RESUMO
RATIONALE: Cystic fibrosis (CF) patients are known to produce cyanide (CN-) although challenges exist in determinations of total levels, the precise bioactive levels, and specificity of its production by CF microflora, especially P. aeruginosa. Our objective was to measure total CN- levels in CF sputa by a simple and novel technique in P. aeruginosa positive and negative adult patients, to review respiratory tract (RT) mechanisms for the production and degradation of CN-, and to interrogate sputa for post-translational protein modification by CN- metabolites. METHODS: Sputa CN- concentrations were determined by using a commercially available CN- electrode, measuring levels before and after addition of cobinamide, a compound with extremely high affinity for CN-. Detection of protein carbamoylation was measured by Western blot. MEASUREMENTS AND MAIN RESULTS: The commercial CN- electrode was found to overestimate CN- levels in CF sputum in a highly variable manner; cobinamide addition rectified this analytical issue. Although P. aeruginosa positive patients tended to have higher total CN- values, no significant differences in CN- levels were found between positive and negative sputa. The inflammatory oxidant hypochlorous acid (HOCl) was shown to rapidly decompose CN-, forming cyanogen chloride (CNCl) and the carbamoylating species cyanate (NCO-). Carbamoylated proteins were found in CF sputa, analogous to reported findings in asthma. CONCLUSIONS: Our studies indicate that CN- is a transient species in the inflamed CF airway due to multiple biosynthetic and metabolic processes. Stable metabolites of CN-, such as cyanate, or carbamoylated proteins, may be suitable biomarkers of overall CN- production in CF airways.
Assuntos
Cianetos/análise , Fibrose Cística/metabolismo , Técnicas Eletroquímicas , Ácido Hipocloroso/química , Processamento de Proteína Pós-Traducional , Escarro/química , Adulto , Cobamidas/química , Cianetos/metabolismo , Fibrose Cística/diagnóstico , Fibrose Cística/microbiologia , Eletrodos , Feminino , Humanos , Ácido Hipocloroso/metabolismo , Cinética , Masculino , Pessoa de Meia-Idade , Oxirredução , Carbamilação de Proteínas , Pseudomonas aeruginosa/metabolismo , Escarro/microbiologiaRESUMO
Urea can be oxidized electrochemically in a chloride solution to carbon dioxide, water, and nitrogen. The microkinetics of this hypochlorite-mediated urea oxidation are elucidated. Based on this kinetic information, the optimal conditions and construction principles for an electrochemical reactor are deduced. The construction of a cheap, disposable oxidation cell and necessary auxiliary equipment are described. In vitro data are reported for urea removal. A 36-L volume was used to simulate a 60-kg patient; 18 L was recirculated through a 0.12-m2 oxidation cell. Within 3 h, 35 g urea could be removed from the system. The technical and economic possibilities as well as safety requirements for hemofiltrate regeneration to a reinfusable substitution solution by anodic urea oxidation are discussed critically. Although the process does not appear to be economically practical for discontinuous hemofiltration, it might be desirable for continuous (24 h/day) treatment.