La Multidrug resistance Protein 1 (MRP1) est impliquée dans le phénotype de résistance multiple aux médicaments (MDR) des cellules cancéreuses. Les substrats physiologiques de MRP1 comprennent notamment le glutathion (GSH). Certains médicaments anti-cancéreux tels que la doxorubicine sont co-transportés avec le glutathion. Pour contourner le phénotype MDR induit par MRP1, nous proposons une nouvelle stratégie thérapeutique basée sur la sensibilité collatérale (SC) des cellules résistantes surexprimant MRP1. Certains composés, comme le vérapamil, provoquent la mort sélective des cellules résistantes (les cellules témoins ne sont pas affectées) en stimulant l'efflux du glutathion médié par MRP1. La déplétion intracellulaire très rapide et très forte du glutathion induit probablement un stress oxydatif déclenchant la mort cellulaire. Nous avons identifié de nouveaux agents de sensibilité collatérale puissants in vitro et nous avons montré l'effet du meilleur composé sur la réduction de la croissance des tumeurs chimiorésistantes chez la souris. Nous avons étudié le mécanisme moléculaire d'action des agents de SC et identifié un résidu, localisé dans une région inattendue, impliqué dans la stimulation de l'efflux de glutathion induit par ces molécules / Multidrug resistance Protein 1 (MRP1) is involved in the multidrug resistance (MDR) phenotype of cancer cells. Physiological substrates of MRP1 include glutathione (GSH) and drugs such as doxorubicin are co-transported with glutathione. To circumvent the MDR phenotype induced by MRP1, we propose a new therapeutic strategy based on collateral sensitivity (CS) of resistant cell expressing MRP1, its overexpression becoming the Achilles heel of the cell. Some compounds, like verapamil, act as MRP1 modulators. They trigger selective death of resistant cells (control cells are not affected) by stimulating MRP1-mediated glutathione efflux. The fast and huge intracellular depletion of glutathione probably induces an oxidative stress triggering cell death. We identified new potent collateral sensitivity agents in vitro and we checked the effect of the strongest compound on reducing resistant tumor growth in nude mice. We studied the molecular mechanism of action of CS agents and identified an unexpected residue involved in the stimulation of glutathione efflux induced by these molecules

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No. of bitstreams: 2 Tese - Lara Stefânia Netto de Oliveira Leão Vasconcelos - 2013.pdf: 3438615 bytes, checksum: 0860846b3615244c0123e230f0e5c789 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-03-28 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The health care environment presents numerous risks making it anharmful place, propitious to colonization and development of infections. Inserted in this scenario are the healthcare workers which are vulnerable to the carrier condition. and the oral cavity represents an important site of colonization. This study aimed to characterize the phenotype of Enterobacteriaceae isolated from the oral cavity of healthcare workers from an oncologic reference hospital in the central area of Brazil. We also aimed to detect co-colonization by Staphylococcus and Pseudomonas, as well as describe the profile socio-demographic, professional, disease/infection and behavioral of individuals colonized by Enterobacteriaceae. It was a descriptive cross-sectional epidemiological study conducted from May/2009 to November/2010 and saliva was collected a from each subjected. A total of 294 individuals participated in the research, being 149 healthcare members and 145 from the support team. The microbiological procedures were performed according to recommended techniques and data collection obtained by a questionnaire and analyzed through descriptive statistics. Among the participants, 55 (18.7%) were colonized by Enterobacteriaceae in the oral cavity. Were isolated 64 bacteria, including species potentially pathogenic. The most prevalent specie was gergoviae Enterobacter (17.2%). The highest rates of resistance were observed to β-lactams and 48.4% of isolates were considered multiresistant. The Extended Spectrum β-lactamases (ESBL) production was negative for the Enterobacteriaceae and none Klebsiella pneumoniae