In vitro expansion of CD34+CD38- cells under stimulation with hematopoietic growth factors on AGM-S3 cells in juvenile myelomonocytic leukemia

advance online publication, August 8, 2014 [Epub ahead of print]ArticleLEUKEMIA. 29(3):606-614 (2015)journal articl

Establishment of sandwich ELISA for soluble alpha-Klotho measurement: Age-dependent change of soluble alpha-Klotho levels in healthy subjects

Background α-Klotho (αKl) regulates mineral metabolism such as calcium ion (Ca2+) and inorganic phosphate (Pi) in circulation. Defects in mice result in clinical features resembling disorders found in human aging. Although the importance of transmembrane-type αKl has been demonstrated, less is known regarding the physiological importance of soluble-type αKl (sαKl) in circulation. Objectives The aims of this study were: (1) to establish a sandwich ELISA system enabling detection of circulating serum sαKl, and (2) to determine reference values for sαKl serum levels and relationship to indices of renal function, mineral metabolism, age and sex in healthy subjects. Results We successively developed an ELISA to measure serum sαKl in healthy volunteers (n = 142, males 66) of ages (61.1 ± 18.5 year). The levels (mean ± SD) in these healthy control adults were as follows: total calcium (Ca; 9.46 ± 0.41 mg/dL), Pi (3.63 ± 0.51 mg/dL), blood urea nitrogen (BUN; 15.7 ± 4.3 mg/dL), creatinine (Cre; 0.69 ± 0.14 mg/dL), 1,25 dihydroxyvitamin D (1,25(OH)2D; 54.8 ± 17.7 pg/mL), intact parathyroid hormone (iPTH; 49.2 ± 20.6 pg/mL), calcitonin (26.0 ± 12.3 pg/mL) and intact fibroblast growth factor (FGF23; 43.8 ± 17.6 pg/mL). Serum levels of sαKl ranged from 239 to 1266 pg/mL (mean ± SD; 562 ± 146 pg/mL) in normal adults. Although sαKl levels were not modified by gender or indices of mineral metabolism, sαKl levels were inversely related to Cre and age. However, sαKl levels in normal children (n = 39, males 23, mean ± SD; 7.1 ± 4.8 years) were significantly higher (mean ± SD; 952 ± 282 pg/mL) than those in adults (mean ± SD; 562 ± 146, P < 0.001). A multivariate linear regression analysis including children and adults in this study demonstrated that sαKl correlated negatively with age and Ca, and positively with Pi. Finally, we measured a serum sαKl from a patient with severe tumoral calcinosis derived from a homozygous missense mutation of α-klotho gene. In this patient, sαKl level was notably lower than those of age-matched controls. Conclusion We established a detection system to measure human serum sαKl for the first time. Age, Ca and Pi seem to influence serum sαKl levels in a normal population. This detection system should be an excellent tool for investigating sαKl functions in mineral metabolism

A giant adrenal pseudocyst presenting with right hypochondralgia and fever: a case report

<p>Abstract</p> <p>Introduction</p> <p>Adrenal pseudocysts are rare cystic masses that arise from the adrenal gland and which are usually non-functional and asymptomatic. Adrenal pseudocysts consist of a fibrous wall without an epithelial or endothelial lining. We report the case of a patient with a giant adrenal pseudocyst presenting with right hypochondralgia and high fever.</p> <p>Case presentation</p> <p>A 52-year-old Japanese man was admitted with right hypochondralgia and a chill. Abdominal computed tomography revealed a well-defined cystic mass measuring 19 cm which was located in the right adrenal region and the contents of which were not enhanced with contrast medium. Abdominal ultrasonography revealed a heterogeneously hypo-echoic lesion with a peripheral high-echoic rim. Serum hormonal levels were almost normal. Despite treatment with antibiotics, the high fever persisted. Based on these findings, we made a preoperative diagnosis of a right adrenal cyst with infection. However, the possibility of malignancy still remained. The patient underwent laparotomy and right adrenal cyst excision with partial hepatectomy in order to relieve the symptoms and to confirm an accurate diagnosis. Histological examination revealed an adrenal pseudocyst with infection. His condition improved soon after the operation.</p> <p>Conclusion</p> <p>We report a case of a giant adrenal pseudocyst with infection. Surgery is required for symptomatic cases in order to relieve the symptoms and in cases of uncertain diagnosis.</p

胆石患者における血清スクアレン

Serum squalene concentration, which represents the degree of cholesterol synthesis,　and other serum and biliary lipids were determined and characteristic abnormalities of　lipid metabolism were discussed in patients with gallstones having high serum squalene. The serum squalene concentration was significantly high in patients with pure　cholesterol and mixed stones. It remained in normal range in patients with combination　and bilirubinate stones. The high serum squalene concentration was accompanied by　a high lithogenic index and serum triglyceride concentration. Patients with high serum　squalene were also accompanied by increased percent cholic and decreased percent　chenodeoxycholic acid. These results may indicate that patients with high serum squalene in our present　study have similar lipid abnormalities as type IV hyperlipoproteinemia which has　increased cholesterol synthesis, hypertriglyceridemia and increased cholic acid pool and　normal chenodeoxycholic acid pool size

Chemically Reactive Supramolecular Hydrogel Coupled with a Signal Amplification System for Enhanced Analyte Sensitivity

Multicomponent supramolecular hydrogels are constructed for sensitive, naked-eye detection of small-molecule biomarkers. A dendritic self-immolative molecule and the corresponding enzyme as a signal amplification system were stably embedded in a hydrogen peroxide (H₂O₂)-responsive supramolecular hydrogel (<b>BPmoc-F</b>_<b>3</b>), together with other enzymes. The nanostructure and mechanical strength of the hybrid <b>BPmoc-F</b>_<b>3</b> gel were not substantially diminished by incorporation of these multiple components in the absence of target biomarkers, but could be destroyed by addition of the biomarker through the multiple enzymatic and chemical cascade reactions operating in combination within the gel matrix. The sensitivity to biomarkers such as H₂O₂, glucose, and uric acid, detected by gel–sol transition, was significantly enhanced by the signal amplification system. An array chip consisting of these multicomponent hydrogels enabled the detection of the level of hyperuricemia disease in human plasma samples