Browsing by Author "Sarfo, Fred Stephen"
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- ItemThe kinetics of mycolactone in relation to the microbiological, clinical and immunological responses to antibiotic therapy for mycobacterium ulcerans disease.(2014-11-20) Sarfo, Fred StephenBackground: Mycobacterium ulcerans is the causative agent for a chronic necrotising skin infection called Buruli ulcer. Pathology of the disease is closely linked with the elaboration of a unique lipid toxin, mycolactone, which has potent cytotoxic and immunomodulatory properties. In this study, assays were developed to detect and quantify mycolactone concentrations in tissues during curative antibiotic therapy in mice and in humans to understand its dynamics in pathogenesis and to explore its potential as a biomarker for diagnosis and monitoring of patients with Buruli ulcer disease on antibiotic therapy. The currently recommended antibiotic regimen for the management of Buruli ulcer is a combination of daily intramuscular injections of streptomycin and oral rifampicin for 8 weeks (RS8). This regimen was compared with streptomycin/rifampicin for 2 weeks followed by clarithromycin/rifampicin (RS2RC6) for 6 weeks in patients to determine the clinical and bacteriological effectiveness in a pilot study. Methods: Biopsies were obtained from infected human skin tissues and BALB/c mouse hind footpads before, during and after 8-weeks of rifampicin-containing combination antibiotic therapy. Lipids were extracted from tissues using organic solvents, mycolactone concentrations were measured using a cytotoxicity assay and mass spectrometry. Trends in mycolactone concentrations and clinical, bacteriological and immuno-histopathological responses were determined. Concentrations of cytokines in supernates of whole blood assays in humans or murine splenocytes after stimulation with mycobacterial antigens/T-cell mitogens were measured using ELISA. iv Results: Eighty-three patients with confirmed Buruli ulcer were randomized to RS8 or RS2RC6 and monitored for recurrence free-healing. Bacterial load in tissue samples before and after treatment for 6 and 12 weeks was monitored in samples obtained by 4mm punch biopsy by semi-quantitative culture. There was no difference in using RS8 or RS2RC6 with respect to healing rate or the proportion healed in each group after 4, 8, 12, 16, 20, 24 and up to 52 weeks. The success rate was 93% in each group and there was no recurrence after 12-month follow-up. There was no difference in the number of bacteria cultured at the different time points for the two regimens. Mycolactone was detectable in 92% and 77% of human samples (n=80) using cytotoxicity assays and mass spectrometry respectively. Antibiotic therapy was associated with a decline in tissue concentration of mycolactone in both human and murine-infected tissues which was paralleled by resolution of clinical lesions, reductions in bacteriological counts and restoration of local and systemic immune responses. Discussions/Conclusions: This study shows that mycolactone concentrations in tissues is closely associated with the presence of M. ulcerans and provides useful proof-of-concept data that mycolactone detection could potentially be used to monitor response to antibiotic therapy as well as for diagnosis of Buruli ulcer disease. The findings from the pilot study indicate that rifampicin combined with clarithromycin can replace rifampicin and streptomycin for the continuation phase after rifampicin-streptomycin treatment for 2 weeks without any apparent loss of efficacy. The implication is that a controlled trial of fully oral therapy using rifampicin and clarithromycin for 8 weeks (RC8) is justified.
