Purification and Some Properties of Debranching Enzyme of Germinating Rice Endosperm

A debranching enzyme was extracted from the endosperm of germinating rice seeds and purified through three steps, namely cyclohexaamylose-coupled Sepharose 6B, Ultrogel AcA-44 and Bio-Gel P-150 column chromatography. This disc-electrophoretically homogeneous enzyme showed a specific activity of 43 units/mg of protein (30°C) with a pH optimum of 5.5. The isoelectric point was 4.9, unlike that (pI 3.5) of debranching enzyme of ungerminated rice seeds. Our enzyme hydrolyzed pullulan rapidly, and glutinous rice starch and waxy corn starch moderately. The enzyme was also able to act on phytoglycogen and glycogen unlike debranching enzymes originating in some plants.     

Syntheses of α and γ-BHC Alkylthio Analogs

Meso-(1245/36)-1,2,4,5,6-pentachloro-3-methylthiocyclohexane, and (124/356)-1,2,4,5,6-pentachloro-3-methylthio and ethylthiocyclohexanes were prepared from (1234/56)-1,4,5,6-tetrachloro-2,3-epoxycyclohexane (α-BTC cis-epoxide).     

Metabolism of Hymexazol, 3-Hydroxy-5-methylisoxazole,in the Rats

Excretion, distribution and metabolism of the fungicide, hymexazol, (3-hydroxy-5-methylisoxazole), labeled with carbon-14 were examined after administration of a single oral dose to Wistar-strain rats. Hymexazol was rapidly absorbed and distributed in the tissues. During 96 hr, 97% of the total radioactivity was excreted in the urine and 0.89% in the feces, and 0.86% was found in the expired gasses for 24 hr. Two metabolites were detected in the urine, whose chemical structures were determined as 3-(β-d-glucopyranuronosyloxy)-5- methylisoxazole and 5-methyl-3-isoxazolyl sulfate.     

In vitro anticancer activity of loquat tea by inducing apoptosis in human leukemia cells

Fresh loquat leaves have been used as folk health herb in Asian countries for long time, although the evidence supporting their functions is still minimal. This study aimed to clarify the chemopreventive effect of loquat tea extract (LTE) by investigating the inhibition on proliferation, and underlying mechanisms in human promyelocytic leukemia cells (HL-60). LTE inhibited proliferation of HL-60 in a dose-dependent manner. Molecular data showed that the isolated fraction of LTE induced apoptosis of HL-60 as characterized by DNA fragmentation; activation of caspase-3, -8, and -9; and inactivation of poly(ADP)ribose polymerase. Moreover, LTE fraction increased the ratio of pro-apoptotic Bcl-2-associated X protein (Bax)/anti-apoptotic myeloid cell leukemia 1 (Mcl-1) that caused mitochondrial membrane potential loss and cytochrome c released to cytosol. Thus, our data indicate that LTE might induce apoptosis in HL-60 cells through a mitochondrial dysfunction pathway. These findings enhance our understanding for chemopreventive function of loquat tea.     

Metabolic stability and inhibitory effect of O-methylated theaflavins on H2O2-induced oxidative damage in human HepG2 cells

Seven new O-methylated theaflavins (TFs) were synthesized by using O-methyltransferase from an edible mushroom. Using TFs and O-methylated TFs, metabolic stability in pooled human liver S9 fractions and inhibitory effect on H₂O₂-induced oxidative damage in human HepG2 cells were investigated. In O-methylation of theaflavin 3′-O-gallate (TF3′G), metabolic stability was potentiated by an increase in the number of introduced methyl groups. O-methylation of TF3,3′G did not affect metabolic stability, which was likely because of a remaining 3-O-galloyl group. The inhibitory effect on oxidative damage was assessed by measuring the viability of H₂O₂-damaged HepG2 cells treated with TFs and O-methylated TFs. TF3,3′G and O-methylated TFs increased cell viabilities significantly compared with DMSO, which was the compound vehicle (p?<?0.05), and improved to approximately 100%. Only TF3′G did not significantly increase cell viability. It was suggested that the inhibitory effect on H₂O₂-induced oxidative damage was potentiated by O-methylation or O-galloylation of TFs.     

Distribution and Properties of NAD Phosphorylating Reaction

Distribution of NAD phosphorylating reactions, phosphorylation through NAD kinase and phosphotransferase, was investigated. NAD kinase activity was distributed rather widely in bacteria, whereas phosphotransferase activity with p-NPP and NAD was limited to a few genera. Proteus mirabilis showed strong activity of phosphotransferase besides NAD kinase activity.

