• Title/Summary/Keyword: Disc assay

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The Effects of TWEAK, Fn14, and TGF-$\beta1$ on Degeneration of Human Intervertebral Disc

  • Huh, Hoon;Lee, Yong-Jik;Kim, Jung-Hee;Kong, Min-Ho;Song, Kwan-Young;Choi, Gun
    • Journal of Korean Neurosurgical Society
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    • v.47 no.1
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    • pp.30-35
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    • 2010
  • Objective: The purpose of this study is to explain the effect and reciprocal action among tumor necrosis factor (TNF) like weak inducer of apoptosis (TWEAK), fibroblast growth factor-inducible 14 (Fn14), and transforming growth factor-$\beta1$ (TGF-$\beta1$) on degeneration of human intervertebral disc (IVD). Methods: Human intervertebral disc tissues and cells were cultured with Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM/F-12) media in $37^{\circ}C$, 5% $CO_2$ incubator. When IVD tissues were cultured with TWEAK, Fn14 that is an antagonistic receptor for TWEAK and TGF-$\beta1$, the level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay and sex determining region Y (SRY)-box 9 (Sox9) and versican messenger ribonucleic acid (mRNA) levels were estimated by reverse transcriptase polymerase chain reaction (RT-PCR). Results: When human IVD tissue was cultured for nine days, the sGAG content was elevated in proportion to culture duration. The sGAG was decreased significantly by TWEAK 100 ng/mL, however, Fn14 500 ng/mL did not change the sGAG production of IVD tissue. The Fn14 increased versican and Sox9 mRNA levels decreased with TWEAK in IVD tissue TGF-$\beta1$ 20 ng/mL elevated the sGAG concentration 40% more than control. The sGAG amount decreased with TWEAK was increased with Fn14 or TGF-$\beta1$ but the result was insignificant statistically. TGF-$\beta1$ increased the Sox9 mRNA expression to 180% compared to control group in IVD tissue. Sox9 and versican mRNA levels decreased by TWEAK were increased with TGF-$\beta1$ in primary cultured IVD cells, however, Fn14 did not show increasing effect on Sox9 and versican. Conclusion: This study suggests that TWEAK would act a role in intervertebral disc degeneration through decreasing sGAG and the mRNA level of versican and Sox9.

A Study on the Antimicrobial Activity and the Pharmacological Activities of matrial Isolated from Coptis Radix (황련(Coptis Radix)으로부터 분리된 물질의 항균효능 및 화장품 약리활성에 대한 연구)

  • Jang, Young-Ah;Kim, Bo-Ae;Chung, Jae-Shik;Hwang, Hye-Jin;Lee, Jin-Tae
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.2
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    • pp.271-279
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    • 2017
  • This study evaluated antimicrobial efficacy and antioxidant effect of fraction isolated from Coptis Radix and confirmed its possibility as a cosmetic material. The extracts of isolated from Coptis Radix conducted an antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Candida albicans by disc diffusion method and measure clear zone. As a result, it was confirmed that antimicrobial activity against S. aureus and candida. A was observed in all samples except Fr 1. The activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and The activity of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) cation radical scavenging were determined by antioxidant assay according to the concentrations (50, 125, 250) ${\mu}g/mL$ of extracts of isolated from Coptis Radix. As a result, DPPH radical scavenging activity of Fr 1, 2, 3, 4 at $250{\mu}g/mL$ was 11.4%, 30.3%, 42.0% and 53.1%, respectively and $ABTS^+$ radical scavenging activity was 28.6%, 96.2%, 98.6% and 97.1% at the same concentration, respectively. Fr. 3 and 4 showed higher radical scavenging activity than the positive the control group BHT at the same concentration. In the WST assay results of measuring the cytotoxicity of Coptis Radix, except for Fr. 4, Fr. 1, 2 and 3 did not show toxicity. As a result, the fractions isolated from Coptis Radix can be regarded as a cosmetic material having antimicrobial activity and antioxidant ability.

