RESUMO
The ceramide (Cer) content of skin and glucosylceramide (GlcCer) intake affect skin moisture conditions, but their mutual relation in skin remains unclear. For clarification of that mutual relation, carbon stable isotopes ((12)C and (13)C) are useful as a tracer. However, carbon isotopic measurement has not been applied to the study of clarifying their skin moisturizing effects. Therefore, we used gas chromatography / combustion / isotope ratio mass spectrometry (GC-C-IRMS) to ascertain the appropriate conditions for carbon isotopic measurements using synthesized Cer (SCer) in substitution for very low concentrations of Cer in skin. SCer was derivatized to trimethylsilylated SCer (TMS-SCer) quantitatively using N-trimethylsilylimidazole (TMSI) depending on the amount of SCer. The derivatization rates were 75-85%. Excess TMSI was removed using three cycles of hexane-water distribution. Under these conditions, carbon isotopic measurements of TMS-SCer conducted using GC-C-IRMS showed high repeatability and good inter-day variation (S.D. < 0.3). The carbon stable isotope ratio value (δ(13)C) of SCer calculated using a mass balance equation was compared with δ(13)C of underivatized SCer, which was regarded as the actual δ(13)C of SCer obtained using sealed tube combustion method. The difference between the calculated δ(13)C of SCer and δ(13)C of the underivatized SCer depended on the TMSI reagent supplier and on the number of hydroxyl groups to be derivatized in SCer. For accurate δ(13)C of Cer in skin using GC-C-IRMS, the measured δ(13)C of a target TMS-Cer must be calculated using a correction factor representing the difference in δ(13)C of underivatized standard SCer from that of TMS-standard SCer having a structure resembling that of the target Cer in skin. In addition, we show that the same lot of TMSI reagent from a specific supplier must be used throughout the experiments.
Assuntos
Ceramidas/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Pele/química , Isótopos de Carbono/metabolismo , Glucosilceramidas/metabolismo , Humanos , Reprodutibilidade dos Testes , Pele/metabolismoRESUMO
A PCR-based method was developed to distinguish between durum/common wheat and common wheat by leveraging slight differences of DNA sequence in Starch Synthase II (SS II) coded on wheat A, B and D genomes. A primer pair, SS II ex7-U/L, was designed to hybridize with a conserved DNA sequence region found in SS II-A, B and D genes. Another primer pair, SS II-D 1769U/1889L, was constructed to recognize a unique sequence in the SS II-D gene. The target region of SS II ex7-U/L with the size of 114 bp was amplified from durum and common wheat DNA, while no amplification was observed from any cereals other than those in the wheat genus. A DNA fragment with the size of 121 bp was specifically amplified from common wheat with SS II-D 1769U/1889L. In blended flour prepared from wheat and other cereals, the developed PCR system composed of two primer pairs effectively detected durum/common wheat and common wheat. These results suggested that PCR using two primer pairs is useful for detecting common and/or durum wheat in blended flour and could be utilized to ensure accurate food labeling.
Assuntos
Farinha , Reação em Cadeia da Polimerase/métodos , Triticum , Sequência de Bases , Triticum/genéticaRESUMO
To develop a method for detecting GM wheat that may be marketed in the near future, we evaluated the proline-rich protein (PRP) gene as an endogenous reference gene of common wheat (Triticum aestivum L.) and durum wheat (Triticum durum L.). Real-time PCR analysis showed that only DNA of wheat was amplified and no amplification product was observed for phylogenetically related cereals, indicating that the PRP detection system is specific to wheat. The intensities of the amplification products and Ct values among all wheat samples used in this study were very similar, with no nonspecific or additional amplification, indicating that the PRP detection system has high sequence stability. The limit of detection was estimated at 5 haploid genome copies. The PRP region was demonstrated to be present as a single or double copy in the common wheat haploid genome. Furthermore, the PRP detection system showed a highly linear relationship between Ct values and the amount of plasmid DNA, indicating that an appropriate calibration curve could be constructed for quantitative detection of GM wheat. All these results indicate that the PRP gene is a suitable endogenous reference gene for PCR-based detection of GM wheat.
Assuntos
Alimentos Geneticamente Modificados , Triticum/genética , DNA de Plantas/análise , Proteínas de Plantas/genética , Reação em Cadeia da Polimerase em Tempo RealRESUMO
We investigated the microbial conversion of curcumin (1) using endophytic fungi associated with the rhizome of Curcuma longa (Zingiberaceae). We found that Diaporthe sp., an endophytic filamentous fungus, converts curcumin (1) into four colorless derivatives, namely (3R,5R)-tetrahydrocurcumin (2), a novel (3R,5S)-hexahydrocurcumin (3) named neohexahydrocurcumin, (3S,5S)-octahydrocurcumin (4) and meso-octahydrocurcumin (5).
Assuntos
Ascomicetos/metabolismo , Curcuma/microbiologia , Curcumina/análogos & derivados , Curcumina/metabolismo , Rizoma/microbiologia , Ascomicetos/químicaRESUMO
[reaction: see text] 13-Hydroxy-14-nordehydrocacalohastine (2) and 13-acetoxy-14-nordehydrocacalohastine (3), two novel modified furanoeremophilane-type sesquiterpenes isolated from Trichilia cuneata, showed inhibitory activities for membrane lipid peroxidation in mitochondria and microsomes. The first, highly convergent total syntheses of new compounds 2 and 3 have also been achieved via a palladium-mediated three-component coupling reaction between 2-iodotoluene (7), 1-penten-4-yn-3-ol (8), and diethyl ethoxymethylenemalonate (9).
