Effect of Cultivation Stages of Hericium erinaceus on the Contents of Major Components and α-Glucosidase Inhibitory Activity

Hericium erinaceus, also known as lion’s mane mushroom, is an edible and medicinal mushroom that belongs to the family Hericiaceae. We previously reported hericene A as an anti-diabetic constituent of H. erinaceus and the effect of cultivation substrates on its content was investigated. As the continuation, the contents of five major compounds such as hericenes A-D, which exerted α-glucosidase inhibitory activity, together with ergosterol were investigated depending on cultivation stages. H. erinaceus was cultured for 25 days (5 stages) to induce fruiting bodies, and the contents of the compounds at each stage were quantified. All the five compounds were detected in fruiting body by HPLC analysis. Among the hericene derivatives in the mushroom, the content of hericene A was the highest, followed by hericene C and the content of hericenes B and D was relative low. All four hericene derivatives present in the highest content at stage 4 whereas the content of ergosterol was highest at stage 5. The highest α-glucosidase inhibitory activity of H. erinaceus was measured at stage 4, which correlates with the contents of hericene derivatives. Conclusively, H. erinaceus with better efficacy and high content of active constituents can be secured by the optimization of cultivation conditions.

Optimization of Cultivation and Extraction Conditions of Pupae-Cordyceps for Cordycepin Production

Cordycepin is a characteristic bioactive compound of Cordyceps militaris with various beneficial effects. Cordyceps grows on both grains and insects, and the content of cordycepin varies depending on the cultivation conditions. In this study, the effect of culture conditions on the cordycepin content was analyzed and the extraction conditions were optimized. Analysis of cordycepin content in Pupae-Cordyceps found that it was highly affected by temperature in culture conditions. In the case of mycelium, it grows well at 20 and 25 oC, but not at 30 oC. However, the content of cordycepin was highest at 30oC and less at 20 oC. The fruiting body also showed a similar tendency: growth was 20 oC > 25 oC > 30 oC, but the cordycepin content was 30 oC > 25 oC > 20 oC. The content of cordycepin decreased after the fruiting bodies were produced. Next, extraction conditions such as solvent and time were optimized for maximum cordycepin content using response surface methodology(RSM). There was a large difference in the content of cordycepin according to the content of ethanol and the extraction temperature. Through RSM, it was confirmed that the optimum condition for extraction of cordycepin was 48.9 oC using 49.0% ethanol, and 160.9 mg/g extract could be obtained under this condition. In conclusion, this study suggested the optimized conditions for the cultivation and extraction of Pupae-Cordyceps for maximizing the content of cordycepin, and this may be applied to the discovery of materials using cordycepin.

Xanthone and Flavonoid Derivatives from the Leaves of Maclura tricuspidata with Antioxidant and Anti-tyrosinase Activity

Masclura tricuspidata, also called as Cudrania tricuspidata, is one of the most common Moraceae family plants in East Asia. Its trivial name follows mulberry due to the similar morphology. Investigation of the bioactive constituents of M. tricuspidata leaves yielded a new xanthone derivative along with twenty known compounds through various chromatographic techniques. A new compound was defined as mascluraxanthone (3), a prenylated xanthone glucoside on the basis of 1D and 2D NMR and MS data. Twenty known compounds were identified as four xanthone derivatives (1-2 and 4-5), two flavans (6-7), six flavanol derivatives (8-13), a flavonone (14) and seven flavonol derivatives (15-21). Among the isolated compounds, flavanol and flavonoid derivatives with 3′,4′-OH groups showed antioxidant and anti-tyrosinase activities. Conclusively, the leaves of M.tricuspidata are rich in aromatic compounds including xanthones and flavonoids. In addition, these constituents showed antioxidant and anti-tyrosinase potentials, which might be useful for oxidative stress related diseases.

