• Title/Summary/Keyword: primary cultures

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Flavonoids Modulate the Proliferation of Neospora caninum in Glial Cell Primary Cultures

  • Matos, Rosan Barbosa De;Braga-de-Souza, Suzana;Pitanga, Bruno Pena Seara;Silva, Victor Diogenes Amaral Da;Jesus, Erica Etelvina Viana De;Pinheiro, Alexandre Morales;Costa, Maria De Fatima Dias;El-Bacha, Ramon Dos Santos;Ribeiro, Catia Suse De Oliveira;Costa, Silvia Lima
    • Parasites, Hosts and Diseases
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    • v.52 no.6
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    • pp.613-619
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    • 2014
  • Neospora caninum (Apicomplexa; Sarcocystidae) is a protozoan that causes abortion in cattle, horses, sheep, and dogs as well as neurological and dermatological diseases in dogs. In the central nervous system of dogs infected with N. caninum, cysts were detected that exhibited gliosis and meningitis. Flavonoids are polyphenolic compounds that exhibit antibacterial, antiparasitic, antifungal, and antiviral properties. In this study, we investigated the effects of flavonoids in a well-established in vitro model of N. caninum infection in glial cell cultures. Glial cells were treated individually with 10 different flavonoids, and a subset of cultures was also infected with the NC-1 strain of N. caninum. All of the flavonoids tested induced an increase in the metabolism of glial cells and many of them increased nitrite levels in cultures infected with NC-1 compared to controls and uninfected cultures. Among the flavonoids tested, 3',4'-dihydroxyflavone, 3',4',5,7-tetrahydroxyflavone (luteolin), and 3,3',4',5,6-pentahydroxyflavone (quercetin), also inhibited parasitophorous vacuole formation. Taken together, our findings show that flavonoids modulate glial cell responses, increase NO secretion, and interfere with N. caninum infection and proliferation.

White Ginseng Saponin Upregulated the Production of -TNFTNF-α, IL-1β and NO in Primary Cultures of Mixed Glial Cells (고려인삼에 의한 신경면역 및 염증반응 조절: 백삼사포닌에 의한 교세포에서의 TNF-α, IL-1β 및 NO 생성 증가)

  • 성정훈;최동희;김동훈;전보권;최상현
    • Journal of Ginseng Research
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    • v.28 no.2
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    • pp.120-126
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    • 2004
  • Glial cells such as astrocytes and microglial cells are the main source of proinflammatory cytokines and nitric oxide(NO) in the central nervous system, which exert neuroimmune and inflammatory functions and other various neurobiologic effects. Though Panax ginseng C.A. Meyer has been known to strengthen the body's defence mechanisms and also to maintain the homeostasis in the central nervous system, the effects of Panax ginseng on the production of immune and inflammatory mediators have not been studied well in the brain. Therefore, this study was designed to study the effects of ginseng saponins on the production of proinflammatory cytokines and NO in the primary cultures of mixed glial cells. White ginseng saponin, 200-500 $\mu$g/ml, showed significant cytotoxicity after 72 hrs and increased TNF-$\alpha$, IL-$\beta$, and NO production. Lower doses of 50-100 $\mu\textrm{g}$/ml showed little cytotoxicity until 72 hrs and also increased the production of TNF-$\alpha$, IL-1$\beta$, and NO. Triple immune staining showed that white ginseng saponin, 200$\mu\textrm{g}$/ml for 72 ks, induced stellation of astrocytes and iNOS expression exclusively in microglial cells. Taken together, the white ginseng saponin increased the production of proinflammatory cytokines such as TNF-$\alpha$ and IL-1$\beta$, and induced iNOS expression and NO production in mixed glial cell cultures, which may be ascribed to the enhancement of central immune responses and the regulation of inflammatory reactions by Panax ginseng.

Establishment of Highly Tumorigenic Human Gastric Carcinoma Cell Lines from Xenograft Tumors in Mice

  • Song, Kyung-A;Park, Jihyun;Kim, Ha-Jung;Kang, Myung Soo;Kim, Sun Young
    • Biomedical Science Letters
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    • v.23 no.3
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    • pp.238-250
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    • 2017
  • Patient's primary tumor-derived tumor cell lines likely represent ideal tools for human tumor biology in vitro and in vivo. Here, we describe eight human gastric carcinoma cell lines derived from established tumors in vivo upon subcutaneous transplantation of primary gastric carcinoma specimens in BALB/c nude mice. These xenografted gastric tumor cell lines (GTX) displayed close similarity with primary gastric tumor tissues in their in vivo growth pattern and genomic alterations. GTX-085 cells were resistant to cisplatin, while GTX-087 was the most sensitive cell line. GTX-085 was the only cell line showing a metastatic potential. Epithelial cell adhesion molecule (EPCAM) expression was especially strong in all tissue samples, as well as in cell cultures. GTX-139, the largest tumor graft obtained after injection, displayed distinct expression of CD44v6, fibroblast growth factor receptor 2 (FGFR2), and prominin 1 (PROM1, also known as CD133). In summary, we established eight xenograft gastric cancer cell lines from gastric cancer patient tissues, with their histological and molecular features consistent with those of the primary tumors. The established GTX cell lines will enable future studies of their responses to various treatments for gastric cancer.