Antioxidant and Skin Whitening Effect of Graviola (Annona muricata) Leaf Extracts (Graviola (Annona muricata) 잎 추출물의 항산화 및 미백 효과)

  • Jo, Eun Hee;Kim, In Hae;Lee, Jae Hwa
    • Applied Chemistry for Engineering
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    • v.28 no.2
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    • pp.198-205
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    • 2017
  • In this study, graviola leaf extracts (GLE) was investigated for the effect of antioxidant, antibacterial, whitening, anti-wrinkle. The antioxidant effect of GLE was measured by an electron donating ability assay. As a result, GLE increased the electron donating ability in a concentration-dependent manner. The antibacterial effect of GLE was found to show the higher antibacterial effect in Methicillin-resistant Staphylococcus aureus CCARM3115 compared with that of ampicillin by a paper disc method. The whitening effect of GLE was also measured by tyrosinase inhibition assay, and it was found that the tyrosinase activity of GLE decreased as the concentration increased. The inhibition activity of tyrosinase involved in hydroxylation reaction which is related to converting L-tyrosine to (DOPA) was higher than that of arbutin's at the concentration ranging from 125 to $250{\mu}g/mL$. In addition, GLE reduced melanin contents of B16-F10 melanoma cells in a dose-dependant manner and decreased to about 76.7% at a concentration of $100{\mu}g/mL$ Regarding wrinkling formation of GLE, an elastase inhibition assay was performed. As a result, GLE and ursolic acid were 10.5% and 56.5%, respectively under the identical concentration. These results suggest that GLE has significant antioxidant and whitening activities, and also may be potentially used as a therapeutic agent for hyperpigmentation treatment as an ingredient of whitening cosmetics.

Evaluation of anti-inflammatory effect of traditional medicinal plants, Gynura segetum

  • Seow, Lay-Jing;Beh, Hooi-Kheng;Sadikun, Amirin;Asmawi, Mohd Zaini
    • CELLMED
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    • v.4 no.1
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    • pp.4.1-4.4
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    • 2014
  • Gynura segetum, family Asteraceae is a cultivated species and can be found growing in the tropical regions of Indonesia and Malaysia. The plant is known for its use for the treatment of cancer, inflammation, diabetes, hypertension and skin afflictions. In the current study, anti-inflammatory effect of Gynura segetum leaf has been investigated. The present study investigated the in vivo anti-inflammatory effect of Gynura segetum leaf by using hen's egg test-chorioallantoic membrane (HET-CAM) assay. Different solvent extract of Gynura segetum leaf were tested, the most active methanol extract was further fractionated and tested. Among the extracts tested, the methanol extract showed a significant good anti-inflammatory effect (76.8% inhibition at 50 ${\mu}g/disc$) on the HET-CAM assay as compared with the anti-inflammatory drug indomethacin (82.1% inhibition). However, the fractionated sample exhibited a significantly lower activity in comparison to crude methanol extract. The results demonstrated that Gynura segetum leaf displayed remarkable anti-inflammatory effects, which support the folkloric uses of this plant for treatment of inflammation.

Nitric Oxide Production Inhibitory Effect and Antibacterial Activity of the Extract and Fractions from Paeoniae Radix (작약 메탄올 추출물 및 분획물의 Nitric Oxide 생성 억제 효과와 피부질환 원인균에 대한 항균활성)

  • Im, Do-Youn;Lee, Kyoung-In
    • Korean Journal of Pharmacognosy
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    • v.43 no.2
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    • pp.173-178
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    • 2012
  • In this study, we investigated antibacterial activity and nitric oxide production inhibitory effect of the methanol extract and its fractions from Paeoniae Radix. In antibacterial activity by the disc diffusion assay against S. aureus, S. epidermidis and P. aeruginosa, the ethyl acetate fraction showed stronger antibacterial activity than other fractions and the extract. Moreover, the ethyl acetate fraction showed strong nitric oxide (NO) production inhibitory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell. However, in NO scavenging ability, the chloroform fraction was higher than the other fractions and the extract. In the lactate dehydrogenase (LDH) assay against RAW 264.7 cell, the extract and fractions were exhibited normal LDH release level as nontoxic result without the ehtyl acetate fraction of 100 ${\mu}g/ml$. As a result, the ethyl acetate fraction and chloroform fraction of the methanol extract from Paeoniae Radix could be applicable to functional materials for antibacterial and anti-inflammatory related fields, respectively.

