Effects of enzyme treatment on volatile and non-volatile compounds in dried green tea leaves.

Green tea contains polyphenols, mainly four catechins, including (-)-epigallocatechin gallate, (-)-epicatechin gallate, (-)-epigallocatechin, and (-)-epicatechin. Enzyme tannase is known to hydrolyze gallated catechins into non-gallated catechins and gallic acid (GA). In this study, dried green tea leaves were treated with tannase to determine changes of volatile and non-volatile compounds by the hydrolysis. The results indicated that (-)-epigallocatechin, (-)-epicatechin, and GA increased, while (-)-epigallocatechin gallate and (-)-epicatechin gallate decreased after the treatment. The GA level increased in the treated samples, which increased titratable acidity significantly, while the pH became lower. Furthermore, the antioxidant activity of the tannase-treated tea leaves increased. The level of glycosidically bound aromas decreased with the concomitant increase of corresponding volatile compounds, while some alcohols derived from fatty acids decreased significantly after the treatment. These results suggest that tannase-treatment influences both volatile and non-volatile compounds in dried green tea leaves.

Grape Seed Extract Supplementation Attenuates the Blood Pressure Response to Exercise in Prehypertensive Men.

We tested the hypothesis that exaggerated pressor responses observed in prehypertensive males (N = 9) during dynamic exercise are attenuated following acute dietary supplementation with grape seed extract (GSE) (i.e., a single dose). Effects of placebo and GSE (300 mg) on systolic blood pressure, diastolic blood pressure, mean arterial pressure (MAP), cardiac output (CO), stroke volume (SV), total vascular conductance (TVC), and rate × pressure product (RPP) in response to two submaximal cycling workloads (40% and 60% VO2peak) were compared 2 h after ingestion of GSE or placebo on different days, 1 week apart. Endothelial function was also evaluated using flow-mediated dilation (FMD). Placebo treatment had no effect on any of the variables. GSE supplementation attenuated MAP at both workloads (40% VO2peak: 115 ± 1 vs. 112 ± 2 mmHg; 60% VO2peak: 126 ± 2 vs. 123 ± 2 mmHg) and RPP at the lower workload. Conversely, SV, CO, and TVC were augmented during both workloads. FMD was augmented by GSE (18.9 ± 2.0 vs. 12.4% ± 2.0%). These findings indicate that in exercising prehypertensive males, a single dose of GSE reduces blood pressure, peripheral vasoconstriction, and work of the heart and enhances O2 delivery; effects that may be due, in part, to endothelium-dependent vasodilation. We propose that acute GSE treatment represents an intervention that may minimize potential increases in the risk of cardiovascular events during dynamic exercise in prehypertensives.

Neuroprotective Effects of Korean Red Pine (Pinus densiflora) Bark Extract and Its Phenolics.

Korean red pine (Pinus densiflora) is one of the major Pinus species in Korea. Red pine bark is removed prior to the chipping process in the wood industry and discarded as waste. However, red pine bark contains a considerable amount of naturally occurring phenolics, including flavonoids, and therefore may have a variety of biological effects. In this study, we investigated if Korean red pine bark extract (KRPBE) could protect neuronal PC-12 cells from oxidative stress and inhibit cholinesterase activity. Analysis of reversed-phase high-performance liquid chromatography results revealed four phenolics in KRPBE: vanillin, protocatechuic acid, catechin, and taxifolin. The total phenolic and flavonoid contents of KRPBE were 397.9 mg gallic acid equivalents/g dry weight (DW) and 248.7 mg catechin equivalents/g DW, respectively. The antioxidant capacities of KRPBE measured using ABTS, DPPH, and ORAC assays were 697.3, 521.8, and 2,627.7 mg vitamin C equivalents/g DW, respectively. KRPBE and its identified phenolics protected against H2O2-induced oxidative cell death in a dose-dependent manner. Acetylcholinesterase and butyrylcholinesterase, which degrade the neurotransmitter acetylcholine to terminate neurotransmission in synaptic clefts, were inhibited by treatment with KRPBE and its identified phenolics. Taken together, these results suggest that KRPBE and its constituent antioxidative phenolics are potent neuroprotective agents that can maintain cell viability under oxidative stress and inhibit cholinesterase activity.

