• Title/Summary/Keyword: MTT method.

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A Study on Antimicrobial Activity of Lysimachia clethroides Duby Root Extracts against Methicillin-resistant Staphylococcus aureus

  • Joung, Dae Ki;Kwon, Dong Yeul;Shin, Dong Won
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.719-724
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    • 2018
  • Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium responsible for a number of infections in humans that are difficult to treat, and as a result, is a substantial contributor to morbidity and mortality. In the present study, in search of natural products capable of inhibiting this multidrug-resistant bacterium, we investigated the antimicrobial activity of Lysimachia clethroides Duby root. The antibacterial activities of EtOH extract of Lysimachia clethroides Duby root and its n-hexane, EtOAc, n-BuOH and water fractions were evaluated against 15 strains of methicillin-resistant staphylococcus aureus (MRSA) and 1 standard methicillin-susceptible S. aureus (MSSA) strain by using the minimal inhibitory concentrations (MICs) assay, colorimetric assay using MTT test, checkerboard dilution test. Antimicrobial activity of n-hexane fraction of Lysimachia clethroides Duby root was remarkable. Against the 16 strains, the minimum inhibitory concentrations (MICs) were in the range of $31.25-62.5{\mu}g/ml$ and FICI values for n-hexane fraction of Lysimachia clethroides Duby root+AM and n-hexane fraction of Lysimachia clethroides Duby root+OX were checkerboard method performed using the MRSA, MSSA and one clinical isolate strains via MICI 0.12-1 and 0.25-0.75, showing the increase of synergistic effect. When combined together, these antibiotic effects were dramatically increased. These effective combinations could be new promising agents in the management of MRSA.

In vitro Skin Irritation Test of Honeypolis using Human Skin Model

  • Woo, SoonOk;Han, Sangmi;Hong, Inpyo;Kim, Sung-kuk
    • Journal of Apiculture
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    • v.33 no.4
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    • pp.277-282
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    • 2018
  • Ethanol extracted propolis (EEP) was mixed with honey (honeypolis) to dissolve well in water and in vitro skin irritation test was conducted. In vitro method is designed to predict and classify the skin irritation potential of a chemical by assessment of its effect on $EpiDerm^{TM}$, a reconstituted three-dimensional human epidermis model. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 60 min exposure period. In this study under the given conditions honeypolis showed no irritant effects. Honeypolis meets acceptance criteria if: mean absolute OD 570 nm of the three negative control tissues is ${\geq}0.8$ and ${\leq}2.8$, mean relative tissue viability of the three positive control tissues is ${\leq}20%$, standard deviation of relative tissue viability obtained from each three concurrently tested tissues is ${\leq}18%$. Honeypolis is therefore classified as "non-irritant" in accordance with UN GHS "No Category".

Nitric Oxide Production and Acetylcholinesterase Inhibitory of Activity Various Extracts from Codonopsis lanceolata by Steaming Times (증숙 더덕 용매별 추출물의 Nitric Oxide 생성 저해 효과 및 Acetylcholinesterase 저해활성)

  • Choi, Hyun-Suk;Choi, DuBok
    • The Korean Journal of Food And Nutrition
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    • v.34 no.3
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    • pp.295-301
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    • 2021
  • Steaming is a method that has traditionally been used for medicinal plant extraction. This study investigated nitrite oxide production, ferrous ion chelating activity, α-glucosidase, xanthine oxidase, and acetylcholinesterase inhibitory activities of ethanol, acetone and hot-water extracts of Codonopsis lanceolata prepared by steaming seven times. MTT assay showed that each extract was non-toxic up to a concentration of 700 ㎍/mL confirming that there was no cytotoxicity in all extracts. The α-glucosidase, xanthine oxidase, and acetylcholinesterase inhibitory activities exhibited by the hot-water extract obtained from steaming seven times were higher (83.1%) than the other extracts. Higher production of nitrite oxide and better ferrous chelating activity was recorded with hot-water extract compared to ethanol and acetone extracts. These results indicated that more steaming of Codonopsis lanceolata extracts would be required to validate the possibility of developing antioxidants. Also, further study is needed to determine if the components present in the tested extracts might be useful in the prevention of Alzheimer's disease. These results showed that hot-water extracts may be useful for their antioxidant and the production inhibitory activity of nitrite oxide. It will be helpful in the investigation of the constituent analysis of the steam-processed product of Codonopsis lanceolata.

Adenine, new anti-wrinkle agent.

