Thesis (Ph. D.)--State University of New York at Binghamton, Department of Chemistry, 2004. / Includes bibliographical references.

Thesis (M.S.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on October 23, 2007) Includes bibliographical references.

Thesis (MTech (Nursing))--Cape Peninsula University of Technology, 2009. / Includes bibliographical references (leaves 80-85). Also available online.

No description available.

HIV-2 is a contemporary human retrovirus with the majority of infections localised to West Africa. Both HIV-1 and HIV-2 are able to cause AIDS; however, in contrast to HIV-1 infection, a common outcome following HIV-2 infection (∼ 37% of patients in this study cohort) is long-term non-progression (LTNP), where patients remain aviraemic and asymptomatic in the absence of treatment, often for decades. HIV-1 and HIV-2 both arose following zoonotic transmission of SIVs from non-human primates at around the beginning of the 20^th century and when patients develop AIDS caused by HIV-2 infection, it is clinically indistinguishable from AIDS following HIV-1 infection. Whilst the estimated number of HIV-2 infections remains small in the context of the global HIV pandemic (HIV-2 ∼ 2 million, HIV-1 group M ∼75 million), the differences in pathogenicity between these two viruses has been a source of great interest, particularly the features of LTNPs that allow control of viral replication in the absence of anti-retroviral treatment. The studies described in this thesis were carried out using samples collected from a well-characterised longitudinal community cohort in Caió, Guinea-Bissau. Chapter 3 of this thesis presents an investigation into the variation and evolution present in the HIV-2 specific accessory gene vpx. The data showed significantly increased signals of positive selection pressure in vpx in viraemic when compared to non-viraemic patients and also allowed the identification of novel variations at high frequencies (up to 22%) in this cohort that were previously un-described. Chapters 4 and 5 present a novel application of shotgun RNA sequencing (RNA- Seq) to HIV ex vitro and ex vivo samples. Chapter 4 demonstrates the divergence seen in a cultured viral isolate at the level of the whole genome, in the absence of many of the biases typically involved in sequencing of RNA viruses. Chapter 5 further extends this method to show the applicability of using RNA-Seq on primary patient HIV samples for the first time. Analysis of diversity estimates over the whole genome in the context of a low bias sequencing method show a high level of diversity in HIV-2 pol and low diversity in vpx. The aim of this work was to combine traditional and novel sequencing methods to facilitate assessment of the variation and evolution acting on vpx and to generate an accurate picture of the genetic diversity over the whole genome of HIV-2.

