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The main challenges for the development of agricultural bio-degradable mulching film concern the degradation during the lifetime of cultivated crops. A set of rice by-product (rice-hull and rice-bran) based bio-degradable mulching films was developed and tested, following the measurement of standard bio-degradability rate and adaptability in a large scale field experiment. The standard bio-degradability of bio-film passed the KS (Korea standard) regulation. The result of mechanical analysis of bio-degradable mulching film presented a higher mechanical strength and elongation rate compared with polyethylene film. In addition, bio-film could be degraded into fragments within 4 months under the field condition of several upland crops. Bio-degradable mulching film indicated great potential as a new source of agricultural bio-degradable material.

Grains of sugary rice were extracted with 80% aqueous methanol, and the concentrated extracts were successively partitioned using ethyl acetate, n-butanol, and water. From the n-butanol fractions, four flavonoid glycosides were isolated through repeated silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies. Based on the nuclear magnetic resonance, mass spectrometry, and infrared spectroscopic data, the chemical structures of the compounds were determined to be taxifolin-7-O-β-d-glucopyranoside (1), hyperin (2), isoquercitrin (3), and quercetin gentiobioside (4). These compounds were isolated from the grains of sugary rice for the first time. All isolated compounds were tested for antioxidant activity and low-density lipoprotein (LDL)-antioxidative activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and LDL assays. Compound 1 exhibited a strong scavenging effect on DPPH, with a 50% inhibition concentration (IC(50)) value of 8.1 μM, and also inhibited LDL oxidation with an IC(50) value of 40.0±20 μM. A simple and efficient high-performance liquid chromatography/diode array detection method for the simultaneous determination of the four bioactive flavonoids (1-4) has been developed and applied to their content determination in the sugary rice. The grains were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to be 1.12±0.045, 0.65±0.011, 0.68±0.032, and 0.89±0.021 mg/g, respectively.

Momilactone B (MB) is a terpenoid phytoalexin present in rice bran that exhibits several biological activities. MB reduced the melanin content in B16 melanocytes melanin content and inhibited tyrosinase activities. Using transcriptome analysis, the genes involved in protein kinase A (PKA) signaling were found to be markedly altered. B16 cells stimulated with MB had decreased concentrations of cAMP protein kinase A activity, and cAMP-response element-binding protein which is a key transcription factor for microphthalmia-associated transcription factor (MITF) expression. Accordingly, the expression of MITF and its target genes, which are essential for melanogenesis, were reduced. MB thus exhibits anti-melanogenic effects by repressing tyrosinase enzyme activity and inhibiting the PKA signaling pathway which, in turn, decreases melanogenic gene expression.

Anthocyanins have been studied as potential antimicrobial agents against Helicobacter pylori. We investigated whether the biosynthesis and secretion of cytotoxin-associated protein A (CagA) and vacuolating cytotoxin A (VacA) could be suppressed by anthocyanin treatment in vitro. H. pylori reference strain 60190 (CagA(+)/VacA(+)) was used in this study to investigate the inhibitory effects of anthocyanins; cyanidin 3-O-glucoside (C3G), peonidin 3-O-glucoside (Peo3G), pelargonidin 3-O-glucoside (Pel3G), and malvidin 3-O-glucoside (M3G) on expression and secretion of H. pylori toxins. Anthocyanins were added to bacterial cultures and Western blotting was used to determine secretion of CagA and VacA. Among them, we found that C3G inhibited secretion of CagA and VacA resulting in intracellular accumulation of CagA and VacA. C3G had no effect on cagA and vacA expression but suppressed secA transcription. As SecA is involved in translocation of bacterial proteins, the down-regulation of secA expression by C3G offers a mechanistic explanation for the inhibition of toxin secretion. To our knowledge, this is the first report suggesting that C3G inhibits secretion of the H. pylori toxins CagA and VacA via suppression of secA transcription.

Seed germination and dormancy of three green manure seeds such as hairy vetch, crimson clover and narrow-leaved vetch (Vicia angustifolia L.) produced in Milyang, southern part of Korea were investigated to provide a basic information on the stability of seedling establishment in green manure seeds. In addition. the effect of seed storage duration on the germinability of imported hairy vetch and crimson seeds was also investigated. To determine the seed maturity time of three green manure seeds, the seeds were harvested manually at May 25, May 30, and June 4, and the 100-seed weight, germinability and percentage of hard seed were evaluated. Freshly harvested seeds of three green manure crops were strongly dormant because of hard seed coat. Germination percentage of the three green manure crops varied depending on the seed harvest time ranging from 30 - 52% in hairy vetch, 16 - 35% in crimson clover and 2 - 61% in V. angustifolia, respectively. Of the three crops, crimson clover and V. angustifolia matured early on May 30 to June 4 (around 35 to 40 days after flower) but hairy vetch seed did not mature until June 4 based on the seed germinability. The matured seed of crimson clover and V. angustifolia germinated less than 20% while scarification on seed coat significantly overcome the innate dormancy. On the other hand, the seed germination of hairy vetch and crimson clover was lower under dark than in the presence of light. The germination percentage of the imported hairy vetch and crimson clover seeds showed high with 77 - 79%, 94 - 95%, respectively, but the seed germination significantly declined by 21 - 32%, 30 - 40% after one-year of storage under natural conditions. The seeds germinated only 8 - 13% for hairy vetch but crimson clover did not germinate at all after two years storage. These results indicate V. angustifolia and crimson clover mature at May 30 to June 4 but hairy vetch did not. The imported hairy vetch and crimson clover seeds should be used within one-year after import for stable seedling establishment in green manure-rice cultivation cropping system.

