>Magister Scientiae - MSc / Background: Despite the effort of scientific research on HIV therapies and to reduce the rate of HIV infection, AIDS still remains one of the major causes of death in the world and mostly in Sub-Saharan Africa. To date, neither a cure, nor an HIV vaccine had been found and the disease can only be managed by using High Active Antiretroviral Therapy (HAART) if detected early. The need for an effective early diagnostic and non-toxic therapeutic treatment has brought about the necessity for the discovery of additional HIV diagnostic methods and treatment regimens to lower mortality rates. Antimicrobial Peptides (AMPs) are components of the first line of defence of prokaryotes and eukaryotes and have been proven to be promising therapeutic agents against HIV. Methods: With the utility of computational biology, this work proposes the use of profile search methods combined with structural modelling to identify putative AMPs with diagnostic and anti-HIV activity. Firstly, experimentally validated anti-HIV AMPs were retrieved from various publicly available AMP databases, APD, CAMP, Bactibase and UniprotKB and classified according to super-families. Hidden Markov Model (HMMER) and Gap Local Alignment of Motifs (GLAM2) profiles were built for each super-family of anti- HIV AMPs. Putative anti-HIV AMPs were identified after scanning genome sequence databases using the trained models, retrieved AMPs and ranked based on their E-values. The 3-D structures of the 10 peptides that were ranked highest were predicted using 1-TASSER. These peptides were docked against various HIV proteins using PatchDock and putative AMPs showing highest affinity and having the correct orientation to the HIV -1 proteins gp 120 and p24 were selected for future work so as to establish their function in HIV therapy and diagnosis. Results: The results of the in silica analysis showed that the constructed models using the HMMER algorithm had better performances compare to that of the models built by the GLAM2 algorithm. Furthermore, the former tool has better statistical and probability explanation compared to the latter tool. Thus only the HMMER scanning results were considered for further study. Out of 1059 species scanned by the HMMER models, 30 putative anti-HIV AMPs were identified from genome scans with the family specific profile models after elimination of duplicate peptides. Docking analysis of putative AMPs against HIV proteins showed that from the 10 best performing anti-HIV AMPs with the highest Escores, molecules 1,3, 8 and 10 firmly binds the gp120 binding pocket at the VIN2 domain and at the point of interaction between gp120 and T cells, with the 1st and 3rd highest scoring anti-HIV AMPs having the highest binding affinities. However, all 10 putative anti-HIV AMPs bind to the N-terminal domain of p24 with large surface interaction, rather than the C-terminal. Conclusion: The in silica approach has made it possible to construct computational models having high performances, and which enabled the identification of putative anti-HIV peptides from genome sequence scans. The in silica validation of these putative peptides through docking studies has shown that some of these AMPs may be involved in HIV/AIDS therapeutics and diagnostics. The molecular validation of these findings will be the way forward for the development of an early diagnostic tool and as a consequence initiate early treatment. This will prevent the invasion of the immune system by blocking the VIN2 domain and thus designing of a successful vaccine with broad neutralizing activity against this domain.