Assuntos
Meliaceae/química , Plantas Medicinais/química , Sesquiterpenos , Técnicas de Química Combinatória , Estrutura Molecular , Sesquiterpenos/síntese química , Sesquiterpenos/química , Sesquiterpenos/farmacologiaRESUMO
Three monoamine oxidase (MAO) inhibitors were isolated from Gentiana lutea. Their structures were elucidated to be 3-3''linked-(2'-hydroxy-4-O-isoprenylchalcone)-(2'''-hydroxy-4''-O-isoprenyldihydrochalcone) (1), 2-methoxy-3-(1,1'-dimethylallyl)-6a,10a-dihydrobenzo(1,2-c)chroman-6-one and 5-hydroxyflavanone. These compounds, and the hydrolysis product of 1, displayed competitive inhibitory properties against MAO-B which was more effective than MAO-A.
Assuntos
Encéfalo/efeitos dos fármacos , Gentiana/química , Mitocôndrias/efeitos dos fármacos , Inibidores da Monoaminoxidase/farmacologia , Monoaminoxidase/metabolismo , Animais , Encéfalo/enzimologia , Encéfalo/metabolismo , Técnicas In Vitro , Masculino , Mitocôndrias/enzimologia , Estrutura Molecular , Inibidores da Monoaminoxidase/isolamento & purificação , Extratos Vegetais/química , Pós , Ratos , Ratos WistarRESUMO
Isoaffinetin (5,7,3',4',5'-pentahydroxyflavone-6-C-glucoside) was isolated from Manilkara indica as a potent inhibitor of lens aldose reductase by bioassay-directed fractionation. This C-glucosyl flavone showed specific inhibition against aldose reductases (rat lens, porcine lens and recombinant human) with no inhibition against aldehyde reductase and NADH oxidase. Kinetic analysis showed that isoaffinetin exhibited uncompetitive inhibition against both dl-glyceraldehyde and NADPH. A structure-activity relationship study revealed that the increasing number of hydroxy groups in the B-ring contributes to the increase in aldose reductase inhibition by C-glucosyl flavones.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Manilkara , Fitoterapia , Extratos Vegetais/farmacologia , Animais , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/química , Inibidores Enzimáticos/uso terapêutico , Flavonoides/administração & dosagem , Flavonoides/química , Flavonoides/uso terapêutico , Humanos , Concentração Inibidora 50 , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Folhas de Planta , Ratos , Relação Estrutura-Atividade , SuínosRESUMO
A meroterpene and four flavonoids were isolated from the seeds of Psoralea corylifolia as antioxidative components. Their structures were elucidated by spectral data and identified as bakuchiol (1), bavachinin (2), bavachin (3), isobavachin (4) and isobavachalcone (5). In particular, meroterpene 1 and flavonoids 4 and 5 showed broad antioxidative activities in rat liver microsomes and mitochondria. They inhibited NADPH-, ascorbate-, t-BuOOH- and CCl(4)-induced lipid peroxidation in microsomes. They also prevented NADH-dependent and ascorbate-induced mitochondrial lipid peroxidation. Bakuchiol (1) was the most potent antioxidant in microsomes and the inhibition of oxygen consumption induced by lipid peroxidation was time-dependent. Furthermore, bakuchiol (1) protected human red blood cells against oxidative haemolysis. These phenolic compounds in P. corylifolia were shown to be effective in protecting biological membranes against various oxidative stresses.
Assuntos
Antioxidantes/farmacologia , Flavonoides/farmacologia , Extratos Vegetais/farmacologia , Psoralea , Animais , Chalcona/análogos & derivados , Chalcona/química , Chalcona/isolamento & purificação , Chalcona/farmacologia , Chalconas , Eritrócitos/efeitos dos fármacos , Flavonoides/química , Flavonoides/isolamento & purificação , Hemólise/efeitos dos fármacos , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Estrutura Molecular , Consumo de Oxigênio/efeitos dos fármacos , Fenóis/química , Fenóis/isolamento & purificação , Fenóis/farmacologia , Extratos Vegetais/química , Ratos , Ratos Wistar , Sementes/químicaRESUMO
Dodecyl (C12) gallate exhibits both potent chain-breaking and preventive antioxidant activity. The pyrogallol moiety is responsible for both activities. Dodecyl (lauryl) gallate prevents generation of superoxide radicals by xanthine oxidase, and this activity comes from its ability to inhibit the enzyme. The inhibition kinetics analyzed by Lineweaver-Burk plots found that dodecylgallate is a noncompetitive inhibitor for the generation of superoxide anion. Dodecyl gallate also inhibits formation of uric acid. The inhibition kinetics analyzed by Lineweaver-Burk plots found that dodecyl gallate is a competitive inhibitor for this oxidation. Mitochondrial lipid peroxidation induced by Fe(III)-adenosine 5'-diphosphate/reduced nicotinamide adenine dinucleotide was inhibited by dodecyl gallate while its parent compound, gallic acid, did not show this inhibitory activity. Dodecyl gallate protected mitochondrial functions and human red blood cells against oxidative stresses, but gallic acid showed little effect. The hydrophobic dodecyl group is largely associated with the preventive antioxidative activity.