Fresh and chopped aerial parts of Oryza sativa were extracted in 80% aqueous mehthanol, and the concentrated extract was successively partitioned using n-hexane, ethyl acetate (EtOAc), nbutanol, and H2O. From the EtOAc fraction, a new flavonolignan (2) and a known flavonolignan, salcolin B (1), were isolated through repeated SiO2 and octadecyl silica gel column chromatography. Based on NMR, mass spectrometer, and IR spectroscopic data, the chemical structure of compound (2) was determined to be tricin-4’-O-[erythro-β-guaiacyl-(7’’-O-methyl)-glyceryl] ether, which has thus far never been reported and was named salcolin C (2). Salcolin B (1) and salcolin C (2) dose-dependently inhibited nitric oxide production in RAW 264.7 cells with IC50 values of 29.8±0.6 and 10.8±1.0 μM, respectively, without visible toxic effect.

Nutritional and physicochemical properties of new giant embryo mutant rice (YR23517Acp79, YR) were analyzed. YR exhibited increased total protein (9.3±0.3%), lipid (3.7±1.1%), amino acid (663.28±1.9 mg/g), and mineral contents (Ca=284.0±6.2, Mg=1417.5±13.6 mg/kg). In YR brown rice four major physicochemicals, including gamma-aminobutyric acid (brown rice= 0.46±0.014 mg/g), γ-oryzanol (0.43±0.021 mg/g), vitamin B1 (6.42±0.3 mg/kg), and tocopherols (alpha= 2.68±0.1, beta=0.11±0.01, gamma=0.05±0.001 mg/100 g) increased in comparison to reported giant embryo (Keunnunbyeo, KB) and normal embryo rice (Ilmibyeo, IB). YR showed higher scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (0.2 g/mL=57.1±2.25) and 2,2′-Azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (0.2 g/mL=50.2±1.45) radicals but also inhibited lipopolysaccharide-induced nitric oxide production without cytotoxicity. These results indicate YR is a high quality functional rice due to its high nutrition content and antioxidant effects of physicochemicals.

Total polyphenolic, protein, lipid, and anthocyanin contents in grains of 12 Korean colored rice varieties were evaluated for their 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl) (DPPH) and 2,2- Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium (ABTS) radical- scavenging activities. Three anthocyanins, cyanidin-3-O-glucoside (Cy-3-G), petunidin-3-O-glucoside (Pt-3-G), and peonidin-3-O-glucoside (Pn-3-G), were also characterized by ultra performance liquid chromatography. Among the rice varieties, ‘Heugjinju’, ‘Heugseol’, and ‘Sintoheugmi’ contained high total phenolic contents compared to other varieties. Anthocyanin content was highest in ‘Heugjinju’; no anthocyanins were detected in ‘Hongjinju’ and ‘Jeogjinju’. Highest antioxidant activity was observed in ‘Heugjinju’ and ‘Jeogjinju’ exhibited the lowest activity. To determine the correlation of total polyphenolic and anthocyanin contents to antioxidant activity, linear regression analysis was carried out. The results showed total polyphenolic content was strongly correlated with antioxidant activity, suggesting total phenolic content as key factor in antioxidant activity of colored rice. ‘Heugjinju’, ‘Heugseol’, and ‘Sintoheugmi’ could have important nutritional value.

The radiosensitizing effects of naturally occurring triterpenes were investigated in human lung cancer cells. Several quinone methide-containing triterpenes (QMTs) enhanced the cytotoxic effect of ionizing radiation (IR) and of these QMTs, celastrol (CE) had the greatest enhancing effect on IR-induced cell death in vitro. Additionally, the quinone methide moiety of CE was shown to be essential for CE-mediated radiosensitization; in contrast, dihydrocelastrol (DHCE), does not contain this moiety. Reactive oxygen species (ROS) production by IR was augmented in combination with CE, which was responsible for CE-mediated radiosensitization. CE induced the thiol reactivity and inhibited the activities of antioxidant molecules, such as thioredoxin reductase and glutathione. In vivo, nude mouse xenografting data also revealed that tumor growth delay was greater in mice treated with CE plus IR, compared with those treated with CE or IR alone. When DHCE, instead of CE, was combined with IR, tumor growth delay was similar to that in IR alone-treated mice. These results demonstrate that CE synergistically enhances the effects of IR and suggest the novel anticancer therapeutic use of CE in combination with radiation therapy.

This study was aimed to develop blend films by rice by-product (rice-hull and rice-bran) and bio-degradable materials. The rice by-product was firstly prepared from the pulverizing for making fine powder. Bio-degradable materials could be prepared by melting at high temperature. The mixture of the fine powder of rice by-product and melted bio-degradable materials was then blended and cast into films. The obtained films were investigated on their morphology, secondary structures and properties by using SEM, ICP and ASTM, respectively. Mechanical properties and degradability of these films were measured and compared to those of the PE films. Mechanical strength of bio-films was higher than that of PE films, however elongation ratio showed lower percent than that of PE film. In addition, bio-film could be degraded into fragments within 3 months under the field condition of normal upland crop cultivation. Bio-degradable mulching film indicated great potential for agronomic use as a new source of bio-degradable material.

