El documento digital no refiere asesor / Estudia los casos de tuberculosis del aparato digestivo para conocer los factores epidemiológicos, estratificar a los pacientes en grupos de riesgo, determinar los parámetros de diagnóstico clínico y hallazgos de exámenes bioquímicos y radiológicos, tener en cuenta las localizaciones más frecuentes de esta patología y los métodos de diagnóstico presuntivo y definitivo, así como las comorbilidades y condiciones asociadas, que han afectado a los pacientes tratados en el centro hospitalario en los últimos 6 años. Mediante una ficha recopila los datos de las historias clínicas de 137 pacientes con diagnóstico de tuberculosis del aparato digestivo. El análisis de estos datos determina que el cuadro clínico sigue siendo bastante inespecífico y de difícil diagnostico, debido a que los exámenes en general solo revelan hallazgo de compromiso crónico, y son pocos los que dan una aproximación al diagnostico. A su vez, el control de respuesta al tratamiento fue ambulatorio evaluándose solo a 67 (48,91 %) luego del alta siendo la respuesta al tratamiento en este grupo evaluado del 71,64 %; el resto fallecieron o fueron controlados en centros de salud desconociéndose la evolución posterior. / Trabajo académico

Thesis (MSc. (Chemistry)) -- University of Limpopo, 2015 / Alkyne-quinoxaline compounds were successfully synthesised form the cross-coupling of 2-benzenesulfonyloxyquinoxaline with terminal alkynes. The current study demonstrates the ability of benzenesulfonyloxy as a good leaving group in Sonogashira coupling reaction, generating a number of alkyne-quinoxaline compounds. The structures of the synthesised compounds were characterised through the comprehensive analysis of spectroscopic data from NMR, HPLC-MS and others selected compounds with IR. Several alkyne-quinoxaline compounds were synthesised in moderate to good yields. During analysis, it was observed that the highest yield was obtained when using 4-(trifluoromethyl)phenylacetylene 4e and the lower yield was obtained when using the 3-ethynylthiophene 4s as a substrate. The alkyne-quinoxaline compounds were investigated for 1,3-dipolar cycloaddition which was done from 2-ethylquinoxaline 6. The 1,3-dipolar cycloaddition reactions were successful and the compounds were obtained in good yields. The in vitro analysis on anti-tubercular screening against H37RvMA strains of M tb reveals that compounds 5e and 5o which contained a trifluoroanisole moiety showed promising activity amongst all the synthesised compounds. The compounds were having MIC90 values 11.8 μM and 12.7 μM respectively, however, they were found to be less active than rifampicin. Compounds 5a, 5b, 5e, 5g, 5h, 5i, 5I, and 5r were evaluated for anti-cancer activity on A549 cells. The results have showed that 5i was able to retard the migration of A549 cells in the Wound-Healing Migration Assay. / National Research Foundation (NRF) Sasol Inzalo Foundation