produced Carbapenemase (KPC). However, among the 43 CESP isolates group (Citrobacter, Enterobacter, Serratia, Providencia), 51.2% produced AmpC β- lactamase by induction and 48.8% were hiper producers. Staphylococcus and Pseudomonas were detected in the saliva of 56.4% individuals. The majory of the healthcare workers colonized by Enterobacteriaceae was men over 40 years. The nursing technician was the professional category most colonized (52.7%), followed by the cleaning staff (23.6%). The cleaning and hygiene sector (23.6%) and nursing stations (18.2%) represented the largest number of workers colonized. Some professionals (27.3%) had second job in another health service, while 41.8% remained in the institution around 40 hours or more a week. Some infection was reported among carriers, as tonsillitis, sinusitis and pharyngitis, being the first the most frequent (52.7%). Enterobacteriaceae carriage (18.2%) reported the use of personal protective equipment (PPE) as unnecessary to their s activity, 7.3% did not use the mask as precautionary measure, 23.6% realized sporadically the exchange of mask and 1.8 % never changed. The use of oral antiseptics was observed in 29.1% of the workers, the recent use of antimicrobials in 16.4% and self-medication in 27.3%. Among the colonized persons was observed lack of knowledge about multiresistant microorganisms. The prevalence Enterobacteriaceae carriage is considered high and the resistance is a problem especially due to multidrug resistance and production of β-lactamase AmpC observed. The results of this research contribute with important subsidies to the programs for nosocomial infections prevention and control of, since knowledge of carrier status reduces the risk of transmission of microorganisms. / O ambiente da assistência à saúde apresenta inúmeros riscos que o torna um local insalubre, propício à colonização e ao desenvolvimento de infecções. Inseridos neste cenário estão os trabalhadores dos serviços de saúde, os quais são vulneráveis à condição de portador. A cavidade bucal representa assim um importante sítio de colonização. Este estudo buscou caracterizar o fenótipo de Enterobacteriaceae isoladas da cavidade bucal de trabalhadores de um hospital oncológico referência no Centro-Oeste brasileiro. Também fizeram parte dos objetivos desta pesquisa detectar co-colonização por Staphylococcus e Pseudomonas, bem como descrever o perfil sócio-demográfico, profissional, doença/infecção e comportamental dos indivíduos colonizados por Enterobacteriaceae. Estudo epidemiológico transversal do tipo descritivo realizado de maio/2009 a novembro/2010. Participaram 294 trabalhadores, sendo 149 membros da equipe de saúde e 145 da equipe de apoio. Os procedimentos microbiológicos foram realizados segundo técnicas padronizadas e a coleta de dados realizada por meio da aplicação de um formulário. Foi coletada uma amostra de saliva de cada sujeito Os dados foram analisados por meio de estatística descritiva. Dentre os participantes, 55 (18,7%) estavam colonizados por Enterobacteriaceae na cavidade bucal. Foram isoladas 64 bactérias, incluindo espécies potencialmente patogênicas. A espécie mais prevalente foi Enterobacter gergoviae (17,2%). As maiores taxas de resistências foram observadas para os β-lactâmicos e 48,4% dos isolados foram considerados multirresistentes. Para as enterobactérias pesquisadas, a produção de β-lactamase de Espectro Ampliado (ESBL) e Klebsiella pneumoniae Carbapenemase (KPC) foi negativa. Porém, dentre os 43 isolados do grupo CESP (Citrobacter, Enterobacter, Serratia, Providencia), 51,2% foram considerados produtores de β-lactamase AmpC por indução e 48,8% mutantes hiperprodutores. Staphylococcus e Pseudomonas estavam presentes na saliva de 56,4% dos sujeitos colonizados por Enterobacteriaceae. O maior número de portadores foi constituído por homens acima de 40 anos. Entre os técnicos de enfermagem, 52,7% estavam colonizados, seguidos dos auxiliares de limpeza (23,6%). O setor de higienização e limpeza (23,6%) e os postos de enfermagem (18,2%) contemplaram o maior número de trabalhadores colonizados. Alguns trabalhadores (27,3%) possuíam segundo emprego em outro serviço de saúde, enquanto que 41,8% permaneciam na instituição 40 horas ou mais na semana. Quadros