The purpose of this study was to analyze and report the nutritional and physicochemical properties of 13 barnyard millet varieties (Echinochloa spp.) in Korea. Among the varieties, IT153600 exhibited the highest total protein (14.75±1.7%), lipid (6.92±1.1%), and amino acids contents (137.10±3.1 mg/g), and had a high germination rate (98.9±1.8%). For fatty acid composition, the highest linoleic acid (67.6±2.5%) content was found in K141285. K141286 varieties exhibited the highest amylose content. The highest mineral content was found in IT153604. Based on excellent agricultural properties, K141285 was selected and investigated for its antioxidant effects using DPPH and ABTS. The 80% methanol extract of K141285 showed significantly high total phenolic (38.45±0.45) and flavonoid (28.71±0.27) contents. These results suggest that all of the selected barnyard millet (E. spp.) varieties can be considered to be new functional crops due to their antioxidative effects and high nutrient contents.

The bio-degradable transplanting pot containing rice by-product (rice-hull and rice-bran) were developed, and tested their ability for agronomic use. Rice by-products were crosslinked with biodegradable aliphatic ally aromatic copolyesters or urea resin for making transplanting pot. Mechanical properties and degradability of these pots were measured and compared to those of the Jiffy pot (commercially used bio-degradable pot). Mechanical strength was higher than that of Jippy pot, and bio-degradability was excellent under the actual field condition. In addition, the pot could be degraded within 3 months under the ground. Our result indicated bio-degradable pot containing rice by-products has a great potential for such agronomic use.

Purpose HSP27 or HSP25 negatively regulates apoptosis pathways after radiation or chemotherapeutic agents. Abrogation of HSP27 function may be a candidate target for overcoming radio- and chemoresistance. Methods and Materials Zerumbone (ZER), a cytotoxic component isolated from Zingiber zerumbet smith . Clonogenic survival assay and flow cytometry after Annexin V staining were performed to determine in vitro sensitization effects of ZER with ionizing radiation. A nude mouse xenografting system was also applied to detect in vivo radiosensitizing effects of ZER. Results ZER produced cross-linking of HSP27, which was dependent on inhibition of the monomeric form of HSP27. ZER was directly inserted between the disulfide bond in the HSP27 dimer and modified normal HSP27 dimerization. Pretreatment with ZER before radiation inhibited the binding affinity between HSP27 and apoptotic molecules, such as cytochrome c and PKCδ, and induced sensitization in vitro and in an in vivo xenografted nude mouse system. Structural analogs lacking only the carbonyl group in ZER, such as α-humulene (HUM) and 8-hydroxy-humulen (8-OH-HUM), did not affect normal cross-linking of HSP27 and did not induce radiosensitization. Conclusions We suggest that altered cross-linking of HSP27 by ZER is a good strategy for abolishing HSP27-mediated resistance.

A new flavonolignan glucoside, tricin-4′-O-(erythro-β-guaiacylglyceryl)ether 9′′-O-β-D-glucopyranoside (9′′-O-glucopyranosyl salcolin B), was isolated from the aerial parts of Oryza sativa L. The structure of the compound was established on the basis of NMR, FAB-MS, and IR spectroscopic data.

Overexpression of c-Myc represents the most frequently deregulated genetic event in cancer, and therefore c-Myc may represent a good molecular target for cancer therapy. The human lung carcinoma cell line, NCI-H1299, shows resistance to conventional cancer treatments, such as ionizing radiation (IR) and cisplatin, while the lung carcinoma cell line, NCI-H460, is sensitive to treatment with these agents. However, when treated with a chalcone compound [toluenesulfonylamido-chalcone, 4'-(p-toluene sulfonyl amino)-3,4-dihydroxy chalcone (TSHDC)], cell death was dramatically induced in NCI-H1299 cells as compared to NCI-H460 cells. TSHDC-mediated cytotoxicity was not dependent on the status of p53 and p21. However, TSHDC exerted increased c-Myc-dependent reactive oxygen species (ROS) production in NCI-H1299 cells in which c-Myc is overexpressed, while increased ROS production did not occur in A549 or NCI-H460 cells with a low c-Myc level. Several colon and brain cancer cells also showed a correlation between c-Myc expression and TSHDC-mediated increased cell death. Tumor regression by TSHDC was more dramatic in NCI-H1299 cells than NCI-H460 cells, when these cells were grafted to nude mice. However, in the case of IR and cisplatin, NCI-H460 cells were more sensitive than NCI-H1299 cells. From these results, c-Myc-mediated ROS production may be a good target for screening of novel cancer drugs and TSHDC might be a good candidate as a cancer drug, specifically in cancer cells that overexpress c-Myc.

The 4'-(p-toluenesulfonylamino)-4-hydroxychalcone (TSAHC), which bears inhibitory chemotypes for both alpha-glucosidase and tyrosinase, was evaluated for tyrosinase activity and depigmenting ability relative to compounds designed to only target tyrosianse activity. TSAHC emerged to be a competitive reversible inhibitor of mushroom tyrosinase. More importantly, it was also able to return the melanin content of alpha-melanocyte stimulated by alpha-MSH to base levels unlike other inhibitors that only targeted tyrosinase. The Western blot for expression levels of proteins involved in melanogenesis showed that TSAHC significantly decreased three main tyrosinase related protein in melanin biosynthesis, tyrosinase, TRP-1 and TRP-2.

Graphical abstract The presence of a quinone-methide moiety appears to play a relatively significant role in 3CLpro inhibition., Quinone-methide triterpenes, celastrol (1), pristimerin (2), tingenone (3), and iguesterin (4) were isolated from Triterygium regelii and dihydrocelastrol (5) was synthesized by hydrogenation under palladium catalyst. Isolated quinone-methide triterpenes (1–4) and 5 were evaluated for SARS-CoV 3CLpro inhibitory activities and showed potent inhibitory activities with IC50 values of 10.3, 5.5, 9.9, and 2.6 μM, respectively, whereas the corresponding 5 having phenol moiety was observed in low activity (IC50 = 21.7 μM). As a result, quinone-methide moiety in A-ring and more hydrophobic E-ring assist to exhibit potent activity. Also, all quinone-methide triterpenes 1–4 have proven to be competitive by the kinetic analysis.

