The current, grim world-TB scenario, with TB being the single largest infectious disease killer, warrants a more effective approach to tackle the deadly pathogen, Mycobacterium tuberculosis. The deadly synergy of this pathogen with HIV and the emergence of drugresistant strains of the organism present a challenge for disease treatment (Russell et al., 2010). Thus, there is a pressing need for newer drugs with faster killing-kinetics which can claim both the actively-multiplying and latent forms of this pathogen causing the oldest known disease to man. This thesis entitled “Mechanistic and Regulatory Aspects of the Mycobacterium tuberculosis Dephosphocoenzyme A Kinase” describes one such potential drug target, which holds promise in future drug development, in detail. The development of efficacious antimycobacterials now requires previously unexplored pathways of the pathogen and cofactor biosynthesis pathways present a good starting point. Therefore, the mycobacterial Coenzyme A (CoA) biosynthesis was chosen for investigation, with the last enzyme of this pathway, dephosphocoenzyme A kinase (CoaE) which was shown to be essential for M. tuberculosis survival, as the focus of the present study (Sassetti et al., 2003). This thesis presents a detailed biochemical and biophysical characterization of the enzymatic mechanism of mycobacterial CoaE, highlighting several hitherto-unknown, unique features of the enzyme. Mutagenic studies described herein have helped identify the critical residues of the kinase involved in substrate recognition, binding and catalysis. Further, a role has been assigned to the UPF0157 domain of unknown function found in the mycobacterial CoaE as well as in several organisms throughout the living kingdom. Detailed insights into the regulatory characteristics of this enzyme from this work further our current understanding of the regulation of the universal CoA biosynthetic pathway and call for the attribution of a greater role to the last enzyme in pathway regulation than has been previously accredited. The thesis begins with a survey of the current literature available on tuberculosis and where we stand today in our fight against this dreaded pathogen. Chapter 1 details the characteristic features of the causative organism M. tuberculosis, briefly describing its unique genome and the cellular envelope which the organism puts forward as a tough shield to its biology. This is followed by a brief description of the infection cycle in the host, the pathogen-host interplay in the lung macrophages, the deadly alliance of the disease with HIV and our current drug arsenal against tuberculosis. Further, emphasizing on the need for newer, faster-acting anti-mycobacterials, Chapter 1 presents the rationale for choosing the mycobacterial coenzyme A biosynthetic pathway as an effective target for newer drugs. A detailed description of our current understanding of the five steps constituting the pathway follows, including a comparison of all the five enzymatic steps between the human host and the pathogen. This chapter also sets the objectives of the thesis, describing the choice of the last enzyme of the mycobacterial CoA biosynthesis, dephosphocoenzyme A kinase, for detailed investigation. As described in Chapter 1, the mycobacterial CoaE is vastly different from its human counterpart in terms of its domain organization and regulatory features and is therefore a good target for future drug development. In this thesis, Rv1631, the probable mycobacterial dephosphocoenzyme A kinase annotated in the Tuberculist database (http://genolist.pasteur.fr/TubercuList), has been unequivocally established as the last enzyme of the tubercular CoA biosynthesis through several independent assays detailed in Chapter 2. The gene was cloned from the mycobacterial genomic DNA, expressed in E. coli and the corresponding recombinant protein purified via a single-step affinity purification method. The mechanistic details of the enzymatic reaction phosphorylating dephosphocoenzyme A (DCoA) to the ubiquitous cofactor, Coenzyme A, have been described in this chapter which presents a detailed biochemical and biophysical characterization of the mycobacterial enzyme, highlighting its novel features as well as unknown properties of this class of enzymes belonging to the Nucleoside Tri-Phosphate (NTP) hydrolase superfamily. The kinetics of the reaction have been biochemically elucidated via four separate assays and the energetics of the enzyme-substrate and enzymeproduct interactions have been detailed by isothermal titration Calorimetry (ITC). Further details on the phosphate donor specificity of the kinase and the order of substrate binding to the enzyme provide a complete picture of the enzymatic mechanism of the mycobacterial dephosphocoenzyme A kinase. Following on the leads generated in Chapter 2 on the unexpected strong binding of CTP to the enzyme but its inability to serve as a phosphate donor to CoaE, enzymatic assays described in Chapter 