ABSTRACT
Pediatric chronic kidney disease (CKD) is currently assessed using glomerular filtration rate (GFR), which is calculated from different equations based on serum creatinine concentration. However, serum creatinine is affected by several factors and is not reliable enough for the diagnosis of CKD, especially at early stages. Recent targeted and untargeted metabolomics studies found 7 new potential biomarkers that could be useful for early pediatric chronic kidney disease diagnosis. Thus, a new LC-QQQ-MS analytical method has been developed and validated aimed at routine analysis of these 7 potential biomarkers: NG,NG'-dimethyl-l-arginine di(p-hydroxyazobenzene-p'-sulfonate) (SDMA), S-adenosyl-l-methionine (SAM), n-butyryl-l-carnitine (nC4), iso-butyryl-l-carnitine (iC4), citrulline (CIT), creatinine (CNN) and d-erytro-sphingosine-1-phosphate (S1P), in addition to creatinine, the classical biomarker for CKD diagnosis. Then, this analytical method has been used for the quantification of plasma samples from a heterogeneous group of CKD pediatric patients and a control pediatric population. Data analysis of these results showed that it is possible to differentiate between CKD and control populations based on the metabolite concentration in plasma.
Subject(s)
Biomarkers/blood , Biomarkers/chemistry , Plasma/chemistry , Renal Insufficiency, Chronic/blood , Renal Insufficiency, Chronic/diagnosis , Arginine/analogs & derivatives , Arginine/blood , Arginine/chemistry , Child , Child, Preschool , Chromatography, Liquid/methods , Citrulline/chemistry , Creatinine/blood , Early Diagnosis , Female , Glomerular Filtration Rate/physiology , Humans , Male , Metabolomics/methods , S-Adenosylmethionine/blood , S-Adenosylmethionine/chemistry , Tandem Mass Spectrometry/methodsABSTRACT
Pediatric chronic kidney disease (CKD) is a clinical syndrome characterized by renal hypofunction occurring due to gradual and irreversible kidney damage that can further progress over time. New biomarkers may help early diagnosis of pediatric patients suffering from CKD and improve the outcome. Untargeted metabolomics based on LC-QTOF-MS has been used to find new biomarkers for the early diagnosis of CKD in plasma from pediatric patients. In order to avoid any bias in the determination of statistically significant entities as a consequence of the data analysis method followed, two different chemometric approaches have been used, Mass Profiler Professional (MPP) software and Matlab R2015a software. Metabolic fingerprints of control and CKD pediatric patients were compared and five metabolites which showed a significant change common to both data analysis procedures were identified. Sphingosine-1-phosphate, n-butyrylcarnitine, cis-4-decenoylcarnitine and an unidentified feature with 126.0930 m/z were found to be increased in plasma from pediatric patients with CKD, whereas bilirubin was significantly decreased. A partial least squares discriminant analysis model built with these 5 entities classified correctly 96% of the samples. In addition, when considering only early CKD patients against controls, a performance of 97% was obtained. Thus, these promising metabolites could be suitable biomarkers for the early diagnosis of pediatric CKD in a clinical setting.
Subject(s)
Biomarkers/blood , Metabolomics/methods , Renal Insufficiency, Chronic/blood , Renal Insufficiency, Chronic/diagnosis , Adolescent , Child , Child, Preschool , Chromatography, Liquid , Female , Humans , Male , Tandem Mass SpectrometryABSTRACT
Chronic kidney disease (CKD) is a progressive pathological condition in which renal function deteriorates in time. The first diagnosis of CKD is often carried out in general care attention by general practitioners by means of serum creatinine (CNN) levels. However, it lacks sensitivity and thus, there is a need for new robust biomarkers to allow the detection of kidney damage particularly in early stages. Multivariate data analysis of plasma concentrations obtained from LC-QTOF targeted metabolomics method may reveal metabolites suspicious of being either up-regulated or down-regulated from urea cycle, arginine methylation and arginine-creatine metabolic pathways in CKD pediatrics and controls. The results show that citrulline (CIT), symmetric dimethylarginine (SDMA) and S-adenosylmethionine (SAM) are interesting biomarkers to support diagnosis by CNN: early CKD samples and controls were classified with an increase in classification accuracy of 18% when using these 4 metabolites compared to CNN alone. These metabolites together allow classification of the samples into a definite stage of the disease with an accuracy of 74%, being the 90% of the misclassifications one level above or below the CKD stage set by the nephrologists. Finally, sex-related, age-related and treatment-related effects were studied, to evaluate whether changes in metabolite concentration could be attributable to these factors, and to correct them in case a new equation is developed with these potential biomarkers for the diagnosis and monitoring of pediatric CKD.
