No abstract available.
Edema*

BACKGROUND: Analysis of urinary organic acids is an important procedure for the diagnosis of inherited disorders of amino acid and organic acid metabolism. Analysis of urinary organic acids by gas chromatography-mass spectrometry (GC-MS) were initially developed for qualitative purposes, and quantitative anlytical procedure have seldom been extensively studied if at all. The purpose of this study is to evaluate a quantitative procedure for profiling organic acids with GC-MS. METHODS: Urine samples (1.5 mL) were extracted with ethylacetate, and derivated to trimethylsilyl derivatives. The compounds were analysed with MD-800 GC-MS (Fisons, Manchester, U.K.). The quantitation was done by establishment of calibration curves with the standard solutions of 74 organic acids. A response factor for internal standard was used to quantify organic acids of which the standards were not available. Extraction efficiencies for 51 organic acids were evaluated. Interassay and intraassay imprecisions were estimated from the analysis of two quality control specimens with the different concentrations of organic acids. RESULTS: Extraction efficiencies varied from 7.9 0.2% to 182.7 4.8% according to organic acids. Interassay imprecisions of specimen I and II were 4.1~60.7% and 10.7~84.9%, respectively. Intraassay imprecisions were 1.7~24.3% and 2.8~38.1%, respectively. But interassay imprecisions for clinically important analytes were below 20%. CONCLUSIONS: The described method for quantification of urinary organic acids with GC-MS is a acceptable routine method for screening of urinary organic acids. The result that imprecisions of clinically significant organic acids were less than 20% suggests that the method would be acceptable not only for diagnosis, but also for follow-up.
Calibration ; Diagnosis ; Gas Chromatography-Mass Spectrometry* ; Mass Screening ; Metabolism ; Quality Control

BACKGROUND AND OBJECTIVES: It is well known that, regardless of whether a person has cardiovascular diseases, the reduction of heart rate after exercise reflects the impairment of the autonomic nervous system. It is also a predictive factor of death rate and it correlates to insulin resistance. Therefore, we assessed these correlations in normoglycemic subjects. SUBJECTS AND METHODS: Exercise stress testing was performed according to the Bruce protocol. Anthropometric indices of adiposity, metabolic variables, blood pressure (BP) and several cardiovascular risk factors were measured. The HOMA index was used as the insulin resistance, and the impairment of the autonomic nervous system was assessed by measuring the reduction of heart rate for 2 minutes after the cessation of exercise. RESULTS: The reduction of heart rate during 2 minutes after the cessation of exercise statistically correlated with the HOMA index, gender, age, body mass index, the waist circumference, heart rate during rest, the maximum heart rate, serum total cholesterol concentration, serum high density lipoprotein cholesterol concentration and serum low density lipoprotein cholesterol concentration, (p<0.05). However, on multiple regression analysis, the HOMA index, gender, heart rate during rest, and the maximum heart rate significantly correlated to the reduction of heart rate during 2 minutes after the cessation of exercise. CONCLUSION: In individuals with normal serum glucose levels, even after adjustment was made for other factors, the reduction of heart rate after the cessation of exercise correlated to insulin resistance. Hence, in individuals with normal serum glucose levels, efforts to improve insulin resistance have to be made, and prospective study on this subject is required.
Adiposity ; Autonomic Nervous System ; Blood Glucose ; Blood Pressure ; Body Mass Index ; Cardiovascular Diseases ; Cholesterol ; Cholesterol, HDL ; Cholesterol, LDL ; Exercise Test ; Heart Rate* ; Heart* ; Humans ; Insulin Resistance* ; Insulin* ; Mortality ; Risk Factors ; Waist Circumference

The molecular mechanism underlying the initiation of somatic cell reprogramming into induced pluripotent stem cells (iPSCs) has not been well described. Thus, we generated single-cell-derived clones by using a combination of drug-inducible vectors encoding transcription factors (Oct4, Sox2, Klf4 and Myc) and a single-cell expansion strategy. This system achieved a high reprogramming efficiency after metabolic and epigenetic remodeling. Functional analyses of the cloned cells revealed that extracellular signal-regulated kinase (ERK) signaling was downregulated at an early stage of reprogramming and that its inhibition was a driving force for iPSC formation. Among the reprogramming factors, Myc predominantly induced ERK suppression. ERK inhibition upregulated the conversion of somatic cells into iPSCs through concomitant suppression of serum response factor (SRF). Conversely, SRF activation suppressed the reprogramming induced by ERK inhibition and negatively regulated embryonic pluripotency by inducing differentiation via upregulation of immediate early genes, such as c-Jun, c-Fos and EGR1. These data reveal that suppression of the ERK-SRF axis is an initial molecular event that facilitates iPSC formation and may be a useful surrogate marker for cellular reprogramming.

Endothelium, particularly pulmonary endothelium, is predisposed to injury by reactive oxygen species (ROS) and their derivatives. Heme oxygenase (HO) has been demonstrated to provide cytoprotective effects in models of oxidant-induced cellular and tissue injuries. In the present study, we investigated the effects of YS 49 against oxidant [tert-butylhydroperoxide (TBH) ]-induced injury using cultured sheep pulmonary artery endothelial cells (SPAECs). The viability of SPAECs was determined by quantifying reduction of a fluorogenic indicator Alamar blue. We found that TBH decreased cell viability in a time- and concentration-dependent manner. YS 49 concentration- and time-dependently increased HO-1 induction on SPAECs. As expected, YS 49 significantly decreased the TBH-induced cellular injury. In the presence of zinc protophorphyrin, HO-1 inhibitor, effect of YS 49 was significantly inhibited, indicating that HO-1 plays a protective role for YS 49. Furthermore, YS 49 showed free radical scavenging activity as evidenced by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and inhibition of lipid peroxidation. However, YS 49 did not inhibit apoptosis induced by lipopolysaccharide (LPS) in SPAECs. Taken together, HO-1 induction along with strong antioxidant action of YS 49 may be responsible for inhibition of TBH-induced injury in SPAECs.
Apoptosis ; Cell Survival ; Endothelial Cells* ; Endothelium ; Heme Oxygenase (Decyclizing) ; Lipid Peroxidation ; Pulmonary Artery* ; Reactive Oxygen Species ; Sheep* ; Zinc