The azuki bean in Korea consists of seven domestic varieties which have been developed and registered for the public during last 25 years. Here, we present a simple but reliable method to screen and identify Korean azuki bean varieties. A method based on simple sequence repeat (SSR) markers is widely used for prominent gene identification and variety discrimination. In molecular biology, real-time polymerase chain reaction (PCR) is a laboratory technique based on the polymerase chain reaction that is used to amplify and simultaneously quantify a targeted DNA molecule. It enables easy detection of a specific sequence in a DNA sample without performing electrophoresis and further processes. For separation of seven Korean azuki bean varieties, 110 unique azuki bean SSR markers from an (AG)n-enriched library were selected, synthesized and used for polymerase chain reaction (PCR). Data were taken through acrylamide gel electrophoresis and automated multi-capillary electrophoresis system for selection of specific markers and then changed into proper formats for data mining analysis. Ten primer pairs that showed high polymorphism were chosen for the indepth study. These ten primers were re-amplified with real-time PCR and checked the cycle threshold (Ct) and temperature (Tm) for comparison of amplification sequence in seven varieties. Consequently, a total of 20 alleles and 6 SSR primers were detected from the standard PCR amplification. Within these 6 primers, 7 alleles of 3 SSR primers were isolated for variety identification. From real-time PCR results, 3 SSR primers were selected as efficient markers for discrimination of seven Korean azuki bean varieties. The approach described here could be applied in monitoring our varieties and can be adapted in the azuki bean breeding program.

Naturally occurring triterpenoid compounds have long been used as anti-inflammatory, antimalarial, and insecticidal agents. It has become evident that some of the natural or synthetic triterpenoids have promising clinical potential as both a therapeutic and chemopreventive agent for cancer. However, the molecular basis for the antitumor activity of triterpenoid has yet to be defined. In this study, we show that pristimerin, a natural triterpenoid, induces mitochondrial cell death in human cervical cancer cells and that reactive oxygen species (ROS)-dependent activation of both Bax and poly(ADP-ribose) polymerase-1 (PARP-1) is critically required for the mitochondrial dysfunction. We also showed that c-Jun N-terminal kinase (JNK) is involved in ROS-dependent Bax activation. Treatment of pristimerin induced an increase in intracellular ROS, JNK activation, conformational change, and mitochondrial redistribution of Bax, mitochondrial membrane potential loss, and cell death. The PARP-1 was also found to be activated by pristimerin treatment. An antioxidant, N-acetyl-l-cysteine (NAC), inhibited pristimerin-induced JNK activation, Bax relocalization, and PARP-1 activation, as well as mitochondrial cell death. Moreover, inhibition of JNK clearly suppressed conformational change and mitochondrial translocation of Bax and subsequent mitochondrial cell death but did not affect PARP-1 activation. Inhibition of PARP-1 with 1,5-dihydroxyisoquinoline (DIQ) or with small interfering RNA of PARP-1 significantly attenuated pristimerin-induced mitochondrial membrane potential loss and cell death but did not affect JNK activation and Bax relocalization. These results indicate that the natural triterpenoid pristimerin induces mitochondrial cell death through ROS-dependent activation of both Bax and PARP-1 in human cervical cancer cells and that JNK is involved in ROS-dependent Bax activation.

The growth of normal cells is arrested when they come in contact with each other, a process known as contact inhibition. Contact inhibition is lost during tumorigenesis, resulting in uncontrolled cell growth. Here, we investigated the role of the tetraspanin transmembrane 4 superfamily member 5 (TM4SF5) in contact inhibition and tumorigenesis. We found that TM4SF5 was overexpressed in human hepatocarcinoma tissue. TM4SF5 expression in clinical samples and in human hepatocellular carcinoma cell lines correlated with enhanced p27Kip1 expression and cytosolic stabilization as well as morphological elongation mediated by RhoA inactivation. These TM4SF5-mediated effects resulted in epithelial-mesenchymal transition (EMT) via loss of E-cadherin expression. The consequence of this was aberrant cell growth, as assessed by S-phase transition in confluent conditions, anchorage-independent growth, and tumor formation in nude mice. The TM4SF5-mediated effects were abolished by suppressing the expression of either TM4SF5 or cytosolic p27Kip1, as well as by reconstituting the expression of E-cadherin. Our observations have revealed a role for TM4SF5 in causing uncontrolled growth of human hepatocarcinoma cells through EMT.

(3R,4S)-3-Hydroxy-4-N-allyl-N-Boc-amino-1-pentene 10, an important precursor for the synthesis of polyhydroxylated piperidines, has been achieved as a single diastereomer without racemization via vinyl Grignard addition to N-Boc-N-allyl aminoaldehyde 9, which was derived from an enantiopure natural amino acid. Having forged a tetrahydropyridine ring scaffold 13 from 10 in 85% yield via RCM using Grubbs II catalyst, we were able to effect its stereodivergent dihydroxylation, via a common epoxide intermediate to yield a range of interesting hydroxylated piperidines, including ent-1,6-dideoxynojirimycin (ent-1,6-dDNJ) 1 (28% overall yield) and 5-amino-1,5,6-trideoxyaltrose 2 (29% over all yield) in excellent dr. To the best of our knowledge, our synthesis of ent-1,6-dDNJ 1 is the most expeditious to date.

