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.
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.
Kim, Jai-Min;Kim, Soon-Ae;Yoo, Geun-Chang;Seo, Eun-Sun
Journal of Korean Ophthalmic Optics Society
/
v.8
no.1
/
pp.65-71
/
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.
Journal of the Korea Organic Resources Recycling Association
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v.8
no.4
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pp.100-110
/
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.
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
/
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.
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 $B_6$.
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.
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.
Purpose - This study reviews the achievements of a pilot project for the revitalization of a commercial district performed for three years after its establishment in 2011. The project for the revitalization of the commercial district was performed to create a new local community space in connection with the traditional market and nearby districts. Although it was a pilot project, the project for the revitalization of the commercial district has been performed for almost three years. Therefore, this seems a proper time to conduct an interim evaluation of the project. This study aims to review and evaluate how the government support policy is influential for the revitalization of the commercial district. In other words, this research aims to identify what projects positively affected consumers' intention to revisit the downtown commercial area among the commercial district revitalization projects-promotion events, promotion activities, education, merchants cooperation system, IT projects, cultural events, and residents' communication. Research design, data, and methodology - This study designated seven management improvement projects affecting commercial district revitalization based on preceding studies. The survey of the degree of satisfaction on seven management improvement projects was executed targeting consumers who visited the commercial areas. Additionally, visitors' revisit intentions regarding currently visited commercial areas were also investigated. Therefore, revisit intention was set as a dependent variable and the satisfaction degrees of the respective management improvement projects were set as the independent variables. A total of 1,209 consumers were examined in six districts in the country. Result - Multiple regression analysis results showed that cultural events, education, the merchants' cooperation system, and IT projects brought statistically significant effects to the revisit intentions of consumers. In contrast, promotion events, resident communication projects, and promotion activities did not affect the revisit intentions of consumers. Particularly, the residents' communication project did not show significant influence because of consumers' recognition that it is similar to a cultural event. Conclusion - The following implications for the revitalization of business districts in the urban central area are drawn. From a general perspective, the businesses of culture, education, and cooperative system among seven businesses play positive roles regarding the intention to revisit so that the project is required to be promoted periodically through unique performances differentiated for each district, the merchant training reinforced for professionalism, and the expansion of joint events of merchants. Moreover, the sales promotion project and public relations activity are shown to be not influential to the intention to revisit. Therefore, while short-term sales promotion such as one-time gift events are required, sales promotion and public relation activities to induce revisits by mileage savings and accumulated gift presentation to attract long-term customers are required. The IT business is positively influential to the intention of revisit. Therefore, detailed information on the revitalized commercial district should be provided and additional functions such as discount coupons for continuous utilization should be included in the mobile app and the website.
A novel glucanhydrolase from a mutant of Lipomyces starkeyi KSM 22 has additional amylase activity besides mutanolytic activity and has been suggested as promising anti-plaque agent. It has been shown effective in hydrolysis of mutan, reduction of mutan formation by Streptococcus mutans and removal pre-formed sucrose-dependent adherent microbial film and has been strongly bound to hydroxyapatitie. These in vitro properties of Lipomyces starkeyi KSM 22 glucanhydrolase are desirable for its application as a dental plaque control agent. In human experimental gingivitis model and 6 month clinical trial, mouthrinsing with Lipomyces starkeyi KSM 22 dextranase was comparable to 0.12% chlorhexidine mouthwash in inhibition of plaque accumulation and gingival inflammation and local side effect was negligible. This study was aimed to evaluate the cytotoxic effect of Lipomyces starkeyi KSM 22 glucanhydrolase on human gingival fibroblasts. Primary culture of human gingival fibroblasts at the 4th to 6th passages were used. Glucanhydrolase solution was made from lyophilized glucanhydrolase powder from a mutant of Lipomyces stakeyi KSM 22 solved in PBS and added to DMEM medium to the final concentration of 0.5, 1, and 2 unit. Cells were exposed to glucanhydrolase solution or 0.1 % chlorhexidine and the cells cultured in DMEM with 10% FBS and 1% antibiotics as control. After exposure, the morphological change, cell attachment, and cell activity by MTT assay were evaluated in 0.5, 1.5, 3, 6, 24 hours after treatment. The cell proliferation and cell activity was also evaluated at 2 and 7 days after 1 minute exposure, twice a day. The cell morphology was similar between the Lipomyces smkeyi KSM 22 glucanhydrolase groups and control group during the incubation periods, while most fibroblasts remained as round cell regardless of incubation time in the chlorhexidine group. The numbers of the attached cells in the glucanhydrolase groups were comparable to that of control and significantly higher than the chlorhexidine group. The numbers of the proliferated cells in the glucanhydrolase groups at 7 days of incubation were comparable to the control group and higher than the chlorhexidine group. The cell activity in glucanhydrolase groups paralleled with the increased cell number by attachment and proliferation. According to these results, Lipomyces starkeyj KSM 22 glucanhydrolase has little harmful effect on attachment and proliferation of human gingival fibroblasts, in contrast to 0.1% chlorhexidine which was cytotoxic to human gingival fibroblasts. Therefore this glucanhydrolase preparation is considered as a safe and promising agent for new mouthwash formula in the near future.
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