frequentes de infecção foram relatados entre os portadores, como amigdalites, sinusites e faringites, sendo o primeiro o mais frequente (52,7%). Alguns portadores de Enterobacteriaceae (18,2%) relataram o uso de EPI como desnecessário à sua atividade, 7,3% não faziam o uso de máscara como medida de precaução, 23,6% realizavam a troca de máscara esporadicamente e 1,8% nunca trocava. O uso de antissépticos bucais foi observado em 29,1% dos trabalhadores, o uso recente de antimicrobianos em 16,4% e a prática da automedicação em 27,3%. A desinformação sobre micro-organismos multirresistentes foi comum entre os colonizados. A prevalência de portadores de Enterobacteriaceae foi elevada e o fenótipo de resistência dos isolados preocupante, especialmente pela multirresistência e produção de β-lactamases AmpC. As variáveis avaliadas revelaram realidades relevantes para o contexto da assistência à saúde e para a saúde do trabalhador. Os resultados desta pesquisa contribuíram com subsídios importantes para os programas de prevenção e controle das infecções nosocomiais, pois o conhecimento do estado portador reduz os riscos de transmissão de micro-organismos.

Enterococos são ubíquos no ambiente e fazem parte da microbiota do trato gastrintestinal de humanos e animais. A importância dessas bactérias tem sido associada com infecções hospitalares e resistência a múltiplas drogas, principalmente à vancomicina. O objetivo do presente estudo foi realizar a caracterização molecular de cepas de Enterococcus spp. resistentes à vancomicina (VRE) isoladas a partir de amostras coletadas de pacientes hospitalizados, água superficial de rios urbanos e carne de frango comercializada no Brasil. A presença do gene vanA foi confirmada em 20 cepas multirresitentes isoladas durante 1997-2011. Dentre os isolados VRE, 12 cepas foram identificadas como E. faecium e oito como E. faecalis. Cepas de E. faecium isoladas de amostras clínicas e águas foram classificadas como clonalmente relacionadas pelo PFGE, com perfil virulência predominante (acm+, esp+). Adicionalmente, enquanto cepas de E. faecium isoladas dos rios pertenceram aos ST203, ST412 e ST478 (previamente caracterizados como endêmicos em hospitais brasileiros), novos STs foram identificados entre as cepas de E. faecalis (ST614, ST615 e ST616) e E. faecium (ST953 e ST954) isoladas de alimentos. Sequências completas do transposon Tn1546 das cepas clínicas VREfm 320/07 (ST478) e ambiental VREfm 11 (ST412) mostraram Tn1546-like element de ~12800 pb, com um ponto de mutação no gene vanA na posição 7.698 (substituição do nucleotídeo T pelo C) e uma no gene vanX na posição 8.234 (G pelo T). Além disso, uma deleção na extremidade esquerda do Tn1546, e as sequências IS1251 e IS1216E na região intergênica vanHS e vanYX, respectivamente, também foram detectados. A este respeito, a IS1216E na região intergênica vanXY constitui um conjunto de genes previamente relatado em cepas clínicas de VREfm no Brasil, denotando uma característica regional. IS1216E tem sido associada com os genes tcrB e aadE que conferem resistência ao cobre e aminoglicosídeos, em E. faecium e Streptococcus agalactiae, respectivamente. Portanto, essa IS pode contribuir para a rápida aquisição de resistência antimicrobiana entre as espécies de cocos Gram-positivos clinicamente importantes. Os tipos de Tn1546 indistiguíveis que foram identificados no atual estudo isolados de humano e ambientes aquáticos sugerem uma comum partilha de um pool de genes de resistência à vancomicina. / Enterococci are ubiquitous in the environment and in the intestinal tract of humans and animals. The importance of these bacteria has been associated with nosocomial infection and multiple resistance to antimicrobial agents, mainly vancomycin. The aim of the present study was to perform molecular characterization of vancomycin-resistant Enterococcus spp. strains (VRE) isolated from hospitalized patients, surface water of urban rivers and retail chicken meat in Brazil. The presence of the vanA gene was confirmed in 20 multidrug-resistant strains isolated in 1997-2011. Among these VRE isolates, (n = 12) were identified as E. faecium and (n = 8) as E. faecalis. E. faecium strains isolated from water and clinical samples were classified as clonally