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1.
Minerva Chir ; 71(3): 168-72, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26112363

RESUMO

BACKGROUND: The aim of this study was to study the clinical significance of joint detection of mALB and NAG in early kidney damage in burn patients. METHODS: Forty-five burn patients of different degrees were selected and divided into mild, moderate, severe, and heavy burns, and normal healthy controls according to their severity. Their b2- macroglobulin (b2-MG), a1-macroglobulin (a1-MG), mALB and N-acetyl-b-D-NAG were tested for 3 days, 1 week, 2 weeks, 4 weeks after the burn, respectively. RESULTS: The urine concentration change b2-MG, a1-MG, MALB, and NAG are related to the area, depth and degree of the burn. The more serious the burn is, the higher the levels of mALB and NAG (P<0.001 or P<0.01) is. CONCLUSIONS: Early detection of mALB and NAG is helpful for early kidney damage diagnosis in burn patients to prevent further complications.


Assuntos
Queimaduras/complicações , Nefropatias/diagnóstico , Nefropatias/urina , Proteínas de Neoplasias/urina , alfa-N-Acetilgalactosaminidase/urina , Adolescente , Adulto , Biomarcadores/urina , Criança , Pré-Escolar , Feminino , Humanos , Nefropatias/etiologia , Nefropatias/metabolismo , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Sensibilidade e Especificidade , Índice de Gravidade de Doença
2.
Rapid Commun Mass Spectrom ; 29(21): 1929-37, 2015 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-26443390

RESUMO

RATIONALE: Schindler disease is caused by the deficient activity of α-N-acetylgalactosaminidase, which leads to an abnormal accumulation of O-glycopeptides in tissues and body fluids. In this work the Schindler condition is for the first time approached by ion mobility (IMS) tandem mass spectrometry (MS/MS), for determining urine glycopeptide fingerprints and discriminate isomeric structures. METHODS: IMS-MS experiments were conducted on a Synapt G2s mass spectrometer operating in negative ion mode. A glycopeptide mixture extracted from the urine of a patient suffering from Schindler disease was dissolved in methanol and infused into the mass spectrometer by electrospray ionization using a syringe-pump system. MS/MS was performed by collision-induced dissociation (CID) at low energies, after mobility separation in the transfer cell. Data acquisition and processing were performed using MassLynx and Waters Driftscope software. RESULTS: IMS-MS data indicated that the attachment of one or two amino acids to the carbohydrate backbone has a minimal influence on the molecule conformation, which limits the discrimination of the free oligosaccharides from the glycosylated amino acids and dipeptides. The structural analysis by CID MS/MS in combination with IMS-MS of species exhibiting the same m/z but different configurations demonstrated for the first time the presence of positional isomers for some of the Schindler disease biomarker candidates. CONCLUSIONS: The IMS-MS and CID MS/MS platform was for the first time optimized and applied to Schindler disease glycourinome. By this approach the separation and characterization of Neu5Ac positional isomers was possible. IMS CID MS/MS showed the ability to determine the type of the glycopeptide isomers from a series of possible candidates.


Assuntos
Glicopeptídeos/química , Glicopeptídeos/urina , Doenças por Armazenamento dos Lisossomos/urina , Distrofias Neuroaxonais/urina , Espectrometria de Massas em Tandem/métodos , alfa-N-Acetilgalactosaminidase/deficiência , Pré-Escolar , Humanos , Isomerismo , Masculino , alfa-N-Acetilgalactosaminidase/urina
3.
J Proteome Res ; 14(8): 3123-35, 2015 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-26143644

RESUMO

Individuals with type 1 diabetes (T1D) often have higher than normal blood glucose levels, causing advanced glycation end product formation and inflammation and increasing the risk of vascular complications years or decades later. To examine the urinary proteome in juveniles with T1D for signatures indicative of inflammatory consequences of hyperglycemia, we profiled the proteome of 40 T1D patients with an average of 6.3 years after disease onset and normal or elevated HbA1C levels, in comparison with a cohort of 41 healthy siblings. Using shotgun proteomics, 1036 proteins were identified, on average, per experiment, and 50 proteins showed significant abundance differences using a Wilcoxon signed-rank test (FDR q-value ≤ 0.05). Thirteen lysosomal proteins were increased in abundance in the T1D versus control cohort. Fifteen proteins with functional roles in vascular permeability and adhesion were quantitatively changed, including CD166 antigen and angiotensin-converting enzyme 2. α-N-Acetyl-galactosaminidase and α-fucosidase 2, two differentially abundant lysosomal enzymes, were detected in western blots with often elevated quantities in the T1D versus control cohort. Increased release of proteins derived from lysosomes and vascular epithelium into urine may result from hyperglycemia-associated inflammation in the kidney vasculature.


