Paenibacillus konkukensis sp. nov., isolated from animal feed.

A Gram-stain-positive, oxidase- and catalase-positive, aerobic, rod-shaped bacterium, designated strain SK-3146T, was isolated from animal feed. Phylogenetic analysis, based on 16S rRNA gene sequence comparisons, revealed that the strain formed a distinct lineage within the genus Paenibacillus that was closely related to Paenibacillusyunnanensis JCM 30953T (98.6 %), Paenibacillusvulneris CCUG 53270T (98.0 %) and Paenibacilluschinjuensis DSM 15045T (96.9 %). Cells were non-motile, endospore-forming and formed milky colonies on NA and R2A agar media. Growth of strain SK-3146T occurred at temperatures of 18-45 °C, at pH 6.0-9.5 and between 0.5-3.0 % NaCl (w/v). The major menaquinone was MK-7, with lesser amounts of MK-6 present. The cell wall peptidoglycan of strain SK-3146T contained meso-diaminopimelic acid. The major fatty acids were anteiso-C15 : 0 and iso-C16 : 0. The major polar lipids were diphosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 53.8 mol% and the DNA-DNA hybridization relatedness values between strain SK-3146T and P.yunnanensis JCM 30953T and P.vulneris CCUG 53270T were 26.13±0.8 % and 38.7±0.6 %, respectively. The phenotypic, phylogenetic and chemotaxonomic results indicate that strain SK-3146T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus konkukensis sp. nov. is proposed. The type strain is SK-3146T (=KACC 18876T=LMG 29568T).

Cordycepin-mediated transcriptional regulation of human GD3 synthase (hST8Sia I) in human neuroblastoma SK-N-BE(2)-C cells.

In the present study, we firstly found that cordycepin elevated the gene expression of the human GD3 synthase (hST8Sia I) in human neuroblastoma SK-N-BE(2)-C cells. To elucidate the mechanism underlying the upregulation of hST8Sia I gene expression in cordycepin-treated SK-N-BE(2)-C cells, functional characterization of the promoter region of the hST8Sia I gene was performed. Analysis of promoter activity using varying lengths of 5'-flanking region showed a dramatic increase by cordycepin in the -1146 to -646 region, which contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1, and NF-κB. Site-directed mutagenesis for these binding sites and chromatin immunoprecipitation assay revealed that the NF-κB binding site at -731 to -722 is essential for the cordycepin-induced expression of the hST8Sia I in SK-N-BE(2)-C cells. Moreover, the hST8Sia I expression induced by cordycepin was significantly repressed by pyrrolidinedithiocarbamate, an inhibitor of NF-κB. These results suggested that cordycepin induces upregulation of hST8Sia I gene expression through NF-κB activation in SK-N-BE(2)-C cells.

Triptolide induces apoptosis through extrinsic and intrinsic pathways in human osteosarcoma U2OS cells.

Triptolide, a diterpene derived from Tripterygium wilfordii Hook f., a Chinese medicinal herb, has been reported to inhibit cell proliferation and induce apoptosis in various human cancer cells, but its anticancer effects on human osteosarcoma cells have not yet been elucidated. In this study, we investigated whether triptolide induces apoptosis in human osteosarcoma cells and the underlying molecular mechanisms. We firstly demonstrated that triptolide inhibited cell growth and induced apoptosis in U2OS cells. Western blot analysis showed that the levels of procaspase-8, -9, Bcl-2, Bid and mitochondrial cytochrome c were downregulated in triptolide-treated U2OS cells, whereas the levels of Fas, FasL, Bax, cytosolic cytochrome c, cleaved caspase-3 and cleaved PARP were upregulated. These results suggest that triptolide induces apoptosis in U2OS cells by activating both death receptor and mitochondrial apoptotic pathways.

JNP3, a new compound, suppresses PMA-induced tumor cell invasion via NF-κB down regulation in MCF-7 breast cancer cells.

The expression of matrix metalloproteinase (MMPs)-9 is critical for cell migration and can lead to invasion and metastasis of cancer cells. In the present study, we examined the inhibitory effects of JNP3, a new compound which was isolated from traditional Chinese medicine, on cell invasion and MMP-9 activation in phorbol myristate acetate (PMA)-induced MCF-7 cells. Treatment with JNP3 significantly and selectively inhibited PMA-induced MMP-9 secretion, mRNA expression and protein levels, and these results led to reduction of cell invasion and migration in PMA-induced MCF-7 cells. The results of MMP-9 promoter assay and EMSA showed that JNP3 specifically inhibited PMA-induced MMP-9 gene expression by blocking NF-κB-dependent transcriptional activity. In addition, PMA-induced phosphorylation of ERK1/2 and JNK were suppressed by JNP3 treatment, whereas the phosphorylation of p38 MAPK was not affected by JNP3. These results suggest that JNP3 can be potential anti-cancer agents through specific inhibition of NF-κB-dependent MMP-9 gene expression.