related by PFGE, the predominant virulence profile being (acm+, esp+). Additionally, while E. faecium strains isolated from rivers belonging to ST203, ST412 and ST478 (previously characterized as endemic in Brazilian hospitals), new STs were identified among strains of E. faecalis (ST614, ST615 and ST616) and E. faecium (ST953 and ST954) isolated from food. Complete sequences of transposon Tn1546 from VREfm clinical strain 320/07 (ST478) and environmental strain VREfm 11 (ST412) showed a Tn1546-like element of ~12800 bp, with T7698C vanA and G8234T vanX mutations. Moreover, deletion of the Tn1546 left extremity, and the IS1251 and IS1216E sequence inside the vanHS and vanYX intergenic region, respectively, were also detected. In this regard, the IS1216E sequence inside the vanXY intergenic region constitutes a gene array previously reported for Brazilian VREfm clinical strains alone, denoting a regional characteristic. IS1216E has been associated with tcrB and aadE genes, which confer resistance to copper and aminoglycosides, in E. faecium and Streptococcus agalactiae, respectively. Therefore, IS1216E should contribute to rapid acquisition of antimicrobial resistance among species of the clinically important Gram-positive cocci. On the other hand, Tn1546-like elements were identical among clinical and environmental VREfm isolates, suggesting sharing of a common vancomycin resistance gene pool.

Escherichia coli transporter protein AcrB and its homologues are the inner membrane components of the Resistance-Nodulation-Division (RND) family efflux pumps in Gram-negative bacteria. It is well accepted that soluble proteins are only marginally stable, but such insight is missing for membrane proteins. The lack of stability data, including thermodynamic stability and oligomer association affinity is a result of intrinsic difficulties in working with membrane proteins. In addition, the degradation of soluble proteins in E. coli has been extensively studied whereas the degradation process of membrane proteins remains unclear. A focus of my thesis is the validation and development of methods used to measure the thermo- and oligomeric- stability of membrane proteins. I investigated the mechanism of a popular thermal-stability assay developed specifically for the study of membrane proteins uses a thiol-specific probe, 7-diethylamino-3-(4-maleimidophenyl)-4-methylcoumarin (CPM). I found that, contrary to current understanding, the presence of a sulfhydryl group was not a prerequisite for the CPM thermal stability assay. The observed fluorescence increase is likely caused by binding of the fluorophore to hydrophobic patches exposed upon protein unfolding. I then applied these methods in the study of three projects. In the first project, I investigated how suppressor mutations restore the function of AcrBP223G, in which the Pro223 to Gly mutation compromised the function of AcrB via disrupting AcrB trimerization. The results suggested that the function loss resulted from compromised AcrB trimerization could be restored through various mechanisms involving the compensation of trimer stability and substrate binding. In the second project, I created two AcrB fusion proteins, with C-terminal yellow fluorescence protein (YFP) and cyan fluorescence protein (CFP), respectively. YFP and CFP form a fluorescence resonance energy transfer (FRET) pair. Using this pair of fusion proteins, I studied AcrB assembly both in detergent micelles and in lipid bilayers. A positive cooperativity was observed in kinetic studies of association of AcrB trimer. Reconstitution experiment revealed that the association showed a higher FRET efficiency and faster association rate in liposome than in DDM. In the last project, I developed a fluorescence method to study the degradation of AcrB-ssrA by the ClpXP system. Comparing to the degradation of GFP-ssrA, degradation of AcrB-CFP-ssrA showed a lower maximum velocity and tighter binding to the enzymes with a positive cooperativity.