Orientador: Pablo Agustin Vargas / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-10T01:33:44Z (GMT). No. of bitstreams: 1 Leon_JorgeEsquiche_D.pdf: 14373006 bytes, checksum: 92ef59d2c370853b3a5a566177688788 (MD5) Previous issue date: 2008 / Resumo: Os objetivos do presente estudo foram descrever as características histopatológicas, immunoistoquímicas e de hibridização "in situ" (HIS) das lesões encontradas nas glândulas submandibular e sublingual de pacientes autopsiados com AIDS entre 1996 e 1999 no Serviço de Verificação de Óbitos da Capital (SVOC) - Faculdade de Medicina da Universidade de São Paulo (FMUSP). Dados referentes ao gênero, idade, contagem de células TCD4 e história clínica foram obtidos dos prontuários clínicos de 105 pacientes, destes 103 casos corresponderam às glândulas submandibulares e 92 casos às sublinguais. Todas as glândulas foram examinadas macroscopicamente e no estudo histopatológico utilizamos colorações de H&E, Gomori-Grocott; Ziehl-Neelsen e Mucicarmin. Análise imunoistoquímica foi realizada para detectar os agentes infecciosos (CMV, LMP-EBV, HSV-l, HSV-2, p24-HIV e BCG) e caracterizar as sialadenites (CD3, CD4, CD8, CD20 e CD68), já a HIS analizou a presença do VEB (EBER1/2). A média de idade dos pacientes e o nível de células TCD4 nos casos das glândulas submandibular e sublingual foi de 36,62 '+ ou ¿ ¿ 11,18 anos e 74,37 '+ ou ¿ ¿ 112,82 células/¿mu'L, e 35,93 '+ ou ¿ ¿ 0,2 anos e 78,75 '+ ou ¿ ¿ 118,98 células/¿mu'L, respectivamente. Foram considerados microscopicamente normais .51 (49,5%) casos da glândula submandibular e 54 (58,7%) casos da sublingual. As lesões infecciosas foram as mais freqüentes na glândula submandibular (n=35), seguida pela sialadenite crônica inespecífica em ambas as glândulas (n=25). Micobacteriose (11 e 07 casos nas glândulas submandibular e sublingual, respectivamente), citomegalovirose (14 e 02 casos nas glândulas submandibular e sublingual, respectivamente) e criptococose (03 e 04 casos nas glândulas submandibular e sublingual, respectivamente) foram freqüentemente detectadas. A análise imunoistoquímica mostrou que todos os casos em ambas as glândulas foram negativos para LMP-EBV, HSV-l e HSV-2, enquanto BCG mostrou intensa imunopositividade somente nos casos de micobacteriose com padrão macrofágico difuso (n=2). Dos 16 casos de citomegalovirose, apenas 06 casos (05 e 01 caso nas glândulas submandibular e sublingual, respectivamente) foram detectados pela imunoistoquímica. A proteína p24-HIV (02 e 01 caso nas glândulas submandibular e sublingual, respectivamente) e EBER1/2 (09 e 04 casos nas glândulas submandibular e sublingual, respectivamente), este último avaliado somente nos casos de sialadenités, foram expressas somente em escassos histiócitos e linfócitos, respectivamente, indicando que a participação destes vírus na etiopatogênese das sialadenites é pouco provável e precisa ser ainda melhor definida. As sialadenites crônicas inespecíficas foram principalmente compostas por linfócitos TCD8 (p=0,323); enquanto as sialadenites granulomatosas, as sialadenites macrofágicas difusas associada com micobacteriose e as sialadenites macrofágicas difusas inespecíficas apresentaram grande quantidade de macrófagos CD68+ (p=0,99) permeados por numerosos linfócitos TCD8, em ambas as glândulas. Além disso, um caso de linfoma não-Hodgkin difuso de grandes células B afetou ambas as glândulas. Estes resultados mostram que as glândulas submandibular e sublingual foram principalmente acometidas por lesões infecciosas disseminadas e sialadenite crônica inespecífica. Similarmente, as glândulas parótidas, num estudo previamente publicado por nossa equipe, foram acometidas por lesões infecciosas sistêmicas. Sendo assim, os clínicos devem estar atentos à ocorrência destas lesões em glândulas salivares maiores de pacientes com AIDS em fase avançada / Abstract: This study describes the histopathological, immunohistochemical (lHC), and in situ hybridization (ISH) features found in the submandibular (n=103) and sublingual (n=92) glands of autopsied patients who died with AIDS between 1996 and 1999 in the SVOC FMUSP (Medical School of São Paulo University). Sex, age, CD4 cell count, and clinical history were obtained from the clinical records of 105 patients, of them 103 cases corresponded to the submandibular glands and 92 cases to the sublingual glands. All glands were examined for macroscopical alterations and stained using H&E, Gomori-Grocott, Ziehl-Neelsen, and Mucicarmine. IHC analysis to detect infectious agents (CMV, LMP-EBV, HSV-l, HSV-2, HIV-p24, and BCG) and to characterize the sialadenitis (CD3, CD4, CD8, CD20, and CD68) was performed, while the ISH assessed the presence of EBV (EBERl/2). The mean age of the patients and CD4 cell count of the cases of the submandibular and sublingual glands were 36,62 '+ or ¿ ¿ 11,18 years and 74,37 '+ or ¿ ¿ 112,82 cells¿mu¿L POT .-1¿, and 35,93 '+ or ¿ ¿ 10,2 years and 78,75 '+ or ¿ ¿ 118,98 cells¿mu¿¿L POT .-1¿, respectively. There were no histological alterations in 51 (49,5%) and 54 (58,7%) cases of the submandibular and sublingual glands, respectively. The infection conditions were the most common in the submandibular gland (n=35), followed by chronic non-specific sialadenitis in both glands (n=25). Mycobacteriosis (11 and 07 cases of the submandibular and sublingual glands, respectively), cytomegalovirosis (14 and 02 cases of the submandibular and sublingual glands, respectively), and cryptococcosis (03 and 04 cases of the submandibular and sublingual glands, respectively) were found frequently. The IHC analysis did not show immunoreactivity for LMP-EBV, HSV-l, and HSV-2; while BCG displayed strong immunopositivity only in cases of chronic diffuse macrophagic infiltrate associated to mycobacteriosis (n=2). Six out of 16 cases of cytomegalovirosis (05 and 01 case of the submandibular and sublingual glands, respectively) were detected for IHC analysis only. The p24-HIV protein (02 and 01 case of the submandibular and sublingual glands, respectively) and EBERl/2 (09 and 04 cases of the submandibular and sublingual glands, respectively), this latter evaluated only in the sialadenitis cases, were expressed only in macrophages and lymphocytes, respectively, indicating that the participation of these viruses in the etiopathogenesis of the sialadenitis is still unc1ear. Chronic non-specific sialadenitis revealed CD8+ T-lymphocytes predominance (p=0,323), hile the granulomatous, diffuse macrophagic associated to mycobacteriosis, and chronic non-specific diffuse macrophagic sialadenitis showed great amount of CD68+ macrophages (p=0,99) surrounded by numerous CD8+ T-lymphocytes, in both glands. Moreover, one case of diffuse large B-celllymphoma affected both glands. These results indicate that both submandibular and sublingual glands were affected mainly by infectious diseases and chronic non-specific sialadenitis. Similarly, the parotid glands in a study previously published by our team, were mainly affected for systemic infectious diseases. Therefore, c1inicians should consider more carefully the occurrence of these lesions in major salivary glands of patients with advanced AIDS / Doutorado / Patologia / Doutor em Estomatopatologia

