• Title/Summary/Keyword: Activity assay

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Antibacterial and Intracellular Clearance Effect of Saururus chinensis Baill Water Extract against Salmonella typhimurium (삼백초 수용성 추출물의 Salmonella typhimurium 균에 대한 항균 및 숙주세포 내 사멸 효과 규명)

  • Kim, Dong-Hyeok;Lee, Jin-Ju;Lim, Jeong-Ju;Kim, Dae-Geun;Kim, Gon-sub;Lee, Hu-Jang;Min, Won-gi;Rhee, Man-Hee;Chang, Hong-Hee;Kim, Suk
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.111-119
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    • 2010
  • Salmonella spp. are one of major pathogens for zoonosis in worldwide, and can replicate within host cells and generally cause enterocolitis and foodborne poisoning which represents a considerable public health burden. The present study was designated to investigate the safty for host cells, antibacterial effects of Saururus chinensis Baill water extract (SCWE) on pure culture and infection with Salmonella enterica serovar Typhimurium (S. typhimurium) in murine derived macrophage RAW 264.7 cells. The different treatment of SCWE concentration (1, 10 or $100{\mu}g/ml$) did not show any cytotoxic effect to RAW 264.7 cells for 24 h incubation. In determination of antibacterial activity of SCWE against S. typhimurium, bacterial viability was markedly decreased compared to SCWE-untreated control. In RAW 264.7 cells, SCWE significantly induced morphological change (p<0.05). In infection assay of S. typhimurium in RAW 264.7 cells pretreated with $100{\mu}g/ml$ of SCWE, which are non-cytotoxic concentration, bacterial uptake ability of macrophage was increased corresponding with morphological change, whereas bacterial survival rates within macrophage was markedly reduced comparing to that of SCWE-untreated control. Furthermore, nitric oxide (NO) production in SCWE-treated cells was slightly decreased until 24 h post infection. Taken together, these findings demonstrated that SCWE have the antibacterial activity for S. typhimurium and the protective effects against S. typhimurium infection through activating murine macrophage independent on NO, suggesting that SCWE were beneficial on the disease caused by intracellularly replicating pathogens as a safe alternatives of conventional chemotherapies.

Physicochemical properties and antioxidant activities of Streptococcus thermophilus KCCM 3782 strain fermented Cordyceps militaris grown on Tenebrio molitor (갈색거저리 동충하초의 Streptococcus thermophilus KCCM 3782 유산균 발효에 따른 물리화학적 특성과 산화방지 활성 비교)

  • Gim, Do-Woo;Park, Jun-Ki;Kim, Hyun-Jin;Cho, In-Kyung;Cho, Ja-Yong;Kim, Yong-Doo;Huh, Chang-Ki
    • Food Science and Preservation
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    • v.25 no.1
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    • pp.136-144
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    • 2018
  • This study aimed to provide basic evidence regarding the development of materials by analyzing the physicochemical properties and antioxidant activities of Streptococcus thermophilus KCCM 3782 strain-fermented Cordyceps militaris grown on Tenebrio molitor. Among the solvents tested, DPPH radical scavenging activity was the highest in the hot water-extracted sample after 30 min of extraction. Moisture content decreased, whereas crude protein and fat content increased, after lactic acid bacteria-mediated fermentation. Sodium, magnesium, calcium, and zinc contents increased, with potassium levels attaining the maximum value, whereas free amino acid content decreased after the fermentation. Among Hunter's color values, a value increased to $66.7{\rightarrow}149.92$ after fermentation, whereas the L and b values decreased to $15.79{\rightarrow}-15.75$ and $54.45{\rightarrow}0.01$, respectively. Cordycepin content assay increased from 7.02 mg% to 8.66 mg%. The DPPH free radical scavenging activity of the fermented product was 91.92 EDA%, which was higher than that of the extract (84.69 EDA%). The ABTS free radical scavenging and superoxide dismutase (SOD) activities were also higher in the fermented products.