- ItemThe kinetics of mycolactone in relation to the microbiological, clinical and immunological responses to antibiotic therapy for mycobacterium ulcerans disease.(2014) Sarfo, Fred StephenBackground: Mycobacterium ulcerans is the causative agent for a chronic necrotising skin infection called Buruli ulcer. Pathology of the disease is closely linked with the elaboration of a unique lipid toxin, mycolactone, which has potent cytotoxic and immunomodulatory properties. In this study, assays were developed to detect and quantify mycolactone concentrations in tissues during curative antibiotic therapy in mice and in humans to understand its dynamics in pathogenesis and to explore its potential as a biomarker for diagnosis and monitoring of patients with Buruli ulcer disease on antibiotic therapy. The currently recommended antibiotic regimen for the management of Buruli ulcer is a combination of daily intramuscular injections of streptomycin and oral rifampicin for 8 weeks (RS8). This regimen was compared with streptomycin/rifampicin for 2 weeks followed by clarithromycin/rifampicin (RS2RC6) for 6 weeks in patients to determine the clinical and bacteriological effectiveness in a pilot study. Methods: Biopsies were obtained from infected human skin tissues and BALB/c mouse hind footpads before, during and after 8-weeks of rifampicin-containing combination antibiotic therapy. Lipids were extracted from tissues using organic solvents, mycolactone concentrations were measured using a cytotoxicity assay and mass spectrometry. Trends in mycolactone concentrations and clinical, bacteriological and immuno-histopathological responses were determined. Concentrations of cytokines in supernates of whole blood assays in humans or murine splenocytes after stimulation with mycobacterial antigens/T-cell mitogens were measured using ELISA. iv Results: Eighty-three patients with confirmed Buruli ulcer were randomized to RS8 or RS2RC6 and monitored for recurrence free-healing. Bacterial load in tissue samples before and after treatment for 6 and 12 weeks was monitored in samples obtained by 4mm punch biopsy by semi-quantitative culture. There was no difference in using RS8 or RS2RC6 with respect to healing rate or the proportion healed in each group after 4, 8, 12, 16, 20, 24 and up to 52 weeks. The success rate was 93% in each group and there was no recurrence after 12-month follow-up. There was no difference in the number of bacteria cultured at the different time points for the two regimens. Mycolactone was detectable in 92% and 77% of human samples (n=80) using cytotoxicity assays and mass spectrometry respectively. Antibiotic therapy was associated with a decline in tissue concentration of mycolactone in both human and murine-infected tissues which was paralleled by resolution of clinical lesions, reductions in bacteriological counts and restoration of local and systemic immune responses. Discussions/Conclusions: This study shows that mycolactone concentrations in tissues is closely associated with the presence of M. ulcerans and provides useful proof-of-concept data that mycolactone detection could potentially be used to monitor response to antibiotic therapy as well as for diagnosis of Buruli ulcer disease. The findings from the pilot study indicate that rifampicin combined with clarithromycin can replace rifampicin and streptomycin for the continuation phase after rifampicin-streptomycin treatment for 2 weeks without any apparent loss of efficacy. The implication is that a controlled trial of fully oral therapy using rifampicin and clarithromycin for 8 weeks (RC8) is justified.
- ItemMycolactone Diffuses into the Peripheral Blood of Buruli Ulcer Patients - Implications for Diagnosis and Disease Monitoring(PLoS Neglected Tropical Diseases, 2011-07-19) Sarfo, Fred Stephen; Chevalier, Fabien Le; Aka, N’Guetta; Phillips, Richard Odame; Amoako, Yaw; et. alBackground: Mycobacterium ulcerans, the causative agent of Buruli ulcer (BU), is unique among human pathogens in its capacity to produce a polyketide-derived macrolide called mycolactone, making this molecule an attractive candidate target for diagnosis and disease monitoring. Whether mycolactone diffuses from ulcerated lesions in clinically accessible samples and is modulated by antibiotic therapy remained to be established. Methodology/Principal Finding: Peripheral blood and ulcer exudates were sampled from patients at various stages of antibiotic therapy in Ghana and Ivory Coast. Total lipids were extracted from serum, white cell pellets and ulcer exudates with organic solvents. The presence of mycolactone in these extracts was then analyzed by a recently published, fieldfriendly method using thin layer chromatography and fluorescence detection. This approach did not allow us to detect mycolactone accurately, because of a high background due to co-extracted human lipids. We thus used a previously established approach based on high performance liquid chromatography coupled to mass spectrometry. By this means, we could identify structurally intact mycolactone in ulcer exudates and serum of patients, and evaluate the impact of antibiotic treatment on the concentration of mycolactone. Conclusions/Significance: Our study provides the proof of concept that assays based on mycolactone detection in serum and ulcer exudates can form the basis of BU diagnostic tests. However, the identification of mycolactone required a technology that is not compatible with field conditions and point-of-care assays for mycolactone detection remain to be worked out. Notably, we found mycolactone in ulcer exudates harvested at the end of antibiotic therapy, suggesting that the toxin is eliminated by BU patients at a slow rate. Our results also indicated that mycolactone titres in the serum may reflect a positive response to antibiotics, a possibility that it will be interesting to examine further through longitudinal studies.
