• Title/Summary/Keyword: culture-dependent

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Comparison of teratogenecity induced by nano- and micro-sized particles of zinc oxide in cultured mouse embryos

  • Jung, A Young;Jung, Ki Youn;Lin, Chunmei;Yon, Jung-Min;Lee, Jong Geol;Lee, Beom Jun;Yun, Young Won;Nam, Sang-Yoon
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.133-139
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    • 2015
  • The increasing uses of zinc oxide nanoparticles (nZnO) in industrial and personal care products raise possible danger of using nZnO in human. To determine whether ZnO induces size-dependent anomalies during embryonic organogenesis, mouse embryos on embryonic day 8.5 were cultured for 2 days under 50, 100, and $150{\mu}g$ of nZnO (< 100 nm) or micro-sized ZnO (mZnO; $80{\pm}25{\mu}m$), after which the morphological changes, cumulative quantity of Zn particles, and expressions of antioxidant and apoptotic genes were investigated. Although embryos exposed to $50{\mu}g$ of ZnO exhibited no defects on organogenesis, embryos exposed to over $100{\mu}g$ of ZnO showed increasing anomalies. Embryos treated with $150{\mu}g$ of nZnO revealed significant changes in Zn absorption level and morphological parameters including yolk sac diameter, head length, flexion, hindbrain, forebrain, branchial bars, maxillary process, mandibular process, forelimb, and total score compared to the same dose of mZnO-treated embryos. Furthermore, CuZn-superoxide dismutase, cytoplasmic glutathione peroxidase (GPx) and phospholipid hydroperoxidase GPx mRNA levels were significantly decreased, but caspase-3 mRNA level was greatly increased in nZnO-treated embryos as compared to normal control embryos. These findings indicate that nZnO has severer teratogenic effects than mZnO in developing embryos.

Continuous Hydrogen Production by Heterotrophic Growth of Citrobacter amalonaticus Y19 in Trickle Bed Reactor (Citrobacter amalonaticus Y19의 영양종속 성장을 이용한 Trickle Bed Reactor에서의 연속적인 수소생산)

  • Park, Ji-Young;Lee, Tae-Ho;Oh, You-Kwan;Kim, Jun-Rae;Seol, Eun-Hee;Jung, Gyoo-Yeol;Kim, Mi-Sun;Park, Sung-Hoon
    • KSBB Journal
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    • v.20 no.6
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    • pp.458-463
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    • 2005
  • [ $H_2$ ] from CO and water was continuously produced in a trickle bed reactor(TBR) using Citrobacter amalonaticus Y19. When the strain C. was cultivated in a stirred-tank reactor under a chemoheterotrophic and aerobic condition, the high final cell concentration of 13 g/L was obtained at 10 hr. When the culture was switched to an anaerobic condition with the continuous supply of gaseous CO, CO-dependent hydrogenase was fully induced and its hydrogen production activity approached 16 mmol/g cell/hr in 60 hr. The fully induced C. amalonaticus Y19 cells were circulated through a TBR packed with polyurethane foam, and the TBR was operated for more than 20 days for $H_2$ production. As gas retention time decreased or inlet CO partial pressure increased, $H_2$ production rate increased but the conversion from CO to $H_2$ decreased. The maximum $H_2$ production rate obtained was 16 mmol/L/hr at the gas retention time of 25 min and the CO inlet partial pressure of 0.4 atm. The high $H_2$ production rate was attributed to the high cell density in the liquid phase circulating the TBR as well as the high surface area of polyurethane foam used as packing material of the TBR.

Inhibition of LPS-induced nitric oxide production by transduced Tat-arginine deiminase fusion protein in Raw 264.7 cells

  • Lee, Min-Jung;Kim, Dae-Won;Lee, Yeom-Pyo;Jeong, Hoon-Jae;Kang, Hye-Won;Shin, Min-Jae;Sohn, Eun-Jeong;Kim, Mi-Jin;Jang, Sang-Ho;Kang, Tae-Cheon;Won, Moo-Ho;Min, Bon-Hong;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.42 no.5
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    • pp.286-292
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    • 2009
  • Arginine deiminase (ADI), an arginine-degrading enzyme, has anti-proliferative and anti-tumor activities and is capable of inhibiting the production of nitric oxide (NO). Modulation of nitric oxide (NO) production is considered a promising approach for the treatment of various diseases including cancer, inflammation and neuronal disorders. In this study, an ADI gene was fused with an HIV-1 Tat peptide in a bacterial expression vector to produce an genetic in-frame Tat-ADI fusion protein. When added exogenously to the culture media, the expressed and purified Tat-ADI fusion proteins were efficiently transduced into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced Tat-ADI fusion proteins markedly increased cell viability in cells treated with lipopolysaccharide (LPS). This increase in viability was mediated by an inhibition of NO production. These results suggest that this Tat-ADI fusion protein can be used in protein therapies of NO-related disorders such as cancer, inflammation and neuronal diseases.

