AIMTo develop a rapid and sensitive method for monosaccharide composition analysis.

METHODSGlycoprotein was first hydrolyzed to monosaccharides, which were subsequently reacetylated (amino monosaccharides), tagged with 2-aminopyridine and then separated and monitored in selected ion mode by CapGCC-LC/MS. The use of tetradeuterium labeled-aminopyridyl-monosaccharides prepared by tagging monosacahrides with hexadeuterium labeled 2-aminopyridine as internal standards improved the linearity and reproducibility in quantification.

RESULTSThis method was successfully applied to monosaccharide composition analysis of model glycoproteins, fetuin and erythropoietin down to 1 pmol monosaccharides.

CONCLUSIONThis method has been shown to be highly sensitive and is applicable to monosaccharide composition analysis of glycoproteins.

Animals ; Chromatography, Liquid ; methods ; Erythropoietin ; chemistry ; Glycoproteins ; chemistry ; Hydrolysis ; Isotope Labeling ; Monosaccharides ; analysis ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; alpha-Fetoproteins ; chemistry

BACKGROUND: Irisin is a myokine implicated in lipid and glucose metabolism. The objective of this study is to examine the effect of a body weight reduction on the serum irisin level and physical indicators in obese Japanese patients without diabetes. METHODS: The subjects were 22 patients (male/female, 5/17; age, 46.1±16.0 years; body mass index [BMI], 36.9±5.0 kg/m²) who completed a 6-month body weight reduction program at our clinic. The program included diet, exercise therapy and cognitive behavioral therapy. Blood parameters, body composition, exercise tolerance, homeostasis model assessment of insulin resistance (HOMA-IR), and serum irisin were determined before and after intervention, and relationships among changes in these data were examined. RESULTS: There were significant decreases in body weight and BMI after the intervention. Irisin before the intervention was significantly positively correlated with HOMA-IR (r=0.434, P<0.05). The mean irisin level showed no significant change after the intervention in all participants. However, improvements in % body fat, subcutaneous fat area, triglycerides, and fasting glucose were significantly greater in patients with an increase in irisin compared to those with a decrease in irisin after the intervention. Patients with an increase in irisin also had significantly lower fasting insulin (9.7±4.8 vs. 16.4±8.2, P<0.05) and HOMA-IR (2.2±1.1 vs. 3.7±1.6, P<0.05) after the intervention, compared to patients with a decrease in irisin. CONCLUSION: Body weight reduction did not alter irisin levels. However, irisin may play important roles in fat and glucose metabolism and insulin resistance, and the effects of body weight reduction on irisin kinetics may be a key for obesity treatment.
Adipose Tissue ; Asian Continental Ancestry Group ; Body Composition ; Body Mass Index ; Body Weight* ; Cognitive Therapy ; Diet ; Exercise Therapy ; Exercise Tolerance ; Fasting ; Glucose ; Homeostasis ; Humans ; Insulin ; Insulin Resistance ; Kinetics ; Metabolism ; Obesity ; Subcutaneous Fat ; Triglycerides

BACKGROUND: It has recently been suggested that skeletal muscle has an important role in insulin resistance in obesity, in addition to exercise tolerance and the fat index. The aim of this study was to identify body composition factors that contribute to improvement of insulin resistance in female patients with obesity who reduce body weight. METHODS: We studied 92 female obese patients (age 40.9±10.4 years, body mass index 33.2±4.6 kg/m2) who reduced body weight by ≥5% after an intervention program including diet, exercise therapy, and cognitive behavioral therapy. Before and after the intervention, body composition was evaluated by dual-energy X-ray absorptiometry to examine changes in skeletal muscle mass. Homeostasis model assessment of insulin resistance (HOMA-IR) was measured as an index of insulin resistance. Cardiopulmonary exercise was also performed by all patients. RESULTS: There were significant improvements in body weight (-10.3%±4.5%), exercise tolerance (anaerobic threshold oxygen uptake 9.1%±18.4%, peak oxygen uptake 11.0%±14.2%), and HOMA-IR (-20.2%±38.3%). Regarding body composition, there were significant decreases in total body fat (-19.3%±9.6%), total fat-free mass (-2.7%±4.3%), and % body fat (-10.1%±7.5%), whereas % skeletal muscle significantly increased (8.9%±7.2%). In stepwise multiple linear regression analysis with change in HOMA-IR as the dependent variable, the change in % skeletal muscle was identified as an independent predictor (β=-0.280, R2=0.068, P<0.01). CONCLUSION: Improvement of insulin resistance in female obese patients requires maintenance of skeletal muscle mass.
Absorptiometry, Photon ; Adipose Tissue ; Body Composition ; Body Mass Index ; Body Weight* ; Cognitive Therapy ; Diet ; Exercise Therapy ; Exercise Tolerance ; Female* ; Homeostasis ; Humans ; Insulin Resistance* ; Insulin* ; Linear Models ; Muscle, Skeletal ; Obesity* ; Oxygen