Thesis (MPH.) -- University of Limpopo, 2022 / Background: Termination of pregnancy among young women is a public health issue, particularly in South Africa where high prevalence of pregnany terminations has lately been reported. It is estimated that 260000 terminations of pregnancy take place in South Africa every year. Studies in South Africa have reported that risk factors associated with termination of pregnancy such as financial problems, being poorly educated, being young, unemployed, dependent on parents, widowed or single and other relationship problems were most common. Approximately 1200 pregnancies were terminated in the District Hospital of Limpopo Province between 2017 and 2018. There is also an increased rate of unintended pregnancy among HIV positive women which suggest that women with HIV may be more likely to terminate pregnancy but chooses not to terminate due to fear of being judged. Therefore, the primary objective of this study was to investigate the risk factors associated with termination of pregnancy at a District Hospital in Limpopo Province. Methodology: A cross-sectional descriptive retrospective review study in which convenience sampling of the records of women who terminated pregnancies was used in this study. A self-constructed data extraction tool was used to extract the data from patients records. The tool covered variables such as the age of the women, educational status, marital status, year and month of termination of pregnancy, gestational age, parity, and gravidity, HIV status and circumstances leading to termination of pregnancy. Data analysis was done using the STATA statistical software version 12 for Windows (STATA Corporation, College Station, Texas). The independent t-test was used for variables having two categories as it assesses whether the difference between means of two groups are statistically significant. This test was performed at the 95% confidence level. The p-value of less than 0.05 in the study results was used for statistical significant difference in means between the categories which were investigated. vi Results: The mean age was 27.9 years (standard error [Std. Err.] =0.37) and majority of women who terminated pregnancies were in the age group 20 – 24 years, single and had a secondary educational level. There was a statistical significance difference between age groups and the gestational age, parity and gravidity at p=0.004 and p<0.001 respectively. The proportion of women who were at gestational age of 1 to 8 weeks decreased with increasing maternal age from 22.4% in age ≤20 years to 13% in age group 30 – 34 years. There was again a statistical significance difference (p<0.001) in relation to number of pregnancies that have each resulted in the birth of an infant capable of survival (parity) and similarly to gravida. The prevalence of HIV amongst women who terminated pregnancy in the current study was found to be 21.3% and the risk of women who terminated pregnancies being HIV positive increased significantly with age as older women (age 20 years and above) were 6.5 times more likely to be HIV positive as compared to younger ones (p<001). Low educational level, gestational age of more than 13 weeks and parity of 1 – 2 were significantly associated with termination of pregnancy. The association of gravida of women who terminated pregnancies and HIV revealed that women who were in their second or third pregnancies (gravida) while HIV positive were 3.9 times more likely to terminate pregnancies as compared to those who were first pregnancy (p<0.001). Marital status was not significantly associated with termination of pregnancy. Conclusion: Termination of pregnancies among adolescents and youth is a major public health issue and the findings of this study highlight the need to address the structural socio-economic drivers of family planning which results in high number of termination of pregnancy amongst the youth. Structural interventions, such as increasing contraceptive use which may be useful for reducing the burden of unplanned pregnancies. These findings suggest the need for targeted interventions for women of child-bearing age to access reproductive health interventions to prevent unintended pregnancies and the associated risk of termination.

Thesis (MPH. (Epidemiology)) -- University of Limpopo, 2022 / Background: Tuberculosis (TB) is a fatal disease globally, if not managed well, with a million or more people dying by the disease annually in low and middle-income countries (LMIC). Around two billion people are thought to be asymptomatically (latently) infected with Mycobacterium tuberculosis, putting them at risk of acquiring active tuberculosis. Tests that identify immunoreactivity to mycobacterial antigens rather than live bacteria, as well as mathematical modelling, are used to estimate the prevalence of latent tuberculosis infection. According to reports, tuberculosis (TB) was the cause of 1.3 million fatalities among HIV-negative people in 2016, surpassing the global number of HIV/acquired immune deficiency syndrome (AIDS) deaths. In addition, TB was a factor in 374,000 HIV-related deaths. Despite the effectiveness of chemotherapy over the last seven decades, tuberculosis remains the world's leading infectious killer. In 2016, 10.4 million new cases were reported, a number that has remained constant since the dawn of the twenty-first century, confounding public health specialists tasked with designing and implementing measures to lessen the global burden of tuberculosis disease. As a result, the current study aims to look into the epidemiology of tuberculosis in Lesotho in order to help policymakers make decisions on TB control in the country. Methodology:. In the current investigation, a cross-sectional, retrospective descriptive study design was used, as well as a probability sampling strategy. The National TB-Database from the Ministry of Health in Lesotho was used as the source of data for this quantitative investigation, which was analyzed using STATA statistical software version 12 for Windows (STATA Corporation, College Station, Texas). A Chi-Squared test was used to compare categorical variables, while a t-test was used to examine continuous variables. A statistically significant P-value of 0.05 was used. Results: A total of 18 836 TB patient records were recovered, with 45 percent of the TB patients being females. The average age of the TB patients was 35.9 years, with a standard deviation of 12.7%, and the ages ranged from one year to 84 years. There vi was a statistically significant difference between the age groups (p value 0.001), with 33.1 percent of TB patents being in the age group 25–34 years, followed by 29 percent, 15.4 percent, 11.2 percent, and 5.5 percent in the age groups 35–44 years, 45–55 years, 15–24 years, and 55–64 years 65 years.. There has been a fluctuating treatment outcome of TB from 63.5% for cured patients in 2012 to 57.2% in 2013 and this rose to 60.4% in 2014 then eventually reached 76.7% in 2019. The TB treatment success rate in Lesotho also showed a similar trend as the cure rate. The overall TB death rates in the current study was found to be increasing on an annual basis from 7.4% in 2012 to 9.2% in 2018 then dropped to 8.5% in 2019. The TB patients who have not been evaluated for treatment outcomes have been decreasing annually from 4.4% in 2012 to 0.8% in 2019. The proportion of TB patients with known HIV status increased from 22.3% in 2015 to 90.5% in 2019 and similarly to the proportion of TB patients with HIV status positive increased from 15.1% in 2015 to 60.4% in 2019. The proportion of TB patients with HIV status positive increased with increasing age group all age groups. Conclusion: TB is still a concern in Lesotho, where treatment target goals have not yet been fulfilled, the findings of this study underline the importance of addressing the underlying socio-economic causes of TB. The most important goal in TB control is to detect 70% and cure at least 85% of sputum smear positive cases. If these goals are met, the prevalence, incidence, transmission, and medication resistance to tuberculosis (TB) could all decrease. Despite the National Tuberculosis Control Programme's attempts to enhance TB patients' access to treatment and adherence to therapy, the percentage of patients who have good treatment outcomes remains low. Despite having an 84 percent detection rate and using the DOTS technique, the available data did not identify the types of tuberculosis, therefore we were unable to forecast multidrug-resistant tuberculosis (MDR-TB).