Optimization of Cultivation and Extraction Conditions of Pupae-Cordyceps for Cordycepin Production

Cordycepin is a characteristic bioactive compound of Cordyceps militaris with various beneficial effects. Cordyceps grows on both grains and insects, and the content of cordycepin varies depending on the cultivation conditions. In this study, the effect of culture conditions on the cordycepin content was analyzed and the extraction conditions were optimized. Analysis of cordycepin content in Pupae-Cordyceps found that it was highly affected by temperature in culture conditions. In the case of mycelium, it grows well at 20 and 25 oC, but not at 30 oC. However, the content of cordycepin was highest at 30oC and less at 20 oC. The fruiting body also showed a similar tendency: growth was 20 oC > 25 oC > 30 oC, but the cordycepin content was 30 oC > 25 oC > 20 oC. The content of cordycepin decreased after the fruiting bodies were produced. Next, extraction conditions such as solvent and time were optimized for maximum cordycepin content using response surface methodology(RSM). There was a large difference in the content of cordycepin according to the content of ethanol and the extraction temperature. Through RSM, it was confirmed that the optimum condition for extraction of cordycepin was 48.9 oC using 49.0% ethanol, and 160.9 mg/g extract could be obtained under this condition. In conclusion, this study suggested the optimized conditions for the cultivation and extraction of Pupae-Cordyceps for maximizing the content of cordycepin, and this may be applied to the discovery of materials using cordycepin.

Two New Caffeoyl Threonate Esters from the Leaves of Toxicodendron vernicifluum

Toxicodendron vernicifluum, also called as Rhus verniciflua is a deciduous tree belonging to Anacardiaceae family. Two new caffeoyl threonate esters, rhuseols A (1) and B (2), together with 5-O-(E)-caffeoylquinic acid methyl ester (3) were isolated from the leaves of T. vernicifluum. The structures of isolated compounds were established by using 1D and 2D NMR in combination with HR-ESI-MS. Compounds 1 – 3 showed DPPH radical scavenging effects with IC₅₀ values of 47.9, 107.8 and 15.4 µM, respectively. Taken together, these compounds might contribute to the antioxidant properties of the leaves of T. vernicifluum, which will be useful for various oxidative stress mediated diseases.

Antioxidant Activity and Phenolic Content of Different Parts of Lotus and Optimization of Extraction Condition using Response Surface Methodology

Nelumbo nucifera Gaertn. (Nymphaeaceae) is commonly called lotus and its leaves are widely been used as functional ingredients due to its antioxidant activity. For maximum efficacy, optimized extraction condition was established using response surface methodology. The high F-values, low p-values and insignificant p-value for lack-of-fit supported the fitness of the model and yielded the second-order polynomial regression for the antioxidant activity. The optimized extract was obtained by the extraction of 1 g of lotus leaves with 40 mL of 50% MeOH at 10.0℃, which exerted 70.1% antioxidant activity. Close correlation between phenolic content and antioxidant activity suggested phenolic compounds as active constituents of lotus leaves. In addition, comparison of different parts of lotus demonstrated the most potent antioxidant activity of flowers, followed by leaves and roots. Taken together, these results provide useful information about lotus leaves for the development as antioxidant ingredients. In addition, flowers and roots as well as leaves are suggested as good sources for antioxidant activity.

Nitric Oxide Inhibitory Constituents from the Fruits of Amomum tsao-ko

Bioactivity-guided fractionation of MeOH extract of the dried fruits of Amomum tsao-ko led to isolation of nine compounds (1 – 9). Their structures were elucidated by spectroscopic methods including extensive 1D and 2D-NMR, as alpinetin (1), naringenin-5-O-methyl ether (2), naringenin (3), hesperetin (4), 2′,4′,6′-trihydroxy-4-methoxy chalcone (5), tsaokoin (6), boesenbergin B (7), 4-hydroxyboesenbergin B (8), and tsaokoarylone (9). Of these, compound 8 was isolated from a natural source for the first time, which was previously reported as a synthetic product. The isolated compounds (1 – 9) were tested for their inhibitory effects on LPS-induced nitric oxide production in RAW 264.7 macrophages. Among them, three chalcone derivatives (compounds 5, 7, and 8) and a diarylheptanoid (compound 9) exhibited significant inhibitory activity on the NO production with IC₅₀ values ranging from 10.9 to 22.5 µM.