Effects of Curcuma longa L.on Human Stomach Cancer Cells (울금(鬱金)이 위암세포(胃癌細胞)에 미치는 영향(影響))

  • Cho, Yu-Kyung;Yoon, Song-Ryub;Kim, Beong-Woo;Kim, Jin-Sung;Ryu, Ki-Won;Ryu, Bong-Ha
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.9 no.1
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    • pp.15-37
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    • 2003
  • Objective: We are aimed to identify anti-tumor effects of Curcuma longa L. on the stomach cancer cells through molecular biologic methods. Material & Methods: We used AGS as human stomach cancer cells obtained from American Type Culture Collection. The boiled extract of Curcuma longa L. $5{\mu}l$ (Sample I), $10{\mu}l$ (Sample II) was treated to cultural media(ml) for 0, 6, 12, 24, 48 hours. We measured the killing effect on stomach cancer cells through Trypan blue exclusion test and the suppressive effect on viability of stomach cancer cells via MTT assay. For identification of its anticancer mechanism, the revelation of Bcl-2, Bcl-XL, and Bax which are genes related to apoptosis using the quantitative RT-PCR, change of mitochondria membrane permeability and membrane potential via flow cytometry, the cycle of cell mitosis, caspase cleavage and annexin V staining were examined. Results: 1. showed significant killing effect on stomach cancer cell than the control group with a time(6 hours later) and density dependent manner, which was statistical significance. 2. Extract of Curcuma longa L. showed suppressive effect on viability of stomach cancer cells that each test groups had more suppressive effects on viability of stomach cancer cells than the control group with a time(6 hours later), which was statistical significance.(p<0.05) 3. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 and Bcl-XL decreased with a density manner which was statistical significance. but the revelation of Bax was not changed with statistical significance. 4. Extract of Curcuma longa L. caused apoptosis by decreasing the absorbance of mitochondria with statistical significance, and also induced apoptosis by decreasing the membrane potential of mitochondria. 5. Extract of Curcuma longa L. destructed the cell cycle of cell mitosos. 6. Cell apoptosis was induced by extract of Curcuma longa L. certificated by method of caspase cleavage and annexin V staining. Conclusion: This experiment showed that Curcuma longa L. has anti-tumor effect with statistical significance. This is in vitro experiment and basic experiment on Curcuma longa L.. We hope more progressive research on Curcuma longa L. will go on and its anti-tumor effects will be more practically identified.

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Apoptotic response to various apoptotic inducers on cultured HCE cells (여러 가지 apoptosis 유도 물질의 각막 상피세포에 대한 apoptosis 유도 반응)

  • Kim, Jai-Min;Kim, Soon-Ae;Yoo, Geun-Chang;Seo, Eun-Sun
    • Journal of Korean Ophthalmic Optics Society
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    • v.8 no.1
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    • pp.65-71
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    • 2003
  • The corneal epithelium is constantly being shed. The mechanism of corneal desquamation is not fully understood. Apoptosis, programmed cell death, may play a role. Apoptosis can be induced by a number of factors and different mechanisms. The study was performed to examine the apoptotic index induced in human corneal epithelial cells maintained in tissue culture by various apoptotic inducers. Various inducers, recombinant human cytokines($INF{\gamma}$, $TNF{\alpha}$, FASAb), actinomycin D. camptothecin, cycloheximide, dexamethasone and etoposide, were purchased from commercial suppliers. Inducers at manufacturer-recommended concentration were added to the corneal epithelial cells for 48 hours. Cell viability was measured using MTT assay. The cells were then assessed for the level of apoptosis. Morphologic changes and quantification of apoptotic cells were determined and counted under fluorescence microscope after inducers-treated human corneal epithelial (HCE) cells for 48 hours with Hoechst 33342 staining. Annexin V-FITC/PI staining and DePsipher assay. The expression of Fas protein was studied by immunocytochemistry. All inducers induced apoptosis in HCE cells in a dose dependent manner. Actinomycin D. camptothecin and etoposide induced apoptosis at lower than manufacturer-recommended concentration, while cytokines, cycloheximide and dexamethasone induced apoptosis at higher concentrations at the end of 48 hours. All inducers elicited typical apoptotic morphologic changes (chromatin condensation, nucleus fragmentations non-orange-red colored mitochondria) and expresses Fas protein highly. Apoptotic index of HCE cells by these inducers was different from the other cell lines. RNA synthesis inhibitor and topoisomerase inhibitors induced apoptosis at lower concentration than manufacturer-recommended concentration. Cytokines, cycloheximide and dexamethasone were able to produce apoptosis at 10 times higher concentrations. RNA synthesis inhibitor and topoisomerase inhibitors are more sensitive than intracellular receptor-activators in apoptotic induction of HCE cells.

