Apigenin Induces Apoptosis through a Mitochondria/Caspase-Pathway in Human Breast Cancer MDA-MB-453 Cells

In this study, we investigated the mechanistic role of the caspase cascade in extrinsic and intrinsic apoptosis induced by apigenin, which has been targeted as a candidate in the development of noncytotoxic anticancer medicines. Treatment with apigenin (1–100 µM) significantly inhibited the proliferation of MDA-MB-453 human breast cancer cells in a dose- and time-dependent manner with IC50 values of 59.44 and 35.15 µM at 24 and 72 h, respectively. This inhibition resulted in the induction of apoptosis and the release of cytochrome c in cells exposed to apigenin at its 72 h IC50. Subsequently, caspase-9, which acts in mitochondria-mediated apoptosis, was cleaved by apigenin. In addition, apigenin activated caspase-3, which functions downstream of caspase-9. The apigenin-induced activation of caspase-3 was accompanied by the cleavage of capases-6, -7, and -8. These results are supported by evidence showing that the activity patterns of caspases-3, -8, and -9 were similar. The present study supports the hypothesis that apigenin-induced apoptosis involves the activation of both the intrinsic and extrinsic apoptotic pathways

Robust Co-catalytic Performance of Nanodiamonds Loaded on WO3 for the Decomposition of Volatile Organic Compounds under Visible Light

Proper co-catalysts (usually noble metals), combined with semiconductor materials, are commonly needed to maximize the efficiency of photocatalysis. Search for cost-effective and practical alternatives for noble-metal co-catalysts is under intense investigation. In this work, nanodiamond (ND), which is a carbon nanomaterial with a unique sp(3)(core)/sp(2)(shell) structure, was combined with WO3 (as an alternative co-catalyst for Pt) and applied for the degradation of volatile organic compounds under visible light. NDs-loaded WO3 showed a highly enhanced photocatalytic activity for the degradation of acetaldehyde (similar to 17 times higher than bare WO3), which is more efficient than other well-known co-catalysts (Ag, Pd, Au, and CuO) loaded onto WO3 and comparable to Pt-loaded WO3. Various surface modifications of ND and photoelectochemical measurements revealed that the graphitic carbon shell (sp(2)) on the diamond core (spa) plays a crucial role in charge separation and the subsequent interfacial charge transfer. As a result, ND/WO3 showed much higher production of OH radicals than bare WO3 under visible light. Since ND has a highly transparent characteristic, the light shielding that is often problematic with other carbon-based co-catalysts was considerably lower with NDs-loaded WO3. As a result, the photocatalytic activity of NDs/WO3 was higher than that of WO3 loaded with other carbon-based co-catalysts (graphene oxide or reduced graphene oxide). A range of spectroscopic and photo(electro)chemical techniques were systematically employed to investigate the properties of NDs-loaded WO3. ND is proposed as a cost-effective and practical nanomaterial to replace expensive noble-metal co-catalysts.1124Ysciescopu

STEM OF Elsholtzia splendens INHIBITS ADIPOCYTE DIFFERENTIATION BY REGULATING PPARγ and C/EBPα GENE EXPRESSION IN 3T3-L1 CELLS

ABSTRACT Elsholtzia splendens (E. splendens), is a wild herb found in Korea that has been used in traditional medicine. In this study, the effects of E. splendens ethanolic extraction lipid accumulation in 3T3-L1 preadipocytes were investigated. Morphological changes and the degree of lipid accumulation were measured by Oil Red O staining and an intracellular triglyceride (TG) assay. Four parts of E. splendens (stem, leaf, flower, roots) were decreased lipid accumulation in adipocytes in a concentration -dependent manner. Results of the study demonstrated that100μg/mL of E. splendens stem extract (ESS) inhibit intracellular lipid accumulation by 56.4%. In addition, treatment with ESS decreased intracellular TG contents.The levels of peroxisome proliferators-activated receptor γ (PPARγ) and CCAAT/enhancerbinding proteins (C/EBPα) mRNA were decreased by the E. splendens extract. These findings suggest that E. splendens inhibits intracellular lipid accumulation by regulating the transcription of PPARγ and C/EBPα. KEYWORDS Elsholtzia splendens Adipocyte Lipid accumulation PPARγ C/EBP

Strategies For Manufacturing Servitization Of Korean SMEs: By Using Data Envelopment Analysis

