• Title/Summary/Keyword: Cell Activity

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Isolation, identification, and probiotic characteristics of Bacillus strains affecting the biogenic amine content in fermented soybean paste (발효 된장의 바이오제닉 아민 함량에 영향을 미치는 바실러스균의 분리 동정 및 프로바이오틱 특성)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.131-142
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    • 2019
  • The primary objective of this study was to determine the content of biogenic amines in Korean traditional fermented soybean pastes (doenjang) and to isolate potential probiotic Bacillus sp. with the ability to inhibit biogenic amines accumulation. There were significant differences in the bacterial cell counts, pH value, titratable acidity, salinity, and biogenic amine content between the samples. Among Bacillus strains isolated from doenjang, Bacillus (B.) licheniformis DB102, B. subtilis DB203, B. stearothermophilus DB206, Bacillus sp. DB209, Bacillus sp. DB310, B. coagulans DB311, B. cereus DB313, B. amyloliquefaciens DB714, Bacillus sp. DB917, B. cereus DB 915, B. subtilis DB1020, and Bacillus sp. DB1022 were found to be able to produce biogenic amines. On the other hand, biogenic amine-degrading strains were identified as Bacillus sp. DB403, Bacillus sp. DB407, B. subtilis DB517, B. licheniformis DB612, and B. subtilis DB821. In particular, Bacillus sp. DB407 and B. subtilis DB821 showed probiotic properties including tolerance to artificial digestive juices, adherence to intestinal epithelial cells, resistance to antibiotics, and antibacterial activity against biogenic amine-producing strains. In conclusion, the two probiotic Bacillus strains may be considered as the suitable starter for manufacture of fermented soybean foods with low biogenic amines content.

Anti-inflammatory Effects of the Extracts of Prunus pendula for. ascendens (Makino) Ohwi Leaves and Identification of Active Constituents (올벚나무 잎 추출물의 항염 효능 및 유효성분 규명)

  • Hong, Hye Jin;Ko, Ha Na;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.2
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    • pp.117-129
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    • 2019
  • In this study Anti-inflammatory activity was investigated for the extract of Prunus pendula for. ascendens (Makino) Ohwi by monitoring nitric oxide (NO) production in LPS-stimulated RAW 264.7 murine macrophage cells. The P. pendula ethyl acetate (EtOAc) fraction showed to decrease the NO synthesis by 76.3% at $100{\mu}g/mL$ concentration. The inhibition occurred in a dose-dependent manner without causing cell toxicity. The EtOAc fraction also inhibited the production of $PGE_2$, $IL-1{\beta}$, IL-6 and expression of iNOS, COX-2 protein in dose-dependent manner. From the phytochemical study to isolate the active constituents, five known compounds were identified, which are ursolic acid (1), prunasin (2), methyl p-coumarate (3), kaempferol (4), astragalin (5). All of the compounds 1 - 5 were isolated for the first time from the P. pendula. Among the isolates, the flavonoids 4 and 5 were verified to inhibit NO production with high efficiency. These results suggested that extract of P. pendula leaves could be useful as anti-inflammatory agents in pharmaceutical or cosmetic applications.

Anti-inflammatory Activities of Ethanol Extracts from Leaf, Seed, and Seedpod of Nelumbo nucifera (연잎, 연자육, 연자방 에탄올 추출물의 항염증 활성)

  • Lee, Eun-Joo;Seo, Yu-Mi;Kim, Yong-Hyun;Chung, Chungwook;Sung, Hwa-Jung;Sohn, Ho-Yong;Park, Jong-Yi;Kim, Jong-Sik
    • Journal of Life Science
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    • v.29 no.4
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    • pp.436-441
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    • 2019
  • Nelumbo nucifera, also known as sacred lotus, has mainly been used as a food throughout the Asian countries. In the present study, we prepared ethanol extracts from leaf (NL), seed (NS), and seedpod (NSP) of Nelumbo nucifera and investigated their anti-inflammatory activities in mouse macrophage RAW 264.7 cells. To evaluate the anti-inflammatory activities of NL, NS, and NSP, nitric oxide (NO) production was measured in LPS-stimulated RAW 264.7 cells. NL, NS, and NSP significantly reduced NO production in a dose-dependent manner without affecting cell viabilities. NL, NS, and NSP dramatically decreased the protein expression of pro-inflammatory genes such as iNOS and COX-2. NL, NS, and NSP also suppressed phosphorylation of MAPKs and the nuclear translocation of $NF-{\kappa}B$ p65 indicating they have their anti-inflammatory activities via regulating mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B ($NF-{\kappa}B$) pathways. In addition, we analyzed the production of reactive oxygen species (ROS) by the treatment of NL, NS, and NSP. All extracts reduced ROS production in a dose-dependent manner. And also, they increased heme oxygenase-1 (HO-1) protein expression and the nuclear translocation of nuclear respiratory factor 2 (Nrf2). In conclusion, our results suggest that Nelumbo nucifera has its anti-inflammatory activity via regulating MAPKs, $NF-{\kappa}B$, and Nrf2/HO-1 pathways.