Effects of freeze-drying on antioxidant and anticholinesterase activities in various cultivars of kiwifruit (Actinidia spp.).

Oxidative stress contributes to neurodegenerative disorders such as Alzheimer's disease. Phenolic antioxidants can efficiently reduce oxidative stress. In this study, we evaluated the effects of the freeze-drying process on phenolics, antioxidants, and cholinesterase inhibition in five cultivars of kiwifruits grown in Korea, Actinidia chinensis cv. Hort16A, cv. Happygold, and cv. Haegeum; A. deliciosa cv. Hayward; and A. eriantha cv. Bidan, by comparing them with their fresh counterparts. Among the five cultivars of both fresh and freeze-dried kiwifruits tested in this study, cv. Bidan had the highest levels of total phenolics, total flavonoids, and antioxidants, and cv. Hayward had the lowest. Freezedried kiwifruits inhibited acetylcholinesterase and butyrylcholinesterase that catalyze the breakdown of acetylcholine (neurotransmitter). On sensory evaluation, cv. Happygold had the highest overall preference scores among the freeze-dried kiwifruits. The results suggest that freeze-dried kiwifruit could serve as a good source of antioxidants and cholinesterase inhibitors.

Crossing the Border between Laboratory and Field: Bacterial Quorum Quenching for Anti-Biofouling Strategy in an MBR.

Quorum quenching (QQ) has recently been acknowledged to be a sustainable antifouling strategy and has been investigated widely using lab-scale membrane bioreactor (MBR) systems. This study attempted to bring this QQ-MBR closer to potential practical application. Two types of pilot-scale QQ-MBRs with QQ bacteria entrapping beads (QQ-beads) were installed and run at a wastewater treatment plant, feeding real municipal wastewater to test the systems' effectiveness for membrane fouling control and thus the amount of energy savings, even under harsh environmental conditions. The rate of transmembrane pressure (TMP) build-up was significantly mitigated in QQ-MBR compared to that in a conventional-MBR. Consequently, QQ-MBR can substantially reduce energy consumption by reducing coarse bubble aeration without compromising the effluent water quality. The addition of QQ-beads to a conventional MBR substantially affected the EPS concentrations, as well as microbial floc size in the mixed liquor. Furthermore, the QQ activity and mechanical stability of QQ-beads were well maintained for at least four months, indicating QQ-MBR has good potential for practical applications.

Toxic metal content in 52 frequently prescribed herbal medicines on the Korean market.

This study was conducted to investigate the toxic metal content (Pb, As, Cd and Hg) of 52 frequently prescribed herbal medicines and to identify herbal medicines that exceed the Korea Food and Drug Administration (KFDA) maximum limits. A total of 3534 samples, including 1966 domestic samples and 1568 imported samples, were analysed using an Inductively Coupled Plasma-Mass Spectrometer (ICP-MS). Total amounts of Pb, As, Cd and Hg were significantly different between domestic (0.63 mg kg(-1)) and imported (0.81 mg kg(-1)) medicines (p < 0.05). Among the 52 kinds of samples, 4 kinds of herbs required quality control for Pb and 12 kinds of herbs required quality control for Cd. No sample contained As and Hg above the limits.

Neutralization of ocular surface TNF-α reduces ocular surface and lacrimal gland inflammation induced by in vivo dry eye.