Effects of Cytokinins on Secondary Embryogenesis and Plant Regeneration from Somatic Embryos of Aralia cordata Thunb. (땅두릅의 체세포배로부터 2차배 발생과 식물체 재생에 미치는 싸이토카이닌의 영향)

  • 이종천;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.149-154
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    • 2000
  • Embryogenic suspension cultures were initiated using embryogenic callus from immature inflorescence explants (Aralia cordata Thunb.) cultured on solid MS medium containing 1 mg/L 2,4-D for 8 weeks and then the embryogenic callus was proliferated in liquid MS medium containing 1 mg/L 2,4-D. After sieving the suspensions (pore size 270$\mu$m), embryogenic cells were cultured in liquid MS medium with cytokinins (kinetin, BA, zeatin) for two weeks. When the embryogenic cells were transferred to liquid MS basal medium, primary somatic embryos were developed after 5 weeks of culture. Secondary embryos were developed directly from the primary torpedo and cotyledonary embryos cultured in solid MS basal medium. Frequency of secondary embryogenesis was higher on medium containing 2 mg/L kinetin than the other cytokinins. Plant regeneration was highly recorded by placing secondary cotyledonary embryos induced from primary cotyledonary embryos in MS medium containing 2 mg/L kinetin or 2 mg/L zeatin (25.4% and 28.6%, respectively). The plant regeneration from secordary embryos was prohibited by tertiary embryogenesis.

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Vitellogenin and Its mRNA Induction by $Estradiol-17\beta$ in the Primary Culture of Hepatocytes in the Rainbow Trout, Oncorhynchus mykiss

  • Hwang Un-Gi
    • Fisheries and Aquatic Sciences
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    • v.4 no.4
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    • pp.186-191
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    • 2001
  • Vitellogenin (VTG) and VTG mRNA induction by $estradiol-17\beta\;(E_2)$ were examined in the primary cultures of hepatocyte in the rainbow trout. Hepatocytes were precultured for 2 days, then $E_2$ was added and cultured for another 5 days. Media and hepatocytes were then analyzed by electrophoresis and Northern blotting for VTG and VTG mRNA, respectively. The hepatocytes were formed a few aggregates within 5 days without further spreading to a monolayer. Cell viability and high DNA content were maintained during the incubation. The hepatocyte culture with E2 induced a weak VTG band at a molecular weight of 175kDa on Day 2 after $E_2$ addition. The relative amount of VTG was expressed in percentage of total protein concentrations. VTG was gradually increased as $1.9\%$ on Day 2, $6.3\%$ on Day 4 and $7.3\%$ on Day 5. VTG mRNA band was detected at about 6.6 kb in the culture with $E_2$ at day 1 of culture. The level of VTG mRNA expression linearly increased with time until Day 5 (r=0.97).

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Effects of Aqueous Extract Isolated from Platycodon grandiflorum Against t-Buty lhydroperoxide-induced Oxidative Stress in Rat Primary Hepatocytes (일차배양 간세포에서 t-Buty lhydroperoxide에 의해 유발된 산화적 스트레스에 대한 길경 열수 추출물의 보호효과)

  • 최철웅;이경진;정혜광
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.466-471
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    • 2002
  • Oxidative stress is considered to be associated with many diseases, such as inflammatory and cardiovascular diseases, aging and cancer. An important etiological mechanism of these diseases may be a causal relationship between the presence of oxidants and the generation of lipid hydroperoxides derived from enzymatic reactions or xenobiotic metabolism. The hydroperoxides can be decomposed to alkoxy- (ROㆍ) and peroxy- (ROOㆍ) free radicals that can oxidize other cell components, resulting in changes in enzyme activity or the generation of mediators, which can cause further cell damage. The aim of this study was to evaluate the ability of aqueous extract from the roots of Platycodon grandiflorum A. DC (Campanulaceae), Changkil (CK), to affect cellular response in primary cultures of rat hepatocytes to t-butyl hydroperoxide (t-BHP) induced oxidative stress and hepatotoxicity. CK-treated cells showed an increased resistance to oxidative challenge, as revealed by a higher percent of survival capacity in respect to control cells. CK reduced t-BHP-enhanced lipid peroxidation measured as production of malondialdehyde and enhanced intracellular reduced glutathione depletion by t-BHP. Furthermore, CK protected from the t-BHP-induced intracellular generation of reactive oxygen species assessed by monitoring dichlorodihydrofluorescein fluorescence. It can be concluded that CK exerts an antioxidant action inside the cell, responsible for the observed modulation of the cellular response to oxidative challenge, and CK have a marked antioxidative and hepatoprotective potency.

