• Journal of Life Science
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• v.29 no.4
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• pp.410-420
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• 2019

Citrus unshiu peel extracts possess a variety of beneficial effects, and studies on their anticancer activity have been reported. However, the exact mechanisms underlying this activity remain unclear. In the current study, the apoptotic effect of ethanol extract of C. unshiu peel (EECU) on human breast adenocarcinoma MDA-MB-231 cells and related mechanisms were investigated. The results showed that the survival rate of MDA-MB-231 cells treated with EECU was significantly inhibited in a concentration-dependent manner, which was associated with the induction of apoptosis. EECU-induced apoptosis was associated with the activation of caspase-8 and caspase-9, which initiate extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3, a representative effect caspase. EECU suppressed the expression of the inhibitor of apoptosis family of proteins, leading to an increased Bax/Bcl-2 ratio and proteolytic degradation of poly (ADP-ribose) polymerase. EECU also enhanced the loss of the mitochondrial membrane potential and cytochrome c release from the mitochondria to the cytosol, along with truncation of Bid. In addition, EECU activated AMP-activated protein kinase (AMPK), and compound C, an AMPK inhibitor, significantly weakened EECU-induced apoptosis and cell viability reduction. Furthermore, EECU promoted the generation of reactive oxygen species (ROS), which acted as upstream signals for AMPK activation as pretreatment of cells, with the antioxidant N-acetyl cysteine reversing both EECU-induced AMPK activation and apoptosis. Collectively, these findings suggest that EECU inhibits MDA-MB-231 adenocarcinoma cell proliferation by activating intrinsic and extrinsic apoptotic pathways, which was mediated through ROS/AMPK-dependent pathways.

• Journal of Life Science
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• v.29 no.3
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• pp.332-336
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• 2019

Obesity is associated with an increased risk of many diseases including type 2 diabetes mellitus, hypertension, and hyperlipidemia. The flowers of Chrysanthemum boreale have been used as traditional medicines for the treatment of diseases such as obesity and type 2 diabetes mellitus. This study aimed to evaluate the effect of C. boreale Makino flower essential oil (CFEO) on adipocyte differentiation using preadipocyte cell line 3T3-L1. CFEO at concentrations between 0.1 and $5{\mu}g/ml$ did not affect 3T3-L1 cell viability. A CFEO concentration of between 0.1 and $1{\mu}g/ml$ significantly inhibited lipid accumulation during MDI-induced differentiation in 3T3-L1 cells in a dose-dependent manner, reaching a maximal level at $1{\mu}g/ml$ ($28.94{\pm}2.01%$; approximately 30% of control treated with MDI alone). Western blot analysis revealed that CFEO concentrations between 0.1 and $1{\mu}g/ml$ suppressed the activations of three adipogenic transcription factors in the MDI-stimulated 3T3-L1 cells: peroxisome proliferator-activated receptor ${\gamma}$; CCATT/enhancer binding protein ${\alpha}$; and sterol regulatory element binding protein-1. Moreover, the expressions of lipogenic enzymes, acetyl-CoA carboxylase, and fatty acid synthase were also inhibited by treatment with CFEO between 0.1 and $1{\mu}g/ml$. CFEO may therefore be a promising functional material for obesity prevention.

• Journal of the Korea Organic Resources Recycling Association
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• v.27 no.2
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• pp.19-30
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• 2019