Optimization of Extraction Condition of Methyl Jasmonate-treated Wild Ginseng Adventitious Root Cultures using Response Surface Methodology

The usage of wild ginseng (Panax ginseng C.A. Meyer) has been limited due to short supply and high price. Therefore, sufficient production as well as efficient extraction of mountain ginseng are required for the development as products. In this study, wild ginseng adventitious root cultures were prepared for efficient production with advantages of fast growth and stable production. Treatment of methyl jasmonate (MJ) to wild ginseng adventitious root cultures increased the extraction yield and antioxidative activity. Further investigation on effect of extraction conditions suggested the importance of ethanol concentration on antioxidative activity and extraction yield of MJ-treated wild ginseng adventitious root cultures. Optimized extraction condition of MJ-treated wild ginseng adventitious root cultures for maximum extraction yield and antioxidative activity was determined using response surface methodology with three-level-three-factor Box-Behnken design (BBD). Extraction of 1 g MJ-treated wild ginseng adventitious root culture with 30 ml of 9% ethanol at 30 ℃ produced 310.2 mg extract with 71.0% antioxidative activity at 100 µg/ml. Taken together, MJ-treated wild ginseng adventitious root culture is valuable source for wild ginseng usage and optimized extraction condition can be used for the development of functional products or folk remedies.

Chemical Constituents from Buddleja officinalis and Their Inhibitory Effects on Nitric Oxide Production

Bioactivity-guided fractionation of a methanolic extract of Buddleja officinalis led to the isolation of two monoterpenes, crocusatin M (1), crocusatin C (2), a flavonoid, acacetin (3), three lignans, lariciresinol (4), pinoresinol (5), and syringaresinol (6), and two triterpenoidal saponins, mimengoside B (7) and songarosaponin A (8). The structures of isolates were identified based on 1D-, 2D-NMR, and MS data analysis. All isolates were tested for their inhibition on LPS-induced NO production in RAW 264.7 cells. As a result, mimengoside B (7) and songarosaponin A (8) showed a mild inhibitory activity of NO production.

Effect of Extraction Conditions of Green Tea on Antioxidant Activity and EGCG Content: Optimization using Response Surface Methodology

Green tea, the leaves of Camellia sinsneis (Theaceae), is generally acknowledged as the most consumed beverage with multiple pharmacological functions including antioxidant activity. This study was performed to analyze the effect of extraction conditions of green tea on its antioxidant effects using DPPH assay. Three extraction factors such as extraction solvent (EtOH, 0 – 100%), extraction time (3 – 15 min) and extraction temperature (10 – 70℃) were analyzed and optimized extraction condition for antioxidant activity of green tea extract (GTE) was determined using response surface methodology with three-level-three-factor Box-Behnken design (BBD). Regression analysis showed a good fit of data and the optimal conditions of extraction were found to be 57.7% EtOH, 15 min and 70℃. Under this condition, antioxidant activity of experimental data was 88.4% which was almost fit to the ideal value of 88.6%. As epigallocatechin gallate (EGCG) is known for the major ingredient for antioxidant activity of green tea, we investigated the effect of EGCG on antioxidant activity of GTE. EGCG showed antioxidant activity with the IC50 value of 4.2 µg/ml and a positive correlation was observed between EGCG content and the antioxidant activity of GTE with R2 = 0.7134. Interestingly, however, GTE with 50 – 70% antioxidant activity contain less than 1.0 µg/ml of EGCG, which is much lower than IC50 value of EGCG. Therefore, we suppose that EGCG together with other constituents contribute to antioxidant activity of GTE. Taken together, these results suggest that green tea is more beneficial than EGCG alone for antioxidant ability and optimal extraction condition of green tea will be useful for the development of food and pharmaceutical applications.