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Staphylococcus aureus is a Gram-positive coccus belonging to the Stapylococcaeae family. S. aureus causes a wide range of infections that range from skin infections to lifethreatening infections such as pneumonia and endocarditis and is the major cause of hospital and community-acquired infections. Despite antibiotics being available for the treatment of S.aureus infections, resistance to a number of antibiotics has developed over the years due to their improper and continuous use. S. aureus develops resistance to various drugs via different mechanisms, one of which is the extrusion of the antibiotics through efflux pumps that play a role in its acquisition of multidrug resistance. The ability of methicillin-resistant S.aureus to develop resistance to a variety of antibiotics is causing global concern as treatment options are being limited. Various antimicrobial studies carried out on purified plant-based flavonoids have shown that flavonoids enhance the antibacterial effect of antibiotics. This study analysed antibacterial effects of the antibiotics; tetracycline, ampicillin, methicillin and vancomycin and three flavonoids; chrysin, naringenin and 7-hydroxyflavone, against methicillin-sensitive ATCC 25923 (MSSA) and methicillin-resistant ATCC 33591 (MRSA) S. aureus strains, using the Kirby-Bauer disk diffusion and microtitre microdilution assays. In the Kirby- Bauer assay, the antibiotics demonstrated inhibitory effects on the growth of MSSA ATCC 25923. However MRSA ATCC 33591 was only susceptible to vancomycin, with minimal inhibition zones observed with ampicillin. The flavonoids did not enhance or reduce the antibacterial activities of the antibiotics as the zones of inhibition sizes remained unchanged in the combination studies. Microtitre assay results revealed that naringenin enhanced the antibacterial activities of the antibiotics tetracycline and ampicillin, against MSSA ATCC 25923 and MRSA 33591. This was evident as calculated synergistic ratios by the Abbot formula showed that naringenin had an additive effect. The presence of the efflux pump genes in MSSA ATCC 25923 and MRSA ATCC 33591 was compared using polymerase chain reaction (PCR). The mepA and gyrA genes were identified in both strains whereas sepA was identified in MRSA ATCC 33591. The presence of efflux pump genes in both MSSA ATCC 25923 and MRSA ATCC 33591 also confirmed that the presence or absence of the genes may contribute to antibiotic resistance. The presence of sepA in the MRSA and not the MSSA confirmed that this gene plays a role in conferring drug resistance.

Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Magister Pharmaceuticae - MPharm / Many studies have reported low cure rates for multidrug-resistant tuberculosis (MDRTB) patients and MDR-TB patients co-infected with human immunodeficiency virus (HIV). However, little is known about the effect of HIV infection and antiretroviral therapy on the treatment outcomes of MDR-TB in South Africa. Therefore, the objectives of the study are: to find out whether HIV infection and interactions between ARVs and second line anti-TB drugs have an impact on the following MDR-TB treatment outcomes: cure rate and treatment failure at Brewelskloof Hospital. MDR-TB patients were treated for 18-24 months. The study was designed as a case-control retrospective study comparing MDR-TB treatment outcomes between HIV positive (cases) and HIV negative patients (controls). Patients were included in the study only if they complied with the following criteria: sensitivity to second line anti-TB drugs, MDR-TB infection, co-infection with HIV (for some of them), male and female patients, completion of treatment between 1 January 2006 and 31 December 2008. Any patients that presented with extreme drug-resistant tuberculosis (XDR-TB) were excluded from the study. Data were retrospectively collected from each patient’s medical records. There were a total of 336 patients of which 242 (72%) were MDR-TB patients and 94 (27.9%) MDRTB co-infected with HIV patients. Out of the 242 MDR-TB patients, 167 (69.2%) were males and 75 (30.7%) were females. Of the 94 patients with MDR-TB co-infected with HIV, 51 (54.2%) males and 43 (45.7%) females. Patients with multidrug-resistant tuberculosis co-infected with HIV who qualify for antiretroviral therapy were treated with stavudine, lamivudine and efavirenz while all MDR-TB patients were given kanamycin, ethionamide, ofloxacin, cycloserine and pyrazinamide. The cure rate of MDR-TB in HIV (+) patients