RESUMO
BACKGROUND/AIM: In this study, we compared the dialysis efficiency, oxidative stress, and nutritional conditions between predilution on-line hemodiafiltration (pre-OL-HDF) and conventional hemodialysis (HD) using a super-flux dialyzer (CHD). METHOD: This was a crossover study of 38 maintenance HD patients. All patients were treated with CHD for the first 4 months (1st CHD period), then were switched to pre-OL-HDF for 4 months (pre-OL-HDF period), and were returned to CHD for the next 4 months (2nd CHD period). RESULTS: We found no significant difference in the removal ratio of small uremic substances or the indices of inflammation or nutritional states between the pre-OL-HDF and CHD periods. However, we found higher removal of ß2 micro-globulin in the pre-OL-HDF period, and the human mercapto-albumin (Alb)/human serum Alb ratio was significantly higher in the pre-OL-HDF period. CONCLUSION: Treatment with pre-OL-HDF enabled enhanced removal of middle molecule uremic toxins and better Alb redox than did CHD.
Assuntos
Hemodiafiltração/métodos , Estresse Oxidativo , Albumina Sérica Humana/isolamento & purificação , Compostos de Sulfidrila/isolamento & purificação , Toxinas Biológicas/isolamento & purificação , Idoso , Estudos Cross-Over , Feminino , Hemodiafiltração/economia , Hemodiafiltração/instrumentação , Humanos , Inflamação/sangue , Nefropatias/sangue , Nefropatias/terapia , Masculino , Pessoa de Meia-Idade , Oxirredução , Albumina Sérica Humana/análise , Compostos de Sulfidrila/análise , Toxinas Biológicas/sangueRESUMO
Several diseases are associated with perturbations in redox signaling and aberrant hydrogen sulfide metabolism, and numerous analytical methods exist for the measurement of the sulfur-containing species affected. However, uncertainty remains about their concentrations and speciation in cells/biofluids, perhaps in part due to differences in sample processing and detection principles. Using ultrahigh-performance liquid chromatography in combination with electrospray-ionization tandem mass spectrometry we here outline a specific and sensitive platform for the simultaneous measurement of 12 analytes, including total and free thiols, their disulfides and sulfide in complex biological matrices such as blood, saliva and urine. Total assay run time is <â¯10â¯min, enabling high-throughput analysis. Enhanced sensitivity and avoidance of artifactual thiol oxidation is achieved by taking advantage of the rapid reaction of sulfhydryl groups with N-ethylmaleimide. We optimized the analytical procedure for detection and separation conditions, linearity and precision including three stable isotope labelled standards. Its versatility for future more comprehensive coverage of the thiol redox metabolome was demonstrated by implementing additional analytes such as methanethiol, N-acetylcysteine, and coenzyme A. Apparent plasma sulfide concentrations were found to vary substantially with sample pretreatment and nature of the alkylating agent. In addition to protein binding in the form of mixed disulfides (S-thiolation) a significant fraction of aminothiols and sulfide appears to be also non-covalently associated with proteins. Methodological accuracy was tested by comparing the plasma redox status of 10 healthy human volunteers to a well-established protocol optimized for reduced/oxidized glutathione. In a proof-of-principle study a deeper analysis of the thiol redox metabolome including free reduced/oxidized as well as bound thiols and sulfide was performed. Additional determination of acid-labile sulfide/thiols was demonstrated in human blood cells, urine and saliva. Using this simplified mass spectrometry-based workflow the thiol redox metabolome can be determined in samples from clinical and translational studies, providing a novel prognostic/diagnostic platform for patient stratification, drug monitoring, and identification of new therapeutic approaches in redox diseases.
Assuntos
Dissulfetos/isolamento & purificação , Metaboloma , Estresse Oxidativo , Compostos de Sulfidrila/isolamento & purificação , Cromatografia Líquida , Dissulfetos/sangue , Dissulfetos/urina , Glutationa/sangue , Glutationa/isolamento & purificação , Glutationa/urina , Humanos , Espectrometria de Massas , Oxirredução , Compostos de Sulfidrila/sangue , Compostos de Sulfidrila/urinaRESUMO
Quantitative phytochelatin (PC) analysis is, due to oxidation sensitivity of the PCs, matrix effects, and time consuming sample preparation, still a challenging analytical task. In this study, a rapid, simple, and sensitive method for precise determination of native PCs in crude extracts of the green alga Chlamydomonas reinhardtii was developed. Algae were exposed 48 h to 70 µM Cd. Coupling of ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry with multi-reaction mode transitions for detection permitted the required short-time, high-resolution separation and detection specificity. Thus, under optimized chromatographic conditions, 10 thiol peptides were baseline-separated within 7 min. Relative detection limits in the nanomolar range in microliter sample volumes were achieved (corresponding to absolute detection limits at femtomole level). Next to glutathione (GSH), the most abundant cadmium-induced PCs in C. reinhardtii, namely CysGSH, PC(2), PC(3), CysPC(2), and CysPC(3), were quantified with high reproducibility at concentrations between 15 and 198 nmol g(-1) fresh weight. The biological variation of PC synthesis of nine independently grown alga cultures was determined to be on average 13.7%.
Assuntos
Chlamydomonas reinhardtii/química , Fitoquelatinas/química , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida de Alta Pressão/economia , Cromatografia Líquida de Alta Pressão/métodos , Fitoquelatinas/isolamento & purificação , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização por Electrospray/métodos , Compostos de Sulfidrila/química , Compostos de Sulfidrila/isolamento & purificação , Espectrometria de Massas em Tandem/economia , Fatores de TempoRESUMO
This work presents the performances of a new odour scrubber. The reactor is packed with a new structure which enables co-current operations at high gas velocities. Energy consumption and removal efficiency of sulphur compounds by oxidative alkaline scrubbing were studied. The influence of both superficial gas (U(SG)) and liquid (U(SL)) velocities, ranging from 5.6 to 28 m.s(-1) and 0.016 to 0.055 m.s(-1) respectively, were quantified. Thus, the range of 0.5 to 5 liquid-to-gas mass ratio (L/G) was studied. A comparison has been made with a previous study on static mixers (SM) and with classical random packed towers (PT). It has been shown that superficial liquid and gas velocities have a significant influence on these parameters. Hydrogen sulphide (H2S) abatement reached values up to 99%. As concerns methylmercaptan (CH3SH), the maximal removal efficiency was 87%. As a result, if well scaled-up, our reactor can be a small single stage efficient apparatus for the elimination of low concentrations of sulphur compounds as H2S and CH3SH in high flow rates of polluted gas effluents.