Valine and alanine were converted into the corresponding α-hydroxy-β-amino acids through intramolecular S N 2' cyclization. The novel cyclization protocol relied upon the use of N-benzyl-protected carbamates derived from α-amino acids.

Young barley leaves (YLB) have been no studies comparing the antioxidant activities of various cultivars, or differences the antioxidant activity at different harvest times. The objective of this study was to evaluate the differences in the antioxidant activities of YLB by different cultivars and growth stages. The 4 Korean YLB, Nakyoung, Dahyang, Saegang, and Boanchal, were sprouted, and their total polyphenols, total flavonoids, and ABTS radical cation (ABTS.+) scavenging activity were measured. The 4 cultivars exhibited high total polyphenol contents (TPC) and total flavonoid contents (TFC), with a range of 1047.8-1263.2 GAE mg.100 g-1 and 443.7-550.7 CE mg.100 g-1, DW, respectively. In addition, they exhibited high levels of ABTS.+ (RC50 value range of 41.8-80.8 ug.mL-1) scavenging activity. After sprouting, the antioxidant activity of the 4 cultivars increased. That is, in these YLB, the TPC, TFC and radical scavenging ability were enhanced by sprouting. Lutonarin (LN) and saponarin (SN) are the major compounds of YLB. LN exhibited stronger radical scavenging activity than SN. Over time, LN contents increased and SN contents decreased. As a result, the antioxidant ability of YLB was improved with increased growing time.

Mangosenone F (MSF), a natural xanthone, was isolated form Carcinia mangotana, and a few studies have reported its glycosidase inhibitor effect. In this study we investigated the anti lung cancer effect of MSF both in vitro and in vivo. MSF inhibited cancer cell cytotoxicity and induced and induced apoptosis via reactive oxygen species (ROS) generation in NCI-H460. MSF treatment also showed in pronounced release of apoptogenic cytochrome c from the mitochondria to the cytosol, downregulation of Bcl-2 and Bcl-xL, and upregulation of Bax, suggesting that caspase-mediated pathways were involved in MSF-induced apoptosis. ROS activation of the mitogen-activated protein kinase signaling pathway was shown to play a predominant role in the apoptosis mechanism of MSF. Compared with cisplatin treatment, MSF treatment showed significantly increased inhibition of the growth of NCI-H460 cells xenografted in nude mice. Together, these results indicate the potential of MSF as a candidate natural anticancer drug by promoting ROS production.

The enantiomerically pure (2R,3S)- and (2R,3R)-3-amino-2-hydroxy-4-phenylbutanoic acids (AHPBA) 1 and 3 are readily obtained from D-glucono-δ-lactone. Both AHPBAs are the structural key units of KMI derivatives which are the potent inhibitors of BACE 1 (β-secretase) and HIV protease. Additionally, the obtained AHPBAs 1 and 3 are converted to dipeptides of bestatin stereoisomers 2 and 4.

In Korea, Orostachys japonicus has been used traditionally as a drug and health food. The aim of this study was to investigate possible inhibitory effects of O. japonicus extracts on the formation of N-nitrosodimethylamines (NDMA). Chloroform extraction was the most effective method for recovering the highest number of phenolic compounds and flavonoids; in these extracts the greatest nitrite-scavenging activity and inhibition of NDMA formation occurred at pH 2.5. The chloroform extract was separated into 10 fractions (J1−J10); fraction J4 inhibited NDMA formation by 90.1 ± 0.4%. This fraction was then separated into five subfractions (J4-1−J4-5) using a silica gel column. Subfractions J4-2 [(+)-catechin] and J4-4 (3,4-dihdroxybenzoic acid) inhibited NDMA formation by 89.5 ± 0.9 and 77.6 ± 0.8%, respectively. Keywords: NDMA; nitrite-scavenging ability; Orostachys japonicus; phenolics

The high diastereoselectivity of the base-catalyzed epimerization of oxazolidin-2-ones 7 and 8 is shown to depend on the nature of the N-substituent (R group); when R = Bn, the 4,5-trans-product (4S,5R)-9 is formed, whereas when R = H the 4,5-cis-product (4S,5S)-10 is formed, both with >99:1 dr. The successful hydrolysis of the oxazolidin-2-one group in both cis- and trans-derivatives show this to be a stereodivergent route to enantiopure alpha-hydroxy-beta-amino isopentanoic acids (2R,3S)-1 and (2S,3S)-2.

The aggregation pheromone of the bean bug, Riptortus clavatus, consists of three components; (E)-2-hexenyl (Z)-3-hexenoate (E2HZ3H), (E)-2-hexenyl (E)-2-hexenoate (E2HE2H), and myristyl isobutyrate (MI). The role of each component to attract the bug was tested in the field using synthetic chemicals. The effectiveness of using E2HZ3H, E2HE2H, or MI alone to attract R. clavatus did not vary from that of the control. Blends of E2HZ3H+E2HE2H showed no attractiveness to R. clavatus, however, the addition of MI to E2HZ3H+E2HE2H (tertiary blend) showed significantly higher attractiveness to the bug than that of the control. The E2HE2H+MI blend was also equally attractive to the tertiary blend. A mixture of 7 mg MI with E2HZ3H (7 mg) and E2HE2H (36 mg) rivaled the single use of 100 mg MI in attractiveness. These findings indicate that MI can exert full attractiveness to the bug only in the presence of E2HZ3H and E2HE2H.

Intramolecular nucleophilic attack onto allylsulfonates promoted by silica gel acting as an acid catalyst provides expedient stereoselective access to 4,5-difunctionalized oxazolidin-2-ones. Precursors were prepared efficiently from enantiopure α-amino acids and subsequent manipulation of the oxazolidin-2-ones yielded enantiopure α-hydroxy-β-amino acids.