  • Kim, Y. J.;Kim, Y. S.;S.Y. Eom;Kim, J. H.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.804-819
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    • 2003
  • It has been known that adenine is a very important material in living cells. Because, adenine is a member of nucleotide base, so it takes part in DNA, RNA and ATP synthesis. There are many reports that adenine participated in ingredients, especially DNA, RNA, NADH and ATP, affect on the cell. As well adenosine, conjugated adenine to glycoside, was known to anti-wrinkle compound. But there is no report whether adenine shows a good effect on the skin, especially anti-wrinkle. So, in this study, we tested whether adenine affects cell proliferation, collagen synthesis, collagenase synthesis inhibition in human dermal fibroblasts. In addition, we performed clinical study with adenine cream. Cell proliferation effect was tested by MTT assay. Collagen and collagenase synthesis were measured by Immunoassay with ELISA kit. Clinical study was performed by IECK according to KFDA Functional Cosmetic method. The results of cell proliferation show that 10$^{-6}$ ~10$^{-8}$ % of adenine increases cell proliferation about 50 % compare with non-treated control. At 10$^{-7}$ ~10$^{-10}$ %, adenine increases type I collagen synthesis about 50%, decreases type I collagenase about 22% compare with non-treated control. The results of clinical study show that 0.05% adenine treated group reduces wrinkle significantly compare with placebo treated group. Therefore adenine may be a new anti-wrinkle candidate, through increases cell proliferation and collagen synthesis dramatically. And it decreases collagenase synthesis. So adenine could be used as a new anti-wrinkle agent.

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Effect of Water Extract from Artemisiae Argi Folium on Mouse Macrophage Stimulated by LPS (Lipopolysaccharide로 활성화된 마우스 대식세포에서 애엽(艾葉) 물추출물의 면역활성 연구)

  • Park, Wan-Su
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.151-157
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    • 2009
  • Objectives : The purpose of this study is to investigate the effect of Water Extract from Artemisiae Argi Folium (WAAF) on mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods : Cell viabilities were measured by MTT assay. And the intracellular productions of hydrogen peroxide (H2O2) were measured by dihydrorhodamine 123 assay. TNF-$\alpha$ and IL-6 production from Raw 264.7 were measured by ELISA method. Results : The results of the experiment are as follows. 1. WAAF significantly increased the cell viability compared to the control group (treated with LPS only) at the concentrations of 10, 50, 100, 200, 400 ug/mL. 2. WAAF significantly increased the intracellular production of H2O2 compared to the control group at the concentrations of 50, 100, 200 ug/mL. 3. WAAF significantly decreased the production of TNF-$\alpha$ compared to the control group at the concentrations of 100, 200 ug/mL. 4. WAAF significantly decreased the production of IL-6 compared to the control group at the concentrations of 50, 100, 200 ug/mL. Conclusions : WAAF could be supposed to have the immune-modulating activity related with the macrophage's immunoactivity.

Electrochemical Monitoring of NADH Redox with NPQD-modified Electrodes for Cell Viability Assessment

  • JuKyung Lee;Hye Bin Park;Chae Won Seo;Chae Won Seo;SangHee Kim
    • Journal of Sensor Science and Technology
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    • v.32 no.6
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    • pp.412-417
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    • 2023
  • There is increasing interest in the rapid and highly sensitive monitoring of cell viability in biological and toxicological research. Conventional methods depend on optical assays using Water Soluble Tetrazolium-8 (WST-8) or 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay, which requires a large volume of samples and special instruments, necessitating shipment of clinical samples to laboratories. This paper reports on the development of a rapid and sensitive electrochemical (EC) sensor using screen printed electrode (SPE) and surface modification using 4'-mercapto-N-phenylquinone diamine (4'-NPQD), as double electron mediators, for monitoring cell viability via the measurement of nicotinamide adenine dinucleotide (NADH). We used the sensor to observe the viability of MCF-7 and doxorubicin (Dox)-treated cells. The oxidation current of NADH was measured via chronoamperometry (CA), and the EC results showed a good linear relationship when compared with NADH quantification using WST-8 assay. The analysis time was only 10 s and limit of detection (LOD) of NADH was 1.78 µM. Our EC method has the potential to replace conventional WST assays for cell viability and cytotoxicity experiments.

Effect of Radiation Dosage Changes on the Cell Viability and the Apoptosis Induction on Normal and Tumorigenic Cells (방사선의 선량변화가 수종의 정상세포와 종양세포주의 세포활성도와 apoptosis 유발에 미치는 영향)

  • Park In-Woo;Lee Sam-Sun;Heo Min-Suk;Choi Soon-Chul
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.29 no.2
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    • pp.435-449
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    • 1999
  • Purpose : The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. Materials and Methods : The study. that was generated for two human normal cells(RHEK, HGF-l) and two human tumor cells(KB. HT-1080). was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5. 1, 2. 4. and 8Gy were applied to the cells. The two fractions of 1. 2. 4. and 8Gy were separated with a 4-hour time interval. The irradiation was done with 5.38Gy/min dose rate using Cs-137 irradiator at room temperature. Results and Conclusions : 1. In 3-day group. the cell viability of HGF-1 cell was significantly decreased at 2. 4 and 8Gy irradiation, the cell viability of KB cell was significantly decreased at 8Gy irradiation and the cell viability of HT-I080 cell was significantly decreased at 4 and 8Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2. 4 and 8Gy on HGF-1 cell. at 4 and 8Gy on HT-I080 cell. at 8Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.

