RESUMO
PURPOSE: The dietary flavonoids epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECG) and epigallocatechin gallate (EGCG) have been shown to interact with circulating albumin for transport in blood to different body tissues. This interaction may modulate their bioavailability and effectiveness. METHODS: Using affinity capillary electrophoresis to assess binding constants (K b), we investigated whether posttranslational modification of human serum albumin (HSA) through N- and S-homocysteinylation, commonly observed in hyperhomocysteinemia, may modify its interaction with catechins. RESULTS: S-Hcy HSA had lower Kb values toward EC (14 %), EGC (18 %), ECG (24 %) and EGCG (30 %). Similarly, N-Hcy HSA had lower Kb values toward EC (17 %), EGC (22 %), ECG (23 %) and EGCG (32 %). No differences were observed in the affinity between catechins, albumin and mercaptalbumin. CONCLUSION: Therefore, HSA posttranslational modifications typical of hyperhomocysteinemia reduce its affinity to catechins, potentially affecting their pharmacokinetics and availability at the active sites.
Assuntos
Catequina/metabolismo , Homocisteína/metabolismo , Processamento de Proteína Pós-Traducional , Albumina Sérica/metabolismo , Aminoacilação , Catequina/análogos & derivados , Cisteína/metabolismo , Eletroforese Capilar , Homocisteína/análogos & derivados , Humanos , Hiper-Homocisteinemia/sangue , Hiper-Homocisteinemia/metabolismo , Cinética , Ligantes , Lisina/metabolismo , Albumina Sérica HumanaRESUMO
Phenylalanine, tyrosine, and tryptophan, also known as aromatic amino acids, are involved in many physiological and pathophysiological conditions and are indicative of the liver and kidney function. In this work, we describe a simple and accurate method for their simultaneous quantification, in a single capillary electrophoresis run. This method requires minimal sample manipulation, no derivatization procedures, and methyl tryptophan as internal standard. The human blood plasma sample was precipitated using sulfosalicylic acid and the supernatant was used for the analysis. All the analytes were baseline resolved within 16 min and detected at 200 nm using Tris phosphate 80 mmol/L at pH 1.4 as the background electrolyte. The proposed method showed good linearity (r = 0.998) and repeatability (intra-assay RSD < 2.78%, interassay RSD < 5.4%) for all the analytes. The limit of quantification was 13 µmol/L for phenylalanine and 5 µmol/L for tyrosine and tryptophan. The method suitability was tested measuring aromatic amino acids level in 20 chronic kidney disease patients at basal level and after simvastatin/ezetimibe treatment.
Assuntos
Aminoácidos Aromáticos/sangue , Eletroforese Capilar/métodos , Espectrofotometria Ultravioleta/métodos , Idoso , Animais , Humanos , Pessoa de Meia-IdadeRESUMO
AIM: Since an increase in kynurenine (Kyn) plasma concentrations has been proposed as marker of immune system activation, we studied the associations between the Kyn levels and presence and severity of chronic obstructive pulmonary disease (COPD). METHODS & RESULTS: Plasma Kyn, tryptophan (Trp) and Kyn/Trp ratio were measured in 43 COPD patients with clinically defined mild (n = 29) or moderate (n = 14) disease and 43 age- and sex-matched healthy controls. When compared with controls, COPD patients had significantly higher plasma Kyn concentrations and Kyn/Trp ratios. In multiple logistic regression analysis, after adjusting for clinical and demographic confounders, the Kyn/Trp ratio was independently associated with COPD severity. DISCUSSION & CONCLUSION: Kyn and Kyn/Trp ratio might represent a new, sensitive, biomarker of systemic inflammation in COPD patients.
Assuntos
Biomarcadores/sangue , Cinurenina/sangue , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Triptofano/sangue , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Modelos Logísticos , Masculino , Estresse Oxidativo , Doença Pulmonar Obstrutiva Crônica/patologia , Sensibilidade e Especificidade , Índice de Gravidade de DoençaRESUMO
BACKGROUND: Two precolumn fluorescence derivatization procedures by two different sulfhydryl-reactive iodoacetyl reagents were established to measure simultaneously glutathione and l-ergothioneine in human whole blood by means of CE and LC. MATERIALS & METHODS: Separations were achieved in <5 min on a reverse-phase column (100 mm × 4.6 mm Zorbax Eclipse Plus C18 3.5 µm) for LC analysis, and on an uncoated fused-silica capillary (60 cm × 50 µm) for CE analysis, monitoring the fluorescence of derivatives. RESULTS: Performance of the assays was good in terms of linearity, recovery, intra- and inter-day precision and LOD and LOQ. CONCLUSION: This novel approach allows rapid assessment of circulating glutathione and l-ergothioneine concentrations for clinical and research purposes.
Assuntos
Cromatografia Líquida/métodos , Eletroforese Capilar/métodos , Ergotioneína/sangue , Glutationa/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Antioxidantes/farmacocinética , Feminino , Humanos , Limite de Detecção , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos TestesRESUMO
Hypertension, a common feature in chronic kidney disease (CKD), is an independent risk factor for CKD progression and cardiovascular disease. Although inhibitors of the renin-angiotensin system (RAS) exert salutary effects on blood pressure control and proteinuria in CKD patients, their activity towards traditional and novel oxidative markers is largely unknown. We studied the effects of 6-month treatment with telmisartan versus a combination of telmisartan and ramipril on plasma concentrations of low molecular mass (LMW, including homocysteine and cysteine) and protein thiols (PSH) plasma concentration and their relationships with carotid intima media thickness (IMT), in 24 hypertensive CKD patients (age 60 ± 12 years, 8 females and 16 males). Pretreatment PSH concentrations were independently associated with IMT (r = -0.42, p = 0.039). Neither treatment affected plasma LMW thiols, in both reduced and total form. By contrast, both treatments increased PSH plasma concentrations and reduced IMT, although significant differences were only observed in the combined treatment group. Our results suggest that the beneficial effects of combined RAS inhibitor treatment on IMT in hypertensive CKD patients may be mediated by a reduction of oxidative stress markers, particularly PSH.
Assuntos
Anti-Hipertensivos/uso terapêutico , Benzimidazóis/uso terapêutico , Benzoatos/uso terapêutico , Ramipril/uso terapêutico , Insuficiência Renal Crônica/tratamento farmacológico , Idoso , Anti-Hipertensivos/administração & dosagem , Anti-Hipertensivos/farmacologia , Benzimidazóis/administração & dosagem , Benzimidazóis/farmacologia , Benzoatos/administração & dosagem , Benzoatos/farmacologia , Artérias Carótidas/efeitos dos fármacos , Cisteína/sangue , Combinação de Medicamentos , Feminino , Homocisteína/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Ramipril/administração & dosagem , Ramipril/farmacologia , Insuficiência Renal Crônica/sangue , Telmisartan , Túnica Íntima/efeitos dos fármacosRESUMO
L-ergothioneine levels were measured in amniotic fluid of pregnant sheep after natural mating and transfer of vitrified/thawed in-vitro produced embryos. Amniotic fluids were collected between 60 and 65 and 80-85 days of gestation and analysed by an ultra-performance liquid chromatographic (UPLC)method with fluorescence detection. L-Ergothioneine concentrations ranged between 0.23 and 9.36 µmol/L and were significantly higher in pregnancy obtained by the transfer of vitrified/thawed in-vitro produced embryos. Conversely, no significant changes in amniotic fluid L-ergothioneine concentrations were observed according to the stages of pregnancy considered in this study. These findings suggest that L-ergothioneine concentrations, are not affected as much by the gestational age, but rather by the method used to induce the pregnancy. On the whole, the measurement of L-ergothioneine in amniotic fluid could serve as a useful biomarker of oxidative stress and/or inflammatory state in pregnancy.