Repeated SiO 2 and octadecyl silica gel (ODS) columnchromatographies of the EtOAc fraction from Magnolia obovatafruits, 10 neolignans, named magnolol (1), honokiol (2),isoobovatol (3), isomagnolol (4), obovatol (5), obovatal (6), 9-methoxyobovatol (7), magnobovatol (8), obovaaldehyde (9), and2-hydroxyobovaaldehyde (10) were isolated and identified. Allisolated compounds were evaluated for in vitro cytotoxicityagainst seven human cancer cell lines.Keywords cancer cell · cytotoxicity · honokiol · Magnolia obovata · magnolol · neolignanMagnolia obovata Thunb. is a deciduous tree that is distributedthroughout Korea, China, and Japan. This plant has been used forthe treatment of fever, headache, diarrhea, anxiety, and relief ofasthma in Chinese medicine. M. obovata has been reported tohave anti-platelet (Pyo et al., 2002a; 2002b), anti-gastritic (Cho etal., 2008), anti-inflammatory (Tzeng and Liu, 2004; Seo et al.,2013) and cytotoxic (Min et al., 2008; Youn et al., 2008) activities.So far, magnolol, honokiol, and obovatol have been reported toshow cytotoxic activities against human cancer cells (Kim andRyu, 1999; Lin et al., 2001; Yang et al., 2003; Youn et al., 2008;Patrick et al., 2011). However cytotoxic effects of neolignans fromM. obovata fruits have never been reported. Therefore, in thepresent study, the neolignans from M. obovata fruits wereevaluated for cytotoxic effects against human cancer cell lines invitro using the micro culture tetrazolium (MTT) assay. We isolated10 neolignans from M. obovata fruits using SiO

The indium-mediated allylation of u-aminoaldehyde 3 from D-glucono-δ-lactone provided the syn-aminoalcohol with excellent diastereoselectivity. syn-Aminoalcohol was easily applied to the total syntheses of the potent glycosidase inhibitors; (+)-castano-spermine and (+)-6-epicastanospermine.

Neuraminidase (NA) is one of the key enzymes responsible for bacterial infection and pathogenesis. This study aimed to gain deeper insights into the inhibitory effects of flavone-glucosides (1-9) isolated from barley sprouts (BS) on neuraminidase activity. The isolated compounds were identified as, lutonarin (1), saponarin (2), isoorientin (3), orientin (4), isovitexin (5), isoscoparin-7-O-[6-sinapoyl]-glucoside (6), isoscoparin-7-O-[6-feruloyl]-glucoside (7), isovitexin-7-O-[6-sinapoyl]-glucoside (8), and isovitexin-7-O-[6-feruloyl]-glucoside (9). Among them, compounds 1-5 exhibited neuraminidase-inhibitory activities in a dose-dependent manner, with IC50 values ranging from 20.1 to 32.7 μM, in a non-competitive inhibition mode according to kinetic studies. Moreover, the individual flavone-glucoside levels differed notably, in particular, lutonarin (1) and saponarin (2) were shown to be present in the greatest amounts, according to UPLC analysis. Consequently, our results suggest that BS may be utilized as an effective NA inhibitor in human health food, additives, and feed.

Osteoporosis, an age associated skeletal disease, exhibits increased adipogenesis at the expense of osteogenesis from common osteoporotic bone marrow cells. In this study, black rice (Oryza sativa L.) extracts (BRE) were identified as osteogenic inducers. BRE stimulated the alkaline phosphatase (ALP) activity in both C3H10T1/2 and primary bone marrow cells. Similarly, BRE increased mRNA expression of ALP and osterix. Oral administration of BRE in OVX rats prevented decreases in bone density and strength. By contrast, BRE inhibited adipocyte differentiation of mesenchymal C3H10T1/2 cells and prevented increases in body weight and fat mass in high fat diet fed obese mice, further suggesting the dual effects of BRE on anti-adipogenesis and pro-osteogenesis. UPLC analysis identified cyanidin-3-O-glucoside and peonidin-3-O-glucoside as main anti-adipogenic effectors but not for pro-osteogenic induction. In mechanism studies, BRE selectively stimulated Wnt-driven luciferase activities. BRE treatment also induced Wnt-specific target genes such as Axin2, WISP2, and Cyclin D1. Taken together, these data suggest that BRE is a potentially useful ingredient to protect against age related osteoporosis and diet induced obesity.

Chalcones 1-20, a new class of glycosidase inhibitors, were synthesized, and their glycosidase inhibitory activities were investigated. Non-aminochalcones 1-12 had no inhibitory activity, however, aminochalcones 13-20 had strong glycosidase (alpha-glucosidase, alpha-amylase, and beta-amylase) inhibitory activities. In particular, sulfonamide chalcones 17-20 had more potent alpha-glucosidase inhibitory activity than aminated chalcone 13-16. 4'-(p-Toluenesulfonamide)-3,4-dihydroxy chalcone 20 (IC(50)=0.4microM) was the best inhibitor against alpha-glucosidase, and these sulfonamide chalcones showed non-competitive inhibition.

Column chromatographic technology was applied to isolate six purified ursane triterpenoids from the calyx of Fragaria ananassa and they were identified on the basis of spectroscopic methods to be ursolic acid (1), pomolic acid (2), 2-oxo-pomolic acid (3), 3-O-acetyl pomolic acid (4), fupenzic acid (5) and euscaphic acid (6). This is the first study in which these compounds have been isolated from the calyx of F. ananassa. Compared to a well-known inhibitor, α-arbutin, compounds 2-6 showed a significant decrease in intracellular melanin content in B16-F10 cells, and in culture media melanin.