- ItemStroke Among Young West Africans Evidence From the SIREN (Stroke Investigative Research and Educational Network) Large Multisite Case–Control Study(Stroke, 2018) Sarfo, Fred Stephen; Ovbiagele, Bruce; Gebregziabher, Mulugeta; Wahab, Kolawole; Akinyemi, Rufus; Akpalu, Albert...et.al.Background and Purpose—Stroke in lower and middle-income countries affects a young and productive age group. Data on factors associated with stroke in the young are sorely lacking from lower and middle-income countries. Our objective is to characterize the nature of stroke and its risk factors among young West Africans aged <50 years old. Methods—The SIREN (Stroke Investigative Research and Educational Network) is a multicenter, case–control study involving 15 sites in Nigeria and Ghana. Cases included adults aged ≥18 years with computed tomography/magnetic resonance imaging-confirmed stroke. Controls were age-and gender-matched stroke-free adults recruited from the communities in catchment areas of cases. Comprehensive evaluation for vascular, lifestyle, and psychosocial factors was performed. We used conditional logistic regression to estimate odds ratios and population attributable risks with 95% confidence intervals. Results—Five hundred fifteen (24.3%) out of 2118 cases enrolled were <50 years old. Among subjects <50 years old, hemorrhagic stroke proportion was 270 (52.5%) versus 245 (47.5%) for ischemic strokes. Etiologic subtypes of ischemic strokes included large artery atherosclerosis (40.0%), small vessel disease (28.6%), cardioembolism (11.0%), and undetermined (20.4%). Hypertension (91.7%), structural lesions (3.4%), and others (4.9%) were causally associated with hemorrhagic stroke. Six topmost modifiable factors associated with stroke in descending order of population attributable risk (95% confidence interval) were hypertension: 88.7% (82.5%–94.8%), dyslipidemia: 48.2% (30.6%–65.9%), diabetes mellitus: 22.6% (18.7%–26.5%), low green vegetable consumption: 18.2% (−6.8%–43.2%), stress: 14.5% (4.9%–24.1%), and cardiac disease: 8.4% (5.8%–11.1%). Conclusions—The high and rising burden of stroke among young Africans should be curtailed via aggressive, populationwide vascular risk factor control.
- ItemStroke in Indigenous Africans, African Americans, and European Americans Interplay of Racial and Geographic Factors(Stroke, 2017) Owolabi, Mayowa; Sarfo, Fred Stephen; Howard, Virginia J.; Irvin, Marguerite R.; Gebregziabher, Mulugeta; Akinyemi, Rufus; Bennett, Aleena; Armstrong, Kevin; Tiwari, Hemant K.; Akpalu, Albert...et.al.Background and Purpose—The relative contributions of racial and geographic factors to higher risk of stroke in people of African ancestry have not been unraveled. We compared stroke type and contributions of vascular risk factors among indigenous Africans (IA), African Americans (AA), and European Americans (EA). Methods—SIREN (Stroke Investigative Research and Educational Network) is a large multinational case–control study in West Africa—the ancestral home of 71% AA—whereas REGARDS (Reasons for Geographic and Racial Differences in Stroke) is a cohort study including AA and EA in the United States. Using harmonized assessments and standard definitions, we compared data on stroke type and established risk factors for stroke in acute stroke cases aged ≥55 years in both studies. Results—There were 811 IA, 452 AA, and 665 EA stroke subjects, with mean age of 68.0±9.3, 73.0±8.3, and 76.0±8.3 years, respectively (P<0.0001). Hemorrhagic stroke was more frequent among IA (27%) compared with AA (8%) and EA (5.4%; P<0.001). Lacunar strokes were more prevalent in IA (47.1%), followed by AA (35.1%) and then EA (21.0%; P<0.0001). The frequency of hypertension in decreasing order was IA (92.8%), followed by AA (82.5%) and then EA (64.2%; P<0.0001) and similarly for diabetes mellitus IA (38.3%), AA (36.8%), and EA (21.0%; P<0.0001). Premorbid sedentary lifestyle was similar in AA (37.7%) and EA (34.0%) but lower frequency in IA (8.0%). Conclusions—Environmental risk factors such as sedentary lifestyle may contribute to the higher proportion of ischemic stroke in AA compared with IA, whereas racial factors may contribute to the higher proportion of hypertension and diabetes mellitus among stroke subjects of African ancestry.