PURPOSE: The purpose of this study was to investigate the effectiveness of tumor necrosis factor (TNF)-α blocker for treatment of dry eye (DE)-induced inflammation and determine a more effective method to suppress lacrimal gland inflammation. Efficacy of topical versus systemic administration of TNF-α blockers was determined using a murine dry eye (DE) model. METHODS: The TNF-α blocker HL036 was developed by modification of the TNF receptor I. Protein purity, binding affinity, and clearance of TNF-α was compared with etanercept. Using DE-induced C57BL/6 mice, corneal erosion and goblet cell counts were measured after subcutaneous or topical treatment with etanercept or HL036. Inflammatory cytokines in cornea and lacrimal glands were determined by quantitative RT-PCR and ELISA. RESULTS: HL036 showed TNF-α binding affinity comparable to etanercept, as measured by surface plasmon resonance. HL036 concentration was significantly higher in cornea and anterior segment than etanercept and effectively eliminated TNF-α on ocular surfaces. Etanercept was preferentially concentrated in the posterior segment. Corneal erosion and goblet cell counts were improved only with topically applied etanercept and HL036. Ocular surface IFN-γ, IL-6, and IL-21 were significantly decreased by topical HL036. DE-induced lacrimal gland IFN-γ and IL-6 expression was effectively suppressed by topical etanercept and HL036. CONCLUSIONS: Topical TNF-α blockers effectively suppressed lacrimal gland and corneal inflammation by suppressing IFN-γ, IL-21, and IL-6. Differences in TNF-α affinity, clearance, and local concentration of blockers may account for the anti-inflammatory effects in different ocular regions.

Comparison of different methods to quantify fat classes in bakery products.

The definition of fat differs in different countries; thus whether fat is listed on food labels depends on the country. Some countries list crude fat content in the 'Fat' section on the food label, whereas other countries list total fat. In this study, three methods were used for determining fat classes and content in bakery products: the Folch method, the automated Soxhlet method, and the AOAC 996.06 method. The results using these methods were compared. Fat (crude) extracted by the Folch and Soxhlet methods was gravimetrically determined and assessed by fat class using capillary gas chromatography (GC). In most samples, fat (total) content determined by the AOAC 996.06 method was lower than the fat (crude) content determined by the Folch or automated Soxhlet methods. Furthermore, monounsaturated fat or saturated fat content determined by the AOAC 996.06 method was lowest. Almost no difference was observed between fat (crude) content determined by the Folch method and that determined by the automated Soxhlet method for nearly all samples. In three samples (wheat biscuits, butter cookies-1, and chocolate chip cookies), monounsaturated fat, saturated fat, and trans fat content obtained by the automated Soxhlet method was higher than that obtained by the Folch method. The polyunsaturated fat content obtained by the automated Soxhlet method was not higher than that obtained by the Folch method in any sample.

Impact of fermentation rate changes on potential hydrogen sulfide concentrations in wine.

The correlation between alcoholic fermentation rate, measured as carbon dioxide (CO2) evolution, and the rate of hydrogen sulfide (H2S) formation during wine production was investigated. Both rates and the resulting concentration peaks in fermentor headspace H2S were directly impacted by yeast assimilable nitrogenous compounds in the grape juice. A series of model fermentations was conducted in temperature-controlled and stirred fermentors using a complex model juice with defined concentrations of ammonium ions and/or amino acids. The fermentation rate was measured indirectly by noting the weight loss of the fermentor; H2S was quantitatively trapped in realtime using a pre-calibrated H2S detection tube which was inserted into a fermentor gas relief port. Evolution rates for CO2 and H2S as well as the relative ratios between them were calculated. These fermentations confirmed that total sulfide formation was strongly yeast strain-dependent, and high concentrations of yeast assimilable nitrogen did not necessarily protect against elevated H2S formation. High initial concentrations of ammonium ions via addition of diammonium phosphate (DAP) caused a higher evolution of H2S when compared with a non-supplemented but nondeficient juice. It was observed that the excess availability of a certain yeast assimilable amino acid, arginine, could result in a more sustained CO2 production rate throughout the wine fermentation. The contribution of yeast assimilable amino acids from conventional commercial yeast foods to lowering of the H2S formation was marginal.

Vardenafil increases cell proliferation in the dentate gyrus through enhancement of serotonin expression in the rat dorsal raphe.