Subject(s)
Chromatography, High Pressure Liquid/methods , Metabolomics/methods , Renal Insufficiency, Chronic/diagnosis , Tandem Mass Spectrometry/methods , Adolescent , Age Factors , Arginine/analogs & derivatives , Arginine/blood , Arginine/metabolism , Biomarkers/blood , Child , Child, Preschool , Chromatography, High Pressure Liquid/instrumentation , Citrulline/blood , Citrulline/metabolism , Creatinine/blood , Creatinine/metabolism , Early Diagnosis , Female , Glomerular Filtration Rate , Humans , Male , Metabolic Networks and Pathways , Metabolomics/instrumentation , Multivariate Analysis , Renal Insufficiency, Chronic/blood , Renal Insufficiency, Chronic/metabolism , S-Adenosylmethionine/blood , S-Adenosylmethionine/metabolism , Sex Factors , Tandem Mass Spectrometry/instrumentationABSTRACT
An automated thermal desorption-gas chromatography-mass spectrometry method for the determination of triazin herbicides in aqueous solution with excellent sensitivity was developed. The method is based on the use of stir bar sorptive extraction. The main parameters such as extraction time, sample volume, the addition of salt and organic modifiers, desorption temperature, desorption flow and desorption time which affect the efficiency of the proposed methodology are fully discussed. The proposed method is sensitive and shows a good linearity within a range of 10-10000 ng L(-1) with correlation coefficients higher than 0.998. Quantitation limits are, in all cases, below the limits accepted by European legislation for human waters consumption and ranging between 11.3 ng L(-1) and 0.7 ng L(-1). The repetitivity, expressed as a relative standard deviation, has values of lower than 8% for all analytes. Using this method, determination of 10 triazines in underground water samples was successfully performed. The average concentrations obtained in the analysis of the spiked samples at two different levels of concentration correspond to mean recoveries ranging from 94.4+/-5.1% to 106.0+/-6.3% for a significance level of 0.05.
Subject(s)
Gas Chromatography-Mass Spectrometry/methods , Pesticide Residues/analysis , Triazines/analysis , Water Pollutants, Chemical/analysis , Adsorption , Chemical Fractionation/methods , Hot Temperature , Pesticide Residues/isolation & purification , Reproducibility of Results , Sensitivity and Specificity , Triazines/isolation & purification , Water Pollutants, Chemical/isolation & purificationABSTRACT
The aim of this study was to develop a methodology for the analysis of the insecticide fenitrothion and its two main environmental metabolites, fenitrooxon and 3-methyl-4-nitrophenol. For this purpose, a solid-phase microextraction (SPME) method coupled to high performance liquid chromatography (LC) was optimized. Two on-line detectors, diode array (DAD) and direct current amperometrical (DCAD) were used in order to determine sensitivity and selectivity. The effects of the extraction parameters, including exposure and desorption time, pH, temperature, salt concentration and desorption mode on the extraction efficiency were studied. A satisfactory reproducibility for extractions from samples at 20 ppb-level with RSD < 12.5% (n = 10) was obtained. The calibration graphs were linear in the range of 10-1000 microg l(-1) and detection limits for the target compounds were between 1.2 and 11.8 microg l(-1) depending on which detector was used. The method was applied for determining fenitrothion and both its metabolites in river waters which run through forest areas near to aerial application of the pesticide.
Subject(s)
Chromatography, High Pressure Liquid/methods , Electrochemistry/methods , Fenitrothion/analysis , Insecticides/analysis , Water Pollutants, Chemical/analysis , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
The selective monoalkylation at the smaller (lower) rim of the p-tert-butylcalix[4]- and -[6]arenes using bis(butyltin)oxide and different alkylating agents is described. The procedure is remarkable for the mild conditions used allowing an efficiently access to monoalkylated calixarene derivatives in moderate to good yields. Monoalkynylcalix[4]arene and monoalkynylcalix[6]arene have been synthetically exploited for the synthesis of bis-calix[n]arenes (n = 4, 6) with a diyne bridge by oxidative coupling of alkynes. In addition, intermolecular methathesis of the obtained monoalkenyl-calix[4]arene allowed the preparation of bis-calix[4]arene that are single bridged at the smaller (lower) rim with a 2-butenyl moiety.
ABSTRACT
Utilizando un sistema desarrollado para producir factor inhibidor de la migración de leucocitos (LIF) en altos títulos, hemos encontrado que cuando activamos linfocitos humanos con concanavalina-A, obtenemos LIF en 55% de los experimentos y factor estimulador de la migración en 38%. Cuando los linfocitos fueron activados por medio de cultivos mixtos de linfocitos encontramos resultados muy parecidos. Se seleccionaron sobrenadantes de linfocitos humanos activados con concanavalina-A que tuvieron una alta actividad de LIF. Al bloquear el LIF con N-acetil-D-glucosamina se pudo observar que estos sobrenadantes contenían también factor estimulador de la migración. En pruebas de LIF directo usando como antígeno a la estreptocinasa-estreptodornasa, encontramos que cuando el ensayo se realiza en presencia de N-acetil-D-glucosamina el valor de índice de migración se incrementa de manera importante. Concluimos que cuando los linfocitos humanos son estimulados ya sea por mitógenos, aloantígenos o antígenos, ellos producen tanto factor inhibidor como estimulador de la migración de leucocitos. Finalmente se discute la importancia de este hallazgo en la determinación de immunidad celular in vitro en humanos por medio de la prueba de LIF