ABSTRACT
Mushrooms of the Omphalotus genus are known to be rich in secondary metabolites. In the quest for new bioactive compounds, we analyzed the compounds isolated from the mycelium of the poisonous mushroom Omphalotus japonicus. As a result, a new polyisoprenepolyol, which was named omphaloprenol A, was identified, along with known substances such as hypsiziprenol A10 and A11, illudin S, and ergosterol. The chemical structure of omphaloprenol A was elucidated by nuclear magnetic resonance and infrared spectroscopies and mass spectrometry, and its bioactivity was investigated. Omphaloprenol A showed growth promoting activity against the root of lettuce seeds and cytotoxicity against HL60 cells. To the best of our knowledge, this is the first report on the isolation of a polyisoprenepolyol compound from Omphalotaceae mushrooms.
Subject(s)
Agaricales/chemistry , Mycelium/chemistry , HL-60 Cells , Humans , Lactuca/drug effectsABSTRACT
Six new sesquiterpenes, tsukiyols A-C, neoilludin C, and 4-O-methylneoilludins A and B, were isolated from the fruiting body of Omphalotus japonicus (Kawam.) Kirchm. & O. K. Mill. Additionally, six known compounds, illudin S, neoilludins A-B, 5-hydroxydichomitol, ergosterolperoxide, and 3ß,5α,9α-trihydroxyergosta-7,22-diene-6-one, were also obtained. Their chemical structures were determined with MS, IR, and NMR spectra and the absolute configurations of neoilludins A-C, 4-O-methylneoilludins A, and B were determined with electronic circular dichroism (ECD). Illudin S and 3ß,5α,9α-trihydroxyergosta-7,22-diene-6-one showed cytotoxicity against human acute promyelocytic leukemia HL60 cells. Illudin S, 4-O-methylneoilludin A, B, and tsukiyol C showed growth-restoring activity against mutant yeast via Ca2+-signal transduction.
Subject(s)
Agaricales/chemistry , Antifungal Agents/pharmacology , Antineoplastic Agents/pharmacology , Fruiting Bodies, Fungal/chemistry , Saccharomyces cerevisiae/drug effects , Sesquiterpenes/pharmacology , Antifungal Agents/chemistry , Antifungal Agents/isolation & purification , Antineoplastic Agents/chemistry , Antineoplastic Agents/isolation & purification , Calcium Signaling/drug effects , Cell Proliferation/drug effects , Density Functional Theory , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , HL-60 Cells , Humans , Microbial Sensitivity Tests , Molecular Conformation , Mutation , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Sesquiterpenes/chemistry , Sesquiterpenes/isolation & purification , Structure-Activity RelationshipSubject(s)
Diglycerides/metabolism , Enzyme Assays/methods , Lipase/blood , Pancreas/enzymology , Catalysis , Humans , Lipase/metabolismABSTRACT
BACKGROUND: There is a need for a pancreatic lipase (LIP) reference assay to provide an accurate base to which routine methods can be traceable. METHODS: This study developed a novel LIP assay method in which 1,2-dioleoylglycerol (DODG) is the substrate and LIP activity is measured in a coupled enzymatic reaction from the increase in absorbance at 340 nm with production of NADPH. RESULTS: With this method, LIP activity was linear up to 440 U/L (8-times expected upper limit of physiological concentration). When assayed manually, the between-laboratory variation for six samples surveyed at five laboratories was 3.80-26.4% (CV) for samples containing about 20-290 U/L LIP activity; when assayed using an automated analyzer, the range was 1.86-4.86% (four laboratories). Interference by >5 mmol/L glycerol and low specificity with post-heparin samples were noted, but in practice these are avoidable. Precision analyzed by automated assay of 49 samples twice in random order produced a covariance of 2.27 U/L, which is comparable to routine methods, and good correlations were obtained with five routine methods. CONCLUSIONS: Although further studies are required, the DODG method may be likely applicable as one candidate reference method.
Subject(s)
Biocatalysis , Diglycerides/metabolism , Enzyme Assays/methods , Lipase/blood , Lipase/metabolism , Pancreas/enzymology , Humans , Linear ModelsABSTRACT
The medical criteria for initiating insulin therapy, based on clinical profiles of type 2 diabetic patients, have not yet been clearly established. We explored various parameters with 48 type 2 diabetic patients who were taking oral hypoglycemic medication. Among parameters, body mass index (BMI), the fasting plasma glucose level (FPG), and plasma chloride concentration were identified by forward-stepwise discriminant analysis as parameters that can discriminate between patients who were and those who were not undergoing insulin therapy. In combination, these parameters correctly diagnosed 86.4% of the patients who were undergoing insulin therapy, and 84.6% of those who were not undergoing insulin therapy. Further, we observed significant correlations between plasma chloride concentrations and either plasma sodium or organic acid concentrations, suggesting that impaired insulin action may reduce plasma chloride concentrations through changes in plasma sodium and organic acid metabolism. Our results suggest that plasma chloride concentration is a possible new indicator of insulin insufficiency.
