Thesis (doctoral)--Ludwig-Maximilians-Universität, 1996.

Thesis (doctoral)--Ludwig-Maximilians-Universität, 1996.

Thesis (M.N.)--Memorial University of Newfoundland, 1996. / Typescript. Bibliography: leaves 94-104. Also available online.

Thesis (Ph. D.)--Université Laval, 2005. / Title from PDF title page (viewed on Oct. 5, 2006). Includes bibliographical references (p. [299]-320).

A implementação da integralidade é um grande desafio à consolidação do Sistema Único de Saúde e a vulnerabilidade na Atenção Básica às DST/HIV/aids foi o objeto deste estudo, cujos objetivos foram: caracterizar as Unidades Básicas de Saúde (UBS) segundo as Coordenadorias Regionais de Saúde, as Supervisões Técnicas de Saúde, os modelos de organização da atenção à saúde, as Organizações Sociais de Saúde em contratos de gestão na cogestão da saúde com a Secretaria Municipal de Saúde de São Paulo e a formação profissional do gerente; identificar o grau de Vulnerabilidade Programática das UBS e discuti-lo segundo os componentes: acessibilidade, porta de entrada, vínculo, enfoque familiar, profissionais da saúde e coordenação/integração. A perspectiva conceitual utilizada foi da vulnerabilidade, em sua dimensão programática. Realizou-se estudo descritivo transversal com abordagem quantitativa, tendo sido utilizado um questionário com 51 questões, aplicado online, na plataforma FormSUS, com gerentes das 442 UBS do Município de São Paulo. Foram obtidas respostas de 328 gerentes das UBS; 40,9% eram exclusivas \"tradicionais\"; 55,8% das OSS eram do tipo \"Associação\" e mais da metade dos gerentes das UBS eram Enfermeiros. Há graus diferenciados de vulnerabilidade programático, mas de um modo geral nas UBS é baixo. Há vulnerabilidade na efetivação da integralidade: falta de materiais para atividades educativas; baixa oferta de testes de detecção de sífilis à gestante e teste anti- HIV no pré-natal; demora no retorno do resultado do exame anti-HIV; baixa indicação do tratamento com Penicilina Benzatina ao parceiro da gestante com diagnóstico de sífilis; não realização da abordagem consentida para solicitação de teste para HIV à gestante ou para a população geral; falta de capacitação para realização da abordagem sindrômica das DST e aconselhamento na oferta do teste do HIV; falta de contrarreferência às UBS, DST/aids. Conclui-se que a integralidade traduzida em práticas de saúde na atenção às DST/HIV/aids possibilitou visualizar as vulnerabilidades programáticas nas UBS e apontou desafios para sua efetivação. / Implementation of comprehensive attention is a huge challenge in the consolidation of the Brazilian Unified Health System. The vulnerability to STD/HIV/Aids in Primary Care was focused on this study, aiming to provide tools to improve healthcare. The objectives of the study were: 1) to characterize Primary Care Units (PCU) according to selected variables such as: administrative hierarchy position (Regional Health Coordination, Healthcare Technical Supervision), administrative issues (models healthcare attention of organization, Healthcare Social Organization (HSO) in management or co-management with the Township Healthcare Office in São Paulo), and manager formal professional qualification; 2) to identify the level of Programmatic Vulnerability; and 3) to assess the Programmatic Vulnerability according to the following components: accessibility, point of entry, links, family approach, health professional and integration / coordination. The conceptual approach was the vulnerability in its programmatic dimension. This was a transversal descriptive and quantitative study based on an online survey with 51 questions applied in the FormSUS platform and involving managers from 442 PCU of the city of São Paulo. Responses were received from 328 PCU managers; among them more than half were Nurses. PCU were exclusively \"conventional in 40.9% and 55.8% of the HSO were classified as \"association\". There were different levels of programmatic vulnerability; in general however, the vulnerability was low. There was vulnerability in making comprehensive attention effective: lacking of material for educational activities; insufficiency of availability of tests to detect syphilis in pregnant women and anti-HIV test in the pre-natal period; excessive delay in results of anti-HIV tests causing them to be not time-opportune; low practice of indication of syphilis treatment with Benzathine Penicillin for the pregnant partner; lacking of informed consent to request HIV testing to pregnant women as well general population; lacking of training for the syndromic approach to STDs; lacking of counseling when offering HIV testing; lacking of counter-reference to PCU, STD/Aids. It was concluded that the translation of the comprehensiveness concept into health practices regarding STD/HIV/aids enabled us to assess programmatic vulnerabilities in the PCU, and pointed out challenges to achieve a true comprehensive healthcare attention.

