Non-invasive urinary metabolomic profiles discriminate biliary atresia from infantile hepatitis syndrome.

INTRODUCTION: Neonatal cholestatic disorders are a group of hepatobiliary diseases occurring in the first 3 months of life. The most common causes of neonatal cholestasis are infantile hepatitis syndrome (IHS) and biliary atresia (BA). The clinical manifestations of the two diseases are too similar to distinguish them. However, early detection is very important in improving the clinical outcome of BA. Currently, a liver biopsy is the only proven and effective method used to differentially diagnose these two similar diseases in the clinic. However, this method is invasive. Therefore, sensitive and non-invasive biomarkers are needed to effectively differentiate between BA and IHS. We hypothesized that urinary metabolomics can produce unique metabolite profiles for BA and IHS. OBJECTIVES: The aim of this study was to characterize urinary metabolomic profiles in infants with BA and IHS, and to identify differences among infants with BA, IHS, and normal controls (NC). METHODS: Urine samples along with patient characteristics were obtained from 25 BA, 38 IHS, and 38 NC infants. A non-targeted gas chromatography-mass spectrometry (GC-MS) metabolomics method was used in conjunction with orthogonal partial least squares discriminant analysis (OPLS-DA) to explore the metabolomic profiles of BA, IHS, and NC infants. RESULTS: In total, 41 differentially expressed metabolites between BA vs. NC, IHS vs. NC, and BA vs. IHS were identified. N-acetyl-D-mannosamine and alpha-aminoadipic acid were found to be highly accurate at distinguishing between BA and IHS. CONCLUSIONS: BA and IHS infants have specific urinary metabolomic profiles. The results of our study underscore the clinical potential of metabolomic profiling to uncover metabolic changes that could be used to discriminate BA from IHS.

Latent distal renal tubular acidosis (dRTA) in primary biliary cirrhosis (PBC) and chronic autoimmune hepatitis (CAH).

In an attempt to evaluate the latent distal renal tubular acidosis (dRTA7) in patients with primary biliary cirrhosis (PBC), and chronic autoimmune hepatitis (CAH), and differences between them in relation to the sodium urinary excretion ([Na]u), thirty four patients divided in two groups were studied. Group A: 17 patients who fulfilled criteria for PBC diagnosis (clinical and humoral evidence antimitochondrial antibody titles of 1/80 or above by indirect immunofluorescence technique, and liver biopsy). Group B: 17 patients who fulfilled criteria for CAH diagnosis (clinical and humoral evidence, antinuclear and smooth muscle antibody titles of 1/80 or above and liver biopsy). Patients with ascitis and/or edema were excluded from the study. Ability to acidify urine was evaluated by gradient between pC02 in urine and blood (U-BpC02) after alkali infusion. Five patients in Group A (29.4%7) and six in Group B(35.2%) had dRTA, (p = 0.49). When analyzing patients with dRTA in both groups, the mean [Na]u in Group A was 152.2 +/- 33.8, versus 50.8 +/- 8.1 mEq/l, in Group B. (p = 0.00016). We concluded that the prevalence of dRTA was similar in patients with PBC and CAH but the urinary acidifications impairment of the former did not correlate with [Na]u, as it did with the latter.

Latent distal renal tubular acidosis (dRTA) in primary biliary cirrhosis (PBC) and chronic autoinmune hepatitis (CAH)

In an attempt to evaluate the latent distal renal tubular acidosis (dRTA7) in patients with primary biliary cirrhosis (PBC), and chronic autoinmmune hepatitis (CAH), and differences between them in relation to the sodium urinary excretion ([a]u), thirty four patients divided in two groups were studied. Group A: 17 patients who fullfilled criteria for PBC diagnosis (clinical and humoral and liver biopsy). Group B: 17 patients who fullfilled criteria for CAH diagnosis (clinical and humoral evidence, antinuclear and smooth muscle antibody tiles of 1/80 or above and liver biopsy). Patients with ascitis and/or edema were excluded form the study. Ability to acidify urine was evaluated by gradient between pCO2 in urine and blood (U-BpC02) after alkali infusion. Five patients with dRTA in both groups, the mean [Na]u in Group A was 152.2 ñ 33.8, versus 50.8 ñ 8.1 mEq/l, in Group B. (p=0.00016). We concluded that the prevalence of dRTA was similar en patiens with PBC and CAH but the urinary acidifications impairment of the former did not correlate with [Na]u, as it did whit the latter

Determinación de substancias reductoras en la orina como método inicial para el diagnóstico temprano de galactosemia e intolerancia hereditaria a la fructuosa en niños con hepatitis / Determination of the reducing substances in the urine as a diagnostic method for galactosemia and fructose intolerance in children with neonatal hepatitis

La determinación de substancias reductoras en orina, se realizó en 40 niños con hepatitis neonatal utilizando, para ello, la prueba de Clinitest y Benedict. Sólo en dos niños se obtuvo como resultado una reacción falsa positiva. Se concluye que el clinitest es un método sencillo, práctico, económico y de gran utilidad para la detección temprana e inicial de galactosemia e intolerancia a la fructuosa. Las que deberán corroborarse en caso necesario posteriormente con un estudio metabólico completo

The identification of urinary bile alcohols by gas chromatography-mass spectrometry in patients with liver disease and in healthy individuals.