3 helped in the identification of a hitherto unknown, novel regulator of the last enzyme of CoA biosynthesis, the cellular metabolite CTP. This chapter outlines the remarkable interplay between the regulator, CTP and the leading substrate, dephosphocoenzyme A, possibly employed by the cell to modulate enzymatic activity. The interesting twist to the regulatory mechanisms of CoaE added by the involvement of various oligomeric forms of the enzyme and the influence of the regulator and the leading substrate on the dynamic equilibrium between the trimer and the monomer is further detailed. This reequilibration of the oligomeric states of the enzyme effected by the ligands and its role in activity regulation is further substantiated by the fact that CoaE oligomerization is not cysteine-mediated. Further, the effects of the cellular metabolites on the enzyme have been corroborated by limited proteolysis, CD and fluorescence studies which helped elucidate the conformational changes effected by CTP and DCoA on the enzyme. Thus, the third chapter discusses the novel regulatory features employed by the pathogen to regulate metabolite flow through a critical biosynthetic pathway. Results presented in this chapter highlight the fact that greater importance should be attributed to the last step of CoA biosynthesis in the overall pathway regulation mechanisms than has been previously accorded. The availability of only three crystal structures for a critical enzyme like dephosphocoenzyme A kinase (those from Escherichia. coli, Haemophilus influenzae and Thermus thermophilus) is indeed surprising (Obmolova et al., 2001; O’Toole et al., 2003; Seto et al., 2005). In search of a structural basis for the dynamic regulatory interplay between the leading substrate, DCoA and the regulator, CTP, a computational approach was adopted. Interestingly, the mycobacterial enzyme, unlike its other counterparts from the prokaryotic kingdom, is a bi-domain protein of which the C-terminal domain has no assigned function. Thus both the N- and C-terminal domains were independently modeled, stitched together and energy minimized to generate a three-dimensional picture of the mycobacterial dephosphocoenzyme A kinase, as described in Chapter 4. Ligand-docking analyses and a comprehensive analysis of the interactions of each ligand with the enzyme, in terms of the residues interacted with and the strength of the interaction, presented in this chapter provide interesting insights into the CTP-mediated regulation of CoaE providing a final confirmation of the enzymatic inhibition effected by CTP. These homology modeling and ligand-docking studies reveal that CTP binds the enzyme at the site overlapping with that occupied by the leading substrate, thereby potentially obscuring the active site and preventing catalysis. Further, very close structural homology of the modeled full-length enzyme to uridylmonophosphate/cytidylmonophosphate kinases, deoxycytidine kinases and cytidylate kinases from several different sources, with RMSD values in the range of 2.8-3 Å further lend credence to the strong binding of CTP detailed in Chapter 2 and the regulation of enzymatic activity described in Chapter 3. Computational analyses on the mycobacterial CoaE detailed in this chapter further threw up some interesting features of dephosphocoenzyme A kinases, such as the universal DXD motif in these enzymes, which appears to play a crucial role in catalysis as has been assessed in the next chapter. It is interesting to note that the P-loop-containing nucleoside monophosphate kinases (NMPK), with which the dephosphocoenzyme A kinases share significant homology, have three catalytic domains, the nucleotide-binding domain, the acceptor substrate-binding domain and the lid domain. Computational analyses detailed in Chapter 4 including the structural and sequential homology studies, helped in the delineation of the three domains in the mycobacterial enzyme as well as highly conserved residues potentially involved in crucial roles for substrate binding and catalysis. Therefore important residues from all three domains of the mycobacterial CoaE were chosen for mutagenesis to study their contributions to catalysis. Conservative and non-conservative replacements of these residues detailed in Chapter 5 helped in the identification of crucial residues involved in phosphate donor, ATP binding (Lys14 and Arg140); leading substrate, DCoA binding (Leu113); stabilization of the phosphoryl transfer reaction (Asp32 and Arg140) and catalysis (Asp32). Thus, the results reported here present a first attempt to identify the previously unknown functional roles of highly conserved residues in dephosphocoenzyme A kinases. Chapter 5 also delineates the dependence of this kinase on the divalent cation, magnesium, for catalysis, describing a comparison of the kinetic activity by the wild type and the mutants, in the presence and absence of Mg2+. Therefore, this chapter presents a thorough molecular dissection of the roles played by crucial amino acids of the protein and the results herein can serve as a good starting point for targeted drug development approaches. As described above, another unusual characteristic of the mycobacterial CoaE is the fact that it carries a domain of unknown function, UPF0157, C-terminal to the N-terminal dephosphocoenzyme A kinase domain. The function of this unique C-terminal domain carried by the mycobacterial CoaE has been explored in Chapter 6. The failure of the Nterminal domain (NTD) to be expressed and purified in the soluble fraction in the absence of a domain at its C-terminus (either the mycobacterial CoaE CTD or GST from the pETGEXCT vector) pointed out a possible chaperonic activity for the CTD. A universal chaperonic activity by this domain in the cell was ruled out by carrying out established chaperone assays with insulin, abrin and -crystallin. In order to delineate the CTD sequence involved in the NTD-specific chaperoning activity, deletion mutagenesis helped establish the residues 35-50 (KIACGHKALRVDHIG) of the CTD in the N-terminal domain-specific assistance in folding. Chapter 6 further details the several other potential roles of the mycobacterial CTD probed, including the 4’-phosphopantethienyl transfer, SAM-dependent methyltransferase activity, activation of the NTD via phospholipids among others. Thus the results presented in this chapter are a first attempt at investigating the role of this domain found in several unique architectures in several species across the living kingdom. Chapter 7 is an attempt to stitch together and summarize the results presented in all the preceding chapters, giving an overview of our present understanding of the mycobacterial CoaE and its novel features.

Migrant workers in the Deciduous Fruit Industry are part of the marginalised communities in South Africa. They are often voiceless in the communities they find themselves. They are historically displaced, often prone to xenophobia and very vulnerable in terms of HIV. Not only do they have a high infection rate but they also struggle in isolation to carry the burden of HIV and AIDS affection or infection. They will face double jeopardy when a partner becomes ill, in the homeland and they have to continue with employment. The main aim of this research was to reach a holistic understanding through interdisciplinary investigation. The important question that I aim to answer is; “What is the experience of God in the lives of persons affected or infected by HIV and AIDS.” I have looked at Postfoundationalism and the Seven Movements as proposed by Muller to present the research undertaken among migrant workers with HIV and AIDS. The Practical Theology, which I explore, develops out of a very specific praxis, HIV and AIDS. I have also made used of Transversal Rationality as a practical way of doing interdisciplinary work with the stories of my co-researchers affected with HIV AIDS as a case study. I understand that Christian belief has its own integrity, which is exclusive, but if valid, is vital to be able to incorporate the different dimensions of our modern practise to give it the maximum level of meaning and significance. I hope to demonstrate this possibility through my thesis. / Thesis (PhD)--University of Pretoria, 2010. / Practical Theology / unrestricted

Thesis (MPH.) -- University of Limpopo, 2017. / The present study presents data about the prevalence and determinants of sputum smear non-conversion in smear positive tuberculosis patients. Despite the intervention by the Sekhukhune District Department of Health through continual training and workshops of professional nurses in respect of the NTCP, there are still more challenges observed in terms of TB management. Aim: To investigate the prevalence and determinants of sputum smear non-conversion in smear positive PTB patients after intensive phase of treatment. Method: Quantitative, descriptive retrospective study of TB records was conducted. Data collection was done by extracting data from ETR.net and exporting it to excel. Data cleaning was done before analysis. Data analysis was done using the computer Statistical Package Software for Social research (SPSS) volume 23.1. Findings: 834 TB patients’ records were extracted from the ETR.net database. 34% of records were available at 2 – months; 57% of the patients were males; also, 81% of the patients were diagnosed/treated at PHC facilities; 52% of the patients were HIV positive; 69% percent of the patients who were smear positive grading p+++ failed to convert after two months. In the univariate logistic regression patients with age 20 – 29 were observed to be 4.9 times likely (O.R. = 4.97) to be sputum positive (P = 0.142).Sputum grade 3(p+++) at the time of diagnosis was found to be significantly associated (P = 0.031) with sputum non – conversion after intensive phase of treatment. Conclusion: Two month sputum smear non-conversion is associated with pre-treatment sputum smear grading.

