• Title/Summary/Keyword: in vitro activity test

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Antimicrobial activity and cytotoxicity test of Scrophularia ningpoensis hemsl extracts against Klebsiella pneumoniae

  • Yook, Keun-Dol
    • Journal of the Korea Society of Computer and Information
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    • v.21 no.5
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    • pp.135-139
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    • 2016
  • Scrophularia ningpoensis hemsl has been traditionally used in China and Vietnam for treatment of bacteria, atopy, pimple, tonsillitis, angina and encephalitis for a long time. The main objectives of this study were to evaluate the antibacterial activity of the Scrophularia ningpoensis hemsl extract on biofilm formation of Klebsiella pneumoniae. Antibacterial activity was conducted using disc diffusion assay and minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) were determined using the broth micro dilution method in accordance to Clinical and Laboratory Standards Institute guidelines(CLSI). Furthermore, cytotoxicity on L929 were assessed using animal cell culture for the proliferation test(MTT cell assay) and the biofilm forming capacity of the K. pneumoniae were determined using the colony forming unit (CFU) assay. The extract exhibited considerable antibacterial activity. K. pneumoniae was susceptible to the extract with the MIC and MBC of 0.1875 and $1.5mg/m{\ell}$ respectively. Cytoxicity test in L929 showed no sign of toxicity at the concentration of $0.75mg/m{\ell}$ and at the same concentration the extract caused inhibition of bacterial biofilm formation. The extract of Scrophularia ningpoensis hemsl possesses an in vitro antibacterial antibiofilm activities against K. pneumoniae, with no sign of cytoxicity on L929.

Phototoxicity: Its Mechanism and Animal Alternative Test Methods

  • Kim, Kyuri;Park, Hyeonji;Lim, Kyung-Min
    • Toxicological Research
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    • v.31 no.2
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    • pp.97-104
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    • 2015
  • The skin exposure to solar irradiation and photoreactive xenobiotics may produce abnormal skin reaction, phototoxicity. Phototoxicity is an acute light-induced response, which occurs when photoreacive chemicals are activated by solar lights and transformed into products cytotoxic against the skin cells. Multifarious symptoms of phototoxicity are identified, skin irritation, erythema, pruritis, and edema that are similar to those of the exaggerated sunburn. Diverse organic chemicals, especially drugs, are known to induce phototoxicity, which is probably from the common possession of UV-absorbing benzene or heterocyclic rings in their molecular structures. Both UVB (290~320 nm) and UVA (320~400 nm) are responsible for the manifestation of phototoxicity. Absorption of photons and absorbed energy (hv) by photoactive chemicals results in molecular changes or generates reactive oxygen species and depending on the way how endogenous molecules are affected by phototoxicants, mechanisms of phototoxcity is categorized into two modes of action: Direct when unstable species from excited state directly react with the endogenous molecules, and indirect when endogeneous molecules react with secondary photoproducts. In order to identify phototoxic potential of a chemical, various test methods have been introduced. Focus is given to animal alternative test methods, i.e., in vitro, and in chemico assays as well as in vivo. 3T3 neutral red uptake assay, erythrocyte photohemolysis test, and phototoxicity test using human 3-dimensional (3D) epidermis model are examples of in vitro assays. In chemico methods evaluate the generation of reactive oxygen species or DNA strand break activity employing plasmid for chemicals, or drugs with phototoxic potential.

In vitro response of rat microglia and human polymorphonuclear cells (PMN) to immunoactive compounds

  • Lombardi, Valter RM;Eetcheverria, Ignacio;Fernandez-Novoa, Lucia;Diaz, Joaquin;Seoane, Silvia;Cacabelos, Ramon
    • Advances in Traditional Medicine
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    • v.5 no.3
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    • pp.216-230
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    • 2005
  • Although the field of study in immune enhancing compounds is relatively new, natural products from plants represent a rich and promising source of novel molecules with immunomodulating properties, Microglial cells, the main immune effector cells of the brain, usually display a ramified morphology and low expression levels of immunologically relevant antigens such as MHC class I and class II. Since any compound which participates in activation of phagocytic cells contributes to the production of potentially toxic factors, the search for convenient in vitro test-systems and study of mechanisms of action of these agents are of great interest. Human blood polymorphonuclear (PMN) cells and primary microglial cells isolated from Sprague-Dawley rats were used as cellular screening tests for study of phagocytosis-stimulating action of immunomodulating agents. Numbers of phagocytic activity were evaluated by the phagocyte ingestion of yeast cells and NO-synthase activity, nitrite production, and nitroblue tetrazolium test were determined after phagocyte stimulation. It was possible to demonstrate that indexes of phagocytic activity can be used as quantitative indicators for measurement immunomodulating activity. As a positive control, Zymosan A-induced phagocytosis in both PMN cells and primary microglial cells was used. $IFN-{\gamma}$ (0.1 -1 U/ml) stimulated phagocytosis in PMN cells 1.2 times after 2 - 3 h incubation, although at higher concentrations (10 - 100 U/ml) it strongly inhibited phagocytosis. In a similar way, at higher concentrations, $IFN-{\gamma}$ (100 - 500 U/ml) suppressed phagocytosis in zymosan-A stimulated microglial cells. When Polypodium leucotomus, cambricum and vulgare extracts were tested alone, increased levels of phagocytosis were observed in PMN. In addition, microglial cells showed both increased phagocytosis and MHC class-II antigen expressions. Surprisingly, when PMN and microglia were treated with a combination of Polypodium and $IFN-{\gamma}$, phagocytosis was not inhibited. We did not find changes in NO-synthase activity and nitrite production in both microglia and PMN cells activated by different immunomodulating agents. These results indicate that primary microglial cell cultures as well as human PMN cells can provide reproducible quantitative results in screening phagocytic activity of different immunoactive compounds. Furthermore, both inhibitory or activation mechanisms might be studied using these in vitro experimental approaches.

