with an AR inheritance. The candidate mutation is a nucleotide substitution localized in an acceptor splice site; is not reported in available public databases; and is predicted to be damaging in silico. The mutation disrupts mRNA splicing and is loss-of expression. The patient shows normal counts of lymphoid subsets, with the exception of diminished frequencies of memory CD4+ T, Th2, Th1*, and memory B cells. The patient’s T cells fail to proliferate in response to recall antigens. Naïve CD4+ T cells produce little IL-2 and respond poorly to polyclonal stimulation, a phenotype reverted by exogenous IL-2. Memory CD4+ T cells also produce little amounts of IL-2, and strongly diminished amounts of key effector cytokines (IFN-γ, IL-4, IL-17A and IL-21). The patient exhibited with no detectable specific antibody response following vaccination. Survival and therefore proliferation of naïve B cells are compromised leading to poor generation of plasma cell, and immunoglobulins secretion. The patient shows normal counts of myeloid cells, and frequencies of dendritic cell subsets. IL-12 production is abolished in whole blood in response to BCG+IFN-γ and B-EBV cells in response to mitogens. Although further investigation is needed to fully characterize the patient’s phenotype, these results strongly suggest that the patient suffers from AR complete c-REL deficiency.

Este estudo discute a AIDS enquanto um fenômeno simbólico para indivíduos a partir da experiência clínica e do referencial teórico da psicologia analítica.A técnica utilizada foi a análise de registros de sonhos. Durante 22 meses foram registrados 125 sonhos de 9 de pessoas infectadas pelo HIV, e os sintomas relacionados à AIDS. Destes foram selecionadas três séries oníricas totalizando 74 sonhos, segundo os critérios: idade, sexo, orientação sexual, escolaridade, nível sócio-econômico, período de infecção, período de coleta de dados, local de atendimento e número e qualidade dos sonhos registrados. A análise foi realizada primeiro, a partir das associações e dos dados coletados durante as sessões, procurando verificar as relações entre os sonhos e o momento de vida do indivíduo. A seguir, identificou-se a constelação arquetípica mais presente em cada série, amplificando-a a nível arquetípico e relacionando-a com a situação psicológica do sujeito. Procurou-se também averiguar o movimento de compensação e/ou adaptação do inconsciente à experiência de vida marcada pela AIDS. E por último, analisou-se as três séries em conjunto procurando destacar os pontos relevantes para a compreensão do fenômeno da AIDS como um todo. A análise dos dados indica que a AIDS, enquanto fenômeno simbólico, pode ser inserida em um movimento de reestruturação amplo da personalidade. A compensação parece dar-se dentro deste processo e não em relação ao evento da AIDS isoladamente. Porém, foram registradas constelações do Self durante os eventos somáticos mais graves, o que pode indicar um esforço compensatório frente a uma situação de ameaça à vida. Os sintomas menos graves foram acompanhados pela emersão de conteúdos da sombra com os quais parecem ter uma relação simbólica. As várias referências ao mito de Dioniso ao longo das séries pode indicar, que a nível coletivo, a epidemia da AIDS faz parte de um movimento compensatório à cultura judaico-cristã-apolínea, que visa redimensionar a consciência, através da integração dos elementos instintivos da psique. / This study focuses on AIDS while a symbolic phenomenon in individuals, based on clinical experience supported by the analytical psychology theory. The technique used in this work was dream analysis. In a period of 22 months it was registered 125 dreams of 9 people infected with HIV, as well as the symptoms related to AIDS. It was selected three series of dreams totaling 74 dreams according to: age, sex, sex orientation, scholarship, social-economic level, period of infection and data registration, counseling place and amount of registered dreams. First, the analysis was made, from the associations and therapy sessions' data, in order to verify the relations between dreams and the individual situation. Next it was identified the archetypal constellation present in each series. This archetypal constellation was amplified at archetypal level and related to the psychological situation of the individual. It was also verified the compensation and the adaptation movement in relation to AIDS. Finally, the three series were analyzed all together and the significant aspects to the comprehension of AIDS phenomena were discussed. The data analysis suggests that AIDS, while a symbolic phenomenon, can be understood as part of a great movement of personality change. The compensation movement seems to take place in this process and not in relation to AIDS. However, Self constellations were registered during the most serious somatic events, what can denote a compensation in face of a threatening life situation. Less serious somatic events were followed by the emergence of shadow contents, with which they seem to have a symbolic relation. Many references to Dioniso myth can indicate at a collective level, a compensatory movement against the Judaic-Christian-Apolinean culture that aims the consciousness transformation by integrating the instinctive elements of the psyche.