Studies on the anti-inflammatory action of steamed Rehmannia glutinosa in central nervous system (중추신경계(中樞神經系)에서 숙지황(熟地黃)의 항염증작용(抗炎症作用)에 관(關)한 연구(硏究))

  • Jung Young-Sik;Kang Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.10 no.2
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    • pp.59-70
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    • 1999
  • We investigated whether an aqueous extract of Rehmannia glutinosa steamed root (RGAE) inhibits secretion of inflammatory cytolanes from primary cultures of mouse astrocytes. RGAE dose-dependently inhibited the $TNF-{\alpha}$ secretion by astrocytes stimulated with substance P (SP) and lipopolysaccharide (LPS). Interleukin-1 (IL-1) has been shown to elevate $TNF-{\alpha}$ secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore also investigated whether IL-1 mediated inhibition of $TNF-{\alpha}$ secretion from primary astrocytes by RGAE. Treatment of RGAE to astrocytes stimulated with both LPS and SP decreased IL-1 secretion to the level observed with LPS alone. Moreover, incubation of astrocytes with IL-1 antibody abolished the synergistic cooperative effect of LPS and SP. These results suggest that RGAE has an antiinflammatory activity on the central nervous system curing some pathological disease states.

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참당귀 뿌리 배양에 있어서 root segmentation과 식물생장조절제가 뿌리 생장과 decursinol angelate 생산에 미치는 영향

  • Kim, Ji-Yeon;Jo, Ji-Suk;Jo, Jong-Mun;Lee, Yong-Il;Kim, Ik-Hwan;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.305-308
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    • 2002
  • Decursinol angelate, a new anticancer agent, was produced by root cultures of Angelica gigas Nakai. In addition, difference of specific yields between primary and secondary root was investigated. It was found that specific yield of secondary root was much higher than that of primary root at various conditions, so that it was thought that the formation and growth of secondary root were feasible. From this point of view, effects of root segmentation and plant growth regulators (NAA, IBA) on root morphology and decursinol angelate production were examined. Root segmentation increased secondary root mass and product formation. On the other hand, addition of NAA or IBA at various concentrations promoted secondary root formation and production of decursinol angelate significantly. Five-fold increase of production was obtained at 4 mg/L of IBA compared to control without NAA and IBA.

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Effects of Cholinesterase Inhibitors on Neuronal Injuries in Primary Cultured Rat Cortical Cells (배양한 대뇌피질세포에서 유발한 신경손상에 대한 콜린에스테라제 억제제의 영향)

  • 독고향;이광헌;조정숙
    • YAKHAK HOEJI
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    • v.46 no.3
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    • pp.185-191
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    • 2002
  • Alzheimer's disease (AD) involves neuronal degeneration with impaired cholinergic transmission, particularly in areas of the brain associated with learning and memory. Several cholinesterase inhibitors are widely prescribed to ameliorate the cognitive deficits in AD patients. In an attempt to examine if tacrine and donepezil, two well-known cholinesterase inhibitors, exhibit additional pharmacological actions in primary cultured rat cortical cells, we investigated the effects on neuronal injuries induced by glutamate or N-methyl-D-aspartate (NMDA), $\beta$-amyloid fragment ( $A_{{beta}25-35)}$), and various oxidative insults. Both tacrine and donepezil did not significantly inhibit the excitotoxic neuronal damage induced by glutamate. However, tacrine inhibited the toxicity induced by NMDA in a concentration-dependent fashion. In addition, tacrine significantly inhibited the $A_{{beta}25-35)}$-induced neuronal injury at the concentration of 50 $\mu$M. In contrast, donepezil did not reduce the NMDA- nor $A_{{beta}25-35)}$-induced neuronal injury. Tacrine and donepezil had no effects on oxidative neuronal injuries in cultures nor on lipid peroxidation in vitro. These results suggest that, in addition to its anticholinesterase activity, the neuroprotective effects by tacrine against the NMDA- and $A_{{beta}25-35)$-induced toxicity may be beneficial for the treatment of AD. In contrast, the potent and selective inhibition of central acetylcholinesterase appears to be the major action mechanism of donepezil.

A Study on Developing Cultural Education Program for Female Marriage Immigrants in Korea (여성 결혼이민자 문화교육 프로그램 개발을 위한 시론적 연구)

  • Oh, Yoon-Ja
    • Journal of Family Resource Management and Policy Review
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    • v.12 no.4
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    • pp.63-77
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    • 2008
  • This study was performed to develop the basic framework of multicultural art education programs for female foreign immigrants married to Korean males. First, these programs aim to lessen their cultural variance in acculturizing in Korea, and thereafter enhance their emotional stability and improve their creative ability to integrate cultural diversity into family life. The primary objective of this study was to offer a suitable education program and evaluation criteria for the program; the research has been done through person-to person interviews with 53 subjects using a series of questionnaires composed of 8 five-point Likert-Scaled items. As for the process of the interview, the subjects were asked about their satisfaction with each program item twice in order to comparetheir impressions before and after program participation. The satisfaction scores were analyzed using frequency distribution. In addition, the program effects were also evaluated by coordinators and developers from their subjective comments through periodical evaluation meetings. From the results, it is observed that art education programs are beneficial for improving female immigrants' emotionaland cultural integration. In addition, experts determined that the programs were helpful for improving their human relationship and acculturization in Korea. Finally, it is concluded that multicultural education programs are a primary fundamental infra-structural way of integrating diverse cultures and improving communication exchanges, and therefore, improve family stability among the multicultural families in Korea.

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