Anti-Helicobacter pylori Compounds from Maackia amurensis

Eight isoflavonoid compounds were isolated from the EtOAc fraction of Maackia amurensis which had shown the highest anti-Helicobacter pylori activity among the fractions, using medium pressure liquid chromatography and recrystallization. Based on the spectroscopic data including 1H-NMR, 13C-NMR, HMBC and MS data, the chemical structures of the isolates were determined to be (-)-medicarpin (1), afromosin (2), formononetin (3), tectorigenin (4), prunetin (5), wistin (6), tectoridin (7) and ononin (8). Anti-H. pylori activity of each compound was evaluated with broth dilution assay. As a result, (-)-medicarpin (1), tectorigenin (4) and wistin (6) showed anti-H. pylori activity. (-)-Medicarpin (1) exhibited the most potent growth inhibitory activity against H. pylori with the minimal inhibitory concentration (MIC)90 of 25 microM, and tectorigenin (4) with MIC90 of 100 microM ranked the second. This is the first study to show the anti-H. pylori activity of M. amurensis, and it is suggested that the stem bark of M. amurensis or the EtOAc fraction or the isolated compounds can be a new natural source for the treatment of H. pylori infection.

Optimization of Extraction Condition of Hesperidin in Citrus unshiu Peels using Response Surface Methodology

Hesperidin, which is the most abundant flavonoid of Citrus unshiu (Rutaceae), has been reported to possess diverse activities and widely used as functional foods and cosmetics. For the development of functional products, extraction procedure is indispensable. Extraction conditions affect the composition of extract as well as its biological activity. Therefore, we tried to optimize extraction conditions such as extraction solvent, extraction time and extraction temperature for maximum yield of hesperidin using response surface methodology with threelevel-three-factor Box-Behnken design (BBD). Regression analysis showed a good fit of the experimental data and the optimal condition was obtained as ethanol concentration, 59.0%; temperature 71.5degrees C and extraction time, 12.4 h. The hesperidin yield under the optimal condition was found to be 287.8 microg per 5 mg extract, which was well matched with the predicted value of 290.5 microg. These results provides optimized extraction condition for hesperidin and might be useful for the development of hesperidin as functional products like health supplements, cosmetics and medicinal products.

Poria cocos ethanol extract and its active constituent, pachymic acid, modulate sleep architectures via activation of GABA(A)-ergic transmission in rats / 동물의과학연구지

Poria cocos is a well-known traditional Chinese traditional medicine (TCM) that grows around roots of pine trees in China, Korea, Japan, and North America. Poria cocos has been used in Asian countries to treat insomnia as either a single herb or part of an herbal formula. In a previous experiment, pachymic acid (PA), an active constituent of Poria cocos ethanol extract (PCE), increased pentobarbital-induced sleeping behaviors. The aim of this experiment was to evaluate whether or not PCE and PA modulate sleep architectures in rats as well as whether or not their effects are mediated through GABA(A)-ergic transmission. PCE and PA were orally administered to individual rats 7 days after surgical implantation of a transmitter, and sleep architectures were recorded by Telemetric Cortical encephalogram (EEG) upon oral administration of test drugs. PCE and PA increased total sleep time and non-rapid eye movement (NREM) sleep as well as reduced numbers of sleep/wake cycles recorded by EEG. Furthermore, PCE increased intracellular chloride levels, GAD65/67 protein levels, and alpha-, beta-, and gamma-subunits of GABA(A) receptors in primary cultured hypothalamic neuronal cells. These data suggest that PCE modulates sleep architectures via activation of GABA(A)-ergic systems. Further, as PA is an active component of PCE, they may have the same pharmacological effects.