Tuberculosis (TB), a transmissible disease caused by Mycobacterium tuberculosis (Mtb), is a global health threat. To design an effective vaccine, we need to better understand how different elements of our immune system collaborate to fight against Mtb. CD4 T cells are crucial in protective immunity to Mtb because they produce cytokines including interferon-γ. In contrast, CD8 T cells are thought to play a modest role. Whether CD4 T cells act as “helper” cells to promote optimal CD8 T cell responses during TB is unknown. We argue CD8 T cells’ role are likely underestimated because CD8 T cell functions are compromised without CD4 T cells. Here, using two independent models, I show that CD4 T cell help promotes CD8 T cell effector functions and prevents CD8 T cell exhaustion. I demonstrate CD4 and CD8 T cells synergistically enhance the survival of infected mice. Purified helped, but not helpless, CD8 T cells effectively restrict intracellular Mtb growth. Thus, CD4 T cell help is indispensable for generating protective CD8 T cell responses. In addition, I investigate the mechanisms of CD4 T cell help. Signals from CD4 T cells, and signals relayed by antigen presenting cells collectively shape CD8 T cell responses. We infer that vaccines aimed for eliciting both CD4 and CD8 T cells, in which CD8 T cells are properly helped by CD4 T cells, are more likely to be successful.

This work deals with the issue of supply restoring for heating plant Teplárny Brno, a. s. Špitálka division, after a system-wide failure of the “blackout”. The thesis solves the “black start” of the Špitálka Teplárny Brno operation by delivering a voltage from the hydroelectric power Vír I, proposes a methodology for commissioning of individual devices, up to phasing the generators with subsequent possibility of creating an “island mode” in the area of Brno city, which the work already does not deal with. The work also deals with the preparation of the operational test, while starts of the selected devices are tested. The schedule of commissioning of each device is supported by calculations of the parameters for the entire system with regard to voltage drops while connecting the high load. The result of this work is the proposal of methodological process of supply restoring for power plant Teplárny Brno – Špitálka division, by submission a voltage from the hydroelectric power Vír I, which can be used both in any system-wide failure of the “blackout”, or during the operational test.

Thesis (MPH.) -- University of Limpopo, 2019 / Background: Tuberculosis still continues to be a global public health problem and leads to many deaths. In an effective TB control strategy, TB patients are allocated to community care workers who provide care to these patients in their homes. It is important to understand the experiences of community care workers in order to strengthen TB control in the country. Objective(s): The purpose of this study was to explore lived experiences of community care workers of supporting patients taking Tuberculosis treatment. Methods: The design of the study was phenomenological, exploratory, descriptive, and contextual. The study site was Hlogotlou area in Limpopo Province. The target population was all community care workers supporting patients on Tuberculosis treatment. Purposive sampling was used with a sample of 13 participants, which was determined by the saturation of data. Semi-structured interviews were conducted using an interview guide and all sessions were audio recorded. The data were analysed using Interpretative Phenomenological Analysis. Results: The results highlighted certain challenges met by community care workers. Patients thought that community care workers are there to kill them with treatment, they had mood swings during treatment and this caused them to use vulgar words and become aggressive to their community care workers. Most community care workers did not have enough information about Tuberculosis. This made it difficult for them to support patients on tuberculosis treatment. Conclusions: A good relationship with patients enhances treatment compliance. The researcher recommends that intensive training about tuberculosis should be provided to community care workers.

Tuberculosis, an infectious disease caused by Mycobacterium tuberculosis, has been a global health problem for years. The emergence of drug resistance in this organism generates the necessity of exploring novel targets and developing new drugs. Topoisomerases are enzymes found in all kingdoms of life responsible for overcoming the topological barriers encountered during essential cellular processes. The genomes of mycobacteria encode only one type IA topoisomerase (MtopI), which has been validated as a novel TB drug target. The goal of this study is to obtain new information on the mechanism and resistance of endogenous and synthetic inhibitors of MtopI. Rv1495 is a M. tuberculosis toxin that belongs to the MazEF family (MazE is the antitoxin and MazF is the toxin), with endoribonuclease activity. Rv1495 (MazF homolog in M. tuberculosis) toxin has been shown to interact directly with the C-terminal domain of MtopI for mutual inhibition. In this study the interaction of Rv1495 with the positively charged C-terminal tail in Mtop I is reported. This new information is useful for rational design and discovery of antibiotics for mycobacteria. Ethacridine, an FDA approved drug has shown activity against MtopI. In this project we studied the mechanisms of resistance associated with this drug as well the use of Ethacridine in combination with Moxifloxacin, to potentiate the bactericidal effect of this current second line drug for TB treatment. Results from sequencing of the genomic DNA isolated from the resistant mutants suggested the involvement of the Holliday-junction Ruv resolvase. Further studies showed that co-treatment with Ethacridine can enhance the moxifloxacin-mediated killing of M. smegmatis. FP-11g, a novel fluoroquinophenoxazine inhibitor of bacterial topoisomerase I, has shown promising activity against M, tuberculosis. We explored the bactericidal activity and resistance mechanisms associated to FP-11g using M. smegmatis as model organism. Additionally, the inhibitory effect of FP-11g was also evaluated on M. abscessus. FP-11g at concentration 4X MIC showed complete bactericidal activity against M. smegmatis after 24 hours. Inhibitory activity against M. abscessus was also observed. Results from sequencing of the genomic DNA isolated from the M. smegmatis resistant mutants revealed mutations in genes associated with general drug resistance.

[pt] O presente trabalho apresenta os resultados obtidos com simulações computacionais de interferência entre sistemas LTE e TV Digital na faixa de 700 MHz e de medições de campo. Os resultados da campanha de medições foram usados para validar uma ferramenta de simulação que permite a avaliação de interferência entre os sistemas SBTVD e LTE em diferentes cenários. A ferramenta foi, então, utilizada para analisar a probabilidade de interferência que depende da distância entre os sistemas, da potência de transmissão, do número de interferentes e outros parâmetros que permitem definir as condições para a coexistência harmoniosa dos sistemas. A partir da validação da ferramenta, cenários de interferência com múltiplos receptores LTE e DTV foram simulados, com o propósito de reproduzir problemas mais realistas. Na última etapa do trabalho, como estudo de caso, foi realizada uma avaliação de interferência do sistema LTE nos receptores de TV Digital em cenário representativo da região da zona sul do município do Rio de Janeiro. / [en] This work presents the results of computer simulations of the interference between LTE system and ISDB-TB Digital TV system in the 700 MHz band and field measurements. The results of the measurement campaign were used to validate a simulation tool that runs different scenarios between the SBTVD systems and LTE. The software tool was used to analyze the probability of interference and its dependence on the distance between systems, transmission power, number of interferers and other parameters, aiming to estimate the conditions for the coexistence of the systems. Once the software tool was validated, different interference scenarios with multiple LTE and DTV receivers were analyzed, in order to reproduce more realistic problems. In the last part of this study, an interference evaluation from LTE system on Digital TV receivers was performed in the scenario which represents a region within the South Zone of the city of Rio de Janeiro.