This study was conducted to evaluate the effects of vardenafil (Levitra), a phosphodiesterase-5 (PDE-5) inhibitor, on cell proliferation in the hippocampal dentate gyrus and on 5-hyroxytryptamine (5-HT, serotonin) synthesis and tryptophan hydroxylase (TPH) expression in the rat dorsal raphe nucleus. Male Sprague-Dawley rats were divided into 6 groups (n=5 in each group): a control group, a 0.5 mg/kg-1 day vardenafil-treated group, a 1 mg/kg-1 day vardenafil-treated group, a 2 mg/kg-1 day vardenafil-treated group, a 1 mg/kg-3 day vardenafil-treated group, and a 1 mg/kg-7 day vardenafil-treated group. 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry was then performed to evaluate cell proliferation in the dentate gyrus. In addition, 5-HT and TPH immunohistochemistry was conducted to evaluate serotonin expression in the dorsal raphe. The results revealed that treatment with vardenafil increased cell proliferation in the dentate gyrus and enhanced 5-HT synthesis and TPH expression in the dorsal raphe in a dose- and duration-dependent manner. The findings demonstrate that the increasing effect of vardenafil on cell proliferation is closely associated with the enhancing effect of vardenafil on serotonin expression under normal conditions.

Development of a method to measure hydrogen sulfide in wine fermentation.

A hydrogen sulfide (H2S) detecting tube was developed for the quantitative determination of H2S produced by yeast during laboratory scale wine fermentations. The detecting tube consisted of a small transparent plastic tube packed with an H2S-sensitive color-indicating medium. The packed medium changed color, with the color change progressing upward from the bottom of the tube, upon exposure to H2S produced by yeast during fermentation. A calibration study using a standard H2S gas showed that the length of the portion that darkened was directly related to the quantity of H2S (microg) with a high correlation coefficient (r2=0.9997). The reproducibility of the H2S detecting tubes was determined with five repetitive measurements using a standard H2S solution [5.6 microg/200 ml (28 ppb)], which resulted in a coefficient of variation of 3.6% at this level of H2S. With the sulfide detecting tubes, the production of H2S was continuously monitored and quantified from laboratory scale wine fermentations with different yeast strains and with the addition of different levels of elemental sulfur to the grape juice. This sulfide detecting tube technology may allow winemakers to quantitatively measure H2S produced under different fermentation conditions, which will eventually lead winemakers to better understand the specific factors and conditions for the excessive production of H2S during wine fermentation in a large production scale.

Discrimination of teas with different degrees of fermentation by SPME-GC analysis of the characteristic volatile flavour compounds.

As tea is traded all over the world, it is necessary for both customs officers and business investigators to develop an easy and reliable method to discriminate teas from each other. A total of 56 kinds of various green, Oolong, and black teas were collected from different countries and markets, and their catechin contents and volatile flavour compounds (VFC) were compared by analyses, using HPLC and solid-phase microextraction-gas chromatograph (SPME-GC). It was found that neither total catechin nor individual catechin contents in green and Oolong teas were significantly different among the samples investigated, but the fermentation processes altered the profiles of tea VFC. Because many of the individual VFC did not change in response to the fermentation levels, several VFC in combination might be more reliable than a single compound to identify broader ranges of teas. A total concentration of five VFC, trans-2-hexenal, benzaldehyde, methyl-5-hepten-2-one, methyl salicylate, and indole, was shown to be able to discriminate clearly unfermented and fermented teas, while that of trans-2-hexenal and methyl salicylate together supplied an index to differentiate semi- and fully-fermented teas. In addition, the SPME-GC analysis was also able to distinguish real jasmine teas from fake jasmine teas based on the disappearance of some grassy/green odorants.

Shade distribution of commercial resin composites and color difference with shade guide tabs.

PURPOSE: To determine the shade distribution of varied shades of contemporary resin composites, and to measure the color difference (deltaE*ab) between individual shades of resin composites and the nearest shade tabs, which showed the smallest color difference with each shade of resin composite, in the VITA shade guide. METHODS: Eight light-curing resin composites, with a total of 41 shades, were studied. Color of specimens was measured on a reflection spectrophotometer over a white background. Ranges and distributions of CIE L*, C*ab, a* and b* values of each brand of resin composites were determined. Color difference between each shade of resin composites and each shade of the shade guide tabs were calculated, and the nearest shade guide tab was selected. RESULTS: The range of CIE L* value for eight brands of resin composites was 3.2-9.0, that of C*ab was 2.5-11.6, that of CIE a* value was 1.1-5.8, and that of CIE b* value was 5.9-11.5. Color differences (deltaE*ab) between each shade of resin composites and the nearest shade tab of the shade guide was 0.9-12.8.