4-Hydroxybenzaldehyde, One of Constituents from Gastrodiae Rhizoma Augments Pentobarbital-induced Sleeping Behaviors and Non-rapid Eye Movement (NREM) Sleep in Rodents

In the previous experiments, we reported that ethanol extract of Gastrodiae Rhizoma, the dried tuber of Gastrodia ElataBlume (Orchidaceae) increased pentobarbital-induced sleeping behaviors. These experiments were undertaken to know whether 4-hydroxybenzaldehyde (4-HBD), is one of the major compounds of Gastrodiae Rhizoma increases pentobarbital-induced sleeping behaviors and changes sleep architectures via activating GABA(A)-ergic systems in rodents. 4-HBD decreased locomotor activity in mice. 4-HBD increased total sleep time, and decreased of sleep onset by pentobarbital (28 mg/kg and 40 mg/kg). 4-HBD showed synergistic effects with muscimol (a GABA(A) receptor agonist), shortening sleep onset and enhancing sleep time on pentobarbital-induced sleeping behaviors. On the other hand, 4-HBD (200 mg/kg, p.o.) itself significantly inhibited the counts of sleep-wake cycles, and prolonged total sleep time and non-rapid eye movement (NREM) in rats. Moreover, 4-HBD increased intracellular Cl- levels in the primary cultured cerebellar cells. The protein levels of glutamic acid decarboxylase (GAD) and GABA(A) receptors subunits were over-expressed by 4-HBD. Consequently, these results demonstrate that 4-HBD increased NREM sleep as well as sleeping behaviors via the activation of GABA(A)-ergic systems in rodents.

Pachymic Acid Enhances Pentobarbital-Induced Sleeping Behaviors via GABA(A)-ergic Systems in Mice

This study was investigated to know whether pachymic acid (PA), one of the predominant triterpenoids in Poria cocos (Hoelen) has the sedative-hypnotic effects, and underlying mechanisms are mediated via gamma-aminobutyric acid (GABA)-ergic systems. Oral administration of PA markedly suppressed locomotion activity in mice. This compound also prolonged sleeping time, and reduced sleep latency showing synergic effects with muscimol (0.2 mg/kg) in shortening sleep onset and enhancing sleep time induced by pentobarbital, both at the hypnotic (40 mg/kg) and sub-hypnotic (28 mg/kg) doses. Additionally, PA elevated intracellular chloride levels in hypothalamic primary cultured neuronal cells of rats. Moreover, Western blotting quantitative results showed that PA increased the amount of protein level expression of GAD65/67 over a broader range of doses. PA increased alpha- and beta-subunits protein levels, but decreased gamma-subunit protein levels in GABA(A) receptors. The present experiment provides evidence for the hypnotic effects as PA enhanced pentobarbital-induced sleeping behaviors via GABA(A)-ergic mechanisms in rodents. Taken together, it is proposed that PA may be useful for the treatment of sleep disturbed subjects with insomnia.

Combined Genome-Wide Linkage and Association Analyses of Fasting Glucose Level in Healthy Twins and Families of Korea

This study was undertaken to identify genetic polymorphisms that are associated with the risk of an elevated fasting glucose (FG) level using genome-wide analyses. We explored a quantitative trait locus (QTL) for FG level in a genome-wide study from a Korean twin-family cohort (the Healthy Twin Study) using a combined linkage and family-based association analysis approach. We investigated 1,754 individuals, which included 432 families and 219 pairs of monozygotic twins. Regions of chromosomes 2q23.3-2q31.1, 15q26.1-15q26.3, 16p12.1, and 20p13-20p12.2, were found to show evidence of linkage with FG level, and several markers in these regions were found to be significantly associated with FG level using family-based or general association tests. In particular, a single-nucleotide polymorphism (rs6138953) on the PTPRA gene in the 20p13 region (combined P = 1.8 x 10(-6)) was found to be associated with FG level, and the PRKCB1 gene (in 16p12.1) to be possibly associated with FG level. In conclusion, multiple regions of chromosomes 2q23.3-2q31.1, 15q26.1-15q26.3, 16p12.1, and 20p13-20p12.2 are associated with FG level in our Korean twin-family cohort. The combined approach of genome-wide linkage and family-based association analysis is useful to identify novel or known genetic regions concerning FG level in a family cohort study.