Effect of antioxidation and antibacterial activity on crude extract and Characterization of American Cockroaches (Periplaneta americana L.) in Korea (국내 서식 미국바퀴(Periplaneta americana L.)의 특성 및 추출물의 항산화·항균 효과)

  • Kim, Jung-Eun;Kim, Seon-Gon;Kang, Sung-Ju;Kim, Chun-Sung;Choi, Yong-Soo
    • Journal of Sericultural and Entomological Science
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    • v.53 no.2
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    • pp.135-142
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    • 2015
  • The American cockroaches, Periplaneta americana L. was the most important worldwide pest species. It has been an public health problems. We were determinated life cycle and extraction of crude extracts by chemical reagents from cockraches (P. americana L.). The extracted crude solution has been antibacterial activity to gram negative bacteria (Pseudomonas aeruginosa, $6.44{\pm}1.03mm$), gram positive bacteria (Bacillus subtilis, $1.88{\pm}0.40mm$), and fungus (Candida albicans, $5.61{\pm}0.57mm$) using radial diffusion assay. We were analysed of up-regulation of Glutathione-S-transferases (GSTs) stimulation, indicating that antioxidantial protein from various classes are simultaneously expressed in a single insect upon infection or injury. The gene from Periplaneta americana L. were cloned, analysed sequence, and measured protein expression by Real Time PCR (Polymerase Chain Reaction).

Anti-proliferation, Cell Cycle Arrest, and Apoptosis Induced by Natural Liquiritigenin from Licorice Root in Oral Squamous Cell Carcinoma Cells (구강편평세포암종 세포에서 감초 유래 Liquiritigenin의 항증식, 세포주기 정지 및 세포사멸 유도)

  • Kwak, Ah-Won;Yoon, Goo;Chae, Jung-Il;Shim, Jung-Hyun
    • Journal of Life Science
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    • v.29 no.3
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    • pp.295-302
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    • 2019
  • Liquiritigenin (LG) is a chiral flavonoid isolated from the roots of licorice. It exhibits multiple biological activities including anti-oxidant, anti-cancer, and anti-inflammatory effects. In particular though, the anti-cancer activity of LG in oral squamous cell carcinoma has yet to be elucidated, and LG-induced apoptosis in oral squamous cell carcinoma remains poorly understood. In the present study, we tested the role of LG in inducing apoptosis in oral squamous cell carcinoma cells. LG treatment of HN22 cells resulted in a dose-dependent inhibition of cell viability as detected by a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay. The induction of apoptosis in terms of Annexin V/7-Aminoactinomycin D staining, sub-G1 population, and multi-caspase activity were assessed with a $Muse^{TM}$ Cell Analyzer. Flow cytometric analysis revealed that LG treatment resulted in G2/M arrest in cell cycle progression and downregulation of cyclin B1 and CDC2 expression in a concentration-dependent manner. It also resulted in significant upregulation of p27. In addition, LG was seen to trigger the generation of reactive oxygen species and induce CCAAT/enhancer-binding protein homologous protein and 78-kDa glucose-regulated protein in concentration-dependent upregulation. The LG treatment of HN22 cells led to a loss of mitochondrial membrane potential (${\Delta}{\Psi}m$); it also reduced the levels of anti-apoptotic protein and increased the expression of apoptotic protease activating factor-1, cleaved poly (ADP-ribose)polymerase and Bax. Overall, our results indicate that the pro-apoptotic effects of LG in HN22 cells depend on the activation of both intrinsic and extrinsic signaling pathways. Thus, our results suggest that LG constitutes a natural compound with a potential role as an anti-tumor agent in oral squamous cell carcinoma.

Anti-oxidative and Anti-cancer Activities of Ethanol Extract of Litsea populifolia (인체 폐암 세포주 A549에서 Litsea populifolia 추출물의 항산화 및 항암활성 분석)

  • Jin, Soojung;Oh, You Na;Jeong, Hyun Young;Yun, Hee Jung;Park, Jung-ha;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.29 no.6
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    • pp.679-687
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    • 2019
  • Litsea populifolia, a plant species of the Lauraceae family, is widely distributed in the tropical and subtropical areas of Asia. The phylogenetic relationships and botanical characteristics of L. populifolia have been reported; however, its anti-oxidative and anti-cancer activities remain unclear. In this study, we evaluated the anti-oxidative and anti-cancer effects of ethanol extracts of L. populifolia (EELP) together with the molecular mechanism of its anti-cancer activity in human lung adenocarcinoma A549 cells. EELP showed significant anti-oxidative effects with a 50% inhibitory concentration at $11.71{\mu}g/ml$, which was measured by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. EELP exhibited cytotoxic activity and induced cell cycle arrest at the G1 phase in A549 cells in a dose-dependent manner, whereas EELP did not have the cytotoxic effect on the normal human lung cell line IMR90. Treatment with EELP also resulted in a decreased expression of G1/S transition-related molecules-including cyclin-dependent kinase (CDK) 2, CDK6, cyclin D1, and cyclin E-both for the transcription and translation levels. EELP-induced G1 arrest was associated with the phosphorylation of checkpoint kinase 2 (CHK2), p53, cell division cycle 25 homolog A (CDC25A), and the reduction of CDC25A expression in A549 cells. Collectively, these results suggest that EELP may exert an anti-cancer effect by cell cycle arrest at the G1 phase through both p53-dependent and p53-independent (ATM/CHK2/CDC25A/CDK2) pathways in A549 cells.

Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

  • Ding, Yueyun;Zhu, Shujiao;Wu, Chaodong;Qian, Li;Li, DengTao;Wang, Li;Wan, Yuanlang;Zhang, Wei;Yang, Min;Ding, Jian;Wu, Xudong;Zhang, Xiaodong;Gao, Yafei;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.922-929
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    • 2019
  • Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

Antioxidant and Anticancer Activities of Euonymus porphyreus Extract in Human Lung Cancer Cells A549 (인체 폐암 세포주 A549에서 Euonymus porphyreus 추출물의 항산화 및 항암활성 분석)

  • Jin, Soojung;Oh, You Na;Son, Yu Ri;Bae, Soobin;Park, Jung-ha;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.31 no.2
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    • pp.199-208
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    • 2021
  • Euonymus porphyreus, a species of plant in the Celastraceae family, is widely distributed in East Asia, especially in Southern China. The botanical characteristics of E. porphyreus have been reported, but its antioxidative and anticancer activities remain unclear. In this study, we evaluated the antioxidative and anticancer effects of ethanol extracts of E. porphyreus (EEEP) and the molecular mechanism of its anticancer activity in human lung adenocarcinoma A549 cells. The total polyphenol and flavonoid compound contents from EEEP were 115.42 mg/g and 23.07 mg/g, respectively. EEEP showed significant antioxidative effects with a concentration at 50% of the inhibition (IC50) value of 11.09 ㎍/ml, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. EEEP showed cytotoxic activity by increasing the SubG1 cell population of A549 cells in a dose-dependent manner. Apoptosis in A549 cells treated with EEEP was evident due to increased apoptotic cells and apoptotic bodies, as detected by Annexin V and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. EEEP-induced apoptosis resulted in increased expression of the First apoptosis signal (Fas), p53, and Bax, with decreased expression of Bcl-2 and subsequent activation of caspase-8, -9, and caspase-3, leading to cleavage of poly (ADP-ribose) polymerase (PARP). Collectively, these results suggest that EEEP may exert an anticancer effect by inducing apoptosis in A549 cells through both intrinsic and extrinsic pathways.

Evaluation of Anti-wrinkle Effect of Peanut Shell Extract and Stability of the Extract in Cosmetic Products (땅콩 겉껍질 추출물의 주름개선 효능 및 화장품 제형에서 추출물의 안정성 평가)

  • Narae Han;Jin Young Lee;Mihyang Kim;Eun Young Choi;Bong-Jeun An;Yu-Young Lee;Moon Seok Kang;Hyun-Joo Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.203-212
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    • 2023
  • This study was conducted to investigate the possibility of peanut shell, a by-product of peanut, as a functional cosmetic ingredient. Peanut shell extract showed high antioxidant activity with IC50 values of 75.00, 46.33, and 472.83 ㎍/mL for DPPH and ABTS radical scavenging and SOD-like activity, respectively. Furthermore, peanut shell extract was efficiently decreased the MMP-1 and MMP-3 protein level in the UVB treated-HaCaT cell and maintained procollagen protein level similar to normal control. Similar to anti-wrinkle related protein expression assay, the IC50 value of elastase and collagnease inhibition in peanut shell extract was lower as 0.30 and 0.09 mg/mL, respectively, than that of the positive control. Additionally, eriodictyol and luteolin, which are isolated from peanut shell extract, showed 53.8 and 98.0% elastase inhibition rate, respectively, and 60.1 and 72.5% collagenase inhibition rate, respectively, at a concentration of 0.1 mg/mL. Thus, luteolin was assumed to be the effective ingredient for wrinkle inhibition in peanut shell extract. As a result of stability evaluation of lotion and cream formulations containing peanut shell extract, it was confirmed to be a stable formulation with no significant changes. Therefore, it is considered that peanut shell extract can be applied as a cosmetic ingredient for wrinkle inhibition.