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Effects on Chemical Compositions and Digestibilities of the Bulking Agents as a Moisture Control and fermentation Methods of food Waste (음식찌꺼기의 발효사료화시 수분조절제와 발효방법이 화학적 조성분 및 소화율에 미치는 영향)

  • Bae, Dong-Ho
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.4
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    • pp.100-110
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    • 2000
  • Studies were conducted to know effects of the bulking agents (saw dusts, mushroom waste, wheat bran coconut meal, rice hulls) adding o moisture control, fermentation methods (aerobic and anaerobic) and periods (1 to 20 days) of food waste fermentation for animal feeds on chemical compositions and in vitro DDM (digestibility of dry matter). Experiment designs were focussed basically to obtain extension service data. The NDF (neutral detergent fiber) composition in the oak and pine saw dust were 93.5% and 95.4% (DM basis) in respectively. Thus, the fermented food waste feeds using saw dust (50%) increased NDF(12%), and decreased in vitro DDM(48%) compared to those of raw materials before aerobic fermentation. The oak saw dust showed higher DDM compared to pine. Mushroom wastes which is a residues of mushroom culture mixed originally willow saw dust (80%) and wheat bran (20%) showed quite higher feed value compared to both saw dusts. It was found that an in vitro DDM and NDF composition in fermented feeds appeared highly dependent or the NDF composition in bulking agents. With an increase wheat bran ratio substitute mushroom waste showed linearly decreased NDF, and increased in vitro DDM in the fermented food waste feeds. The fermented feeds added bottling agents composed higher NDF resulted in higher NDF and lower in vitro DDM with prolonged fermentation time. The feeds from anaerobic fermentation appeared lower NDF and higher in vitro DDM compared to those of aerobic fermentation.

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Interleukin-$1{\beta}$ induces bone resorption by regulation of prostaglandin $E_2$ synthesis and plasminogen activator activity, and TGF-$\beta$ inhibits bone resorption of rat bone cells (쥐의 골세포에서 $PGE_2$ 합성과 plasminogen activator 활성 조절에 의한 IL-$1{\beta}$의 골 흡수유도와 TGF-$\beta$에 의한 골 흡수 억제 기전에 관한 연구)

  • Kim, Young-Hun;Lee, Young-Jun;Chung, Kyu-Rhim;Park, Young-Guk
    • The korean journal of orthodontics
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    • v.30 no.6 s.83
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    • pp.713-721
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    • 2000
  • Bone cells produce multiple growth factors and cytokines that have effects on bone metabolism and can be incorporated into the bone matrix. The present study was designed to extend these observations by examining the interactions between transforming growth factor-$\beta$(TGF-$\beta$) or interleukin-$1\beta$(rhIL-$1\beta$) and bone cells in a rat long bone culture model. IL-$1\beta$ regulates several activities of the osteoblast cells derived from rat long bone explants in vitro. IL-$1\beta$ stimulated cellular proliferation as well as the synthesis of prostaglandin $E_2$ and Plasminogen activator activity in the cultured cells in a dose-dependent manner. TGF-$\beta$ is present in the bone matrix and potentially released during bone resorption. TGF-$\beta$ reduced basal bone resorption and inhibited vitamin $D_3[1,25(OH)_2D_3]$-induced bone resorption in rat long bone cells. These results support the role of IL-$1\beta$ in the pathological modulation of bone cell metabolism, with regard to implication in the Pathogenesis of osteoporosis by IL-$1\beta$, and that TGF-$\beta$ positively inhibits the bone resorption.

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Human brain pyridoxal-5'-phosphate phosphatase (PLPP): protein transduction of PEP-1-PLPP into PC12 cells

  • Lee, Yeom-Pyo;Kim, Dae-Won;Lee, Min-Jung;Jeong, Min-Seop;Kim, So-Young;Lee, Sun-Hwa;Jang, Sang-Ho;Park, Jin-Seu;Kang, Tae-Cheon;Won, Moo-Ho;Cho, Sung-Woo;Kwon, Oh-Shin;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.41 no.5
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    • pp.408-413
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    • 2008
  • Pyridoxal-5'-phosphate phosphatase (PLPP) catalyzes the dephosphorylation of pyridoxal-5'-phosphate (PLP). A human brain PLPP gene was fused with a PEP-1 peptide and produced a genetic in-frame PEP-1-PLPP fusion protein. The purified PEP-1-PLPP fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced PEP-1-PLPP fusion protein was stable for 36 h. The concentration of PLP was markedly decreased by the addition of exogenous PEP-1-PLPP to media pretreated with the vitamin $B_6$ precursors; pyridoxine, pyridoxal kinase and pyridoxine-5'-phosphate oxidase into cells. The results suggest that the transduction of the PEP-1-PLPP fusion protein can be one mode of PLP level regulation, and to replenish this enzyme in the various neurological disorders related to vitamin $B_6$.

The Anticancer Effects and Drug Metabolic Enzyme Change by Intraperitoneal Injection of Agrimonia Pilosa Ledeb (선학초 (짚신나물) 복강주사의 항암효과 탐색 및 약물 대사효소의 변화)

  • Choi, Jung-Won;Jang, Bo-Hyung;Lee, Ju-Ah;Ko, Ho-Yeon;Jung, Hee;Jun, Chan-Yong;Park, Jong-Hyung;Kim, Ji-Hye;Ko, Seong-Gyu;Choi, You-Kyung
    • The Journal of Korean Medicine
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    • v.30 no.4
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    • pp.129-141
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    • 2009
  • Objective: This study was to investigate the anti-tumor effect, safety, safety, mechanism and metabolizing enzyme of Agrimonia pilosa LEDEB (APL) in female C57B/L mouse tumor (in vivo). Method: First, to evaluate the antitumor activity of APL, we divided the mice into four groups: normal, control, APL50 (50mg/kg), and APL100 (100mg/kg). LLC-obtained American Type Culture Collection was used. LLC had been inoculated to induce tumors. To measure the anti-tumor effect of APL, we calibrated tumor size and weight. To analyze the mechanism of anti-tumor in APL, we used western blotting and to observe metabolizing enzyme in APL we used to real-time PCR. Result: APL50 and APL100 significantly inhibited tumor growth from 12 days after medicine injected. APL did not induce caspase-dependent apoptosis in LLC-bearing mouse tumor. In APL100, it decreased 41% and 71% in CYP2D22 and CYP3A11, respectively. Conclusion: These results suggest that APL has some anti-tumor effects in female C57B/L mouse tumor. APL should be used carefully with other drugs related with CYP2D22 and CYP3A11.

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Customer Participation Driven Sustainable Business Ecosystems (고객참여 기반의 지속가능한 비즈니스 생태계 조성)

  • Joo, Jae-Hun;Shin, Matthew Min-Suk
    • Journal of Distribution Science
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    • v.12 no.12
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    • pp.83-92
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    • 2014
  • Purpose - A business ecosystem refers to mutually dependent systems interconnected by a loose foundation of various ecosystem members such as customers, suppliers, partners, and other stakeholders. The ecosystem-based strategy attempts to achieve competitive advantage for firms by enriching a business ecosystem or building a sustainable business ecosystem through the collaboration and co-evolution of its members. A sustainable business ecosystem is a source of competitiveness for firms anda manageable resource for gaining a competitive advantage. Customers represent the core membership of the business ecosystem and play a pivotal role in building a sustainable business ecosystem. This study examines the effects of customer participation on economic and social value in the business ecosystem and suggests a course of action for building a sustainable business ecosystem. Research design, data, and methodology - Two business cases of South Korea are selected from two different business types: business-to-business (B2B) and business-to-customer (B2C) firms. Business ecosystems for B2B and B2C firms reflect contrasting characteristics. Data was collected from in-depth interviews with four representatives of four firms. Results - The study suggested seven propositions for the relationships between customer participation and a sustainable business ecosystem through multiple case studies based on in-depth interviews. The results reveal the following four strategic actions for building sustainable business ecosystems based on the suggested propositions: alignment, systemization, socialization, and co-evolution. Alignment refers to achieving a harmonic balance or virtuous circle among the firm's mission, investment, and value creation. Systemization refers to building and implementing management and infrastructure systems rooted in the corporate culture. Socialization of customers in the business ecosystem reinforces the harmony or virtuous cycle. Finally, co-evolution is associated with the relationship between firms and customers as buyer firms in a restricted business ecosystem. Conclusions - This study considers multiple cases for the execution of a sustainable business ecosystem in collaboration with customers and suggests seven propositions and four strategic actions. The results are based on qualitative data from interviews with business associates from two firms in an open business ecosystem and two firms in a restricted business ecosystem, both in South Korea. Our research results regarding two contrasting business ecosystems shed light on business issues and policy making in Asian business environments, which are in the transition stages from a traditional conglomerate-driven to an inclusive growth-driven economy. The business ecosystem itself should be considered a manageable resource for firms' competitive positions in the market. A customer is a member of the business ecosystem and should thus be viewed not only as a purchasing entity and an object of relationship management but also as a co-creator of value. Therefore, firms should collaborate with customers to build sustainable business ecosystems. For this, firms must create social value, which cannot be created by customers alone, within the business ecosystem. Then, customers participate in a business ecosystem and build it to be favorable to them. Implications for academics and practitioners were suggested.