Objective of this study was to develop an organic fertilizer utilizing domestic livestock horn meal and to investigate the application effect of rice and eggplant. The possibility of utilization of livestock horn meal as an organic resource to replace imported expeller cake fertilizer was examined. In order to select domestic organic resources with high nitrogen content, 8 kinds of organic matter such as chicken manure, fish meal, soybean meal, sesame meal, perilla meal, blood meal, livestock horn meal, and beer sludge were analyzed and organic resources with high nitrogen content were selected. In addition, the conditions for the production of organic fertilizers that can be used in organic agriculture were established by mixing of the rice husk biochar and the rice bran as the supplements with the raw materials for mixing ratios. The content of total nitrogen (T-N) in the livestock horn meal was 12.0 %, which was the next low in 13.5 % blood meal. The content of total nitrogen was 5.9 ~ 7.9 % in fish meal and oil cakes. Total nitrogen content of non-antibiotic chicken manure for organic farming was 3 % and nitrogen content in beer sludge was 3.5 %. Organic fertilizer was produced by using biochar, rice bran as a main ingredient of non-antibiotic chicken manure, livestock horn meal and beer sludge. Compared to nitrogen content (4.0 to 4.2 %) of imported expeller cake fertilizer (ECF), the nitrogen content of organic fertilizer utilizing domestic livestock horn meal is as high as 7.5 %. The developed organic fertilizer is met as Zn 400 mg/kg, Cu 120 mg/kg the quality of organic agricultural materials such as or less. To investigate the effect of fertilizer application on the crops, prototypes of developed organic fertilizer were used for the experiment under selected conditions. As a result of application the developed organic livestock horn meal fertilizer (LHMF) for cultivation of the rice and eggplant, the application quantity of the developed organic LHMF 100 % was decreased by 40 % compared to that of the mixed expeller cake fertilizer (MECF). The application of LHMF, which refers to the application rate corresponding to the nitrogen fertilization recommended by the soil test, was reduced by 40% compared to the application rate of MECF, but the same results were obtained in crop growth and yield. The selection of a new high concentration nitrogen source utilizing domestic organic resources and the development of organic fertilizer is the starting point of the research for substitution of imported ECF using domestic local resources at the present time that the spread of eco-friendly agriculture is becoming increasingly important. If it is expanded in the future, it is expected to contribute to the stable production of eco-friendly agricultural products.

• Journal of Life Science
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• v.28 no.11
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• pp.1301-1315
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• 2018

We purified an antimicrobial peptide from the acidified hemocyte extract of Mytilus coruscus by $C_{18}$ reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was 4041.866 Da based on matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS) and the 25 amino acids of the N-terminus sequence were identified. Comparison of this sequence of the purified peptide with the N-terminus sequences of other antimicrobial peptides revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We also identified a 312 bp open-reading frame (ORF) encoding 103 amino acids based on the obtained amino acid residues. The nucleotide sequence of this ORF and the amino acid sequence also revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We synthesized two antimicrobial peptides with an alanine residue in the C-terminus, and designated them mytilin B1 and B2. These two antimicrobial peptides showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus and Streptococcus parauberis (minimal effective concentration, MECs $41.6-89.7{\mu}g/ml$), gram-negative bacteria, including Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, and Vibrio ichthyoenteri (MECs $7.4-39.5{\mu}g/ml$), and the fungus Candida albicans (MECs $26.0-31.8{\mu}g/ml$). This antimicrobial activity was stable under heat and salt conditions. Furthermore, the peptides did not exhibit significant hemolytic activity or cytotoxic effects. These results suggest that mytilin B could be applied as alternative antibiotic agent, and they add to the understanding of the innate immunity of hard-shelled mussels.

• Journal of Life Science
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• v.28 no.11
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• pp.1361-1368
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• 2018

Inflammation is the most common condition in the human body. Tissue damage triggers inflammation, together with vasodilation and increased blood flow at the inflamed site, resulting in edema. Inflammatory responses are also triggered by lipopolysaccharide (LPS), a Toll-like receptor Enterococcus faecalis, a gram-positive organism, has been reported to possess immunomodulatory and preventive activities; however, its use may present risks of sepsis and other systemic infections. Heat-killed Enterococcus faecalis (EF-2001) has been reported to induce antitumor activity, but its effects on inflammation are not known. In the present study, we investigated the effect of EF-2001 on LPS-induced macrophage inflammatory responses. EF-2001 treatment reduced nitric oxide (NO) production, indicating suppression of inflammatory reactions. EF-2001 showed no cytotoxicity in macrophages. Further investigation of the anti-inflammatory mechanism of EF-2001 indicated that EF-2001 reduced the LPS-induced expression of inducible nitric oxide synthase and cyclooxygenase-2. EF-2001 also reduced f the LPS induction of several inflammatory molecules involved in the nuclear factor-${\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinase pathways, including ERK, JNK, and p38 phosphorylation, in a concentration-dependent manner. Additionally, EF-2001 inhibited Akt phosphorylation and increased the expression of the inhibitory ${\kappa}B$ ($I{\kappa}B$) protein, an inhibitor of $NF-{\kappa}B$. EF-2001 also inhibited the nuclear translocation of p65. These results suggest that EF-2001 has anti-inflammatory properties and may be useful for treating inflammatory diseases.