Thesis (MSc. (Medical Sciences)) -- University of Limpopo, 2016. / The reference phenotypic methods for Mycobacterium tuberculosis drug susceptibility testing are qualitative and based on drug critical concentrations. Limitations include lack of standardization and variations in laboratory preparation of drug stock solutions. The recommended critical concentrations are determined by consensus and experience rather than scientific data. Consequently incorrect and inadequate susceptibility breakpoints are used and patients receive ineffective antimicrobial therapy. The determination of wild-type minimal inhibitory concentration distribution is an important tool used by European Committee for antimicrobial susceptibility Testing (EUCAST) to establish clinical breakpoints in Europe. This could be applicable in South Africa. Aim To determine wild-type minimal inhibitory concentration distributions of first and secondline drugs against Mycobacterium tuberculosis complex clinical isolates and compare these with the recommended critical concentration in Limpopo province. Methods A sample of 101 Mycobacterium tuberculosis complex positive cultures were collected from National Health Laboratory Services in Polokwane (Limpopo province) and subcultured on BACTEC MGIT 960 system. The isolates were inoculated on MYCOTB MIC plates to determine the wild-type MIC distributions of first and second-line drugs. The data were compared with currently recommended critical concentrations. DNA was extracted and amplified by PCR. Genotypic drug susceptibility testing was performed using GenoType MTBDRplus version 2.0 and GenoType MTBDRsl version 2.0 for the first- and second-line drugs, respectively. Genotyping of clinical isolates was performed to determine M. tuberculosis strain families using spoligotyping. vi Results Wild-type MIC distributions range reported in this study are as follows rifampin (≤ 0.12 - 0.5 μg/μg/ml), isoniazid (≤ 0.3 - 2.00 μg/ml), rifabutin (≤ 0.12 - 0.25 μg/ml), ethionamide (≤ 0.12 - 5 μg/ml), ethambutol (≤ 0.5 - 2 μg/ml), streptomycin (≤ 0.25 - 0.5 μg/ml), paraaminosalicylic (≤ 0.5 - 4.0 μg/ml), cycloserine (≤ 2 -16 μg/ml), amikacin (≤ 0.12 - 0.5 μg/ml), kanamycin (≤ 0.6 -2.5 μg/ml), moxifloxacin (≤ 0.6 - 0.5 μg/ml), ofloxacin (≤ 0.25 - 1 μg/ml). GenoType MTBDRplus detected (n= 68, 67%) rifampin resistance (MUT 3=26, MUT 2=18, MUT 2B=8) on the rpoB gene. Isoniazid resistant (n=20, 19.8%) was detected katG MUT (n=20, 19.8%) on katG gene (S315T1). Genotypic resistance to second-line drugs determined by GenoType MTBRsl detected no mutations in (n= 98, 97%) isolates on gyrA, gyrB rrs and eis gene and (n=3, 2.9%) isolates non mycobacterium tuberculosis complex were detected. The frequency and percentage of Mycobacterium tuberculosis family strain were identified in (n= 81, 80%) of the clinical isolates which matched 18 pre-existing shared types. The results showed high genotype diversity with the Beijing strain (n= 30, 29.7%) and T family (n= 19, 18.8%) dominating. Twenty isolates (19.8%) had no shared types thus reported as orphan. Conclusion The findings obtained in this study suggest wild-type Minimal Inhibitory Concentration distributions may be considered when setting clinical breakpoints. Discordant results were observed between phenotypic and genotypic DST for rifampin, isoniazid, streptomycin, rifabutin and ethambutol, suggesting that breakpoint concentrations for some drugs are set too high while others are too low. The Mycobacterium tuberculosis clinical isolates displayed diverse family strain with Beijing and T strain predominate breakpoints for first-line and second-line drugs used in Mycobacterium tuberculosis treatments. Poster Presentations Poster presented at faculty of Health science first annual research day on Second-line drug susceptibility breakpoints for Mycobacterium tuberculosis using MYCOTB MIC plate. University of Limpopo Tiro hall 16th to 17th September 2014. Poster presented at National Health Laboratory Service Pathology Research and Development Congress (PathReD) on Determination of families strains of Mycobacterium tuberculosis circulating in Limpopo Province, South Africa. Emperors Palace 14th April-16th April 2015.

No description available.

Introduction: Biomarkers of tuberculosis (TB) diagnosis and treatment response in patients coinfected with human immunodeficiency virus (HIV) are a necessity to ensure early diagnosis and adequate monitoring of TB treatment response. We conducted 3 sub-studies: study 1 was a bioavailability study; study 2 was a PK study in HIV-TB co-infected persons, and study 3 evaluated a WHO-recommended treatment algorithm in TB-HIV co-infected persons. Study 1 and 2 contributed to the study of 2 (NAT2) polymorphisms. Study 1 was leveraged to evaluate Quantiferon Gold in tube (QFT-GIT) and a quality of life instrument as a longitudinal biomarker in smear and culture positive TB-HIV co-infected patients. Study 3 was leveraged to study urine lipoarabinomannan (LAM) as a diagnostic adjunct in smear-negative HIV-infected patients treated for TB. Methods: Blood was collected from participants with HIV-infection only and TB-HIV coinfection for NAT2 polymorphisms at baseline, and for QFT-GIT at baseline, month 3, 6 and 12; a health-related quality-of-life (HRQOL) instrument was applied at the same timepoints to monitor treatment response in Study 1. An additional 40 TB-HIV co-infected participants (Study 2) were included in the analysis for the assessment of NAT2 polymorphisms and its effect on isoniazid plasma levels and hepatotoxicity. Urine was collected from seriously ill HIV-infected patients with confirmed smear-negative presumptive-TB (Study 3) prior to anti-TB treatment and tested using a commercially available LAM-ELISA. Blood and sputum were collected and processed for TB culture. Results: One hundred and twenty participants (100 TB-HIV co-infected and 20 non-TB but HIVinfected) from Study 1 and Study 2 with genotype results and were evaluated. Percentage of metabolisers in each category were: slow 52.5% (63/120), (NAT2*5/*5); intermediate 35.8% (43/120), (NAT2*4/*5 and NAT2*5/12); and rapid 11.7% (14/120), (NAT2*4/*11, NAT2*11/12 and NAT2*12/12). In general, isoniazid area under the concentration curve (AUC)0-∞ and maximum concentration (Cmax) were lower amongst the study 1 compared to study 2 participants. INH and AcINH PK parameters across genotypes were not statistically significantly different within each study. The log AcINH: log INH ratio, calculated as a measure of acetylation at two and four hours post-dose, showed no statistically significant difference between genotypes.

Evaluation of resistance to antituberculosis drugs is routinely performed with genotypic or phenotypic methods; however, discordance can be seen between these different methodologies. Our objective was to identify mutations that could explain discordant results in the evaluation of susceptibility to rifampicin and isoniazid between molecular and phenotypic methods, using whole genome sequencing (WGS). Peruvian strains showing sensitive results in the GenoType MTBDRplus v2.0 test and resistant results in the proportions in the agar-plaque test for isoniazid or rifampin were selected. Discordance was confirmed by repeating both tests, and WGS was performed, using the Next Generation Sequencing methodology. Obtained sequences were aligned "through reference" (genomic mapping) using the program BWA with the algorithm "mem", using as a reference the genome of the M. tuberculosis H37Rv strain. Discordance was confirmed in 14 strains for rifampicin and 21 for isoniazid, with 1 strain in common for both antibiotics, for a total of 34 unique strains. The most frequent mutation in the rpoB gene in the discordant strains for rifampicin was V170F. The most frequent mutations in the discordant strains for isoniazid were katG R463L, kasA G269S, and Rv1592c I322V. Several other mutations are reported. This is the first study in Latin America addressing mutations present in strains with discordant results between genotypic and phenotypic methods to rifampicin and isoniazid. These mutations could be considered as future potential targets for genotypic tests for evaluation of susceptibility to these drugs. / Revisión por pares

Publicación a texto completo no autorizada por el autor / Describe las características clínico-epidemiológicas, tipo de diagnóstico y pronóstico de la tuberculosis extrapulmonar en pacientes hospitalizados en el Hospital Nacional Arzobispo Loayza entre julio de 2013 y junio de 2015. Es un estudio descriptivo y retrospectivo de historias clínicas de pacientes internados en un hospital general durante dos años. Se incluyen adultos diagnosticados de tuberculosis extrapulmonar con historias completas. Las variables cualitativas se expresan en frecuencias/porcentajes y las cuantitativas como media/desviación estándar o mediana/rango intercuartilar según normalidad. Se reportan 211 casos de tuberculosis extrapulmonar, encontrándose 181 historias clínicas. La mediana de edad es 34 (26 - 51), predomina el sexo masculino. La comorbilidad más frecuente es la infección por HIV, seguida de diabetes mellitus. 26.5% de los pacientes tiene contacto tuberculosis pulmonar y 12.2% tuberculosis pulmonar activa. Los órganos más afectados son el sistema nervioso central y pleura. Hubo compromiso multisistémico en 21% de casos. El modo diagnóstico predominante es bioquímico seguido del histopatológico. Se logra el diagnóstico definitivo en 35.9% de casos. 10.9% de pacientes fallece. Concluye que la tuberculosis extrapulmonar es un tercio de los casos de tuberculosis diagnosticados en hospitalización, la forma pleural y del sistema nervioso son las más frecuentes en hospitalizados. La mayoría se trata en base a diagnóstico probable. La comorbilidad más común es la infección por HIV. / Tesis

No description available.

The primary purpose of this investigation is not only to improve the present technique of culture of Mycobacterium tuberculosis, but also to make a comparative study of the media use, namely, Veterans Administration modification of Trudeau's medium, Lowenstein's egg medium, and that developed by the author, in order to find which, if any, of these will most easily and effectively meet the needs of the hospital laboratory.

published_or_final_version / Community Medicine / Master / Master of Public Health