The neutral steroid fractions in the urine of eleven patients suffering from various forms of liver disease with cholestasis and of ten healthy individuals were studied by glass capillary gas chromatography-mass spectrometry. The steroid conjugates in urine were enzymatically solvolysed, the liberated steroids extracted and transformed into the trimethylsilylether for measurements. The excretion rates of androstane and pregnane metabolites of patients with liver disease were far lower than those of healthy persons. The main compounds in the urine of the former were the bile alcohols 27 - nor - 3 alpha, 7 alpha, 12 alpha, 24 xi, 25 xi - pentahydroxy - 5 beta - cholestane and 3 alpha, 7 alpha, 12 alpha, 25 xi, 26 - pentahydroxy - 5 beta - cholestane. Our data suggest a correlation between the excretion rates of these bile alcohols and the serum levels of bilirubin. While the excretion rate of the two bile alcohols in the urine of healthy individuals was approximately 0.24 mg/24 h (0.6 mumol/24 h) a patient with a serum bilirubin of 841 mumol/l excreted 4 mg/24 h (9 mumol/24 h). The accumulation of bile alcohols described in this study possibly indicates alternative pathways of cholic acid formation in liver disease.

Urinary excretion of methylarginine in human disease.

Methylated amino acids are excreted in urine upon degradation of some tissue proteins. The urinary excretion ratios of NG,N'G-dimethylarginine (syn-DMA) and NG,NG-dimethylarginine (unsym-DMA) were studied in healthy adults and in patients with various diseases. The normal ratio of sym- to unsym-DMA in urine was 0.98 and ranged from 0.71 to 1.33; ratios were not significantly different in multiple sclerosis, cerebrovascular accident, cancer, and systemic lupus erythematosus. However, patients with liver, disease, including chronic active hepatitis, were found on average to have a significantly altered ratio of 0.79, range 0.49-1.30, owing to an increase in the excretion of unsym-DMA. Hence measurements of the urinary excretion of dimethylarginine could become a useful aid in assessing recovery of liver cells in patients with chronic liver disease.

Ratio of urinary 6beta-hydroxycortisol to 17-hydroxycorticosteroids in patients with liver disease.

The ratio of 6beta-hydroxycortisol to 17-hydroxycorticosteroids in the urine of 73 patients with liver disease, and 53 controls has been measured. The mean ratio was significantly greater in the liver disease group (p less than 0.001), this elevation being most marked among patients with liver metastases and patients with acute hepatitis. We feel that the use of the ratio as a test of early liver cell dysfunction requires further evaluation, in particular in the early recognition of metastatic liver disease.

Cellular elements in the urine in health and in acute infectious diseases, especially with respect to the presence of haematuria. A study with application of millipore procedure and Papanicolaou staining.

The excretion of cellular per litre of urine amounted in healthy persons to, in round figures, one million epithelial cells (2.5 cells per visual field) in both sexes, one million leukocytes in males, one million erythrocytes in females and 0.5 million in males. The maximal excretion was calculated to be 5-6 million per litre. In acute infections the number of epithelial cells and leukocytes in the urine rose to more than the double. Pathological microscopic haematuria, judged by exceeding of the maximal value for normal excretion during the acute phase (24 or more erythrocytes per visual field), occurred in no case of mycoplasma infection, in about 4% of measles, mononucleosis, serous meningitis and hepatitis cases, in about 8% of mumps and streptococcal infections, and in more than 20% of influenza A2 cases. Statistical significance or probable significant existed between influenza and other diseases. The haematuria was unrelated either to the general degenerative or to the specific inclusion-provocative reaction within the renal and urinary tract epithelium. The cause is sought in an involvement of glomeruli with increased diapedesis. The special position of influenza may be explained by the marked haemorrhagic reactions produced by this infection. In one case persistent haematuria combined with increased content of inclusion-bearing cells occurred after influenza. Immunoglobulin deposition in glomerular mesangium may perhaps be one explanation of this haematuria.

Measurement of sulfated and nonsulfated bile acids in human serum and urine.

Amberlite XAD-2 was used to extract bile acids from urine or diluted serum of patients with hepatobiliary diseases. Columns containing Sephadex LH-20 were then used to separate the sulfated and nonsulfated bile acids. Thin-layer chromatography of the sulfated bile acid fraction obtained from urine revealed several spots with R(F) values different from those of the taurine or glycine conjugates. According to thin-layer chromatographic mobilities, gas-liquid chromatographic analyses, infrared spectra, and elementary analysis of the sulfated material, one of these sulfated bile acids was identified as glycochenodeoxycholic acid monosulfate, and the others were presumed to be taurochenodeoxycholic acid sulfate and glycocholic acid sulfate. A large amount of bile acid sulfate was found in urine of patients with hepatobiliary diseases. They accounted for 35.5-93.3% of total urinary bile acids and consisted of both di- and trihydroxycholanoic acids, with chenodeoxycholic acid as the major acid. Sulfated bile acids were also found in serum, and accounted for 1.8-21.2% of the total bile acids. Only dihydroxycholanoic acids (mainly chenodeoxycholic) were identified.