and in HIV (-) patients is 34.5% and 30 % respectively. There is no significant difference between both artes (pvalue = 0.80). The MDR-TB cure rate in HIV (+) patients taking antiretroviral drugs and in HIV (+) patients without antiretroviral therapy is 35% and 33% respectively. The difference between both rates is not statistically significant. The study shows that 65 (28.0%) patients completed MDR-TB treatment but could not be classified as cured or failure, 29 (12.5%) patients failed, 76 (32.7%) defaulted, 18 (7.7%) were transferred out and 44 (18.9%) died. As far as treatment completed and defaulted is concerned, there is no significant statistical difference between HIV (+) and HIV (-) The number of patients who failed the MDR-TB treatment and who were transferred out is significantly higher in the HIV (-) group than in the HIV (+) group. Finally the number of MDR-TB patients who died is significantly higher in the HIV (+) group). The median (range) duration of antiretroviral therapy before starting anti-tuberculosis drugs is 10.5 (1-60) months. According to this study results, the MDR-TB treatment cure rate at Brewelkloof hospital is similar to the cure rate at the national level. The study also hows that HIV infection and antiretroviral drugs do not influence any influence on MDR-TB treatment outcomes. / South Africa

Magister Pharmaceuticae - MPharm / The aim of the study was to determine firstly, kanamycin plasma concentrations using liquid chromatography coupled with mass spectrometry (LC/MS); secondly, to investigate the PK parameters of kanamycin in patients infected with MDR-TB and in patients co-infected with MDR-TB and HIV; thirdly, to assess the influence of HIV infection and renal impairment on the PK of kanamycin and fourthly, to find out whether there is any interaction between antiretroviral drugs and kanamycin.

The emergence and spread of multi-drug resistant (MDR-TB) and extensively drugresistant tuberculosis (XDR-TB) are a major medical and public problem threatening the global health. The objectives of this study were to (i) determine the prevalence of MDR-TB and XDR-TB in the Eastern Cape; (ii) analyze patterns of gene mutations in MDR-TB and (iii) identify gene mutations associated with resistance to second line injectable drugs in XDR-TB isolates. A total of 1520 routine sputum specimens sequentially received within a period of 12 months i.e. February 2012 to February 2013 from all MDR-TB and XDR-TB patients treated by Hospitals and clinics in the Eastern Cape were included in this study, of which 1004 had interpretable results. Samples were analyzed with the Genotype MTBDRplus VER 2.0 assay kit (Hain Lifescience) for detection of resistance to Rifampicin and Isoniazid while solid and liquid culture drug susceptibility tests were used for ethambutol, streptomycin, ethionamide, ofloxacin, capreomycin and amikacin. PCR and sequence analysis of short regions of target genes gyrA, (encode subunit of DNA topoisomerase gyrase), rrs (16S rRNA) and tlyA (encodes a 2’-O-methyltransferase) were performed on 20 XDR-TB isolates. MTBDRplus kit results and drug susceptibility tests identified 462 MDR-TB, 284 pre-XDR and 258 XDR-TB isolates from 267 clinics and 25 hospitals in the Eastern Cape. There was a high frequency of resistance to streptomycin, ethionamide, amikacin, ofloxacin and capreomycin. Mutation patterns indicated differences between the health districts as well as differences between the facilities within the health districts. The most common mutation patterns observed were: (i) ΔWT3, ΔWT4, MUT1 [D516V+del515] (rpoB), ΔWT, MUT1 [S315T1] (katG), ΔWT1 [C15T] (inhA) [39 MDR, 204 XDR-TB and 214 pre XDR-TB isolates], (ii) ΔWT8, MUT3 [L533P+S531L] (rpoB), ΔWT, MUT1 [S315T1] [145 MDR, 18 pre-XDR and 3 XDR-TB solates] and (iii) ΔWT3, WT4 [D516Y+del515] (rpoB), ΔWT, MUT1 [S315T1] (katG) [75 MDR, 1 pre-XDR and 7 XDR-TB isolates]. Mutations in inhA promoter regions were strongly associated with XDR-TB isolates. Two thirds (66.6 percent (669/1004) of the isolates had inhA mutations present with 25.4 percent (170/669) found among the MDR isolates, 39.2 percent (262/669) among the pre-XDR isolates and 35.4 percent (237/669) among the XDR-TB isolates, which implies that these resistant isolates