Effects of (1R,9S)-beta-hydrastine on intracellular calcium concentration in PC12 cells.

(1R,9S)-beta-hydrastine (BHS) decreases the basal intracellular Ca(2+) concentration ([Ca(2+)](i)) in PC12 cells.(5) This study examined the effects of (1R,9S)-BHS on [Ca(2+)](i) in PC12 cells. (1R,9S)-BHS at 10-100 microM in combination with a high extracellular K+ level (56 mM) inhibited the release of dopamine in a concentration-dependent manner with an IC(50) value of 66.5 microM. BHS at 100 microM inhibited the sustained increase in [Ca(2+)](i) induced by a high K+ level (56 mM), and had an inhibitory effect on the 2 microM nifedipine-induced blockage of the K+ -stimulated sustained increase in [Ca(2+)](i). In addition, (1R,9S)-BHS at 100 microM prevented the rapid and sustained increase in [Ca(2+)](i) elicited by 20 mM caffeine, but did not have an effect on the increase induced by 1 microM thapsigargin, in the presence of external Ca(2+). These results suggest that the active sites of (1R,9S)-BHS are mainly L-type Ca(2+) channels and caffeine-sensitive Ca(2+)-permeable channels in PC12 cells.

Effects of tributyltin acetate on dopamine biosynthesis and L-DOPA-induced cytotoxicity in PC12 cells.

The effects of tributyltin acetate (TBTA) on dopamine biosynthesis and L-3,4-dihydroxyphenylalanine (L-DOPA)-induced cytotoxicity in PC12 cells were examined. TBTA at concentrations of 0.1-0.2 microM inhibited dopamine biosynthesis by reducing tyrosine hydroxylase (TH) activity and TH gene expression in PC12 cells. TBTA at 0.1-0.4 microM also reduced L-DOPA (20-50 microM)-induced increases in dopamine content for 24 h in PC12 cells. TBTA at concentrations up to 0.3 microM did not affect cell viability. However, TBTA at concentrations higher than 0.4 microM caused apoptotic cytotoxicity. Exposure of PC12 cells to non-cytotoxic (0.1 and 0.2 microM) or cytotoxic (0.4 microM) concentrations of TBTA with L-DOPA (20, 50 and 100 microM) significantly increased the cell loss and the percentage of apoptotic cells after 24 or 48 h compared with TBTA or L-DOPA alone. These data suggest that TBTA inhibits dopamine biosynthesis and enhances L-DOPA-induced cytotoxicity in PC12 cells.

Isolation and structural determination of squalene synthase inhibitor from Prunus mume fruit.

Squalene synthase plays an important role in the cholesterol biosynthetic pathway. Inhibiting this enzyme in hypercholesterolemia can lower not only plasma cholesterol but also plasma triglyceride levels. A squalene synthase inhibitor was screened from Prunus mume fruit, and then purified via sequential processes of ethanol extraction, HP-20 column chromatography, ethyl acetate extraction, silica gel column chromatography, and crystallization. The squalene synthase inhibitor was identified as chlorogenic acid with a molecular mass of 354 Da and a molecular formula of C16H18O9 based on UV spectrophotometry, 1H and 13C NMRs, and mass spectrometry. Chlorogenic acid inhibited the squalene synthase of pig liver with an IC50 level of 100 nM. Since chlorogenic acid was an effective inhibitor against the squalene synthase of an animal source, it may be a potential therapeutic agent for hypercholesterolemia.

Effects of tributyltin chloride on L-DOPA-induced cytotoxicity in PC12 cells.