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

The development of a safe and effective HIV vaccine remains challenging due to its high antigenic variability. Poxviruses are large, stable, and have a track record of use as human vaccine candidates. Recombinant fowlpox virus 9 (rFP9), a highly attenuated host range-restricted poxvirus strain, has been safely administered to humans with no ill effects, and is known to be immunogenic. This thesis describes the construction of complex rFP9 encoding various sequences of SIVmac239. The SIVmac239/macaque model is widely used for HIV vaccine development. The ultimate aim of this work was to combine the advantages of FP9 with those of live attenuated SIV to produce a safe yet hopefully effective model HIV vaccine candidate. Transfer plasmids for five different insertion sites within the FP9 genome were designed and constructed. Homologous recombination (HR) of adjacent FP9 sequences was employed to facilitate the integration of SIVmac239 sequences into the FP9 genome. Positive rFP9 were identified by blue colouration in presence of X-gal using a transient colour selection (TCS) technique, and the final markerless pure recombinants were confirmed by PCR. Expression of the target SIV proteins in the presence of T7 polymerase has been demonstrated by immunocytochemical (ICC) staining and Western blotting (WB) assays. Expression was also quantified by enzyme-linked immunosorbent assay (ELISA) in various cell lines at multiple time points. Five different unique rFP9 have been constructed through this project. All SIVmac239 open reading frames (ORFs) save nef have been integrated into the FP9 genome, and protein expression demonstrated where possible. Moreover, a single rFP9 vector expressing the defective SIVmac239 genome driven by T7 RNA polymerase has been successfully constructed and validated using a green fluorescent protein marker.rFP9 showed appropriate transgene expression in both avian and mammalian cells, although at different levels. The expression efficiency of rFP9 was finally compared to another attenuated poxvirus vector, modified vaccinia Ankara (MVA). Comparing the protein expression levels between rFP9 and rMVA was quite difficult because different poxvirus promoters (early/late in rFP9; intermediate in rMVA) were used to direct the transcription of the T7 RNA gene. Given this limitation, although generally higher levels of expression were seen with rFP9, this cannot be attributed to the FP9 with any certainty.

