• Title/Summary/Keyword: In vitro generation

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Effects of Nitrofurantoin on Lipid Peroxidation and Reactive Oxygen Radical Generation in Porcine Lung Microsome (Nitrofurantion이 폐장 미크로솜 지질과산화와 반응성 산소 라디칼 생성에 미치는 영향)

  • Paick, Jae-Seung;Kim, Si-Whang;Kim, Hae-Won;Chung, Myung-Hee;Kim, Myung-Suk
    • The Korean Journal of Pharmacology
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    • v.21 no.1
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    • pp.34-48
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    • 1985
  • In vitro effects of nitrofurantoin, an antimicrobial agent for acute and chronic urinary tract infection, on the lung microsomal lipid peroxidation and the generation of reactive oxygen radicals were investigated to elucidate the biochemical mechanisms of its in vivopulmonary toxicity. The interaction of nitrofurantoin with porcine lung microsome resulted in significant lipid peroxidation. In addition, nitrofurantoin stimulated the generation of reactive oxygen radicals, $O^{-}_{2}{\cdot},\;H_2O_2$ as well as a highly reactive secondary oxygen species, $OH{\cdot}$. The stimulation of lipid peroxidation was inhibited not only by superoxide dismutase and catalase, but also by hydroxyl radical scavengers, mannitol and thiourea. Neither singlet oxygen $({^1}O_{2})$ was detected during the incubation of microsome with nitrofurantoin, nor lipid peroxidation was inhibited by singlet oxygen scavengers. When incubated anaerobically under the nitrogen atmosphere, the ability of nitrofurantoin to stimulatle lipid peroxidation was abolished. It appears that NADPH-dependent metaboliam of nitrofurantoin in pulmonary microsome under aerobic condition is accompanied by the stimulation of lipid peroxidation through the mediation of reactive oxygen radicals, particularly hydroxyl radical. It is strongly suggested from these results that the stimulation of pulmonary microsomal lipid peroxidation by the reactive oxygen radical may be a in vivo mechanism of pulmonary toxicity caused by nitrofurantoin.

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Study on Anti-thrombotic Activity, Superoxide Generation in Human Neutrophils and Platelet Aggregation in Human Blood of Hwao-tang

  • Park Won Hwan;Park Soo Young;Park Tae Woo;Kim Jong Gu;Kim Seog Ha;Kim Cherl Ho
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1494-1504
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    • 2004
  • The present paper reports the effects of Hwaotang an atherosclerosis using a spontaneous experimental model, We have also investigated the pharmacological effect of Hwaotang on collagen- and ADP-induced blood platelet aggregation, thrombin-induced conversion of fibrinogen and fibrinolysis in in vitro experiments, and various effects on stimuli-induced superoxide generation in human neutrophils. Hwao-tang was shown to have inhibitory effect on collagen- and ADP-induced blood platelet aggregation, on thrombin-induced conversion of fibrinogen to fibrin and on the activity of plasminogen or plasmin. Hwao-tang also significantly inhibited fMLP-induced superoxide generation in a concentration-dependent manner, but not that induced by arachidonic acid. Hwao-tang inhibited neutrophil functions, including degranulation, superoxide generation, and leukotriene B4 production, without any effect on 5-lipoxygenase activity. In conclusion, the protection of extracts of Hwao-tang on the ischemic infarction induced artificially might be involved to their inhibition of thrombotic action. The results also indicate that Hwao-tang exerts the effects on superoxide generation related to the inhibition of neutrophil functions.

In vitro Ccovalent Binding of SC-42867, PGE2 Antagonist, to Rat Liver Microsomal Proteins

  • Lee, Kyung-Tae
    • Archives of Pharmacal Research
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    • v.18 no.6
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    • pp.381-384
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    • 1995
  • Covalent binding of the reactive metabolites of SC_42867 to microsomal proteins has been examined. In the absence of inhibitor of cytochrome oxydase (.alpha.-naphtyl-isothiocyanate) or a radical scavenger (3-terthiobuty-4-hydroxyanisol), up to 4.0% of total redioactivity used in the assay could irreversibly bind to proteins. In the presence of an inhibitor, the highest percentage of covalent binding observed is 0.7% a significant decrease of the metabolism of SC42876 was observed. These results suggest in a cytochrome P-450 dependent generation of SC_42867 metabolites significantly take part in the covalent binding process.