Antioxidative and Antibacterial Activity and Tyrosinase Inhibitory Activity of the Extract and Fractions from Taraxacum coreanum Nakai (흰민들레 에탄올 추출물 및 분획물의 항산화 및 항균활성과 Tyrosinase 저해활성)

  • Im, Do-Youn;Lee, Kyoung-In
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.4
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    • pp.238-245
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    • 2011
  • In this study, we investigated antioxidative activity, antibacterial activity against pathogenic strains including methicillin resistant Staphylococcus aureus (MRSA), and tyrosinase inhibitory activity in 75% ethanol extract of Taraxacum coreanum and its fractions. The total polyphenol and flavonoid contents of the extract were 238.59mg/g and 33.18mg/g and the total polyphenol and flavonoid contents of the ethyl acetate fraction were 427.81mg/g and 148.90mg/g as the highest content of fractions. In DPPH radical scavenging ability, $SC_{50}$ values of the ethyl acetate and butanol fraction were 38.40${\mu}g/ml$ and 82.28 ${\mu}g/ml$, respectively. In antibacterial activity by the disc diffusion assay against S. aureus, S. epidermidis and MRSA, the ethyl acetate fraction showed stronger antibacterial activity than other fractions and the extract. Especially, the ethyl acetate fraction was exhibited effective antibacterial activity against MRSA. In the cytotoxicity measurement by MTT assay, the extract and fractions were exhibited Raw 264.7 cell viabilities of 96.32~143.21% as nontoxic result in concentration of 5~100 ${\mu}g/ml$. As a result, the ethyl acetate fraction of the 75% ethanol extract from T. coreanum could be applicable to functional materials for related fields.

Spectrophotometric microtitre assay for rapid screening of membrane-disrupting herbicides (분광분석을 이용한 막과산화작용 제초제의 신속한 검정법)

  • Kwon, Ok-Kyung;Cho, Kwang-Yun;Kim, Jin-Seog
    • The Korean Journal of Pesticide Science
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    • v.4 no.1
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    • pp.11-18
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    • 2000
  • This study was conducted to establish an efficient screening method of membrane-disrupting herbicides, based on the result that membrane leakage leading to evolution of carbonyl compounds and aldehydes could be colorimetrically assessed by formation of stable chromophores with MBTH and ferric chloride. Assay conditions including plant material, light intensity, irradiation time, concentrations of reagents for colorization, and reaction time were optimized, and activities of different types of known herbicides were investigated with cucumber cotyledon discs. That result indicated that this assay was sensitive to membrane-disrupting herbicides, such as protoporphyrinogen oxidase inhibitors (acifluorfen, oxyfluorfen, oxadiazon) and an electron interceptor from photosystem I(paraquat). With seven compounds newly synthesized, highly positive correlation was observed between herbicidal activities measured by greenhouse bioassay and MBTH method. Because only a single disc(4 mm diameter) from cucumber cotyledon in 60 ${\mu}L$ of incubation solution was required to assay, a large number of herbicides could be tested simultaneously using 96-well microtitre plates. But the assay efficiency is not likely to significantly increase because of the boiling step during the reaction.

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Antibacterial Effect of Ipyo-san against Methicillin-Resistant Staphylococcus aureus (입효산(立效散)의 Methicillin-Resistant Staphylococcus aureus에 대한 항균활성에 관한 연구)

  • Yoon, Jae-Hong;Choi, Yeun-Ju;Jeong, Seung-Hyun;Shin, Gil-Cho
    • The Journal of Internal Korean Medicine
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    • v.34 no.3
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    • pp.278-288
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    • 2013
  • Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.