Oleifolioside A, a New Active Compound, Attenuates LPS-Stimulated iNOS and COX-2 Expression through the Downregulation of NF-κB and MAPK Activities in RAW 264.7 Macrophages.

Oleifolioside A, a new triterpenoid compound isolated from Dendropanax morbifera Leveille (D. morbifera), was shown in this study to have potent inhibitory effects on lipopolysaccharide (LPS-)stimulated nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in RAW 264.7 macrophages. Consistent with these findings, oleifolioside A was further shown to suppress the expression of LPS-stimulated inducible nitric oxide synthase (iNOS) and cyclooxigenase-2 (COX-2) in a dose-dependent manner at both the protein and mRNA levels and to significantly inhibit the DNA-binding activity and transcriptional activity of NF-κB in response to LPS. These results were found to be associated with the inhibition of the degradation and phosphorylation of IκB-α and subsequent translocation of the NF-κB p65 subunit to the nucleus. Inhibition of NF-κB activation by oleifolioside A was also shown to be mediated through the prevention of p38 MAPK and ERK1/2 phosphorylation. Taken together, our results suggest that oleifolioside A has the potential to be a novel anti-inflammatory agent capable of targeting both the NF-κB and MAPK signaling pathways.

Inhibitory effects of organic solvent extracts from Korean local plants on the complement classical pathway.

The study evaluated the anticomplement effects from organic solvent extracts of five Compositae plants (Ligularia fischeri (Ledeb.) Turez, Ligularia taquetii (H.Lev. & Vaniot) Nakai, Ainsliaea acerifolia Sch.Bip, Aster scaber Thunb, Aster koraiensis Nakai, Synurus deltoides Aiton) from South Korea on the classical pathway complement system. We have evaluated organic solvent extracts from five Compositae with regard to its anticomplement activity. Chloroform extracts from L. taquetii showed inhibitory activity against complement system with 50% inhibitory concentrations (IC50) values of 73.2 µg/mL. This is the first report of anticomplement activity from L. taquetii.

Isolated compounds from Sorghum bicolor L. inhibit the classical pathway of the complement.

The present study evaluated the anticomplement effects from isolated compounds of Sorghum bicolor in classical pathway complement system. Using column chromatograph, three compounds; Sorgoleone-362 (1), Sorgoleone-360 (2) and Sorgoleone-386 (3) were isolated and evaluated for in vitro anticomplement activity. Sorgoleone-386 showed inhibitory activity against complement system with 50% inhibitory concentrations (IC(50)) values of 148.3µg/ml. This is the first report of anticomplement activity of isolated compounds from Sorghum bicolor.

Inhibitory effects of three oleanolic acid glycosides from Achyranthes japonica on the complement classical pathway.

The present study evaluated the anticomplement effects of isolated compounds from Achyranthes japonica in the classical pathway of the complement system. Using column chromatography, three compounds: achyranthoside C dimethyl ester, achyranthoside C butyl dimethyl ester, and achyranthoside E dimethyl ester were isolated and evaluated for in vitro anticomplement activity. Achyranthoside C dimethyl ester showed the most potent inhibitory activity against the complement system, with 50% inhibitory concentrations (IC(50)) values of 26.2 µg/mL. This is the first report of anticomplement activity of isolated compounds from A. japonica.

Anticomplement activity of organic solvent extracts from Korea local Amarantaceae spp.

The study evaluated the anticomplement activity from various solvent extracts of nine Amarantaceae plants (Achyranthes japonica (Miq.) Nakai, Amaranthus mangostanus L., Amaranthus retroflexus L., Amaranthus spinosus L., Celosia argentea var. spicata., Amaranthus lividus L., Celosia cristata L., Amaranthus viridis L., Gomphrena globosa L.) from South Korea on the classical pathway. We have evaluated various organic solvent extract from nine Amarantaceae plants with regard to its anticomplement activity on the classical pathway. Achyranthes japonica chloroform extracts showed inhibitory activity against complement system with 50% inhibitory concentrations (IC(50)) value of 73.1µg/ml. This is the first report of anticomplement activity from Amarantaceae plants.

Anticomplement activity of various solvent extracts from Korea local Artemisia spp.