M.Sc. / It is generally believed that the development of a completely effective vaccine for the human immunodeficiency virus (HIV) will likely require neutralizing antibodies that react with the diverse strains of cell-free forms of this virus, as well as induce cellular responses in the form of cytotoxic T-lymphocytes (CTL), to eliminate cell-associated virus. Vaccines based on viral envelope proteins attempt to induce the former response, whilst DNA/vector based approaches aim to induce CTL. The membrane proximal external region (MPER) of HIV-1 gp41 is a target of two broadly neutralizing human monoclonal antibodies, 2F5 and 4E10, and is an important lead for vaccine design. It is conserved among several strains of HIV-1, except for subtype C where restricted mutations are found, especially in the epitopes of 2F5 and 4E10. Mono- and polyvalent (homologous and heterologous) synthetic peptide constructs of the epitopes recognised by 2F5 and 4E10, based on HIV-1 subtype C, have been designed and their immunogenicity compared in this study. The peptide constructs, designated MPER 1 / 2, a / b, induced humoral immune responses in mice and rabbits with the use of adjuvants. The homologous constructs (designated a) induced better humoral immune responses than the heterologous versions (designated b) in small animals. However the antibodies generated in rabbits were not potent enough to neutralize isolates of HIV- 1. The induction of neutralizing antibodies may be addressed by further conformational considerations, as conjugation to an octameric lysine core was insufficient. The peptide constructs did induce proliferative and inflammatory immune responses in a murine model. Additionally, the peptide constructs were highly antigenic as neutralizing anti-HIV antibodies present in naturally infected sera were able to recognise and bind to the MPER peptides as antigen in ELISAs. This suggests that the peptide constructs may be of value for characterizing anti-MPER antibody responses in infected individuals. The constructs were further able to mimic the true representation of these regions in vivo, as human monoclonal antibodies 2F5 and 4E10 were able to recognize and bind 3 of the 4 constructs. The human anti-MPER antibodies as well as the recombinant monoclonal antibodies had a higher binding affinity for the heterologous constructs. The MPER constructs exhibited many beneficial characteristics and may therefore hold application as a component in HIV-1 preventative and therapeutic vaccination following further modification.