Background: There has been tremendous progress in the use of genomics1 in biomedical research and medical care since the launch of the Human Genome Project in 1990. However, it has also introduced new ethical challenges regarding the feedback of findings generated in genomic sequencing. While some would argue in support of the return of individual findings generated from genomics research, participants' preferences regarding which findings should be fed back differs. Most literature discusses feedback of findings in high income countries and very few address this issue in lower and middle-income countries (LMICs). As a result, it remains unclear whether and how individual findings from genomic studies in Africa should be fed back, who should provide these results and when. Methods: In order to contribute to addressing this gap, an empirical study was conducted to explore expectations and preferences for feedback of individual genetic findings in an HIV-TB genomics research project in Botswana. A qualitative study methodology involving deliberative focus group discussions (dFGDs) and in-depth interviews (IDIs) was used. Participants for this study were adolescents involved in an HIV-TB genomics study being conducted at the Botswana-Baylor Children's Clinical Centre of Excellence (BBCCCE). Parents and caregivers of children enrolled in that same genomic study were also enrolled in this study. A total of 93 participants (44 adolescents and 49 parents and caregivers) were enrolled in 12 dFGDs (6 groups of adolescents and 6 groups of parents and caregivers). Each group of participants met twice within a week, resulting in a total of 24 dFGD meetings. Participants of the dFGDs and in-depth interviews were selected purposively. Additionally, indepth interviews were conducted with 12 dFGD participants (6 adolescents and 6 parents or caregivers). The dFGDs and IDIs were conducted in Setswana, audio-recorded, transcribed and translated into English. Data were imported into NVivo 12 and analysed using the framework approach for qualitative data analysis. Results: The study findings revealed that participants' desire to receive individual genetic results is underpinned by their cultural values, mainly solidarity and reciprocity. Participants viewed research participation as a mutual relationship and considered the return of research results to be one way of reciprocating their efforts. This seems to be underpinned by the principle of Ubuntu which advocates for solidarity and reciprocity within communities. Participants noted that when reciprocity obligations are respected, participants feel valued and expressed that not respecting reciprocity expectations could undermine participants' trust and participation in future studies. Almost all participants wanted to receive individual genetic results. While parents and caregivers wanted to receive individual genetic results regardless of their severity, preventability or actionability, adolescents were reluctant to receive results for genetic conditions that are severe and non-preventable, especially if they are also unactionable. Participants advanced different reasons for feedback of results including for awareness, improving lifestyle, accepting one's' situation, and preparing for the future. The findings also reveal the importance of taking into account participants' context, relations and empowerment when making decisions about whether and which results ought to be fed back. When asked about practical considerations for feedback of results, both adolescents and parents expressed that they would prefer to receive individual genetic results in person, with adolescents preferring researchers to provide feedback, while parents preferred feedback from doctors associated with the study. Adolescents and parents both expressed that feedback should be supported by counselling, but they differed on the timing of feedback. Most participants shared that they would like to be informed about the possibility of discovering individual genetic results during the consent process and that consent be obtained for feedback during the enrolment process. They further expressed that in cases where prior consent to feedback was not obtained, then participants should be re-contacted where lifesaving genetic information is discovered. Participants emphasized the need for researchers to ensure that participants' decisions regarding feedback of results are well-informed. Autonomy, transparency, and communication were identified as key values to uphold during the consent process. Conclusion: In conclusion, expectations of solidarity and reciprocity could translate into an obligation to feedback selected individual genetic results in African genomics research. Decisions on practicalities for feedback of results should take into account participants' context and considerations of participants' preferences. For example, in settings like BBCCCE it might be feasible for the study team to relay participants' results to treating doctors in the same centre, while also organising counselling services if necessary. However, in cases where a study is done in a public facility with limited resources, that could be difficult to implement. Consequently, researchers may have to take up the responsibility of feeding back individual results as well as providing genetic counselling in such settings. To make these decisions, researchers should engage with relevant stakeholders including policymakers and local Institutional Review Boards (IRBs) so as to make informed decisions regarding the feasibility and acceptability of their approach to feedback of results. Obtaining participants' consent for feedback of results is important to ensure that their rights and wellbeing are protected in research. This is critical in building trust relationships between participants and researchers. Lastly, although this study is focused in Botswana, these findings could also be generalised to similar contexts in Africa and provide an authoritative voice to H3Africa to be able to mandate projects with potential to generate individual genetic results to make provisions to feedback these results to study participants.

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).