INTRODUÇÃO: A deficiência de IgA (DIgA) é a imunodeficiência primária mais comum e caracteriza-se pela presença de concentrações de IgA sérica abaixo de 7 mg/dL e níveis normais de IgM e IgG. A maioria dos indivíduos acometidos não apresenta doença aparente embora alguns possam apresentar infecções recorrentes ou crônicas de mucosas, atopia e/ou doenças autoimunes (DAIs). Presumivelmente, a doença resulta de um defeito na troca de isótipo para IgA ou de falha na maturação de linfócitos produtores de IgA. A imunodeficiência comum variável (ICV) constitui uma deficiência primária de anticorpos caracterizada por níveis séricos baixos de IgG, IgA e/ou IgM, ao lado de valores normais ou diminuídos de linfócitos B e/ou T, levando a infecções crônicas ou recorrentes principalmente dos tratos respiratório e gastrintestinal. Embora a fisiopatologia da ICV ainda não esteja esclarecida, em muitos pacientes ela pode ser decorrente de algum defeito intrínseco de linfócitos B. De modo especial, as células B de memória (CD27+) têm sido correlacionadas com alguns aspectos clínicos da doença. Números elevados de células B de memória com persistência de IgM (CD27+IgM+) parecem estar correlacionados com a presença de infecções, enquanto valores diminuídos de células B de memória clássicas ou class-switched (CD27+IgG-IgM-) parecem estar associados a baixos níveis de IgG e presença de autoimunidade. A progressão de DIgA para ICV tem sido descrita em alguns pacientes embora não constitua regra geral. Uma hipótese é a de que uma base genética comum e a associação com DAIs possam constituir fatores de risco para a progressão de DIgA para ICV. Há relato anterior de que a persistência de células B imaturas IgM+ IgD+ em alguns pacientes com DIgA estava associada à progressão para ICV. Adicionalmente, há evidências de que a diminuição de células B de memória em uma proporção de pacientes com ICV esteja associada à presença de autoimunidade. OBJETIVOS: comparar em pacientes com DIgA e ICV várias subpopulações de células B e analisar a relação entre estas populações celulares e a presença de DAIs em ambos grupos. MÉTODO: Este estudo incluiu 56 pacientes adultos de ambos sexos com DIgA e ICV, distribuídos em grupos de acordo com a associação com DAI: grupo DIgA sem DAI (14 pacientes), grupo DIgA com DAI (14 pacientes), grupo ICV sem DAI (14 pacientes) e grupo ICV com DAI (14 pacientes). As seguintes subpopulações de células B foram determinadas por citometria de fluxo de quatro-cores: células B naive (CD19+IgM+), células B de memória clássicas ou class-switched (CD27+IgM-IgD-) e células B de memória imaturas (CD27+IgM+ or CD27+IgD+). Na análise estatística foi aplicado o teste de ANOVA; valores significativos foram determinados pela correção de Bonferoni. RESULTADOS: os grupos analisados foram homogêneos quanto à idade e distribuição de gêneros. Os valores de linfócitos totais e de células B naive foram similares nos quatro grupos estudados. Os pacientes com deficiência de IgA e ICV com DAIs associadas apresentaram valores igualmente aumentados de células B de memória imaturas CD27+IgM+ e CD27+IgD+ quando comparados a pacientes sem doenças autoimunes. CONCLUSÕES: estes resultados sugerem que a persistência de células B de memória imaturas possa estar relacionada à presença de autoimunidade em pacientes com DIgA e ICV. Especula-se se a persistência destas células em pacientes com DIgA e DAI associada possa constituir fator preditivo da progressão de DIgA para ICV. / INTRODUCTION: IgA deficiency (IgAD) is the most common primary immunodeficiency disorder and is characterized by serum IgA concentration below 7 mg/dL and normal serum IgM and IgG levels. Most of the affected individuals have no apparent disease, whereas selected patients suffer from recurrent or chronic mucosal infections, atopy and/or autoimmune diseases (AIDs). This defect is presumed to result from impaired class-switching to IgA or from maturational failure of IgA-producing lymphocytes. Common variable immunodeficiency (CVID) is a primary antibody deficiency disease characterized by low serum levels of IgG, IgA and/or IgM, and normal or decreased B and/or T cell numbers, leading to chronic or recurrent infections, noted mostly in the respiratory and gastrointestinal tract. While the pathophysiology of CVID remains elusive, in many patients it may be due to an intrinsic B cell defect. Memory B cells (CD27+) in particular, have been noted to correlate with certain clinical aspects of the disease. High numbers of IgM+ memory B cells (CD27+IgM+) appear to correlate with the presence of infections, whereas decreased numbers of classic (class-switched) memory B cells (CD27+IgG-IgM- ) correlate with lower serum IgG levels and increased rates of autoimmune features. Progression from IgAD to common variable immunodeficiency (CVID) has been reported in some patients, but is not a general rule. It is postulated that a common genetic base and association with AIDs could be risk factors for progression from IgAD to CVID. OBJECTIVES: The aim of this study was to compare B cell subpopulations of patients with IgAD and with CVID, and to assess the relationship between these populations and the presence of autoimmune diseases in both group of patients: . METHOD: The study included 56 adult patients of both genders with IgAD or CVID. Patients were grouped, according to the association with autoimmune disease,as follows: group IgAD with AID (14 patients), group IgAD without AID (14 patients), group CVID with AID (14 patients) and group CVID without AID (14 patients). We determined by immunophenotyping of lymphocytes by four-colour cytometry the following subpopulations of B cells: naïve B cells (CD19+IgM+), class-switched memory B cells (CD27+IgM-IgD-) and immature B memory cells (CD27+IgM+ or CD27+IgD+). Statistical analysis was performed by the ANOVA test; significant P-values were determined by means of Bonferonis correction. RESULTS: there is no statistically significant difference between the average ages and the gender of patients between the groups. the distribution of the sample values seem to indicating that, there is no statistically significant difference in the CD19 levels between the groups. patients with AID represent greater values of CD27 IgM+ and CD27+ IgD+ than patients without AID, independent of the group studied. CONCLUSIONS: These results suggest that the persistence of immature memory B cells in patients with IgAD and CVID can be related to autoimmune diseases. We speculate if the persistence of immature B cells can constitute risk factor to progression of IgAD for CVID.