The study evaluated the anticomplement activity from various solvent extracts of eight Artemisia plants (Artemisia capillaris Thunb., Artemisia fukudo Makino., Artemisia japonica Thunb., Artemisia montana (Nakai) Pamp., Artemisia keiskeana Miq., Artemisia rubripes Nakai., Artemisia stolonifera (Maxim.) Kom., and Artemisia sylvatica Max.) from South Korea on the classical pathway (CP). We have evaluated various organic solvent extract from eight Artemisia plants with regard to its anticomplement activity on the CP. A. rubripes and A. montana chloroform extracts showed inhibitory activity against complement system with 50% inhibitory concentrations (IC50) values of 54.3 and 64.2 µg/mL. This is the first report of anticomplement activity from Artemisia plants.

Anticomplement activity of isolated compounds from Artemisia montana.

The study evaluated the anticomplement activity from isolated compounds from Artemisia montana (Nakai) Pamp. from South Korea on the classical pathway. In a previous work, A. montana (Nakai) Pamp. chloroform extracts showed inhibitory activity against complement system. The chromatographic separation of a chloroform chloride extract of A. montana (Nakai) Pamp. led to the isolation of four compounds. Their structures were characterized to be ezoartemin, yamayomoginin, ezomontanin and 11,13-dihydroezomontanin by spectroscopic data. This is the first report of anticomplement activity of isolated compounds from A. montana (Nakai) Pamp.

Inhibition effects of isolated compounds from Artemisia rubripes Nakai of the classical pathway on the complement system.

The study evaluated the anticomplement activity from isolated compounds from Artemisia rubripes Nakai from South Korea on the classical pathway. In the previous works, Artemisia rubripes chloroform extracts showed inhibitory activity against complement system. The chromatographic separation of a chloroform chloride extract of Artemisia rubripes led to the isolation of three compounds. Their structures were characterized to be scopoletin (1), 11,(13)-triene-6,12-olide (2), and 1ß,6α-dihydroxy-4(15)-eudesmene (3) by spectroscopic data. This is the first report of anticomplement activity of isolated compounds from Artemisia rubripes.

Immunotoxicity activity from various essential oils of Angelica genus from South Korea against Aedes aegypti L.

The leaves of Angelica anomala Lallemant, Angelica cartilagino-marginata var. distans (Nakai) Kitag, Angelica czernevia (Fisch. et Meyer) Kitagawa, Angelica dahurica Benth. et Hooker, Angelica decursiva (Miq.) Franch. & Sav, Angelica fallax Boissieu, Angelica gigas Nakai, Angelica japonica A. gray were essential oil extracted and immunotoxicity effects were studied. The Angelica anomala, A. cartilagino-marginata var. distans, A. czernevia, A. dahurica, A. decursiva, A. fallax, A. gigas, A. japonica essential oil yield were 4.13, 4.83, 4.45, 3.25, 4.11, 4.73, 4.34 and 4.21%. The A. dahurica essential oil had a significant toxic effect against early fourth-stage larvae of Aedes aegypti L with a lethal concentration 50 (LC50) value of 43.12 ppm and an LC90 value of 65.23 ppm. The above indicates that essential oil contents may play a more important role in the toxicity of essential oil.

Branched-chain amino acids complex inhibits melanogenesis in B16F0 melanoma cells.

Present study was investigated the effect of each or complex of three branched-chain amino acids (BCAAs; isoleucine, leucine, and valine) on melanin production in B16F0 melanoma cells treated with various concentrations (1-16 mM) for 72 h. Among the 20 amino acids, lysine and glycine showed the highest activities of DPPH radical scavenging and mushroom tyrosinase inhibition, respectively. Each and combination of BCAAs reduced melanogenesis in a concentration-dependent manner without any morphological changes and cell viability in melanoma cells. Present study was also investigated the inhibitory effects of each or complex of BCAAs at each 10 mM concentration on the 100 µM IBMX-mediated stimulation of melanogenesis in melanoma cells for 72 h and found that IBMX treatment was stimulated to enhance melanin synthesis and that the complex of BCAAs was the most effectively inhibited in the melanin amounts of cellular and extracellular and the whitening the cell pellet. When the inhibitory effect of BCAAs on tyrosinase was examined by intracellular tyrosinase assay, both isoleucine and valine exhibit slightly inhibition, but leucine and combination of BCAAs did not inhibit the cell-derived tyrosinase activity. Present study demonstrated that complex of BCAAs inhibited melanin production without changes intercellular tyrosinase activity. Thus, the complex of BCAAs may be used in development of safe potentially depigmenting agents.

Effect of fermented Angelicae gigantis Radix on carbon tetrachloride-induced hepatotoxicity and oxidative stress in rats.

This study is aimed to evaluate the protective effect of fermented Angelicae gigantis Radix (AGR) with Monascus purpureus strain on carbon tetrachloride (CCl(4))-induced hepatotoxicity and oxidative stress in rats. The activities of liver marker enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and the levels of lipid peroxidation were increased when CCl(4) was treated but these parameters were significantly decreased by fermented AGR treatment. CCl(4) treatment exhibited decrease in serum concentrations of triglyceride, total cholesterol, HDL-cholesterol, and free fatty acids, and these were also decreased by fermented AGR administration. The level of serum leptin was significantly lower in fermented AGR administration than that in normal control group. CCl(4) treatment significantly increased the concentration of liver triglyceride. The current study observed significant elevations of the thiobarbituric acid-reactive substances (TBARS) levels in the liver homogenate, mitochondrial, and microsomal fractions of CCl(4) control group compared with normal control group. CCl(4) treatment resulted in a significant decrease in the levels of plasma and hepatic glutathione, but these reductions were significantly increased by fermented AGR administration. CCl(4) induced the marked hepatocytes necrosis and fatty accumulation around the central veins. Accordingly, fermented AGR may be an ideal candidate for the hepatoprotective effect in animal model.

Hepatoprotective effects on alcoholic liver disease of fermented silkworms with Bacillus subtilis and Aspergillus kawachii.

The purpose of this study was to investigate the protective effect of Bacillus subtilis fermented silkworm powder (BFSP) and Aspergillus kawachii fermented silkworms powder (AFSP) on alcohol-induced hepatotoxicity in Sprague-Dawley rats. Alcohol-feeding rats were fed with diets containing silkworm powder (SP) or both BFSP and AFSP at the 5% (w/w) levels for 4 weeks. Alcohol administration resulted in a significant increase in the activities of liver marker enzymes, aspartate aminotransferase (AST), γ-glutamyl transpeptidase (γ-GTP) and lactate dehydrogenase (LDH). Administration of BFSP markedly prevented alcohol-induced elevation of serum AST, γ-GTP and LDH activities, and the levels of blood alcohol and acetaldehyde. Interestingly, in comparison with both SP and AFSP, BFSP administration drastically increased both hepatic alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities, suggesting that BFSP was more effective in the reduction of blood alcohol and acetaldehyde. BFSP administration showed the highest induction of hepatic ADH expression in alcohol-feeding rats. Also, alcohol treatment resulted in increasing lipid peroxidative index (thiobarbituric acid-reactive substances) and decreasing antioxidant status (reduced glutathione) in the liver. Thus, these results suggest that BFSP treatment improved the antioxidant status of alcoholic rats by decreasing the levels of lipid peroxidative index and by increasing the levels of antioxidant status in the liver and serum. Specially, the concentrations of serum total cholesterol, free fatty acid and hepatic triglyceride were increased, but these parameters were significantly influenced by the BFSP in the alcohol treatment. Unlike the action of alcohol treatment on fatty liver, BFSP administration attenuated lipid droplet accumulation in hepatocytes. A high level of ADH was also observed in AFSP administered rats; on the other hand, a significant change in ALDH was not observed. Therefore, the SP can be a promising candidate in the prevention alcohol-induced hepatotoxicity and oxidative stress.

Neuroprotective effects of triterpene glycosides from glycine max against glutamate induced toxicity in primary cultured rat cortical cells.

To examine the neuroprotective effects of Glycine max, we tested its protection against the glutamate-induced toxicity in primary cortical cultured neurons. In order to clarify the neuroprotective mechanism(s) of this observed effect, isolation was performed to seek and identify active fractions and components. From such fractionation, two triterpene glycosides, 3-O-[α-l-rhamnopyranosyl(1-2)-ß-d-glucopyranosyl(1-2)-ß-d-glucuronopyranosyl]olean-12-en-3ß,22ß,24-triol (1) and 3-O-[ß-d-glucopyranosyl(1-2)-ß-d-galactopyranosyl(1-2)-ß-d-glucuronopyranosyl]olean-12-en-3ß,22ß,24-triol (2) were isolated with the methanol extracts with of air-dried Glycine max. Among these compounds, compound 2 exhibited significant neuroprotective activities against glutamate-induced toxicity, exhibiting cell viability of about 50% at concentrations ranging from 0.1 µM to 10 µM. Therefore, the neuroprotective effect of Glycine max might be due to the inhibition of glutamate-induced toxicity by triterpene glycosides.

Inhibitory effect and mechanism on antiproliferation of isoatriplicolide tiglate (PCAC) from Paulownia Coreana.

Paulownia coreana has traditionally been used as the medicine and health food in the treatment of cancer and infectious diseases. In the present study, a new antiproliferation agent, isoatriplicolide tiglate (PCAC) was isolated from the chloroform soluble fraction of the leaves of Paulownia coreana. The antiproliferation activities of PCAC plant extract was examined in breast and cervical cancer cell lines in a time-and dose-dependent manners. Our in vitro experiments showed that PCAC suppresses the cell growth and proliferation of cancer cells at a relatively low concentration (< 10 µg/mL) and induces apoptosis at a high concentration (> 50 µg/mL). Western blot analysis showed that concentration higher than 50 µg/mL induces a time-dependent increase in the percentage of apoptotic cells. In this case, PCAC uses both extrinsic and intrinsic pathways for the apoptosis. PCAC treatment decreased the expression of pro-caspase 8, 9, and 3, the main regulators of apoptotic cell death, in MDA-MB-231 cells, accompanied by the activation of caspase 8, 9, and 3. More importantly, PCAC inhibited the in vitro proliferation of six other human breast and cervical cancer cell lines. In conclusion, our data strongly suggest that PCAC acts as an antiproliferation agents particularly against breast and cervical cancers by inducing cell cycle arrest in the S/G2 phase and caspase dependent apoptosis at relatively low (< 10 µg/mL) and high (> 50 µg/mL) concentrations, respectively.

Larvicidal activity of major essential oils from stems of Allium monanthum Maxim. against Aedes aegypti L.

The stems of Allium monanthum were extracted, and the major essential oil composition and larvicidal effects were studied. The analyses were conducted by gas chromatography and mass spectroscopy revealed that the essential oils of A. monanthum stems. The A. monanthum essential oil yield was 4.25%, and gas chromatography and mass spectroscopy analysis revealed that its major constituents were dimethyl trisulfide (23.21%), dimethyl tetrasulfide (11.24%) and methlyl propyl trisulfide (8.21%). The essential oil had a significant toxic effect against early fourth-stage larvae of Aedes aegypti L with an LC(50) value of 23.14 ppm and an LC(90) value of 36.31 ppm. Also, dimethyl trisulfide (≥95.0%), dimethyl tetrasulfide (≥95.0%) and methlyl propyl trisulfide (≥95.0%) were tested against the F(21) laboratory strain of A. aegypti. Methlyl propyl trisulfide (≥95.0%) has good activity with an LC(50) value of 19.38 ppm. Also, the above indicates that other major compounds may play a more important role in the toxicity of essential oil.

Effect of betaine on the hepatic damage from orotic acid-induced fatty liver development in rats.

Betaine prevents hepatic damage caused by ethanol and carbone tetrachloride (CCl4) in rats. Present study was to investigate the effect of betaine on the hepatic microsomal triglyceride transfer protein (MTP) mRNA expression in orotic acid (OA)-induced fatty liver in rats. OA feeding was attributed to the significant increase in the hepatic levels of triglyceride and the serum levels of ALT and AST and resulted in typical histology of fatty liver contained numerous largely fat droplets. While concomitant supplementation of betaine to OA diet was slightly reduced the hepatic triglyceride concentrations and was significantly decreased ALT activity. Hepatic MTP mRNA expression by OA treatment increased by 14% despite triglyceride accumulation in the liver in OA treatment rats relative to rats fed a normal diet without OA supplemented, but MTP expression by simultaneous supplementation of OA and betaine was slightly decreased by 7.9% as compared to the OA-feeding rats. A significant elevation of TBARS contents in the liver homogenate, microsome, and mitochondrial fractions of the OA-feeding rats compared with the normal rats, however, these increases were significantly or slightly decreased by simultaneous addition of OA and betaine. The increases of hepatic OA and betaine levels in OA feeding rats was also found when compared to the normal rats, but these increases were significantly lowered in the concomitant supplementation OA and betaine. The content of Fe was significantly increased in the OA feeding rats, but this elevation showed significantly recovered as low as the normal level by concomitant with OA and betaine. Zinc content was also significantly decreased in the OA feeding rats compared with the normal rats, but this reduction was more significantly elevated by concomitant with OA and betaine. Hepatic glutathione content in the OA feeding rats was similar to that of the normal rats, but this content was slightly reduced without statistically significant differences. But, a significant elevation in the hepatic glutathione content was found in the simultaneously administration OA and betaine. The hepatocytes contained numerous largely fat droplets induced by OA administration and was slightly reduced by simultaneous supplementation of OA and betaine. Present study demonstrated that betaine has a weak preventive action on the OA-induced triglyceride accumulation.