are being spread by transmission within the community and circulating in the province. There was good correlation between XDR-TB drug susceptibility test results and sequence analyses of the gyrA and rrs genes. The majority of XDR-TB isolates contained mutations at positions C269T (6/20) and 1401G (18/20) in gyrA and rrs genes respectively. Sequence analysis of short regions of gyrA and rrs genes may be useful for detection of fluoroquinolone and amikacin/ kanamycin resistance in XDR-TB isolates but the tlyA gene is not a sensitive genetic marker for capreomycin resistance. This study highlighted the urgent need for the development of rapid diagnostics for XDR-TB and raised serious concerns regarding ineffective patientmanagement resulting in ongoing transmission of extremely resistant strains of XDRTB in the Eastern Cape suggesting that the Eastern Cape could be fast becoming the epicenter for the development of Totally Drug-resistant Tuberculosis (TDR-TB) in South Africa.

The aim of this study was to assess community members’ knowledge levels regarding Drug Resistant TB and HIV and AIDS. The study was conducted at ward 40 in Green bushes area in Nelson Mandela Metropolitan Municipality (NMMM). A quantitative research method was used in this study. Random sampling is the type of probability sampling method that was used in this study. The sample consisted of 100 respondents above 18 years who were randomly selected from the beneficiary list of for the RDP houses in Green bushes area in the Nelson Mandela Metropolitan Municipality. Data for this study were collected using close ended questions which were administered by the researcher to the selected participants. Data was analysed using bivariate and descriptive statistics according to the identified themes. The study revealed that community members had high knowledge levels regarding Drug Resistant TB and HIV and AIDS prevention, transmission modes and diagnosis and treatment methods. The findings revealed that community members were highly knowledgeable and aware of the fact that abstaining and practising safe sex were means of preventing the spread of HIV and AIDS as it was spread through unprotected sex, while opening of windows and minimisation of close contact with HIV positive people and children with people infected with Drug Resistant TB are infection control measures or methods of preventing the spread of the disease. Additionally, the study indicated that female respondents were more aware and knowledgeable about prevention, transmission modes and diagnosis and treatment of Drug Resistant TB and HIV and AIDS than male respondents. Furthermore, the findings revealed that the respondents were highly knowledgeable and aware about transmission of Drug Resistant TB and HIV and AIDS; knowledgeable about prevention and less knowledgeable about diagnosis and treatment. A high percentage of female respondents knew that there was no vaccine to neither prevent nor cure HIV and AIDS and that antiretroviral drug were used to manage it. The study also showed that female respondents knew that all people irrespective of race and economic class can be infected with Drug Resistant TB and HIV and AIDS. It is important to note that the respondents between 41-60 years possessed more knowledge regarding Drug Resistant TB and HIV and AIDS than the respondents who were between 18-40 years. Lastly, the study showed that there were significant differences in gender and knowledge and no significant differences in age and knowledge of the respondents regarding Drug Resistant TB and HIV and AIDS. It is recommended that in future, research regarding knowledge levels about Drug Resistant TB and HIV and AIDS be extended to other wards in the Nelson Mandela Metropolitan Municipality (NMMM). Accurate knowledge should be provided by ensuring that educational materials that are developed, are appropriate for the various levels of literacy, and that more appropriate and relevant information regarding these diseases is made more accessible to community members in their home languages. The researcher further recommends that during training interventions and educational campaigns more emphasis should be put on prevention, diagnosis and treatment of Drug Resistant TB and HIV and AIDS.