Thesis (M.Med. (Radiation Oncology)) -- University of Limpopo, 2020 / Introduction Invasive cervical cancer (ICC) constitutes almost 50% of all cancer conditions diagnosed and treated at the Polokwane/Mankweng Hospital Complex (PMHC). HIV infection is also a very common condition. There is no consensus on the relationship between the two clinical conditions among patients treated at PMHC. There is a need to describe the simultaneous occurrence of the two clinical conditions among these patients to define a rational approach to these conditions’ clinical management. Methodology This was a retrospective review of medical records of patients diagnosed with ICC who were treated at PMHC in Limpopo Province, South Africa in 2013. Results Three hundred and twenty-nine medical records were reviewed in this study; 64% of the patients were HIV-negative, and only 35% were HIV-positive. Thirty-five percent of the patients were younger than 50 years of age, followed by those aged 50–59 years (23%). Among women in the age group 30–59 years, the most common ICC stages were IIB and IIIB. In women older than 60 years, stages IIB, IIIA, IIIB and IVA were the most common. In the HIV-positive women, 18% had a CD4 cell count of less than 200/μL, compared to 2% in the HIV-negative women (p <0.05). Among the HIV-negative women, stages IIIB (49.8%) and IIB (24.6%) were the most common, where as among those who were HIV-positive, stages IIIB (55.6%) and IIB (22.6%) dominated. Conclusion This retrospective study did not find any relationship between HIV infection and ICC in patients treated at PMHC. However, it indicated that a significant proportion of HIV-positive women with ICC had lower CD4 cell counts compared to those of HIV-negative women. KEY CONCEPTS: Invasive cervical cancer, Human immunodeficiency virus, Stage, Prevalence, CD4 cell count, Age, Polokwane/Makweng Hospital Complex

Philosophiae Doctor - PhD / The continued success of antiretroviral programmes in the treatment of HIV is dependent on access to a cost-effective HIV drug resistance test (HIV-DRT). HIVDRT involves sequencing a fragment of the HIV genome and characterising the presence/absence of mutations that confer resistance to one or more drugs. HIV-DRT using conventional DNA sequencing is prohibitively expensive (~US$150 per patient) for routine use in resource-limited settings such as many African countries. While the advent of ultra deep pyrosequencing (UDPS) approaches have considerably reduced (3-5 fold reduction) the cost of generating the sequence data, there has been an even more significant increase in the volume of data generated and the complexity involved in its analysis. In order to address this issue we have developed Seq2Res, a computational pipeline for HIV drug resistance test from UDPS genotypic data. We have developed QTrim, software that undertakes high throughput quality trimming of UDPS sequencing data to ensure that subsequently analyzed data is of high quality. The comparison of QTrim to other widely used tools showed that it is equivalent to the next best method at trimming good quality data but outperforms all methods at trimming poor quality data. Further, we have developed, and evaluated, a computational approach for the analysis of UDPS sequence data generated using the novel Primer ID that enables the generation of a consensus sequence from all sequence reads originating from the same viral template, thus reducing the presence of PCR and sequencing induced errors in the dataset as well as reducing. We see that while the Primer ID approach does undoubtedly reduce the prevalence of PCR and sequencing induced errors, it artificially reduces the diversity of the subsequently analysed data due to the large volume of data that is discarded as a result of there being an insufficient number of sequences for consensus sequence generation. We validated the sensitivity of the Seq2Res pipeline using two real biological datasets from the Stanford HIV Database and five simulated datasets The Seq2Res results correlated fully with that of the Stanford database as well as identifying a drug resistance mutations (DRM) that had been incorrectly interpreted by the Stanford approach. Further, the analysis of the simulated datasets showed that Seq2Res is capable of accurately identifying DRMs at all prevalence levels down to at least 1% of the sequence data generated from a viral population. Finally, we applied Seq2Res to UDPS resistance data generated from as many as 641 individuals as part of the CIPRA-SA study to evaluate the effectiveness of UDPS HIV drug resistance genotyping in resource limited settings with a high burden of HIV infections. We find that, despite the FLX coverage being almost three times as much as that of the Junior platform, resistance genotyping results are directly comparable between both of the approaches at a range of prevalence levels to as low as 1%. Further, we find no significant difference between UDPS sequencing and the "gold standard" Sanger based approach, thus indicating that pooling as many as 48 patient's data and sequencing using the Roche/454 Junior platform is a viable approach for HIV drug resistance genotyping. Further, we explored the presence of resistant minor variants in individual's viral populations and find that the identification of minor resistant variants in individuals exposed to nevirapine through PMTCT correlates with the time since exposure. We conclude that HIV resistance genotyping is now a viable prospect for resource limited setting with a high burden of HIV infections and that UDPS approaches are at least as sensitive as the currently used Sanger-based sequencing approaches. Further, the development of Seq2Res has provided a sensitive, easy to use and scalable technology that facilitates the routine use of UDPS for HIV drug resistance genotyping.