The in vitro effects on melanogenesis of γ-oryzanol (1), a rice bran-derived phytosterol, were investigated. The melanin content in B16F1 cells was significantly and dose-dependently reduced (-13% and -28% at 3 and 30 μM, respectively). Tyrosinase enzyme activity was inhibited by 1 both in a cell-free assay and when analyzed based on the measurement of cellular tyrosinase activity. Transcriptome analysis was performed to investigate the biological pathways altered by 1, and it was found that gene expression involving protein kinase A (PKA) signaling was markedly altered. Subsequent analyses revealed that 1 stimulation in B16 cells reduced cytosolic cAMP concentrations, PKA activity (-13% for cAMP levels and -40% for PKA activity), and phosphorylation of the cAMP-response element binding protein (-57%), which, in turn, downregulated the expression of microphthalmia-associated transcription factor (MITF; -59% for mRNA and -64% for protein), a key melanogenic gene transcription factor. Accordingly, tyrosinase-related protein 1 (TRP-1; -69% for mRNA and -82% for protein) and dopachrome tautomerase (-51% for mRNA and -92% for protein) in 1-stimulated B16F1 cells were also downregulated. These results suggest that 1 has dual inhibitory activities for cellular melanogenesis by inhibiting tyrosinase enzyme activity and reducing MITF and target genes in the PKA-dependent pathway.

A novel liver X receptor (LXR) modulator, iristectorigenin B isolated from Belamcanda chinensis, stimulated the transcriptional activity of both LXR-α and LXR-β. In macrophages, iristectorigenin B suppressed cholesterol accumulation in a dose-dependent manner and induced the transcriptional activation of LXR-α/-β-responsive genes, ATP-binding cassette transporters A1 and G1. It did not induce hepatic lipid accumulation nor the expression of the lipogenesis genes sterol regulatory element-binding protein-1c, fatty acid synthase, and stearoyl-CoA desaturase-1. Iristectorigenin B thus is a dual-LXR agonist that regulates the expression of key genes in cholesterol homeostasis in macrophage cells without inducing hepatic lipid accumulation.

The objectives of this research were to evaluate antioxidant activities and nutritional components, including phenolic acid, catechin, organic acid, sugar, and amino acid, of persimmon juice from persimmons grown in different regions around Korea. Persimmon (Diospyros kaki) exhibits potent antioxidant effects in DPPH, ABTS, reducing power, and FRAP methods of analysis. The levels of nutritional constituents showed significant differences among all the samples. In particular, tartaric acid, glucose, gallic acid, epicatechin gallate and aspartic acid were observed to be the predominant component for each of their general chemical groups, with total average contents of 1876.51 mg/kg, 62.69 g/kg, 12.73 mg/kg, 208.99 mg/kg, and 31.84 mg/100 g, respectively. Interestingly, persimmons from the Hadong region presented the highest sugar (130.60 g/kg), phenolic acid (42.27 mg/kg), and catechin (527.97 mg/kg) contents in comparison with other regional samples. Moreover, this location exhibited the greatest antioxidant activity with highest total phenolic (298.01 mg GAE/kg) and flavonoid (32.11 mg/kg RE) contents. Our results suggest that strong antioxidant activities of persimmons correlate with high phenolic acid and catechin contents, particularly gallic acid and epicatechin gallate. Additionally, these two compounds may be key factors when considering the useful ingredients of persimmon.

Soybean leaves are eaten as seasonal edible greens in Korea. Analysis of the ethyl acetate extract of these leaves showed that it exhibited potent and selective neuraminidase inhibition, which began at the R3 stage and peaked at R7. Ten pterocarpans, including the new 6a-hydroxypterocarpan 10, were isolated from soybean leaves and their inhibition activities tested against a range of glycosidases. The relationship between structure and enzyme inhibition was investigated: 6a-hydroxypterocarpans exhibited much higher inhibition against neuraminidase (IC(50) = 2.4-89.4 μM) than α-glucosidase (IC(50) = 90.4- 100 μM). Glyceollin VII (7) displayed 40-fold greater activity (IC(50) = 2.4 μM) against neuraminidase than α-glucosidase (IC(50) = 90.4 μM). On the other hand, coumestanes (1-3) were good α-glucosidase inhibitors (IC(50) = 6.0-42.6 μM). In kinetic analysis, the most potent neuraminidase inhibitors (5-10) were noncompetitive. HPLC analysis indicated that most pterocarpan synthesis began from the R3 stage, and a rapid change of pterocarpan concentrations was observed between the R4 and R7 stages.

A combined treatment with conventional chemotherapies can enhance the effectiveness of chemotherapeutic agents against cancers. Here, we have shown that the naturally occurring triterpenoids synergistically enhance the response of cervical cancer cells to taxol. Of the triterpenoid compounds, pristimerin enhanced the anticancer effect of taxol with the highest efficiency by combination. Pristimerin synergizes with taxol to inhibit clonogenic survival and tumor growth in nude mice, and to enhance cell death in cervical cancer cells. A combined treatment with taxol and pristimerin induced cervical cancer cell death by increasing intracellular reactive oxygen species levels, upregulation of death receptor death receptor 5 (DR5), activation of Bax, and dissipation of mitochondrial membrane potential. Treatment with N-acetyl-L-cysteine, a thiol-containing antioxidant completely blocked combined treatment-induced Bax translocation as well as DR5 upregulation. Moreover, inhibition of Jun N-terminal kinase/c-Jun pathway attenuated cell death by blocking DR5 upregulation and Bax activation. These results indicate that the triterpenoid, pristimerin, synergistically enhances taxol response of cervical cancer cells through DR5 expression and Bax activation. Furthermore, the reactive oxygen species-dependent activation of the Jun N-terminal kinase/c-Jun pathway is required for the DR5 upregulation and Bax activation. The molecular mechanism revealed by this study may aid in the design of future combination cancer therapies against cells with intrinsically reduced sensitivity to taxol.