The development of an effective HIV vaccine is hindered by several obstacles. One of the leading challenges is the antigenic variability of HIV-1 that is exhibited throughout all viral gene products but to greatest extent in the viral envelope proteins. This phenomenon is the result of continuous mutations in the HIV genome and is responsible for the immune escape of viral mutants. Many studies have suggested that a multivalent vaccine that elicits broadly cross-reactive antibodies is required to efficiently target antigenic variability. To this end, we have designed and analyzed a synthetic peptide construct that mimicked the major variability exhibited in the V3 loops of HIV-1 subtype C isolates. The peptide construct, described as a multiple epitope immunogen of the V3 loop with 8 branches and termed MEIV3b8, was shown to be non-toxic but highly immunogenic in experimental animals (mice and rabbits) and produced antibodies that were reactive to V3 loop peptides of various subtypes, variant envelope proteins and whole viral isolates [at antibody titers 1000 in enzyme-linked immunosorbent assays (ELISAs)]. Furthermore, functional antibodies were generated in rabbits that mediated neutralization of a neutralization-sensitive HIV-1 isolate and two distinct primary HIV-1 isolates in several different neutralization assays (at antibody titres 1213). Additionally, the MEIV3b8 induced both proliferative and inflammatory immune responses in a murine model.Finally, antibodies in the plasma of individuals (n = 148) infected with HIV-1 subtype C, subtype B and HIV-2 were found to bind to the MEIV3b8 as antigen in ELISAs. Through these findings, this study demonstrated that the variable MEIV3b8 effectively addressed antigenic variability and provided evidence that this peptide construct may hold application in HIV-1 preventative and therapeutic vaccination as well as HIV immunodiagnosis. / Dr. D. Meyer

Magister Curationis / A problem was identified at Aid for AIDS (AfA) whereby some doctors requested a change in treatment within less than a year after their patients started antiretroviral therapy. The requests were normally based on treatment failure. It appears that in most cases where the desired treatment outcome is not achieved is due to poor adherence to therapy. AfA is a HIV / AIDS disease management company offering access to antiretroviral therapy (ART), prevention of opportunistic infections, treatment and blood results monitoring, treatment support through adherence coordinators and expert clinical support and advice to healthcare providers. They monitor treatment adherence through claims history, CD4 and viral load (VL) results as well as telephonic contact with the client. Factors that could contribute to poor adherence are side - effects, barriers e.g. work environment, non - disclosure, lifestyle, lack of client commitment, limited contact between the client and treatment support counsellor, limited funds, stigmatisation and a lack of clear adherence guidelines to improve treatment outcome. Method: A comparative study was done to assess the impact of an intervention to improve patient adherence to ART. The researcher postulates that by the implementation of guidelines to counsellors, client adherence to therapy would increase. A comparative study was used to assess whether structured guidelines can improve client adherence to therapy. Results: The results have proven that guidelines used by treatment support counsellors does improve adherence to ART. Recommendations: It is recommended that treatment support counsellors, to improve their clients’ adherence to ART, should apply adherence guidelines. / South Africa

Magister Legum - LLM / The HIV/AIDS pandemic poses the greatest threat to Africa's efforts to achieve its full potential in the social, economical and political spheres. Cognizant of its devastating consequences, various mechanisms have been designed to address the issue of HIV/AIDS in Africa. This thesis addressed the question: 'Are the legislations and policies of Ethiopia, Botswana and Uganda providing for various modalities of HIV testing consistent with human rights as enshrined under African Human Rights system?' The author of this dissertation critically analyzed the African human rights instruments and the relevant domestic legislation and policies of the three countries. / South Africa