Successful Schooling Rate and Satisfaction of the Inpatient Hospital School Participants among the Child and Adolescent Psychiatric Inpatients in the Closed Ward

OBJECTIVES: The objective of this report is to identify the utilization of hospital school service during hospitalization among patients in their childhood and adolescence with psychiatric disorders. METHODS: We retrospectively reviewed the medical record of child and adolescent psychiatric who were hospitalized during March 2009 through October 2012. We compared the one-year successful schooling and outpatient follow up rate between users and nonusers of the inpatient hospital school service. The hospital schooling experiences of the users were investigated upon follow-up visits to the outpatient clinic. RESULTS: Sixty-three students received hospital school service during hospitalization among total 122 child and adolescent inpatients. Hospital school participants showed a significantly higher school reentry rate (61.9%) than non-participants (40.7%). However, there was no difference on follow up rate between the two groups. More than 60% of the 22 interviewed participants expressed an above-average level of satisfaction about hospital school service. CONCLUSION: Many patients with mental illness experience difficulty in receiving school education during treatment. That induces deterioration in disease, academic failure, poor social skills, low self-esteem, economic difficulties, and future job opportunities. The results of this study emphasize the importance of hospital school service and offer useful guidance for hospital school operation.

Genome-wide linkage analysis for ocular and nasal anthropometric traits in a Mongolian population

Anthropometric traits for eyes and nose are complex quantitative traits influenced by genetic and environmental factors. To date, there have been few reports on the contribution of genetic influence to these traits in Asian populations. The aim of this study was to determine the genetic effect and quantitative trait locus (QTL) of seven traits eyes- and nose-related anthropometric measurements in an isolated Mongolian population. Frontal and lateral photographs were obtained from 1,014 individuals (434 males and 580 females) of Mongolian origin. A total of 349 short tandem repeat markers on 22 autosomes were genotyped for each individual. Heritability estimates of the seven ocular and nasal traits, adjusted for significant covariates, ranged from 0.48 to 0.90, providing evidence for a genetic influence. Variance-component linkage analyses revealed 10 suggestive linkage signals on 5q34 (LOD = 3.2), 18q12.2 (LOD = 2.7), 5q15 (LOD = 2.0), 9q34.2 (LOD = 1.9), 5q34 (LOD = 1.9), 17q22 (LOD = 1.9), 13q33.3 (LOD = 2.7), 1q36.22 (LOD = 1.9), 4q32.1 (LOD = 2.1) and 15q22.31 (LOD = 2.9). Our study provides the first evidence that genetics influences nasal and ocular traits in a Mongolian population. Additional collaborative efforts will further extend our understanding of the link between genetic factors and human anthropometric traits.

Analysis of genetic and non-genetic factors that affect the QTc interval in a Mongolian population: the GENDISCAN study

The QTc interval is a complex quantitative trait and a strong prognostic indicator of cardiovascular mortality in general, healthy people. The aim of this study was to identify non-genetic factors and quantitative trait loci that govern the QTc interval in an isolated Mongolian population. We used multiple regression analysis to determine the relationship between the QTc interval and non-genetic factors including height, blood pressure, and the plasma lipid level. Whole genome linkage analyses were performed to reveal quantitative trait loci for the QTc interval with 349 microsatellite markers from 1,080 Mongolian subjects. Among many factors previously known for association with the QTc interval, age, sex, heart rate, QRS duration of electrocardiogram and systolic blood pressure were also found to have influence on the QTc interval. A genetic effect for the QTc interval was identified based on familial correlation with a heritability value of 0.31. In a whole genome linkage analysis, we identified the four potential linkage regions 7q31-34, 5q21, 4q28, and 2q36.

Erratum

The authors regret an error in discussion, the authors wrote that "We also performed an additional linkage analysis using only the adult population (> or = 20 years old)(Table 7, Figure 2 and 3)." In this sentence, Table 7 should be changed to Table 6.