Tributyltin chloride (TBTC) at concentrations of 0.5-1.0 microM inhibits dopamine biosynthesis in PC12 cells. In this study, the effects of TBTC on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced cytotoxicity in PC12 cells were investigated. TBTC at concentrations up to 1.0 microM neither affected cell viability, nor induced apoptosis after 24 or 48 h in PC12 cells. However, TBTC at concentrations higher than 2.0 microM caused cytotoxicity through an apoptotic process. In addition, exposure of PC12 cells to non-cytotoxic (0.5 and 1.0 microM) or cytotoxic (2.0 microM) concentrations of TBTC in combination with L-DOPA (20, 50 and 100 microM) resulted in a significant increase in cell loss and the percentage of apoptotic cells after 24 or 48 h compared with TBTC or L-DOPA alone. The enhancing effects of TBTC on L-DOPA-induced cytotoxicity were concentration- and treatment time-dependent. These data demonstrate that TBTC enhances L-DOPA-induced cytotoxicity in PC 12 cells.

Altered GABAB receptor immunoreactivity in the gerbil hippocampus induced by baclofen and phaclofen, not seizure activity.

The present study was performed to determine whether the effects induced by GABA(B) receptor-acting drugs would be related with the alteration in GABA(B) receptor expression in the hippocampus using Mongolian gerbil, a genetic epilepsy model. The distribution patterns of both GABA(B) receptor 1A/B and GABA(B)receptor 2 immunoreactivities were similarly detected in the hippocampi of normal and seizure-prone gerbils. Following baclofen (GABA(B) receptor agonist) or phaclofen (GABA(B) receptor antagonist) treatment, GABA(B) receptor immunoreactivities were decreased or increased by dose-dependent manners, respectively. Vigabatrin (GABA transaminase inhibitor) or 3-mercaptopropionic acid (GAD inhibitor) treatment did not affect GABA(B) receptor expressions. These findings suggest that GABA(B) receptor expression in the gerbil hippocampus may be altered by baclofen or phaclofen treatment.

Expression and changes of endogenous insulin-like growth factor-1 in neurons and glia in the gerbil hippocampus and dentate gyrus after ischemic insult.

In the present study, we focused upon expression and changes of endogenous insulin-like growth factor-1 (IGF-1) in the hippocampus of the Mongolian gerbil after ischemic insult. In sham-operated animals, IGF-1 immunoreactivity was absent from the hippocampus. IGF-1-immunoreactive (IR) neurons were detected at 12 h and 1 day after ischemic insult. In the hippocampal CA1 area, the IGF-IR neurons were non-pyramidal cells (GABAergic neurons). In the hippocampal CA2/3 areas, the IGF-1-IR neurons were pyramidal and non-pyramidal cells, and in the dentate gyrus the IGF-1-IR neurons were hilar neurons. Four days after ischemia-reperfusion, IGF-1 immunoreactivity disappeared from neurons, and significantly increased in astrocytes and microglia. These results suggest that the induction of IGF-1 in the CA1 area during the early stage (12-24 h after ischemic insult) is associated with the relative vulnerabilities of pyramidal glutamatergic neurons and non-pyramidal GABAergic neurons. The later increase (4 days after ischemic insult) of IGF-1 expression and protein content was found to promote the activities of astrocytes and microglia. These increases of IGF-1 in astrocytes and in microglia are associated with mechanisms that compensate for the effects of delayed neuronal death.

Altered Na+-K+ ATPase immunoreactivity within GABAergic neurons in the gerbil hippocampal complex induced by spontaneous seizure and vigabatrin treatment.

In the present study, the expression of Na(+)-K(+) ATPase in the gerbil hippocampus associated with various sequelae of spontaneous seizures were investigated in order to identify the roles of Na(+)-K(+) ATPase in the epileptogenesis and the recovery mechanisms in these animals. The population of Na(+)-K(+) ATPase immunoreactive neurons and Na(+)-K(+) ATPase immunodensity were significantly lower in the pre-seizure group of SS gerbils than those in SR gerbils. At 30-min postictal, the Na(+)-K(+) ATPase immunoreactivity was significantly elevated in the hippocampal complex. At 3-h postictal, the Na(+)-K(+) ATPase immunoreactivity in the hippocampus was declined, as compared to the 30-min postictal. At 12h after seizure on-set, Na(+)-K(+) ATPase expression was re-enhanced in the all regions of the hippocampal complex including the dentate hilus. Following administration of vigabatrin Na(+)-K(+) ATPase expression was also increased. The present data suggest that altered Na(+)-K(+) ATPase expression may contribute the regulation of the seizure activity in this animal.