[Case of 5 year-old boy with anaphylaxis due to erythritol with negative prick test and positive intradermal test].

A 5 year-old boy experienced anaphylaxis after eating a jelly product for diet supplement containing erythritol as a major component. Prick test with the jelly product was negative, but the second oral ingestion of the jelly product at home caused another allergic reaction. Prick test with erythritol was negative even at 300 mg/ml, which was almost the solubility limit. Intradermal test was marginally positive at 0.1 mg/ml, and clearly positive at 1 mg/ml or higher concentration. We found subtle dose-response reaction utilizing basophil activation test, examined with 24 hour incubation at the concentration of 40-4000 µg/ml. At the oral challenge test in the hospital, 3 g of erythritol induced remarkable coughing, urticaria, edema, wheezing and hypoxemia. Erythritol is a natural sugar alcohol, with the molecular weight of 122.12, which is recently being widely used for diet supplements, beverages, or drug medicines due to its properties of calorie-free and good-tasting, with easy-to-use physical characteristics. We now have to recognize erythritol as a candidate for food allergen, and to be careful about negative result of prick test.

Detection of Δ4-3-oxo-steroid 5ß-reductase deficiency by LC-ESI-MS/MS measurement of urinary bile acids.

The synthesis of bile salts from cholesterol is a complex biochemical pathway involving at least 16 enzymes. Most inborn errors of bile acid biosynthesis result in excessive formation of intermediates and/or their metabolites that accumulate in blood and are excreted in part in urine. Early detection is important as oral therapy with bile acids results in improvement. In the past, these intermediates in bile acid biosynthesis have been detected in neonatal blood or urine by screening with FAB-MS followed by detailed characterization using GC-MS. Both methods have proved difficult to automate, and currently most laboratories screen candidate samples using LC-MS/MS. Here, we describe a new, simple and sensitive analytical method for the identification and characterization of 39 conjugated and unconjugated bile acids, including Δ(4)-3-oxo- and Δ(4,6)-3-oxo-bile acids (markers for Δ(4)-3-oxo-steroid 5ß-reductase deficiency), using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In this procedure a concentrated, desalted urinary sample (diluted with ethanol) is injected directly into the LC-ESI-MS/MS, operated with ESI and in the negative ion mode; quantification is obtained by selected reaction monitoring (SRM). To evaluate the performance of our new method, we compared it to a validated method using GC-MS, in the analysis of urine from two patients with genetically confirmed Δ(4)-3-oxo-steroid 5ß-reductase deficiency as well as a third patient with an elevated concentration of abnormal conjugated and unconjugated Δ(4)-3-oxo-bile acids. The Δ(4)-3-oxo-bile acids concentration recovered in three patients with 5ß-reductase deficiency were 48.8, 58.9, and 49.4 µmol/mmol creatinine, respectively by LC-ESI-MS/MS.

Diagnostic determination system for high-risk screening for inborn errors of bile acid metabolism based on an analysis of urinary bile acids using gas chromatography-mass spectrometry: results for 10 years in Japan.

BACKGROUND: Some patients with cholestasis of unknown cause may have inborn errors of bile acid metabolism (IEBAM) thus causing abnormalities of bile acid biosynthesis. Although seven types of bile acid synthetic defects have thus far been reported for this disorder, no detailed information on its incidence and so on in Japan is yet available. In order to elucidate the current status of IEBAM in Japan, in July 1996 a diagnostic determination system was established for high-risk screening for IEBAM. METHODS: Urinary bile acids were analyzed on gas chromatography-mass spectrometry (GC-MS) and quantitative analysis was done using selected ion monitoring (SIM). RESULTS AND CONCLUSIONS: In a total of 576 samples analyzed over the 10 year period prior to June 2005, 159 patients were found with cholestasis of unknown etiology. Of these patients, 10 (6.3%) were found to have IEBAM by this system, while 91 (61.1%) had cholestasis without a definitive diagnosis. This diagnostic determination system with GC-MS of urinary bile acids is therefore considered useful for detecting IEBAM.

Bone mineral density in children and adolescent girls with anorexia nervosa in Japan.

BACKGROUND: The correlation between reduced bone mineral density (BMD) and the disease anorexia nervosa (AN) has long been established. The aim of the present study was to examine the relationship in more detail, particularly focusing on the increasing incidence of the disease occurring in adolescent patients. METHOD: Twenty-four girls diagnosed with AN were enrolled in the study. All subjects ranged in age from 11.1 to 15.5 years, with an average age of 13.5 years. The BMD of lumbar spines and femoral necks were measured. All the values for BMD at admission were expressed as means +/- SD and patients with and without menarche were separately investigated. RESULTS: The average BMD of lumbar spines at the time of admission was -0.51 SD in total. However, the average BMD of patients without menarche was -1.28 SD, which was significantly lower than the -0.16 SD on average in patients with menarche. As a whole the BMD of femoral necks at admission tended to be lower than that of lumbar spines. Similarly, it was lower in patients without menarche (-1.7 SD on average) than in those with menarche (-0.77 SD on average). CONCLUSIONS: BMD was lower in children and adolescent AN patients without menarche, and such a tendency was more significant at the femoral neck region. In child AN cases without menarche, the BMD, especially at the femoral neck, needs to be measured, and later recovery should be monitored closely over a long period.

TRRAP as a hepatic coactivator of LXR and FXR function.

TBP-free TAF II-containing-type HAT complex subclasses, which contain hGCN5 HAT and TRRAP, appear to act as common coactivator complexes for nuclear receptors. However, their physiological significance with respect to each nuclear receptor remains to be established. To address this issue, we used hepatic cell lines (HepG2) with reduced endogenous TRRAP expression through antisense RNA expression or with overexpressed TRRAP or other major coactivators. The ligand-induced transactivation function of liver X receptor alpha (LXRalpha) and farnesoid X receptor/bile acid receptor reflected TRRAP expression levels, while that of PPARgamma did not. A GST pull-down assay indicated that TRRAP contains two potential LXRalpha-interacting domains in the C-terminal and central domains. Expression of antisense TRRAP RNA in HepG2 cells abolished the ligand-induced expression of LXRalpha target genes. These results suggested that TRRAP plays an important role as a coactivator, presumably part of a complex, in lipid metabolism through regulation of the LXRalpha-mediated gene cascade in hepatic cells.

Association of gene polymorphisms and bone density in Japanese girls.

Although some studies have reported a relationship between several candidate polymorphic genes and bone mineral density (BMD), little is known concerning the genetic factors influencing BMD in children. This study examined this relationship in healthy Japanese girls (n=125; age, 13.4 +/- 0.89 years; range, 12-15 years). We investigated allelic variants of the vitamin D receptor (VDR) gene, the estrogen receptor (ER) gene, the parathyroid hormone (PTH) gene, the Ca-sensing receptor (CaSR) gene, and the beta3-adrenergic receptor (beta3-AR) gene. The genotype of the VDR gene (Fok I) correlated with lumbar spine, and femoral neck BMD. The PTH polymorphisms (BstB I, Dra II) were also associated with lumbar spine BMD. No relationship was found between genotypes of the ER gene, CaSR gene, or beta3-AR gene and BMD. The age, height, weight, and body mass index did not differ significantly among girls with different VDR and PTH genotypes. These results suggest that the Fok I polymorphism of the VDR gene and the Dra II polymorphism of the PTH gene are risk factors for low bone density in Japanese girls.