Hepatotoxicity and nephrotoxicity of saponin-enriched extract of Asparagus cochinchinensis in ICR mice

  • Sung, Ji Eun;Choi, Jun Young;Kim, Ji Eun;Lee, Hyun Ah;Yun, Woo Bin;Park, Jin Ju;Kim, Hye Ryeong;Song, Bo Ram;Kim, Dong Seob;Lee, Chung Yeoul;Lee, Hee Seob;Lim, Yong;Hwang, Dae Youn
    • Laboraroty Animal Research
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    • v.33 no.2
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    • pp.57-67
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    • 2017
  • The inhibitory effects of Asparagus cochinchinensis against inflammatory response induced by lipopolysaccharide (LPS), substance P and phthalic anhydride (PA) treatment were recently reported for some cell lines and animal models. To evaluate the hepatotoxicity and nephrotoxicity of A. cochinchinensis toward the livers and kidneys of ICR mice, alterations in related markers including body weight, organ weight, urine composition, liver pathology and kidney pathology were analyzed in male and female ICR mice after oral administration of 150, 300 and 600 mg/kg body weight/day saponin-enriched extract of A. cochinchinensis (SEAC) for 14 days. The saponin, total flavonoid and total phenol levels were found to be 57.2, 88.5 and 102.1 mg/g in SEAC, respectively, and the scavenging activity of SEAC gradually increased in a dose-dependent manner. Moreover, body and organ weight, clinical phenotypes, urine parameters and mice mortality did not differ between the vehicle and SEAC treated group. Furthermore, no significant alterations were measured in alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), blood urea nitrogen (BUN) and the serum creatinine (Cr) in the SEAC treated group relative to the vehicle treated group. Moreover, the specific pathological features induced by most toxic compounds were not observed upon liver and kidney histological analysis. Overall, the results of the present study suggest that SEAC does not induce any specific toxicity in the livers and kidneys of male and female ICR mice at doses of 600 mg/kg body weight/day.

H-NS binding on dicA promoter DNA inhibits dicA gene expression (dicA promoter DNA에 붙는 H-NS 단백질에 의한 dicA 유전자의 발현 조절)

  • Yun, Sang Hoon;Lee, Yonho;Lim, Heon M.
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.191-198
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    • 2019
  • H-NS binds to promoter DNA and works as a general transcription silencer. DicA protein, by binding to the promoter DNA of dicA, activates dicA expression and at the same time inhibits expression of dicF and dicB, thus, exerting cell division control in Escherichia coli. H-NS complexed with a nucleoid protein Cnu was known to be involved in dicA expression. However, the exact nature of H-NS binding to dicA promoter DNA and the consequences of H-NS binding in expression of dicA is not clear. In this study, we explored the DNA binding activity of H-NS on the promoter DNA of dicA and found that H-NS binding occurs exclusively to the dicA promoter DNA. We never observed, however, H-NS binding at the vicinity of the dicA promoter. Temperature dependent oligomerization of H-NS was observed during DNA binding and the Cnu protein enhances the oligomerization process of H-NS binding. In vivo measurement of dicA expression in an hns deleted strain showed that dicA expression increased. These results demonstrated that H-NS binds specifically to dicA promoter DNA and functions as a transcription silencer.