Human Immunodeficiency Virus (HIV) is a virus that is transmitted through certain bodily fluids and compromises the immune system of its host. Despite the emergence of antiretroviral therapy (ART) converting human immunodeficiency virus type 1 (HIV-1) infection from a fatal disease to a chronic condition, there is still no cure. ART frequently reestablishes peripheral CD4+ T cell counts, but persistent immune dysfunction and inflammation strongly correlate with increased risks of attaining non-AIDS morbidity and mortality. Elimination of this reservoir may occur by the proposed mechanism of combining latency-reversing agents (LRAs) with immune effectors, such as CD8+ T cells (Meås et al., 2020). Here, our study investigates Toll-like receptor 7/8 (TLR 7/8) superagonists that may act as potent, effective latency reversal agents (LRAs). Whether this will prove to be the case needs to be further studied, and potential adverse toxicities must be identified. Whether comparable results will be observed in peripheral blood mononuclear cells (PBMCs) infected with HIV-1 as in our study using PBMCs infected with simian immunodeficiency virus (SIV) remains to be tested. Our results provide further hope for a potential cure for HIV-infected individuals.

Bakgrund: Humant immunbristvirus (HIV) fortsätter att vara en betydande global hälsoutmaning och påverkar miljontals individer världen över. Inom hälso- och sjukvården möter individer som lever med HIV ofta stora utmaningar, inklusive stigmatisering och diskriminering. Dessa utmaningar förvärras ytterligare av samhälleliga fördomar och en allmän brist på kunskap om HIV. Sjuksköterskor spelar en avgörande roll i att ge etisk och rättvis vård. Det är viktigt att inte bara fokusera på fysisk hälsa utan också belysa den mentala hälsan, eftersom den är lika viktig. Syfte: Att beskriva upplevelser av utmaningar som individer med HIV-infektion har i mötet med hälso-och sjukvården. Metod: En litteraturöversikt med kvalitativ ansats. Artikelsökningen utfördes i databaserna; PubMed, Cinahl och Web of Science. En djupgående granskning ledde till identifieringen av 12 vetenskapliga artiklar med kvalitativ design. Resultat: I litteraturöversikten framkom hinder för individer som lever med HIV i mötet med hälso- och sjukvården såsom nedsättande bemötande, utmaningar samt avsaknad av stödjande vårdmiljö. Slutsats: Stigmatisering, diskriminering, bristfällig kommunikation samt behov av information är utmaningar i mötet med vården för individer som lever med HIV. Förbättring kräver strukturella förändringar, utbildning och policyutveckling för att möjliggöra en mer inkluderande vårdmiljö för individerna. / Background: Human Immunodeficiency Virus (HIV) continues to be a significant global health challenge, affecting millions of individuals worldwide. Within healthcare settings, individuals living with HIV often face substantial challenges, including stigmatization and discrimination. These problems are further exacerbated by societal biases and a general lack of understanding about HIV. Nurses play a crucial role in providing ethical and fair care. It is important to focus not only on physical health but also to highlight mental health, as it is equally important. Purpose: To describe experiences of challenges that individuals with HIV-infection have in encounters with the health care system. Method: A literature review with a qualitative approach. The article search was conducted in the databases; PubMed, Cinahl, and Web of Science. An in-depth review led to the identification of 12 scientific articles with a qualitative-design. Results: In the literature review, obstacles for individuals living with HIV in encounters with healthcare services emerged, such as stigmatizing treatment, challenges, and a lack of supportive care environment. Conclusion: Stigmatization, discrimination, poor communication, and the need for information are challenges in healthcare encounters for these individuals. Improvement requires structural changes, education, and policy development to enable a more inclusive healthcare environment for these individuals.