Assuntos
Diabetes Mellitus Tipo 1/urina , Enzimas/urina , Proteoma/metabolismo , Proteômica/métodos , Irmãos , Molécula de Adesão de Leucócito Ativado/metabolismo , Molécula de Adesão de Leucócito Ativado/urina , Adolescente , Enzima de Conversão de Angiotensina 2 , Western Blotting , Criança , Cromatografia Líquida , Estudos de Coortes , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/metabolismo , Enzimas/metabolismo , Feminino , Humanos , Lisossomos/enzimologia , Lisossomos/metabolismo , Masculino , Peptidil Dipeptidase A/metabolismo , Peptidil Dipeptidase A/urina , Espectrometria de Massas em Tandem , alfa-L-Fucosidase/metabolismo , alfa-L-Fucosidase/urina , alfa-N-Acetilgalactosaminidase/metabolismo , alfa-N-Acetilgalactosaminidase/urina
4.
Carbohydr Res ; 398: 90-100, 2014 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-25243357

RESUMO

In this study an integrative mass spectrometry (MS) approach based on fully automated chip-nanoelectrospray quadrupole time-of-flight was optimized and applied for the discovery and structural characterization of O-glycopeptides in a fraction from the urine of a patient diagnosed with Schindler disease type I. A mixture of O-glycopeptides extracted and purified from an age matched healthy subject served as the control. 49 glycoforms were discovered in the investigated urine fraction from Schindler disease versus only 14 in control urine. Structures with relevant biological significance, previously not described, such as O-fucosylated tetrasaccharides and chains up to pentadecamers O-linked to serine, threonine, or threonine-proline were identified in the pathological urine and characterized by tandem MS (MS/MS). A number of 29 species discovered here, most of which with long chain glycans, were not previously reported as associated to this condition. All glycopeptides were detected in only 1 min analysis time, with a sample consumption situated in the femtomole range.


Assuntos
Glicopeptídeos/urina , Doenças por Armazenamento dos Lisossomos/urina , Distrofias Neuroaxonais/urina , Espectrometria de Massas por Ionização por Electrospray/métodos , alfa-N-Acetilgalactosaminidase/deficiência , Automação , Sequência de Carboidratos , Criança , Glicopeptídeos/química , Humanos , Dados de Sequência Molecular , Nanotecnologia/métodos , alfa-N-Acetilgalactosaminidase/urina
5.
Anal Bioanal Chem ; 406(18): 4337-43, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24788891

RESUMO

The most widely used method for the biochemical screening of oligosaccharidoses is the analysis of the urinary oligosaccharide pattern by thin-layer chromatography on silica gel plates. However, this method is not always sensitive enough, and it is extremely time-consuming and laborious. In this work, the analysis of the urine oligosaccharide pattern was standardized for the first time by using capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection (Beckman P/ACE MDQ) with a 488-nm argon ion laser module. All of the analyses were conducted using the Carbohydrate Labeling and Analysis Kit (Beckman-Coulter), which derivatizes samples with 8-aminopyrene-1,3,6-trisulfonate. Urine samples from 40 control subjects (age range, 1 week to 16 years) and from ten patients diagnosed with eight different lysosomal diseases (six of them included in the Educational Oligosaccharide Kit from ERNDIM EQA schemes) were analyzed. Two oligosaccharide excretion patterns were established in our control population according to age (younger or older than 1 year of age). Abnormal peaks with slower migration times than the tetrasaccharide position were observed for fucosidosis, α-mannosidosis, GM1 gangliosidosis, GM2 gangliosidosis variant 0, Pompe disease, and glycogen storage disease type 3. In conclusion, the first CE-LIF method to screen for oligosaccharidoses and related diseases, which also present oligosacchariduria, has been standardized. In all of the cases, the urine oligosaccharide analysis was strongly informative and showed abnormal patterns that were not present in any of the urine samples from the control subjects. Only urine from patients with aspartylglucosaminuria and Schindler disease displayed normal results.


Assuntos
Eletroforese Capilar/métodos , Doenças por Armazenamento dos Lisossomos/urina , Oligossacarídeos/urina , Adolescente , Aspartilglucosaminúria/urina , Estudos de Casos e Controles , Criança , Pré-Escolar , Eletroforese Capilar/instrumentação , Eletroforese Capilar/normas , Fucosidose/urina , Doença de Depósito de Glicogênio Tipo II/urina , Humanos , Lactente , Recém-Nascido , Lasers , Doenças por Armazenamento dos Lisossomos/diagnóstico , Distrofias Neuroaxonais/urina , Doença de Sandhoff/urina , alfa-N-Acetilgalactosaminidase/deficiência , alfa-N-Acetilgalactosaminidase/urina
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