As ações de prevenção ao HIV/aids na ESF foram objetos desse estudo, cujo objetivo foi analisá-las, sob a ótica da vulnerabilidade na sua dimensão programática, identificar como as equipes de saúde da família reconhecem a vulnerabilidade ao HIV/aids nos seus territórios; descrever as ações de prevenção direcionadas às populações reconhecidas pelas equipes como vulneráveis e caracterizar as dificuldades, as potencialidades e os desafios na prevenção ao HIV/aids na ESF. Para alcançar tais objetivos optou-se por um estudo exploratório, com abordagem qualitativa, utilizando-se a metodologia do Estudo de Caso. O conceito de vulnerabilidade, em sua dimensão programática, foi usado como quadro teórico. A coleta de dados foi realizada por meio de entrevistas semi-estruturadas com gerentes de UBS de Saúde ou pessoas indicadas por eles e mediante técnica de grupo focal com equipes de saúde da família. O estudo foi conduzido no período de junho a dezembro de 2009, em 11 Unidades do distrito de Capão Redondo, região sul do Município de São Paulo, Brasil. Os resultados apontam que as equipes reconhecem a vulnerabilidade ao HIV/aids em seus territórios atrelada à dimensão individual, fundamentada na noção de risco da epidemiologia tradicional, identificando grupos ou comportamentos de risco. Fundamentadas nos mesmos conceitos, percebeu-se o predomínio de intervenções informativas, de caráter prescritivo e normativo, que tinha como principal finalidade a mudança de comportamento do indivíduo. Ações fortemente baseadas no modelo biomédico hegemônico. Como dificuldades encontradas destacaram-se àquelas relacionadas ao processo de trabalho das equipes, que apontam sobrecarga de trabalho pelas demandas dos programas prioritários; alta rotatividade dos profissionais e falta de capacitação dos mesmos para a prevenção ao HIV. A maior potência da ESF na prevenção da aids encontra-se no princípio da longitudinalidade, que permite qualificar o reconhecimento da vulnerabilidade do território e planejar as ações de prevenção de acordo com as necessidades da população. O maior desafio apontado pelas equipes está em reconhecer o processo de produção e reprodução social como determinante da vulnerabilidade ao HIV. Por fim, conclui-se que o modelo técnico-assistencial vigente na atenção básica, apesar de sua reconhecida potencialidade, ainda apresenta práticas embasadas na noção de risco e no modelo biomédico hegemônico, determinando, assim, a vulnerabilidade programática na prevenção ao HIV/aids. / Actions to prevent HIV/AIDS in the FHS were objects of this study, whose objective was to analyze them from the perspective of vulnerability in its programmatic dimension, to identify how the family health teams recognize the vulnerability to HIV/AIDS in their territories; describe the preventive actions directed to the populations recognized by the teams as vulnerable and characterize the problems, potentiality and challenges in the prevention of HIV/AIDS. To achieve these goals was chosen an exploratory study with a qualitative approach, using the methodology of the Case Study. The concept of vulnerability in its programmatic dimension, was used as a theoric framework. Data collection was conducted through semi-structured interviews with managers of Basic Health Units or people appointed from them and through focus group technique with family health teams. The study was conducted from June to December 2009, in 11 units in the district of Capão Redondo, south of São Paulo city, Brazil. The results show that teams recognize the vulnerability to HIV / AIDS in their territories linked to the individual dimension, grounded in the notion of risk in traditional epidemiology, identifying risk groups or risk behaviors. Reasoned on the same concepts, it was noted the predominance of interventions informational, and normative and prescriptive character that had as main purpose to change the behavior of individual. Actions strongly based on the hegemonic biomedical model. As difficulties faced stood out to those linked to the process of work teams, which aim to work overload by the demands of the priority programs, high turnover and lack of professional training to prevent HIV/AIDS. The biggest potential of the FHS is in the principle of longitudinality, which allows to qualify the recognition of the vulnerability of the territory and to plan preventive actions according to the needs of the population. The biggest challenge is appointed by the teams in recognize the process of social production and reproduction as a determinant of vulnerability to HIV/AIDS. Finally, we conclude that the current technical health care model in primary care, despite its recognized potential also presents practices based in the notion of risk and the hegemonic biomedical model, thus determining the programmatic vulnerability in preventing HIV/AIDS.