Partial purification of the phosphotransferase was attempted. The enzyme preparation possessed phosphatase activity as well as phosphotransferase activity. Phosphorylation of NAD proceeded maximally under the conditions below pH 4.0. Cu²⁺ showed stimulating effect on the activity. Besides p-NPP and phenylphosphate, various nucleotides, especially 2′ (or 3′) isomers, served as excellent phosphoryl donors, and various kinds of nucleosides and nucleotides were phosphorylated to form nucleoside monophosphates and nucleoside diphosphates.     

Production of Erythritol by n-Alkane-grown Yeasts

In the course of study on citric acid fermentation by Candida zeylanoides, in which n-alkane (a mixture of C–12 to C–15) was used as the sole source of carbon, we found that a polyol-like substance was accumulated when the medium-pH fell down to below 4.0. This was isolated in crystalline forms and identified as meso-erythritol. Comparing erythritol production among fifty yeast strains, Candida zeylanoides, particularly its glycerol-requiring mutant KY 6166, was found to be an excellent producer.

Erythritol production was also observed with ethanol or acetic acid as the sole carbon source but not with glucose. An efficient condition for large production of erythritol was to keep the medium-pH at low level (2.5 to 4.0) and the concentration of NaCl or KCl at high level (1 to 3%). Under conditions established in this work, more than 55 mg/ml of erythritol was successfully produced in 120 hr incubation in 300-ml flasks, which corresponded to 55% of the alkane used.     

The relationship between the amount of insect-repelling phenolic compounds in pennywort tea and the number of infesting cigarette beetles in the tea bag during one-year storage

Cigarette beetle, Lasioderma serricorne (F.), is a common food-product pest widely found in homes. This paper investigated the relationship between the number of infesting cigarette beetles and the phenolic content of pennywort tea made from young and mature leaves during one-year storage. It was found that cigarette beetles started to appear in tea bags after six months of storage. More adults were found than larvae and pupae. In addition, the number of cigarette beetles infesting pennywort tea made from mature leaves was higher than when it was made from young leaves. The amount of phenolic compounds in pennywort tea was analyzed by Folin–Ciocalteau assay. The results of a regression analysis indicated that the number of adult cigarette beetles correlated negatively with the amount of phenolic compounds in pennywort tea made from young leaves – when the total phenolic content decreased, the number of infesting cigarette beetles increased (r² = 0.80). The number of larvae and pupae showed a weak negative correlation with the amount of phenolic compounds (r² = 0.40 and r² = 0.56, respectively). Similar correlations were found in pennywort tea made from mature leaves, but the number of infesting beetles was higher due to the lower phenolic content of the mature leaves.     

贝那普利联合肾炎四味片对慢性肾小球肾炎患者临床疗效的影响及机制研究

目的:探讨贝那普利联合肾炎四味片对慢性肾小球肾炎患者临床疗效的影响及其作用机制。方法:收集我院收治的慢性肾小球肾炎患者120例,随机分为观察组和对照组,每组各60例。对照组给予肾炎四味片治疗;观察组在对照组的基础上给予贝那普利片。治疗结束后,比较两组患者白细胞诱素1(Lkn-1)、尿酸、TNF-α、收缩压、舒张压、24小时蛋白尿水平的变化及临床总有效率。结果:与治疗前相比,两组患者治疗后Lkn-1、尿酸、TNF-α、收缩压、24小时蛋白尿水平均显著降低(P0.05);与对照组比较,观察组患者Lkn-1、尿酸、TNF-α、收缩压、24小时蛋白尿水平较低(P0.05),临床总有效率较高(P0.05)。结论:贝那普利联合肾炎四味片可有效治疗慢性肾小球肾炎,推测其机制与降低Lkn-1、尿酸、TNF-α、收缩压、24小时蛋白尿水平有关。     

Daily consumption of tea catechins improves aerobic capacity in healthy male adults: a randomized double-blind,placebo-controlled,crossover trial

Our previous studies demonstrated that dietary supplementation with tea catechins combined with exercise improved endurance capacity in mice. This study aimed to demonstrate the effect of daily tea catechin consumption on aerobic capacity in humans. Sixteen Japanese non-athlete male subjects (aged 25–47 years) took 500 mL of a test beverage with or without tea catechins (570 mg) daily for 8 weeks and attended a training program twice a week. Aerobic capacity was evaluated by indirect calorimetry and near-infrared spectroscopy during graded cycle exercise. Catechin beverage consumption was associated with a significantly higher ventilation threshold during exercise and a higher recovery rate of oxygenated hemoglobin and myoglobin levels after graded cycle exercise when compared to subjects receiving the placebo beverage. These results indicate that daily consumption of tea catechins increases aerobic capacity when combined with semiweekly light exercise, which may be due to increased skeletal muscle aerobic capacity.