Antibacterial and Antibiotic Activity Enhancing Effect of Extract and Fractions from the Root of Rumex japonicus Houtt (참소리쟁이 뿌리 추출물 및 분획의 항균 활성과 항생제 증강 활성)

  • Yang, Sun A;Kim, A Young;Pyo, Byoung Sik;Kim, Sun Min
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.6
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    • pp.375-380
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    • 2017
  • Background: The objective of this study were to determine the antibacterial activity and antibiotic activity-enhancing effect of 70% ethanol extract of the root of Rumex japonicus Houtt. and its fractions when used in combination with gentamicin against aerobic skin flora. Methods and Results: The antibacterial activity and antibiotic (gentamicin) activity enhancing effect against aerobic skin flora were determined using the disc diffusion assay. Chloroform fraction (CF) and ethyl acetate fraction (EF) showed higher activities against Staphylococcus aureus and Staphylococcus epidermidis than those shown by other fractions. Regarding the antibiotic (gentamicin) activity-enhancing effect against aerobic skin flora, the n-hexane fraction (HF) and CF showed strong activity. The combination of HF and CF with gentamicin was evaluated using the broth dilution assay to determine the inhibitory effect on the growth of aerobic skin flora. The combination of CF with gentamicin exhibited the highest inhibitory effect on the growth of S. aureus and S. epdermidis. MTT assay performed to determine the viability of L929 cells revealed that EF treatment resulted in viability of 33.96 - 116.76% at the tested concentration. The combination of 70% ethanol extract and its other fractions with gentamicin showed low cell toxicity. Conclusions: Appropriate use of antimicrobial agents is important prior to the development of new antibiotics. The 70% ethanol extract of the root of R. japonicus Houtt. and its fractions showed significant synergism with gentamicin when used in combination against S. aureus and S. epdermidis. Thus, R. japonicus Houtt. could be used as a functional materials in antimicrobial-related fields.

Chemical compositions and biological activities of marine invertebrates from the East Sea of South Korea

  • Kim, Geun-Hyeong;Park, Hyeon-Ho;Chandika, Pathum;Ko, Seok-Chun;Jung, Kyung-Mi;Yoon, Sang Chul;Oh, Taeg-Yun;Kim, Young-Mog;Jung, Won-Kyo
    • Fisheries and Aquatic Sciences
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    • v.22 no.6
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    • pp.13.1-13.9
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    • 2019
  • Background: Marine invertebrates are well known as pivotal bioresources with bioactive substances such as anti-inflammatory sterols, antitumor terpenes, and antimicrobial peptides. However, there are few scientific reports on chemical compositions and bioactivities of marine invertebrates from the East Sea of South Korea. Methods: In this study, chemical compositions and biological activities were evaluated on both 70% EtOH and hot water extracts of 5 species of marine invertebrates (Crossaster papposus japonicus, Actinostola carlgreni, Stomphia coccinea, Actinostola sp., and Heliometra glacialis) collected from the East Sea of South Korea. The antioxidant activities were measured by ABTS radical scavenging assay. The cytotoxicity and anti-inflammatory activity were evaluated using MTT and Griess reagents. Moreover, the antibacterial effect was evaluated using paper disc assay and minimum inhibitory concentration (MIC) assay. Results: In the results of antioxidant activities, 70% EtOH extract of A. carlgreni showed the highest activity ($IC_{50}\;0.19{\pm}0.03mg/ml$) compared to other extracts. Moreover, 70% EtOH extract of A. carlgreni could significantly suppress the nitric oxide (NO) production in lipopolysaccharide-induced RAW 264.7. All extracts treated under $400{\mu}g/ml$ have no cytotoxic effects on RAW 264.7 macrophages. In the antibacterial test, both 70% EtOH extracts of C. papposus japonicus and H. glacialis showed a significant antibacterial effect on Staphylococcus aureus. The MIC values were evaluated at 256 and $512{\mu}g/ml$, respectively. Conclusions: These results suggested the bioactive potentials of marine invertebrates from the East Sea of South Korea in pharmaceutical and nutraceutical applications.