The mechanism of human immunodeficiency virus 1 (HIV-1) nuclear entry, required for productive infection, is not fully understood. Here, we report that in HeLa cells and activated CD4⁺ T cells infected with HIV-1 pseudotyped with VSV-G and native Env protein, respectively, Rab7⁺ late endosomes containing endocytosed HIV-1 promote the formation of nuclear envelope invaginations (NEIs) by a molecular mechanism involving the VOR complex, composed of the outer nuclear membrane protein VAP-A, hyperphosphorylated ORP3 and Rab7. Silencing VAP-A or ORP3 and drug-mediated impairment of Rab7 binding to ORP3-VAP-A inhibited the nuclear transfer of the HIV-1 components and productive infection. In HIV-1-resistant quiescent CD4⁺ T cells, ORP3 was not hyperphosphorylated and neither VOR complex nor NEIs were formed. This new cellular pathway and its molecular players are potential therapeutic targets, perhaps shared by other viruses that require nuclear entry to complete their life cycle.

In den vergangenen Jahren sind Lipide, Membranen und deren Organisationsformen mehr und mehr in den Fokus der biologischen Forschung gerückt. Es wurde vorgeschlagen, dass in zellulären Membranen selbstassemblierende, submikroskopische Aggregate aus Sphingolipiden, Cholesterol und bestimmten Proteinen existieren und man vermutet, dass insbesondere Viren diese “Lipid Rafts” für ihren Zusammenbau nutzen und auf diese Art ihre Proliferationseffizienz erhöhen. Gleichwohl sind die genaue biologische Funktion und auch die molekulare Basis der Assoziation bestimmter Protein mit Lipid Rafts auch weiterhin unbekannt. In der vorliegenden Arbeit wurde Fluoreszenz-Lebenszeit-Mikroskopie genutzt, um die Lipid-Raft-Anreicherung des HIV-1 Glycoproteins gp41 zu untersuchen. Förster-Resonanz-Energietransfer zwischen fluoreszenzmarkierten viralen und Raft-Marker-Proteinen wurde gemessen, um deren gemeinsame, lokale Aufkonzentrierung in Lipid Rafts nachzuweisen. Durch Verwendung verschiedener Deletions- und Mutationsvarianten des Proteins konnte nicht nur seine Lipid-Raft-Präferenz demonstriert, sondern auch das Cholesterol-Bindemotiv (CRAC) als entscheidender Faktor der lateralen Sortierung identifiziert werden. Wir haben in diesem Kontext auch eine systematische Zell-zu-Zell-Variabilität in unseren Daten bemerkt, die einen zugrundeliegenden zellbiologischen Mechanismus der Membranorganisation nahelegt. Mithilfe von Fluoreszenz-Polarisations-Mikroskopie konnte zudem eine klare CRAC-Abhängigkeit der gp41-Oligomerisierung aufgezeigt werden. Die von uns gewonnenen Daten erlauben einen tieferen Einblick in die molekulare Basis und die biologischen Folgen der cholesterol-abhängigen lateralen Proteinorganisation für Virusassemblierungsprozesse an biologischen Membranen. / In recent years, there has been a considerable interest in the molecular organization of biological membranes. It has been hypothesized that self-assembling, freely diffusing, submicroscopic domains consisting of sphingolipids, cholesterol and certain proteins exist and the prevailing view is that those lipid rafts serve as platforms for specific molecular interactions by the preferential exclusion and inclusion of proteins. It was presumed, that in particular viruses make use of plasma membrane lipid rafts to augment the infection process and spread efficiently. However, the exact biological function and physical basis of protein partitioning into microdomains remains an outstanding question in virus biology. In the present study, fluorescence lifetime imaging microscopy was used to study lipid raft partitioning of the HIV-1 glycoprotein gp41 by detecting Foerster Resonance Energy Transfer between fluorescently labeled viral and raft marker proteins in living cells. Plasma membrane microdomain association of gp41 was demonstrated and by introducing systematic mutations and truncations in different gp41 motifs, the cholesterol recognition amino acid consensus (CRAC) was identified as the crucial determinant of the lateral sorting. Interestingly, we observed a systematic cell-to-cell variability in our raft related data that suggests underlying cell-biological mechanisms of membrane organization. Moreover, fluorescence polarization microscopy revealed a distinct CRAC requirement for gp41 oligomerization whereas other properties, such as intracellular distribution and expression efficiency were clearly demonstrated to be CRAC independent. Our data provide further insight into the molecular basis and biological implications of the cholesterol dependent lateral protein sorting for virus assembly processes at cellular plasma membranes.