Anti-tumor Effect of a Combination of Hongyoung Ethanol Extract and Cisplatinin YD-10B Oral Cancer Cells (YD-10B 구강암세포에서 홍영 에탄올 추출물과 시스플라틴 병용에 의한 항암 효과)

  • Eun-Jung Kim;Sung-Hee Hwang;Sangwook Park
    • Journal of Life Science
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    • v.33 no.6
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    • pp.498-505
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    • 2023
  • Solanum tuberosum Linnaeus cv Hongyoung, which represents red potato, was developed in Korea. Hongyoung is known to have anti-oxidant, anti-inflammatory, anti-viral, and anti-tumor properties, but no research has been conducted on the growth inhibition and apoptosis effects of hongyoung in YD-10B oral cancer cells. In this study, the combined treatment of hongyoung ethanol extract (HEE) and cisplatin were examined to determine its ability to inhibit cancer cell growth, induce apoptosis, and inhibit matrix metalloproteinases (MMP)-2 and MMP-9 cancer metastasis. The cell viability was investigated using a 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H- tetrazolium monosodium salt (MTS) assay, and the ability to induce apoptosis was analyzed using an FACS analyzer. The mRNA expression and protein activity of MMP-2 and MMP-9 were measured via RT-PCR and zymography. The YD-10B oral cancer cells showed an increase in growth inhibition as the concentration of HEE increased. The combination of 200 µM cisplatin and 500 ㎍/ml HEE reduced the growth of the YD-10B oral cancer cells by more than 50% compared to cisplatin alone. When phorbol 12-myristate 13-acetate (PMA)-treated YD-10B oral cancer cells were co-treated with 200 µM cisplatin and 500 ㎍/ml HEE, both the mRNA expression and protein activity of the MMP-2 and MMP-9 decreased. In addition, the percentage of the sub-G1 phase, which indicates apoptosis ability, more than doubled when treated in combination with 200 µM cisplatin and 500 ㎍/ml HEE than when cisplatin alone was used. The results of this study therefore suggest the possibility of using a combination of HEE and cisplatin in the development of effective drugs to treat oral cancer.

Application of the Extract of Zanthoxylum piperitum DC to Manufacturing Eco-friendly Nosocomial Infection Control Protective Materials (초피의 항균 활성을 이용한 원내 감염 제어 친환경 방호 소재 개발)

  • Shin Young Park;Ki Yun Kim;Do Youn Jun;Sung Chul Kim;Hyo-Il Jung;Young Ho Kim
    • Journal of Life Science
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    • v.33 no.10
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    • pp.820-827
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    • 2023
  • Since COVID-19 began at the end of 2019, the wearing time of protective clothing used to prevent pathogenic bacteria and virus infection has increased, and the development of safe protective materials that are human-friendly and have antibacterial and antiviral functions has been required. In this study, we investigated the possibility of developing natural antibacterial protection materials using ethanol extract of the medicinal plant Zanthoxylum Piperitum DC. The antibacterial activity assay of the 80% ethanol extract of Z. piperitum DC leaves against various nosocomial infectious bacteria, using the disk diffusion method, showed that Staphylococcus aureus ATCC 25923, Klebsiella pneumoniae ATCC 13883, Salmonella typhimurium, and Aeromonas hydrophila are sensitive to the inhibitory action of the extract. The IC50 values of the ethanol extract against S. aureus, K. pneumoniae, P. vulgaris and A. hydrophila were about 0.59 mg/ml, 0.50 mg/ml, 1.06 mg/ml, and 0.06 mg/ml, respectively. To determine whether the ethanol extract of Z. piperitum DC leaves can be applied to the development of antibacterial protective fabric, the ethanol extract was tested using a protective fabric from the KM Health Care Corp. using the JIS L1902-Absorption method. As a result, the bacteriostatic and bactericidal activity values of S. aureus ATCC 25923 and K. pneumoniae ATCC 13883 appeared to be more than 2.0 when treated with the ethanol extract at a concentration of 1% (w/v). Together, these results suggest that Z. piperitum DC leaves can be applied to develop natural antibacterial functional protective fabrics.