• Journal of Life Science
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• v.28 no.12
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• pp.1507-1515
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• 2018

Benign prostatic hyperplasia (BPH) is characterized by prostatic hypertrophy mainly in the elderly. Corni Fructus is reportedly effective in the prevention and treatment of various diseases, but its efficacy on BPH has not been previously studied. In the present study, we investigated whether or not a Corni Fructus water extract (CF) could prevent testosterone-induced prostatic hyperplasia in rats. To induce BPH, castrated rats were subcutaneously injected with testosterone propionate (TP). CF was administered daily by oral gavage, along with the TP injections, and finasteride, a selective inhibitor of $5{\alpha}$-reductase type 2, was used as a positive control. The results show that CF significantly reduces prostate weight and histopathologic changes while also decreasing levels of serum dihydrotestosterone, similar to the finasteride-treated group. CF also suppresses TP-induced $5{\alpha}$-reductase expression and concentration in prostate tissue and serum, respectively. Furthermore, CF markedly inhibited TP-induced expression of the androgen receptor (AR) and the steroid receptor coactivator 1, an AR coactivator, which was associated with a decrease in prostate-specific antigen levels in both serum and prostate tissue. In conclusion, the results of this study indicate that CF weakens BPH status by inactivation of $5{\alpha}$-reductase and AR.

• Korean Journal of Plant Resources
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• v.32 no.1
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• pp.53-62
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• 2019

The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.

• Asia-Pacific Journal of Business Venturing and Entrepreneurship
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• v.14 no.3
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• pp.59-71
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• 2019

The domestic unemployment rate is now seriously increasing due to the impact of the international trend of incurring high costs using only a small number of manpower in a situation where the business has been being reduced in size. The nation is taking start-up promotion policies as a way to overcome the problem, but with the advent of excessive entrepreneurs and their concentration on specific areas, their survival is in an unclear situation. Thus, this study was intended to provide a useful direction for successful start-ups by identifying the impact of entrepreneurial motivation and entrepreneurship, internal characteristics of entrepreneurs, and of competency characteristics, external characteristics of them, on sustainability through business performance. Thus, the purpose of this study was to provide a useful direction for successful start-ups by identifying the impact of entrepreneurial motivation and entrepreneurship, internal characteristics of entrepreneurs, competency, and external characteristics, on sustainability through business performance. The result of verifying the hypothesis has showed that entrepreneurial motivation, entrepreneurship, and competency characteristics had a positive effect on business performance, and on the other hand, the business performance had a positive effect on sustainability. In addition, in terms of the impact of entrepreneurial motivation, entrepreneurship, and competency characteristics on sustainability, all of them had mediating effects on business performance. It is obvious that studies on the factors affecting business performance and corporate sustainability up to now have been carried out by only collecting the independent variables of this study individually or two of them. so it is judged that if the study, which has verified the practical direction in general through the verification of entrepreneurs's internal and external characteristics from various angles, is performed more comprehensively along with the addition of background characteristics of entrepreneurs, such as characteristics of start-up preparation, etc., in the future, more in-depth results will be able to be obtained.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.384-397
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• 2018

The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.

• Applied Chemistry for Engineering
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• v.30 no.2
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• pp.145-150
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• 2019

In this study, we used extracts obtained from five different solvents (water, ethanol, hexane, ethyl acetate, butanol) of Achyranthes japonica (AJ) and also AJ fermented with Lactobacillus plantarum (LP) to confirm effects on the anti-inflammatory activity in RAW264.7 cells. Experiments of measuring nitric oxide (NO) and cytokine production were performed in lipopolysaccharide (LPS)-induced RAW264.7 cells, and the expression of both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was observed by a western blot method. The cytotoxicity of RAW264.7 was confirmed by the cell counting kit (CCK) assay at a concentration of $100{\mu}g/mL$, which has no toxicity. As a result of the inhibition of NO production, the inhibition rate of AJ-LP extracted with ethanol samples was about 74% higher than that of using the control group. Interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and Interleukin-$1{\beta}$ (IL-$1{\beta}$), which are inflammatory cytokines, also showed an excellent efficacy with inhibition rates of about 57, 70, and 74%, respectively. Comparing to the results of COX-2 and iNOS expression in the AJ group, the inhibition rate of 20-hydroxyecdysone was the highest than others. On the other hand, the COX-2 expression level of AJ-LP group decreased about 16% compared to that of the control group, and the iNOS expression level was also decreased about 7%. These results suggest that the extract of AJ fermented from L. plantarum can be used as an anti-inflammatory natural material.