Registered nurses are exposed to the risk of Human Immunodeficiency Virus (HIV) infection and contracting the Acquired Immunodeficiency Syndrome (AIDS). Qualitative phenomenological research was conducted to explore registered nurses' experiences of working in such a high-risk environment and how their experiences influence the therapeutic relationship. A sample of registered nurses who care for HIV-infected persons or persons who suffer from AIDS was purposefully selected. Focus group interviews were conducted. Qualitative data analysis was performed. Frankl's theory of meaning of life served as a theoretical foundation for interpreting the research findings. The research results revealed that registered nurses experience existential frustration due to the intentional and unintentional risks that they are exposed to. This negatively impacts upon their ability to maintain a healthy therapeutic relationship with patients. However, evidence was obtained indicating that some factors support their quest for finding meaning in life in the workplace. / Health Studies / M.A. (Health Studies)

HIV/AIDS is a global challenge and its impact is evident. This places a burden on hospitals and health professionals. To ease this burden there are home-based care programmes which, through home-based caregivers, provide patients living with HIV/AIDS with physical and palliative care. However, these HBCGs face various challenges such as poverty, discrimination and stigma when caring for PALHIV, and their own emotional strain, which becomes burdensome without sufficient support from the home-based care centre. The qualitative study undertaken investigates the challenges and coping mechanisms of the HBCGs. Thirteen research participants were drawn from a centre in Mamelodi. Data was gathered through interviews and observations, categorised into themes and analysed. Major findings revealed that social challenges faced by HBCGs include poverty, stigmatisation and discrimination. Emotions experienced by HBCGs include guilt, anger, hopelessness, but they have spiritual reliance through prayer as one of their coping mechanisms. / Sociology / M.A. (Social Behaviour Studies in HIV/AIDS)

The purpose of the study was to determine the factors that predict women’s risky sexual behaviour and HIV and AIDS knowledge. A quasi-experimental, pre-test-post-test research design, with a non-equivalent comparison group was conducted to determine if there was an association between young women’s sexual behaviour and HIV/AIDS knowledge on aspects of HIV/AIDS transmission and prevention and behaviour change. The study used both quantitative and qualitative approach. Data collection was done using semi-structured interview schedule and focus group discussion guide. The respondents who participated in the study were women between the ages of 15-25 years. Two groups of respondents participated in the study. Women in the quasi-experimental site (N=200) who received the intervention and women in the control site (N=200) who did not receive any intervention. Quantitative data were analysed with the help of a statistician and the Epi Info statistical package was used. Qualitative data obtained from the focus group discussion were analysed using Tesch’s method of analysis. The major inferences drawn from this study are that young women lack knowledge relating to HIV/AIDS and that some young women were engaged in risky sexual behaviours such as having multiple sexual partners and having unprotected sex. The study indicates that peer education strategy has the potential to make an impact on these young women. It has also shown that peer education can play an important role in increasing knowledge and reducing risky sexual behaviour. / Health Studies / D. Litt. et Phil. (Health Studies)