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Cosmetic Effect of Angelica gigas Nakai Root Extracts (참당귀(Angelica gigas Nakai) 뿌리 추출물의 화장품소재 특성)

  • Park, Suk Kyoung;Hong, Seul-Ki;Kim, Hee Jin;Kim, Bo Young;Kim, Tagon;Kang, Jae Seon;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.47 no.5
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    • pp.553-557
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    • 2009
  • Root extracts of Angelica gigas Nakai were tested to see the possibility for functional cosmetic agents. From ethanol extraction method, 97% of decursin and decursinol angelate was obtained, and concentration ratio of decursin to decursinol angelate was about 3:2. To test possibility as a functional cosmetic agent, DPPH free radical scavenging assay, UVA/B absorption, tyrosinase inhibition assay, melanogenesis inhibition assay, elastase inhibition assay and MTT assay were done. Root extracts of Angelica gigas Nakai showed $45.2{\pm}3.9%$ tyrosinase inhibition of tyrosine, and $24.2{\pm}12.0%$ melanin inhibition at $15{\mu}g/ml$ extract concentration, so that it indicated good whitening effect. DPPH free radical scavenging activity was $40.9{\pm}9.1%$ at $240{\mu}g/ml$ concentration, which is relatively good. Anti-wrinkle effect was poor such that it was $12.7{\pm}6.8%$ at $100{\mu}g/ml$. UVA/B absorption was also negligible. From the research, root extracts of Angelica gigas Nakai showed good potential as a whitening agent.

Antitumor Activity and Nephrotoxicity of the Novel Platinum(II) Coordination Complex (새로운 Platinum (II) Complex [Pt (II)(trans-d-dach)(DPPE)] $(NO_3)_2$의 항암효과 및 신독성에 관한 연구)

  • Jung Jee-Chang;Lee Moon-Ho;Chang Sung-Goo;Rho Young-Soo
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.103-114
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    • 1995
  • Platinum coordination complexes are currently one of the most compounds used in the treatment of solid tumors. However, its use is limited by severe side effects such as renal toxicity. Our platinum-based drug discovery program is aimed at developing drugs capable of diminishing toxicity and improving antitumor activity. We synthesized new Pt (Ⅱ) complex analogue containing 1,2-diaminocyclohexane (dach) as carrier ligand and 1,2-bis(diphenylphosphino) ethane (DPPE) as a leaving group. Furthermore, nitrate was added to improve the solubility. A new series of [Pt(trans-ddach)(DPPE).$2NO_3(PC)$ was synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared (IR), $^{13}carbon$ nuclear magnetic resonance (NMR)]. PC demonstrated acceptable antitumor activity aganist P388, L-1210 lymphocytic leukemia cells and SK=OV3 human ovarian adenocarcinoma cells, and significant. activity as compared with that. cisplatin. The toxicity of PC was found quite less than thar of cisplatin using MTT, $[^3H]$ thymidine uptake and glucose consumption tests in rabbit proximal tubule cells, human kidney cortical cells and human renal cortical tissues. Based on these results, this novel platinum compound represent a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low toxicity.

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EFFECT OF EMD ON HUMAN PERIODONTAL LIGAMENT-DERIVED CELLS AND OSTEOBLAST-LIKE CELLS (MC3T3-E1) IN HIGH GLUCOSE CONDITION (고농도 포도당 환경에서 EMD(Enamel Matrix Derivatives)가 인간 치주인대 세포와 뼈모세포양 세포(MC3T3-E1)에 미치는 영향)

  • Lee, Baek-Soo;Kim, Sun-Wook;Jue, Sung-Sook;Kwon, Yong-Dae
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.5
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    • pp.532-536
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    • 2008
  • Purpose: This study was designed to evaluate effect of EMD on proliferation of HPDLCs and MC3T3-E1 cells in high glucose condition in vitro. Material and method: The Human PDL fibroblasts(HPDLCs) were obtained through typical way and the cells used in this experiment were divided in 4 groups. $1{\times}10^4/ml$ HPDLCs suspension was cultured in typical DMEM and assigned to group 1. The cells cultured in DMEM which included 400mg/dl glucose are allocated to group 3. Group 2 and 4 are established by adding EMD to group 1 and 3 respectively. These control and experimental groups had been cultured for 24 and 48 hours, and MTT assay was conducted. The differences of each group in cellular proliferation was evaluated. The same experiment was conducted for preosteoblast (MC3T3-E1) with adding $25\;{\mu}g/ml$ EMD. Results: EMD had the same effect on both PDL cells and MCT3T3-E1 cells. The experimental group had more meaningful differences and active cellular proliferation than the control group did. The EMD accelerated cellular proliferation not only in normal glucose condition but also in high glucose condition. The same results were observed via MTT assay; EMD-added experimental group had more meaningful differences and showed higher cellular activity than control group did. Each experimental and control group was inspected for statistical significance through Kruskal-Wallis Test. Statistical significances were observed among these groups. (SPSS 12.0 Chicago, IL, USA, p=0.008, p=0.011) Conclusion: EMD is considered to accelerate proliferation of PDL cells and MC3T3-E1 cells in high glucose condition as well as normal glucose condition.