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Preclinical Activity of Lobaplatin as a Single Agent and in Combination with Taxanes for Ovarian Carcinoma Cells

  • Sun, Xu;Lou, Li-Guang;Sui, Dong-Hu;Wu, Xiao-Hua
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9939-9943
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    • 2014
  • Lobaplatin, one of the third - generation platinum compounds, has shown encouraging anticancer activity in a variety of tumor types. However, the efficacy of lobaplatin in ovarian cancer has not been systemically evaluated. In this study, lobaplatin as a single agent and in combination with taxanes was investigated in - vitro and in an in vitro model of ovarian carcinoma. Using the sulforhodamine B (SRB) assay, the cytotoxic effects of lobaplatin alone and in combination with taxanes were compared with cisplatin and carboplatin in seven ovarian cancer cell lines. In addition, in - vitro antitumor activities were evaluated with cisplatin - sensitive and cisplatin - resistant human ovarian cancer xenografts in nude mice. The cytotoxicity of lobaplatin was similar to or higher than that of cisplatin and carboplatin, with $IC_{50}$ values from 0.9 to $13.8{\mu}mol/L$ in a variety of ovarian cancer cells. The combination of lobaplatin with docetaxel yielded enhanced cytotoxic activity in vitro. In addition, in platinum - sensitive ovarian cancer xenografts, lobaplatin alone showed similar antitumor activity to cisplatin and carboplatin. Furthermore, lobaplatin alone or in combination with docetaxel exhibited significant activity in platinum - resistant ovarian cancer xenografts. These results indicate that the use of lobaplatin alone or in combination with docetaxel might be a rational and novel therapeutic strategy for ovarian cancer. Further clinical development of lobaplatin is clearly warranted.

Effects of In Vitro Exposure to Silica on Bioactive Mediator Release by Alveolar Macrophages

  • Lee, Ji-Hee
    • The Korean Journal of Physiology
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    • v.29 no.1
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    • pp.1-11
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    • 1995
  • Alveolar macrophages play a pivotal role in the pathogenesis of silicosis since the macrophages may release a wide variety of toxic and inflammatory mediators as well as mitogenic growth factors. In the present study, the effects of in vitro exposure to silica on release of various mediator such as reactive oxygen species, platelet activating factor(PAF), and interleukin-1 (IL-1) by alveolar macrophages were examined. First, hydrogen peroxide release from alveolar macrophages was monitored by measuring the change in fluorescence of scopoletin in the absence or presence of graded concentration of silica. Significantly enhanced release of hydrogen peroxide was observed at 0.5 mg/ml and above. A maximal enhancement of 10 fold above control was observed at 5 mg/ml silica. Similarly, in vitro exposure to silica also significantly stimulated the generation of chemiluminescence from alveolar macrophages at 0.5 mg/ml and above with n maximal enhancement of 8 fold at 5 mg/ml silica. Second, PAF release from alveolar macrophages after 30 min incubation at $37^{\circ}C$ in absence or presence of zymosan and silica was determined by measuring $^{3}H-serotonin$ release ability of the conditioned macrophage supernates from platelets. 5 mg/ml zymosan as a positive control fur the PAF assay increased PAF release by 19 % of total serotonin release. Furthermore, silica also resulted in significant enhancement of the PAF release compared with that in unstimulated (control) cells, i.e., $17.7{\pm}5.8%$ and $24.0{\pm}4.9%$ of total serotonin release at 5 mg/ml and 10 mg/ml silica, respectively, which represents the release of nanomole levels of PAF. Lastly, IL-1 production by alveolar macrophages was analysed following their stimulation with lipopolysaccharide (LPS) and silica by their capacity to stimulate thymocyte proliferation. $10\;{\mu}g/ml$ LPS resulted in an 11 fold increase in IL-1 production. In comparison, $50\;{\mu}g/ml$ silica resulted in a 4 fold increase in IL-1 release. These data indicate that in vitro exposure of alveolar macrophages to silica activates the release of various bioactive mediators such as reactive oxygen species, PAF and IL-1 which thus contribute to amplification of inflammatory reactions and regulation of fibrotic responses by the lung after inhalation of silica.

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Development of In Vitro Culture System for Male Germline Stem Cells in Porcine (돼지 웅성 생식선 줄기세포의 체외배양기법 개발)

  • Kim, Yong-Hee;Kim, Byung-Gak;Lee, Yong-An;Kim, Bang-Jin;Kim, Ki-Jung;Lee, Myeung-Sik;Im, Gi-Sun;Ryu, Buom-Yong
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.171-177
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    • 2009
  • Spermatogonial stem cells(SSCs) only are responsible for the generation of progeny and for the transmission of genetic information to the next generation in male. Other in vitro studies have cultured SSCs for proliferation, differentiation, and genetic modification in mouse and rat. Currently, information regarding in vitro culture of porcine Germline Stem Cell(GSC) such as gonocyte or SSC is limited and is in need of further studies. Therefore, in this study, we report development of a successful culture system for gonocytes of neonatal porcine testes. Testis cells were extracted from $10{\sim}14$-day-old pigs. These cells were harvested using enzymatic digestion, and the harvested cells were purified with combination of percoll, laminin, and gelatin selection techniques. The most effective culture system of porcine gonocytes was established through trial experiments which made a comparison between different feeder cells, medium, serum concentrations, temperatures, and $O_2$ tensions. Taken together, the optimal condition was established using C166 or Mouse Embryonic Fibroblast(MEF) feeder cell, Rat Serum Free Medium(RSFM), 0% serum concentration, $37^{\circ}C$ temperature, and $O_2$ 20% tension. Although we discovered the optimal culture condition for proliferation of porcine gonocytes, the gonocyte colonies ceased to expand after one month. These results suggest inadequate acquirement of ingredients essential for long term culture of porcine GSCs. Consequently, further study should be conducted to establish a successful long-term culture system for porcine GSCs by introducing various growth factors or nutrients.

MASS PRODUCTION OF ENTOMOPATHOGENIC NEMATODE HETERORHABDITIS BACTERIPHORA IN VIVO AND VITRO CULTURE

  • Yoo, Sun-Kyun;Gaugler, Randy
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.201-207
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    • 2000
  • The strategies of commercial development have been focused on the economy of scale for a process. The design of media has been recognized as a key in assuring mass production of entomopathogenic nematodes. Media optimization was conducted with insect host, proteins, lipids, and symbiotic bacteria mass. G. mellonella (insect host) produced about 290,000 infective juveniles per one. Complex media produced about 250,000 infective juveniles / ml in liquid culture within 8 days (one generation).

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Effect of Apocynin on Acute Lung Injury in Rats Given Interleukin-$1{\alpha}$ Intratracheally

  • Lee, Young-Man
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.6
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    • pp.482-489
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    • 2011
  • Background: Based on the assertion that apocynin diminishes acute lung injury (ALI) by inhibition of NADPH oxidase, the effect of apocynin was tested in interleukin-$1{\alpha}$ (IL-1)-induced ALI in rats. Methods: IL-1 was insufflated into the trachea of Sprague-Dawley rats to induce ALI, and apocynin (8 mg/kg) was given intravenously for inhibition of NADPH oxidase. In addition, we determined whether apocynin inhibited generation of superoxide anions from isolated human neutrophils. Five hours after IL-1 instillation, lung injury parameters, expression of cytosolic phospholipase A2 (cPLA2) by cells from bronchoalveolar lavage (BAL), an index of oxidative stress in lung tissues (${\gamma}$-glutamyltranspeptidase, activity), and ultrastructure of alveolar type II (AT II) cells were evaluated. Results: Apocynin decreased the generation of free radicals from phorbol myristate (PMA)-activated neutrophils in vitro, but did not ameliorate ALI. IL-1 induced enhancement of the expression of cPLA2 on neutrophils was not altered by apocynin. Conclusion: Apocynin induced suppression of the generation of superoxide anions from neutrophils by inhibition of NADPH oxidase does not attenuate IL-1-induced ALI in rats.

Pathogenic variant in NLRP7 (19q13.42) associated with recurrent gestational trophoblastic disease: Data from early embryo development observed during in vitro fertilization

  • Sills, E. Scott;Obregon-Tito, Alexandra J.;Gao, Harry;McWilliams, Thomas K.;Gordon, Anthony T.;Adams, Catharine A.;Slim, Rima
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.1
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    • pp.40-46
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    • 2017
  • Objective: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. Methods: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. Results: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. Conclusion: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.

In Vitro Plant Regeneration from Stolen Node Explant in Eremochloa Ophiuroides (Munro) Hack

  • Barampuram, Shyamkumar;Chung, Byung-Yeoup;Lee, Seung-Sik;An, Byung-Chull;Kim, Jae-Sung
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.161-166
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    • 2007
  • In vitro shoot regeneration and multiple shoot induction has been obtained from the stolen node explants in Eremochloa ophiuroides (Munro) Hack. The highest number of shoots ($10.66{\pm}0.21$) was observed from initial explants after one month culture duration on Murashige and Skoog (MS) medium containing 6-benzyladenine (BA: 0.5 mg/l). First generation shoot was excised and sub-cultured on the same fresh media for further multiplication of shoots. An enhanced number of second round shoots ($15.33{\pm}0.21$) was obtained compared to the initial culture media containing BA (0.5 mg/l). The number of shoots/stolon node was higher among all the concentrations of BA than kinetin (KN). In vitro regenerated shoots were successfully rooted in the phytohormone free MS medium. Plantlets generated with roots were transferred to pots containing compound mixture of soil and kept in green house conditions. Acclimatized plants showed 100% survival rate with normal morphology in green house conditions. The present study demonstrates the effect of explant and different plant growth regulators towards in vitro response in E. ophiuroides. Moreover, the study reveals the effect of cytokinin on induction of shoot number per stolen node explant in E. ophiuroides.