Characterization of Miniimonas sp. S16 isolated from activated sludge (활성슬러지로부터 분리된 Miniimons sp. S16 세균의 특성)

  • Koh, Hyeon-Woo;Kim, Hongik;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.242-247
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    • 2019
  • Biological factors (e.g. microorganism activity) in wastewater treatment plant (WWTP) play essential roles for degradation and/or removal of organic matters. In this study, to understand the microbial functional roles in WWTP, we tried to isolate and characterize a bacterial strain from activated sludge sample. Strain S16 was isolated from the activated sludge of a municipal WWTP in Daejeon metropolitan city, the Republic of Korea. The cells were a Gram-stain-positive, non-motile, facultative anaerobe, and rod-shaped. Strain S16 grew at a temperature of $15{\sim}40^{\circ}C$ (optimum, $30^{\circ}C$), with 0~9.0% (w/v) NaCl (optimum, 1.0~2.0%), and at pH 5.5~9.0 (optimum, pH 7.0~7.5). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S16 was most closely related to the unique species Miniimonas arenae NBRC $106267^T$ (99.79%, 16S rRNA gene sequence similarity) of the genus Miniimonas. The cell wall contained alanine, glutamic acid, serine, and ornithine. Although the isolation source of the type strain NBRC $106267^T$ which considered as a marine microorganism is sea sand, that of strain S16 is terrestrial environment. It might raise an ecological question for habitat transition. Therefore, comparative genome analysis will be valuable investigation for shedding light on their potential metabolic traits and genomic streamlining.

Cudrania tricuspidata Fruit Extract Ameliorates Free Fatty Acid-induced Lipid Accumulation in HepG2 Cells (유리지방산으로 지방축적을 유도한 HepG2 cells 대한 꾸지뽕 열매 추출물의 개선 효과)

  • Lee, Hyo-Jeong;Park, Se-Eun;Kim, Seung
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1144-1151
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    • 2019
  • Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease associated with various metabolic syndromes, such as obesity, dyslipidemia, hypertension, and diabetes. Cudrania tricuspidata is a medicinal plant distributed widely in Asia and has been used in clinical practice to treat various diseases. The aim of this study is to determine the lipid-lowering effects of C. tricuspidata fruit extract (CTE) using a cell model induced by free fatty acids (FFAs). HepG2 cells were exposed to 1mM FFAs (palmitic acid:oleic acid = 2:1) for 24 hr to simulate the conditions of NAFLD in vitro. CTE attenuated the increases of lipid accumulation, intracellular triglyceride, and cholesterol content and inhibited 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) activity in the HepG2 cells in a dose-dependent manner. Also, CTE inhibited the protein expression of lipogenesis-related genes, such as sterol regulatory element-binding protein-1/-2 (SREBP-1/-2), fatty acid synthase (FAS), and stearoyl CoA desaturase-1 (SCD-1) in FFAs-induced lipid accumulation in HepG2 cells. In addition, CTE-induced adenosine monophosphate-activated protein kinase (AMPK) phosphorylation in HepG2 cells. These results suggest that CTE attenuates hepatic lipid accumulation by inhibiting lipogenesis through the modulation of the AMPK signaling pathway on FFAs-induced lipogenesis in HepG2 cells and may potentially prevent NAFLD.

Effect of Evodiae fructus Methanol Extract on Virulence-Related Genes' Expression of Helicobacter pylori (오수유 메탄올 추출물이 Helicobacter pylori 병원성 관련 유전자 발현에 미치는 영향)

  • Yang, Ji Yeong;Lee, Pyeongjae;Kim, Jong-Bae
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.316-322
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    • 2019
  • Helicobacter pylori, a gram-negative bacterium, is one of the risk factors that induces gastritis and gastric cancer. Therefore, much attention has been paid to the compounds that inhibit bacterial growth or eradicate bacteria. Evodiae fructus (EF), the fruit of Evodia rutaecarpa, has been used for treating diarrhea and abdominal pain. EF extract was already found to inhibit the growth of H. pylori. However, to the best of our knowledge, the effect of EF on the virulence factors of H. pylori has not been reported. In this study, when comparing the minimum inhibitory concentration (MIC) of the different methanol concentration extracts, the 95% methanol extract (EF95) showed the lowest MIC value. EF95 extract suppressed the expressions of cagA, vacA and ureB, but interestingly, it up-regulated the expression of ureA. A decrease in production of ammonia in the culture medium and the cell lysates indicated that EF95 inhibited the urease activity in H. pylori, which was the result of EF95 inhibiting the ureB expression. Although the mechanism by which EF95 extract regulates the virulence factors in H. pylori needs further study, EF95 could be used for treatment of gastric troubles induced by H. pylori.

