Potential diagnostic assay for cystinuria by capillary electrophoresis coupled to mass spectrometry

Cystinuria is an autosomal recessive genetic disorder characterized by abnormal intestinal and renal tubular transport of L-cystine as well as of L-lysine, L-arginine and L-ornithine. This leads to excessive urinary excretion of amino acids, with the formation of kidney stones caused by the low solubility of L-cystine in the urine. In this study, an analytical method for simultaneous determination of these four amino acids in urine by capillary electrophoresis coupled to electrospray ionization mass spectrometry (CE-ESI-MS) was developed and validated. Using standard solutions of L-cystine, L-lysine, L-arginine and L-ornithine, the amino acid detection limits by this method were 114.2, 61.3, 72.7 and 86.7 µmol L-1. Standard solutions were injected in a silica capillary column (50 µm i.d. and 70 cm length) under 2 psi of pressure by 10 s. The separation occurred at 300 V cm-1, using 1.0 mol L-1 formic acid in 10% methanol in water as the background electrolyte. The method was applied to the urine of a patient clinically diagnosed as a cystinuria carrier, which revealed the presence of 900.5 ± 5, 600.0 ± 2, 700.2 ± 1 and 500.0 ± 3 µmol L-1 of amino acid, respectively, and 75.3 ± 1 µmol L-1 of creatinine. The CE-ESI-MS method described here for analyzing L-cystine and other cystinuria-related amino acids is a sensitive and reliable diagnostic tool for characterizing and monitoring this disease.Cistinúria é uma alteração genética autossômica recessiva caracterizada por transporte intestinal e renal anormal tubular de L-cistina, assim como de L-lisina, L-arginina e L-ornitina. Esta alteração leva a excreção urinária excessiva destes aminoácidos com a formação de pedras nos rins provocados pela baixa solubilidade de L-cistina na urina. Neste trabalho, um método analítico para a determinação destes quatro aminoácidos por eletroforese capilar acoplada à espectrometria de massas com ionização por electrospray (CE-ESI-MS) foi desenvolvido e validado. Usando soluções padrão de L-cistina, L-lisina, L-arginina e L-ornitina, os limites de detecção dos aminoácidos por este método foram 114,2, 61,3, 72,7 e 86,7 µmol L-1. Soluções padrão foram amostrados em um capilar de sílica (50 µm de diâmetro interno e 70 cm de comprimento total) e injeção de 2 psi de pressão por 10 s. A separação ocorreu a 300 V cm-1, utilizando 1,0 mol L-1 de ácido fórmico em 10% de metanol em água como eletrólito de separação. Aplicação do método para a urina de um paciente diagnosticado clinicamente como portador de cistinúria revelou a presença de 900,5 ± 5, 600,0 ± 2, 700,2 ± 1 e 500,0 ± 3 µmol L-1 de aminoácidos, respectivamente, e 75,3 ± 1 µmol L-1 de creatinina. O método de CE-ESI-MS descrito neste trabalho para a análise de L-cistina e outros aminoácidos associados com cistinúria constitui uma ferramenta para diagnóstico sensível e confiável para caracterização e monitoramento desta doença.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo (UNIFESP) Instituto de Ciências Ambientais, Química e FarmacêuticasUniversidade de São Paulo Instituto de QuímicaUNIFESP, Instituto de Ciências Ambientais, Química e FarmacêuticasSciEL

Efficient degradation of solid yeast biomass from ethanol industry by Fenton and UV-Fenton processes applying multivariate analysis

<div><p>Abstract Organic agro-industrial residues have been successfully used as biosorbents and promoting new uses from agricultural wastes benefits the economy. However, the allocation of a solid waste biosorbent after the sorption of contaminants has limited their effective application on a large scale as an alternative treatment of water and wastewaters. One solution could be degradation to convert the biosorbent material and adsorbed organic contaminants into environmental friendly compounds suitable for discharge. This study used an experimental design to evaluate the Fenton degradation of yeast biomass (YB) from the alcohol industry as a potential biosorbent. The efficiency of degradation was monitored according to the degraded mass (DM) and total organic carbon (TOC) remaining in the solution. The ANOVA showed an error of 9.7% for the effects and the media of interaction for the employed model for DM. Conducting the experiments with the best-predicted conditions (60 min, 25 g of YB, pH 3, 8,000 mg L-1 H2O2 and 40 mg L-1 Fe2+) with 30 W UV irradiation resulted in a YB reduction of 72 ( 2% with a TOC of 30 ( 2%. This suggests that an advanced oxidative process is an alternative for degradation of a biosorbent after sorption.</p></div

Geoeconomic variations in epidemiology, ventilation management, and outcomes in invasively ventilated intensive care unit patients without acute respiratory distress syndrome: a pooled analysis of four observational studies

Background: Geoeconomic variations in epidemiology, the practice of ventilation, and outcome in invasively ventilated intensive care unit (ICU) patients without acute respiratory distress syndrome (ARDS) remain unexplored. In this analysis we aim to address these gaps using individual patient data of four large observational studies. Methods: In this pooled analysis we harmonised individual patient data from the ERICC, LUNG SAFE, PRoVENT, and PRoVENT-iMiC prospective observational studies, which were conducted from June, 2011, to December, 2018, in 534 ICUs in 54 countries. We used the 2016 World Bank classification to define two geoeconomic regions: middle-income countries (MICs) and high-income countries (HICs). ARDS was defined according to the Berlin criteria. Descriptive statistics were used to compare patients in MICs versus HICs. The primary outcome was the use of low tidal volume ventilation (LTVV) for the first 3 days of mechanical ventilation. Secondary outcomes were key ventilation parameters (tidal volume size, positive end-expiratory pressure, fraction of inspired oxygen, peak pressure, plateau pressure, driving pressure, and respiratory rate), patient characteristics, the risk for and actual development of acute respiratory distress syndrome after the first day of ventilation, duration of ventilation, ICU length of stay, and ICU mortality. Findings: Of the 7608 patients included in the original studies, this analysis included 3852 patients without ARDS, of whom 2345 were from MICs and 1507 were from HICs. Patients in MICs were younger, shorter and with a slightly lower body-mass index, more often had diabetes and active cancer, but less often chronic obstructive pulmonary disease and heart failure than patients from HICs. Sequential organ failure assessment scores were similar in MICs and HICs. Use of LTVV in MICs and HICs was comparable (42·4% vs 44·2%; absolute difference -1·69 [-9·58 to 6·11] p=0·67; data available in 3174 [82%] of 3852 patients). The median applied positive end expiratory pressure was lower in MICs than in HICs (5 [IQR 5-8] vs 6 [5-8] cm H2O; p=0·0011). ICU mortality was higher in MICs than in HICs (30·5% vs 19·9%; p=0·0004; adjusted effect 16·41% [95% CI 9·52-23·52]; p&lt;0·0001) and was inversely associated with gross domestic product (adjusted odds ratio for a US$10 000 increase per capita 0·80 [95% CI 0·75-0·86]; p&lt;0·0001). Interpretation: Despite similar disease severity and ventilation management, ICU mortality in patients without ARDS is higher in MICs than in HICs, with a strong association with country-level economic status