In Vitro Inhibitory Activities of Essential Oils from Two Korean Thymus species against Antibiotic-Resistant Pathogens

  • Shin, Seung-Won;Kim, Ji-Hyun
    • Archives of Pharmacal Research
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    • v.28 no.8
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    • pp.897-901
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    • 2005
  • The in vitro inhibitory activities of essential oils from Thymus magnus and T. quinquecostatus as well as their main constituents were evaluated against susceptible and resistant species of Streptococcus pneumoniae, Staphylococcus aureus, Salmonella enteritidis, and Salmonella typhimurium. Notably, the essential oil fraction of T. magnus and its main components displayed significant inhibitory action against both antibiotic-susceptible and resistant strains of S. pneumoniae, S. aureus, and S. typhimurium with minimum inhibitory concentrations (MICs) ranging from 0.125 to 8 mg/mL. The differential MIC values imply that the oil fraction and its main components exhibit distinct patterns of activity against the tested bacterial species. Moreover, the disk diffusion test revealed that the inhibitory activities of oil fraction and components were dose-dependent. Data from the checkerboard titer test confirmed synergism between the antibiotic, norfloxacin, and T. magnus oil or thymol, particularly against the resistant strains of S. aureus.

Insulin secretory activity and mechanism of compound K

  • Choi, Yun-Suk;Han, Gi-Cheol;Sung, Jong-Hwan;Chung, Sung-Hyun
    • Proceedings of the Ginseng society Conference
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    • 2006.05a
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    • pp.69-70
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    • 2006
  • Purpose: Panaxadiols are more potent than panaxatriols as far as insulin secretory activity is concerned. In this study, we examined insulin secretory activity and mechanism of compound K (CK), a major intestinal bacterial metabolite of ginsenosides. Method: Insulin secretory activity of CK was examined using pancreatic beta cells and in Oral Glucose Tolerance Test assay. In addition, insulin secretory mechanism was studied in terms of calcium dependent or independent pathways. Results: In vitro, CK enhanced the insulin secretion concentration-dependently when compared to glucose-stimulated control cells. Insulin secretory mechanism of CK seems to block ATP sensitive K channels, which was confirmed by diazoxide (K channel opener) but, insulin resistance ameliorating activity of CK can't be ruled out. In vivo, CK showed hypoglycemic effect in OGTT.

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Factors Affecting in Vitro Activity of LB20304, a New Flu-oroquinolone

  • Paek, Kyoung-Sook;Ahn, Mi-Jeong;Kim, Mu-Yong;Kim, In-Chull;Kwak, Jin-Hwan
    • Archives of Pharmacal Research
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    • v.19 no.2
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    • pp.143-147
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    • 1996
  • LB20304 is a novel fluoroquinolone that exhibits a potent broad spectrum antibacterial activity against both gram-positve and gram-negative bacteria. The MICs (Minimal Inhibitory Concentration) of LB20304 were determined against both gram-positve and gram-negative bacteria under various conditions including several media, pHs, and inoculum concentrations. The in vitro activity of LB20304 was not significantly affected by the changes in testing conditions such as components of media and inoculum concentrations, but it was slightly reduced by acid condition. The MICs and MBCs (Minimal Bactericidal Concentration) of LB20304 against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were hardly affected by the presence of 50 % human serum, mouse serum, guinea pig serum or horse serum, and the MBCs were equal to or at most four-times higher than the MiCs. The activities of LB20304 were decreased by the presence of high concentraion of $Mg^{++}$ or human urine (pH, 5.5) in the test media. The frequencies of mutants resistant to LB20304 were similar to or lower than those found in ciprofloxacin and sparfloxacin.

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Cytolytic Activities of Taxol on Neural Stem Cells

  • Lee, In-Soo;Han, Hye-Eun;Lee, Hye-Young;Kim, Seung-U.;Kim, Tae-Ue
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.273-278
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    • 2007
  • Stem cells have been the subject of increasing scientific interest because of their utility in numerous biomedical applications. Stem cells are capable of renewing themselves; that is, they can be continuously cultured in an undifferentiated state, giving rise to more specialized cells of the human body. Therefore, stem cells are an important new tools for developing unique, in vitro model systems to test drugs and chemicals and a potential to predict or anticipate toxicity in humans. In the present study, in vitro cultured F3 immortalized human neural stem cell line and in vivo adult Sprague Dawley rats was used to evaluate the cytotoxicity of anticancer drug paclitaxel. In vitro apoptotic activity of paclitaxel was evaluated in F3 cell line by a MTT assay and DAPI test. The cell death was induced with the treatment of 20 nM paclitaxel and chromatin degradation was detected by DAPI staining, which was analyzed by fluorescent microscope. In vivo studies, we also observed nestin immunoreactivity on subventricular zone, which is stem cell rich region in the adult brain of the SD rat. Immunofluorescent staining result shows that pixel intensities of nestin were decreased in a dose dependent manner. These results suggest that paclitaxel is able to induce cytotoxic activity both in F3 neural stem cell line and neural stem cell in SD rat brain.