Anti-inflammatory Effects of Aurantio-obtusin isolated from Cassia tora L. in RAW264.7 Cells (결명자로부터 분리된 Aurantio-obtusin의 항염증 활성)

  • Lee, Ki Ho;Jang, Ji Hun;Woo, Kyeong Wan;Nho, Jong Hyun;Jung, Ho Kyung;Cho, Hyun Woo;Yong, Ju Hyun;An, Byeongkwan
    • Korean Journal of Pharmacognosy
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    • v.50 no.1
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    • pp.11-17
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    • 2019
  • Cassia tora L. have been used as a folk medicine in Korea. This study investigated anti-inflammatory effect of aurantio-obtusin isolated from C. tora. We isolated aurantio-obtusin from 50% ethanol extracts of C. tora L. We investigated the anti-inflammatory effects of aurantio-obtusin on the lipopolysaccharide (LPS)-stimulated inflammatory response in murine macrophage cell line (Raw 264.7). To investigate the cytotoxicity of aurantio-obtusin on RAW 264.7 cells, MTS assay was performed. RAW 264.7 cells were treated with aurantio-obtusin at different concentrations (12.5, 25, 50, $100{\mu}M$) for 30 h. The result showed that aurantio-obtusin had no cytotoxic effect in a concentration range of $12.5-100{\mu}M$. To determine the effect of aurantio-obtusin on LPS-induced NO production, the NO concentration measurement was performed. RAW 264.7 cells were treated with aurantio-obtusin at 12.5, 25, 50 and $100{\mu}M$ for 24 h, and the results showed that the NO production of aurantio-obtusin-treated cells compared to LPS alone treated group was significantly decreased in a dose-dependent manner. Pretreatment of aurantio-obtusin inhibited LPS-induced NO production in a dose-dependent manner. To find out inhibitory mechanisms of aurantio-obtusin on inflammatory mediators, we examined the $PGE_2$ pathways. As a result, $PGE_2$ were decreased in a dose-dependent manner by aurantio-obtusin. The release of interleukin-$1{\beta}$ (IL-$1{\beta}$) and IL-6 were also reduced. Moreover, aurantio-obtusin suppressed LPL-induced $I{\kappa}B-{\alpha}$ degradation. These results suggest that the down regulation of NO, $PGE_2$, IL-$1{\beta}$ and IL-6 expression by aurantio-obtusin are achieved by the downregulation of NF-${\kappa}B$ activity.

Anti-inflammatory and Antioxidant Effects of Hot Water Extracts from Kaempferia Galanga L (삼내자 열수추출물의 항산화 및 항염 효과)

  • Chan, Ching Yuen Venus;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.9 no.6
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    • pp.218-226
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    • 2019
  • In this study, we investigated the possibility of Kaempferia Galanga(KG) hot water extract on the antioxidant, cytotoxic and anti-inflammatory efficacy as a cosmetic ingredient. Antioxidant effects were evaluated based on DPPH and ABTS radical scavenging activity, FRAP assay, and total polyphenol contents. The MTT assay was used to confirm the cell toxicity in mouse macrophage RAW264.7 cells. Anti-inflammatory effects were also investigated in LPS-induced RAW264.7 cells by measuring secretion of NO, $TNF-{\alpha}$ and iNOS, $TNF-{\alpha}$ mRNA expression level. As a result, DPPH and ABTS radical scavenging activities were increased in a concentration-dependent manner. The ferric reducing antioxidant power(FRAP) was the highest at 5 mg/mL as 24.5 uM. The measurements of total polyphenol content was $1.28{\pm}0.064mg\;GAE/g$. The cytotoxicity of the KG extract results showed no cytotoxicity at concentration of 0.625 to 2.5 mg/mL. In addition, the extract of KG significantly suppressed the LPS-induced nitrite, $TNF-{\alpha}$ secretion and the mRNA expression of iNOS, $TNF-{\alpha}$ in RAW264.7 cells. Taken together, these data suggest that the KG hot water extracts can be used as a safe and functional cosmetic raw material.