The radiosensitizing activity of celastrol, a quinone methide triterpene was examined. We found that celastrol treatment of the NCI-H460 lung cancer cell line increased radiation-induced cell killing. The increased radiosensitivity was correlated with decreased levels of Hsp90 clients, such as EGFR, ErbB2 and survivin as well as with increased p53 expression. Celastrol inhibited the ATP-binding activity of Hsp90. Furthermore, celastrol treatment dissociated an Hsp90 client protein, EGFR, and this in turn resulted in degradation of the client protein. These results were not observed with another structurally similar triterpenoid, 6β-acetonyl-22β-hydroxytingenol (TG), suggesting that a specific structural feature of the triterpenoid is required for radiosensitization. Moreover celastrol treatment increased p53 levels by phosphorylating Ser15 and Ser20 residues as well as by inhibiting its proteasomal degradation. Celastrol may be considered an effective radiosensitizer acting as an inhibitor of Hsp90 and a p53 activator. The two activities could be applicable to a broad range of cancer cells with either wild-type or mutant p53 because either activity could be effective for the enhancement of radiation cell killing. Further analysis with other triterpenoids should identify the functional moiety of the structure and additional candidates for effective radiosensitizers, which can be used in combined radiotherapy.

Quinone-methide triterpenes, celastrol (1), pristimerin (2), tingenone (3), and iguesterin (4) were isolated from Triterygium regelii and dihydrocelastrol (5) was synthesized by hydrogenation under palladium catalyst. Isolated quinone-methide triterpenes (1-4) and 5 were evaluated for SARS-CoV 3CL(pro) inhibitory activities and showed potent inhibitory activities with IC(50) values of 10.3, 5.5, 9.9, and 2.6 microM, respectively, whereas the corresponding 5 having phenol moiety was observed in low activity (IC(50)=21.7 microM). As a result, quinone-methide moiety in A-ring and more hydrophobic E-ring assist to exhibit potent activity. Also, all quinone-methide triterpenes 1-4 have proven to be competitive by the kinetic analysis.

Purpose Previous data suggest that the PKCδ catalytic V5 (PKCδ-V5) heptapeptide (HEPT) (FEQFLDI) binds HSP27 and blocks HSP27-mediated radio- or chemoresistance. Here we investigated further the in vivo function of the PKCδ-V5 HEPT. Methods and Materials Labeling of HEPT with Cy5.5 or fluorescein isothiocyanate was performed to evaluate in vitro or in vivo distribution of HEPT. A clonogenic survival assay, flow cytometry, and Western blotting of cleaved caspase-3 were performed to determine in vitro sensitization effects of HEPT plus ionizing radiation (IR) versus IR alone or those of HEPT plus cisplatin(Cis) versus Cis alone. A nude mouse xenografting system was also applied to detect in vivo sensitizing effects of HEPT. Results HEPT efficiently bound to HSP27 and showed sensitization after combined treatment with IR versus treatment with Cis alone in NCI-H1299 lung carcinoma cells, with higher HSP27 expression, which was similar to that of combined treatment with IR or with Cis alone in NCI-H460 lung carcinoma cells with lower HSP27 expression. In vivo image analysis using Cy5.5-labeled HEPT showed that HEPT was retained in HSP27-overexpressing cancer cells after xenografting to nude mice. Combined treatment of HEPT with IR versus that with Cis alone in xenografted mice showed that HEPT increased radio- or chemosensitization in NCI-H1299 cells compared to that in mice xenografted with NCI-H460 cells. Conclusions The novel PKCδ-V5 HEPT may help overcome HSP27-mediated radio- or chemoresistance.

Purpose Previously, we reported that radiation-induced ST6 Gal I gene expression was responsible for an increase of integrin β1 sialylation. In this study, we have further investigated the function of radiation-mediated integrin β1 sialylation in colon cancer cells. Methods and Materials We performed Western blotting and lectin affinity assay to analyze the expression and level of sialylated integrin β1. After exposure to ionizing radiation (IR), adhesion and migration of cells were measured by in vitro adhesion and migration assay. Results IR increased sialylation of integrin β1 responsible for its increased protein stability and adhesion and migration of colon cancer cells. However, for cells with an N -glycosylation site mutant of integrin β1 located on the I-like domain (Mu3), these effects were dramatically inhibited. In addition, integrin β1–mediated radioresistance was not observed in cells containing this mutant. When sialylation of integrin β1 was targeted with a sulfonamide chalcone compound, inhibition of radiation-induced sialylation of integrin β1 and inhibition of radiation-induced adhesion and migration occurred. Conclusion The increase of integrin β1 sialylation by ST6 Gal I is critically involved in radiation-mediated adhesion and migration of colon cancer cells. From these findings, integrin β1 sialylation may be a novel target for overcoming radiation-induced survival, especially radiation-induced adhesion and migration.

Intramolecular nucleophilic attack onto allylsulfonates promoted by silica gel acting as an acid catalyst provides expedient stereoselective access to 4,5-difunctionalized oxazolidin-2-ones. Precursors were prepared efficiently from enantiopure α-amino acids and subsequent manipulation of the oxazolidin-2-ones yielded enantiopure α-hydroxy-β-amino acids.