RESUMO
BACKGROUND: Blood gene expression profiling has been used in several studies to identify patients with a number of conditions and diseases. A blood test with the ability to differentiate Crohn's disease (CD) from ulcerative colitis (UC) and noninflammatory diarrhea would be useful in the clinical management of these diseases. METHODS: Affymetrix U133Plus 2.0 GeneChip oligonucleotide arrays were used to generate whole blood gene expression profiles for 21 patients with UC, 24 patients with CD, and 10 control patients with diarrhea, but without colonic pathology. RESULTS: A supervised learning method (logistic regression) was used to identify specific panels of probe sets which were able to discriminate between UC and CD and from controls. The UC panel consisted of the four genes, CD300A, KPNA4, IL1R2, and ELAVL1; the CD panel comprised the four genes CAP1, BID, NIT2, and NPL. These panels clearly differentiated between CD and UC. CONCLUSIONS: Gene expression profiles from blood can differentiate patients with CD from those with UC and from noninflammatory diarrheal disorders.
Assuntos
Biomarcadores/sangue , Colite Ulcerativa/diagnóstico , Doença de Crohn/diagnóstico , Diarreia/diagnóstico , Perfilação da Expressão Gênica , Adulto , Idoso , Aminoidrolases/sangue , Aminoidrolases/genética , Antígenos CD/sangue , Antígenos CD/genética , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/sangue , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/genética , Proteínas de Ciclo Celular/sangue , Proteínas de Ciclo Celular/genética , Colite Ulcerativa/sangue , Colite Ulcerativa/genética , Doença de Crohn/sangue , Doença de Crohn/genética , Proteínas do Citoesqueleto/sangue , Proteínas do Citoesqueleto/genética , Diarreia/sangue , Diarreia/genética , Proteínas ELAV/sangue , Proteínas ELAV/genética , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Oxo-Ácido-Liases/sangue , Oxo-Ácido-Liases/genética , Receptores Imunológicos/sangue , Receptores Imunológicos/genética , Receptores Tipo II de Interleucina-1/sangue , Receptores Tipo II de Interleucina-1/genética , alfa Carioferinas/sangue , alfa Carioferinas/genéticaAssuntos
Argininossuccinato Liase/genética , Corantes , Erros Inatos do Metabolismo/sangue , Oxo-Ácido-Liases/genética , Erros Inatos do Metabolismo dos Aminoácidos/sangue , Erros Inatos do Metabolismo dos Aminoácidos/diagnóstico , Argininossuccinato Liase/sangue , Acidúria Argininossuccínica , Reações Falso-Positivas , Genótipo , Humanos , Erros Inatos do Metabolismo/diagnóstico , Oxo-Ácido-Liases/sangue , Oxo-Ácido-Liases/deficiência , Reação em Cadeia da Polimerase/métodosRESUMO
Sialate pyruvate-lyases, also known as sialate aldolases (EC 4.1.3.3), reversibly catalyse the cleavage of free N-acetylneuraminic acids to form pyruvate and N-acetylmannosamine. These enzymes are widely distributed and are present in numerous pro- and eukaryotic cells, in which they are localized only in the cytosol. They play an important role in the regulation of sialic acid metabolism by controlling the intracellular concentration of sialic acids of biosynthetic or exogenous origin, thus preventing the accumulation of toxic levels of this sugar. Application of an original colorimetric micromethod for N-acetylmannosamine determination, as well as the use of [4,5,6,7,8,9-14C]N-acetylneuraminic acid, led us to evidence a cytosolic neuraminate aldolase activity in human red blood cells (RBCs) and then to define the main characteristics of this enzyme: Michaelis-Menten type, K(m:) 1.4 +/- 0.05 mM, optimal pH: 7.6 +/- 0.2, optimal temperature: 70 +/- 2 degrees C, inhibition by heavy metals: Ag(+) and Hg(++). These enzyme parameters are close to those of the bacterial and mammalian aldolases described up to now. At the moment, the presence of sialate pyruvate-lyase in the cytosol of red blood cells remains an enigma.
Assuntos
Eritrócitos/enzimologia , Oxo-Ácido-Liases/sangue , Autorradiografia , Radioisótopos de Carbono , Catálise , Cátions/farmacologia , Cromatografia em Camada Fina , Citosol/enzimologia , Humanos , Concentração de Íons de Hidrogênio , Cinética , Metais/farmacologia , Ácido N-Acetilneuramínico/química , Ácido N-Acetilneuramínico/metabolismo , Ácido Pirúvico/farmacologia , Especificidade por Substrato , Temperatura , Fatores de TempoRESUMO
Patients with 3-hydroxy-3-methylglutaric aciduria due to a deficiency of 3-hydroxy-3-methylglutaryl Coenzyme A lyase usually present with a life-threatening crisis of hypoglycemia, metabolic acidosis and hyperammonemia. Diagnosis of this inborn error of leucine degradation is usually based upon gas-chromatographic analysis of organic acids in a patient's urine. In this paper we describe a simple spectrophotometric method allowing the activity of HMG-CoA lyase to be measured in leukocytes or platelets within a few hours, thus contributing to a rapid, unequivocal diagnosis and subsequent treatment. The validity of the method was established by demonstrating a deficient activity of HMG-CoA lyase in two patients with 3-hydroxy-3-methylglutaric aciduria. Furthermore, using this method, heterozygote detection can be done with great reliability.
Assuntos
Plaquetas/enzimologia , Leucócitos/enzimologia , Oxo-Ácido-Liases/deficiência , Pré-Escolar , Feminino , Humanos , Concentração de Íons de Hidrogênio , Lactente , Recém-Nascido , Masculino , Erros Inatos do Metabolismo/sangue , Erros Inatos do Metabolismo/diagnóstico , Oxo-Ácido-Liases/sangue , Espectrofotometria/métodosRESUMO
A girl, now three years old, is reported, in whom at the age of 5 months the diagnosis of 3-HMG-CoA lyase deficiency was established. The characteristic excretion pattern consisted of 3-HMG, 3-CH3-glutaconic acid, 3-CH3-glutaric acid and 3-HIVA. Activity of 3-HMG-CoA lyase in leucocytes was very low. She had compensated metabolic acidosis and mild hypoglycemia. Therapy with a leucine restricted diet decreased excretion of metabolites moderately but did not influence the tendency to metabolic acidosis. Clinically the infant presented with macrocephaly. At the age of 3 years she is severely retarded. CAT scan revealed the picture of progressive demyelination of the white matter.