This study examines the efficiency of Korean manufacturing SMEs through data envelopment analysis. We divide business processes into support activities (e.g., business management and product planning) and primary activities (e.g., production and sales and after-sales services) to measure the efficiency of each process by product module. Moreover, we verify the effect of manufacturing servitization by comparing between the efficiency of servitized and non-servitized firms. The result shows that support activities are generally more efficient than primary activities and that standardized mass production with option (module 2) and totally customized production (module 4) is more efficient than either standardized mass production without option (module 1) or customized assembly after standardized production (module 3). The results suggest that, as SMEs are small, they have an advantage in support activities but also lack production and sales channel infrastructure. Moreover, modules 2 and 4 are relatively efficient because they can increase their product values via product differentiation. In addition, servitized firms are more efficient than non-servitized firms in almost every process and module, implying that servitization in manufacturing is an effective way to improve product value. Finally, module 1 and the production process are relatively inefficient, while servitized firms have higher efficiency scores than do non servitized firms. Likewise, module 3 and sales and after-sales services are relatively inefficient, while servitized firms have higher efficiency scores than do non servitized firms. There results imply that servitization can be used by Korean manufacturing SMEs to develop efficiently and effectively

Preventive effect of Ligularia fischeri on inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages depending on cooking method

BACKGROUND: Ligularia fischeri (common name Gomchwi) is known for its pharmaceutical properties and used in the treatment of jaundice, scarlet-fever, rheumatoidal arthritis, and hepatic diseases; however, little is known about its anti-inflammatory effect. In this study the influence of blanching and pan-frying on the anti-inflammatory activity of Ligularia fischeri (LF) was evaluated. RESULTS: Fresh LF and cooked LF showed no significant effect on the viability of macrophages after 24 h incubation. Fresh LF was found to be the most potent inhibitor of nitric oxide (NO) production at 100 μg/ml, while pan-fried LF showed little inhibitory effect on lipoloysaccharide (LPS) stimulated murine machrophage RAW264.7 cells. In contrast with its effect on NO production, pan-fried LF showed significant attenuation of the expression of inducible nitiric oxide synthase (iNOS) compared with fresh LF. In the cooking method of LF, PGE2 production was not affected in the LPS-induced RAW 264.7 cells. In LPS-induced RAW 264.7 cells, pretreatment by fresh and cooked LF increased COX2 mRNA expression. The 3-O-caffeoylquinic acid content of blanching and pan-frying LF increased by 4.92 and 9.7 fold with blanching and pan-frying respectively in comparison with uncooked LF. CONCLUSIONS: Regardless of the cooking method, Ligularia fischeri exhibited potent inhibition of NO production through expression of iNOS in LPS-induced RAW264.7 cells

Folding machineries displayed on a cation-exchanger for the concerted refolding of cysteine- or proline-rich proteins

<p>Abstract</p> <p>Background</p> <p><it>Escherichia coli </it>has been most widely used for the production of valuable recombinant proteins. However, over-production of heterologous proteins in <it>E. coli </it>frequently leads to their misfolding and aggregation yielding inclusion bodies. Previous attempts to refold the inclusion bodies into bioactive forms usually result in poor recovery and account for the major cost in industrial production of desired proteins from recombinant <it>E. coli</it>. Here, we describe the successful use of the immobilized folding machineries for <it>in vitro </it>refolding with the examples of high yield refolding of a ribonuclease A (RNase A) and cyclohexanone monooxygenase (CHMO).</p> <p>Results</p> <p>We have generated refolding-facilitating media immobilized with three folding machineries, mini-chaperone (a monomeric apical domain consisting of residues 191–345 of GroEL) and two foldases (DsbA and human peptidyl-prolyl <it>cis-trans </it>isomerase) by mimicking oxidative refolding chromatography. For efficient and simple purification and immobilization simultaneously, folding machineries were fused with the positively-charged consecutive 10-arginine tag at their C-terminal. The immobilized folding machineries were fully functional when assayed in a batch mode. When the refolding-facilitating matrices were applied to the refolding of denatured and reduced RNase A and CHMO, both of which contain many cysteine and proline residues, RNase A and CHMO were recovered in 73% and 53% yield of soluble protein with full enzyme activity, respectively.</p> <p>Conclusion</p> <p>The refolding-facilitating media presented here could be a cost-efficient platform and should be applicable to refold a wide range of <it>E. coli </it>inclusion bodies in high yield with biological function.</p

Boosting Up the Low Catalytic Activity of Silver for H2 Production on Ag/TiO2 Photocatalyst: Thiocyanate as a Selective Modifier