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Antimutagenic Activity of Asterina pectinifera (별불가사리의 항돌연변이 활성)

  • 함정혜;한영환;박창훈;이동웅
    • YAKHAK HOEJI
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    • v.43 no.6
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    • pp.771-775
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    • 1999
  • The antimutagenic activities of the total extract and several fractions of starfish, Asterina pectinifera (Asteriidae), were investigated in vitro by SOS chromotest with Escherichia coli PQ37 and Ames test with Salmonella typhimurium TA100. When various fractions was tested, the chloroform and butanol fractions showed low induction factors, which means both fractions increased antigenotoxicity against the base substitution mutagen MNNG. Even though higher antigenotoxic effect of the chloroform fraction, no effective result of Ames test was found in revertant formation of S. typhimurium TA100. The most effective antigenotoxic and antimutagenic fraction was a butanol one: i.e., When 0.5 mg/tube of butanol fraction was applied, the induction factor was 0.68. As the concentration of the fraction was increased the formation of revertants of S. typhimurium TA100 by about 81%. There was no cytotoxic effect of butanol fraction against S.typhimurium TA100. This result might be useful for further study to search a possible anticancer agent from the starfish, Asterina pectinifera.

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New Antibiotics Produced by Streptomyces melanosporofaciens II. Antimicrobial Activities and Isolation, Purification, and Structure Determination of the Active Compound (Streptomyces melanosporofaciens가 생산하는 새로운 항생물질 II. 물질의 항균활성과 황성물질의 분리.정제 및 구조결종)

  • 김시관;김상석;김근수;정영륜;김창한
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.235-241
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    • 1991
  • - A phthalic acid derivative and basic macrolide antibiotics, with antimicrobial activity against Gram positive bacteria and phytopathogenic fungi, respectively, were found to be produced by a strain 88-GT-161 identified as being a variety of Streptomyces melanosporofaciens. This paper describes an isolation procedure of the active compounds produced by this strain, their in vitro and in vivo (pot test) antimicrobial activites, and structure determination of one of the compounds, bis (2-ethylhexyl) phthalate, a phthalic acid derivative antibiotic. This compounds, upon cornparision with authentic bis (2-ethylhexyl) phthalate, dioctyl phthalate, revealed a difference in antimicrobial activity even though physico-chemical properties of these two compounds seemed indentical. This is the first report that dioctyl phthalate is biosynthetically produced by a Streptomyces sp. and shows antimicrobial activity.

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Effect of Gentianae Radix on Neurogenesis and Apoptosis in Hippocampus of Ethanol-induced Newborn Rats (급성 알코올 중독 어린 흰쥐의 해마 치상회에서 용담추출물이 신경세포생성과 세포사멸에 미치는 영향)

  • Lee, Jin-Gyu;Kim, E-Hwa;Lee, Jae-Hyok
    • Journal of Oriental Neuropsychiatry
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    • v.21 no.2
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    • pp.29-44
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    • 2010
  • Objectives : The purpose of this study was to investigate the effect of Gentianae Radix on neurogenesis and apoptosis in ethanol- induced newborn rats hippocampus dentate gyrus. Methods : In vivo, laboratory animals were divided into three groups; Normal group(N), Control group(C) and Treated group (TG)(n=7 for each group). N were treated saline daily for five days. C were treated 1.5 g/kg ethanol and saline daily for five days. TG were treated 1.5 g/kg ethanol and 300 mg/kg Gentianae Radix daily for five days. BrdU(5-bromo-2-deoxyuridine) assay was used to test neurogenesis in the dentate gyrus. And TUNEL(Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay was used to test apoptosis in the dentate gyrus. Three groups were measured body weight, serum ethanol concentration, BrdU-positive cells and TUNEL-positive cells in the dentate gyrus. In vitro, MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to test viability in SK-N-MC cells. BrdU assay was used to test neurogenesis in SK-N-MC cells. DNA fragmentation and caspase-3 enzyme activity assay were used to test apoptosis in SK-N-MC cells. And treated ethanol and Gentianae Radix of all in vitro tests were made various concentration. Results : In vivo, Gentianae Radix modulated ethanol-induced neurogenesis and apoptosis in newborn rats hippocampus dentate gyrus. In vitro, TG 100 ${\mu}g/ml$ have significantly modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells. And only TG 100 ${\mu}g/ml$ have significantly protected SK-N-MC cells from ethanol-induced cytotoxicity. Conclusions : Gentianae Radix may have the effect that modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells.