ABSTRACT
OBJECTIVE: Some, but not all, beta-thalassemia/hemoglobin E (beta-thal/HbE) patients respond to hydroxyurea treatment. It would be helpful if patient responses to hydroxyurea could be screened in vitro to identify responders and nonresponders before beginning in vivo treatment. MATERIALS AND METHODS: Thirteen beta-Thal/HbE patients were treated with hydroxyurea orally for 2 years at a starting dose of 5 mg/kg/day for 5 days/week with escalation to a maximum of 10 mg/kg/day. For comparison, erythroid cells obtained from peripheral blood of the same patients 1 year after they had stopped hydroxyurea treatment were treated with hydroxyurea in vitro. The gamma-globin mRNA was measured by real-time reverse-transcription polymerase chain reaction, fetal hemoglobin (HbF) by high-performance liquid chromatography, (G)gamma- and (A)gamma-globin chains by Triton X-100 acid urea polyacrylamide gel electrophoresis. RESULTS: Treatment of cells in primary culture with 30 microM hydroxyurea for 96 hours significantly increased the fractional HbF content in beta-Thal/HbE patients. The (G)gamma:(A)gamma-globin mRNA was induced 0.30- to 8-fold in vitro and 0.30- to 6-fold in vivo (r(2) = 0.51, p = 0.16 by paired t-test); the fractional HbF content was induced 0.50- to 19-fold in vitro and 0.30- to 12-fold in vivo (r(2) = 0.61, p = 0.20) and the (G)gamma:(A)gamma-globin chain ratio was increased 0.80- to 1.40-fold in vitro and 1- to 1.20-fold in vivo (r(2) = 0.62, p = 0.13). CONCLUSION: The correlation of in vivo and in vitro results of HbF synthesis and globin mRNA suggest that in vitro testing may predict the in vivo response.
Subject(s)
Drug Monitoring/methods , Fetal Hemoglobin/genetics , Hemoglobin E , Hydroxyurea/pharmacology , beta-Thalassemia/diagnosis , beta-Thalassemia/drug therapy , Adult , Cells, Cultured , Erythroid Cells/drug effects , Female , Fetal Hemoglobin/analysis , Gene Expression Regulation/drug effects , Hemoglobinuria/blood , Hemoglobinuria/diagnosis , Hemoglobinuria/drug therapy , Humans , Hydroxyurea/administration & dosage , Male , Middle Aged , Predictive Value of Tests , Prognosis , beta-Thalassemia/bloodABSTRACT
A bacterial strain, SWU-4, capable of using benzothiophene (BT) as a sole carbon and energy source was isolated from a petroleum-contaminated site in Thailand and identified by 16S rRNA gene sequence analysis to be in the genus of Mycobacterium. The strain was Gram-positive, nonspore former, and grew at 50 degrees C. Colonies of the strain on nutrient agar were rod-shaped, smooth with a convex surface, slightly mucoid, and yellow pigmented. The thermophilic Mycobacterium sp. strain SWU-4 rapidly degraded 2% (w/v) BT at 50 degrees C. Interestingly, this strain was able to degrade a wide variety of organosulfur compounds including thiophene, bromo(alpha)thiophene, and 3-methylthiophene in liquid minimum medium at 50 degrees C, which will be beneficial for industrial applications.