Noble metal cocatalysts like Pt have been widely employed as an essential ingredient in many kinds of photocatalytic materials for solar hydrogen production. The high material cost of Pt is the biggest limitation. Silver is far less expensive but much less active than Pt and Au as a hydrogen evolving catalyst. Here we demonstrate a new strategy to boost up the activity of silver in Ag/TiO2 for photocatalytic H-2 production via forming a simple surface complexation of thiocyanate (SCN-) on silver. The addition of thiocyanate in the suspension of Ag/TiO2 markedly enhanced the photocatalytic production of H-2 by about 4 times. Thiocyanate was not consumed at all during the photoreaction, which ruled out the role of thiocyanate as an electron donor. Such a positive role of thiocyanate was not observed with bare TiO2, Pt/TiO2, and Au/TiO2. The selective chemisorption of thiocyanate on silver was confirmed by the analyses of Raman spectroscopy and spot-profile energy-dispersive spectroscopy. In the presence of thiocyanate, the overpotential for water reduction on Ag/TiO2 electrode was slightly reduced, and the interfacial charge transfer resistance on Ag/TiO2 (measured by electrochemical impedance spectroscopy) was significantly decreased, whereas other electrode systems (bare TiO2, Au/TiO2, and Pt/TiO2) showed the opposite effect of thiocyanate. These results indicate that the adsorption of thiocyanate on Ag facilitates the transfer of photogenerated electrons on the Ag/TiO2 electrode. It is proposed that the formation of Ag-SCN surface complex enhances the interfacial electron transfer rate and facilitates the reduction of protons on Ag/TiO2.115640Ysciescopu

Role of Matrix Metalloproteinase 3-mediated alpha-Synuclein Cleavage in Dopaminergic Cell Death

Evidence suggests that the C-terminal truncation of alpha-synuclein is equally important as aggregation of alpha-synuclein in Parkinson disease (PD). Our previous results showed that an endopeptidase, matrix metalloproteinase-3 (MMP3), was induced and activated in dopaminergic (DA) cells upon stress conditions. Here, we report that MMP3 cleaved alpha-synuclein in vitro and in vivo and that alpha-synuclein and MMP3 were co-localized in Lewy bodies (LB) in the postmortem brains of PD patients. Incubation of alpha-synuclein with the catalytic domain of MMP3 (cMMP3) resulted in generation of several peptides, and the peptide profiles of WT alpha-synuclein (WTsyn) and A53T mutant (A53Tsyn) were different. Combined analysis using mass spectrometry and N-terminal determination revealed that MMP3 generated C-terminally truncated peptides of amino acids 1-78, 1-91, and 1-93 and that A53Tsyn produced significantly higher quantities of these peptides. Similar sizes of peptides were detected in N27 DA cells under oxidative stress and RNA interference to knock down MMP3-attenuated peptide generation. Co-overexpression of cMMP3 with either WTsyn or A53Tsyn led to a reduction in Triton X-100-insoluble aggregates and an increase in protofibril-like small aggregates. In addition, overexpression of the 1-93-amino acid peptide in the substantia nigra led to DA neuronal loss without LB-like aggregate formation. The results strongly indicate that MMP3 digestion of alpha-synuclein in DA neurons plays a pivotal role in the progression of PD through modulation of alpha-synuclein in aggregation, LB formation, and neurotoxicity

Modular evolution of glutathione peroxidase genes in association with different biochemical properties of their encoded proteins in invertebrate animals

<p>Abstract</p> <p>Background</p> <p>Phospholipid hydroperoxide glutathione peroxidases (PHGPx), the most abundant isoforms of GPx families, interfere directly with hydroperoxidation of lipids. Biochemical properties of these proteins vary along with their donor organisms, which has complicated the phylogenetic classification of diverse PHGPx-like proteins. Despite efforts for comprehensive analyses, the evolutionary aspects of GPx genes in invertebrates remain largely unknown.</p> <p>Results</p> <p>We isolated GPx homologs via <it>in silico </it>screening of genomic and/or expressed sequence tag databases of eukaryotic organisms including protostomian species. Genes showing strong similarity to the mammalian PHGPx genes were commonly found in all genomes examined. GPx3- and GPx7-like genes were additionally detected from nematodes and platyhelminths, respectively. The overall distribution of the PHGPx-like proteins with different biochemical properties was biased across taxa; selenium- and glutathione (GSH)-dependent proteins were exclusively detected in platyhelminth and deuterostomian species, whereas selenium-independent and thioredoxin (Trx)-dependent enzymes were isolated in the other taxa. In comparison of genomic organization, the GSH-dependent PHGPx genes showed a conserved architectural pattern, while their Trx-dependent counterparts displayed complex exon-intron structures. A codon for the resolving Cys engaged in reductant binding was found to be substituted in a series of genes. Selection pressure to maintain the selenocysteine codon in GSH-dependent genes also appeared to be relaxed during their evolution. With the dichotomized fashion in genomic organizations, a highly polytomic topology of their phylogenetic trees implied that the GPx genes have multiple evolutionary intermediate forms.</p> <p>Conclusion</p> <p>Comparative analysis of invertebrate GPx genes provides informative evidence to support the modular pathways of GPx evolution, which have been accompanied with sporadic expansion/deletion and exon-intron remodeling. The differentiated enzymatic properties might be acquired by the evolutionary relaxation of selection pressure and/or biochemical adaptation to the acting environments. Our present study would be beneficial to get detailed insights into the complex GPx evolution, and to understand the molecular basis of the specialized physiological implications of this antioxidant system in their respective donor organisms.</p