• Microbiology and Biotechnology Letters
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• 제26권5호
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• pp.435-441
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• 1998

Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.

• Radiation Oncology Journal
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• 제21권4호
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• pp.306-314
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• 2003

Purpose : In our Previous study, we have shown the main cel1 death pattern Induced by irradiation or protein tyrosine kinase (PTK) inhibitors in K562 human myeiogenous leukemic cell line. Death of the cells treated with irradiation alone was characterized by mitotic catastrophe and typical radiation-induced apoptosis was accelerated by herblmycin A (HMA). Both types of cell death were inhibited by genistein. In this study, we investigated the effects of HMA and genistein on cell cycle regulation and its correlation with the alterations of radiation-induced cell death. Materials and Methods: K562 cells In exponential growth phase were used for this study. The cells were Irradiated with 10 Gy using 6 MeV Linac (200-300 cGy/min). Immediately after irradiation, cells were treated with 250 nM of HMA or 25 $\mu$N of genistein. The distributions of cell cycle, the expressions of cell cycle-related protein, the activities of cyclin-dependent kinase, and the yield of senescence and differentiation were analyzed. Results: X-irradiated cells were arrested In the G2 phase of the cell cycle but unlike the p53-positive cells, they were not able to sustain the cell cycle arrest. An accumulation of cells in G2 phase of first ceil-cycle post-treatment and an increase of cyclin Bl were correlated with spontaneous, premature, chromosome condensation and mitotic catastrophe. HMA induced rapid G2 checkpoint abrogation and concomitant p53-independent Gl accumulation. HMA-induced cell cycle modifications correlated with the increase of CDK2 kinase activity, the decrease of the expressions of cyclins I and A and of CDK2 kinase activity, and the enhancement of radiation-induced apoptosis. Genistein maintained cells that were arrested in the G2-phase, decreased the expressions of cyclin Bl and cdc25c and cdc25C kinase activity, increased the expression of pl6, and sustained senescence and megakaryocytic differentiation. Conclusion: The effects of HMA and genistein on the radiation-induced cell death of KS62 cells were closely related to the cell cycle regulatory activities. In this study, we present a unique and reproducible model in which for investigating the mechanisms of various, radiation-induced, cancer cell death patterns. Further evaluation by using this model will provide a potent target for a new strategy of radiotherapy.

• Journal of the Korean Society of Food Science and Nutrition
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• 제30권6호
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• pp.1177-1183
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• 2001

This study was done to investigate the effects of ethanol extract of Cassia semen (Cassia tora L.) on the activities of hepatic oxygen free radicals metabolizing enzymes and blood lipid profile in rats of hepatotoxicity induced by ethanol. Sprague-Dawley male rats weighing 100~160 g were divides into 5 groups; control grouts (CON), Cassia semen ethanol extracts (200 mg/kg) treated group (CEL), ethanol (10 mL/kg, 35%) treated group (ETH), Cassia semen ethanol extracts (200 mg/kg) and ethanol treated group (CE1 ) and Cassia semen ethanol extracts (400 mg/kg ) and ethanol treated group (CE2), respectively. Compared with ETH, the growth rate of CE1 and CE2 were to be increased tendency, and in blood levels of total cholesterol, LDL-cholesterol and the activities of alanine aminotranferase and asparate aminotranferase elevated by ethanol were significantly decreased (p<0.05). It was observed that the activities of superoxide dismutase, catalase, xanthine oxidase and glutathione peroxidase of rat liver increased by ethanol, were more decreased by the treatment of Cassia semen ethanol extract than the only ethanol-treated group. The content of glutathione depleted by ethanol treatment was increased in CE1 and CE2. TBA-reactants of liver increased by ethanol were decreased in CE1 and CE2, compared with ethanol-treated group. These results suggested that ethanol extract of Cassia semen may influence upon the ability of oxygen free radical detoxication and lowering of blood lipid level on ethanol-induced hepatotoxicity in rat.

• Journal of Life Science
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• 제13권4호
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• pp.433-440
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• 2003

Osmotic and solid matrix priming treatments enhanced germination performance. We compared osmotic with solid matrix priming to determine the more effective treatment for improving seed germination in pepper and tomato. Seed hydration was immediately observed after osmotic priming and solid matrix priming treatment. The moisture content of solid matrix primed seeds was lower than that of osmotic primed seeds in the two vegetable crops. Osmotic priming and solid matrix priming did not increased percent germination, but showed shorter number of days to 50% of the final germination percentage ($T_{50}$) compared with untreated seeds, regardless of germination temperature. $T_{50}$ value was reduced in osmotic or solid matrix primed pepper seeds about 6.0, 5.0, 4.6 and 4.0 days compared with untreated seeds at 15, 20, 25, and $30^{\circ}C$, respectively. While, that in tomato seeds was reduced about 3.3, 5.0, 4.6 and 4.0 days compared with untreated seeds at 15, 20, 25, and $30^{\circ}C$, respectively. The effectiveness of osmotic priming or solid matrix priming in reducing the $T_{50}$ was greater when the seeds were germinated at $15^{\circ}C$ than at temperature of higher than $20^{\circ}C$. Solid matrix primed seeds germinated faster than osmotic primed seeds at all temperature in pepper. However, there was no significant difference on the percentage germination between solid matrix and osmotic primed seeds in tomato. After priming, dried-bark seeds showed faster germination than surface-dried seeds in pepper. On the other hand, there was no significant difference in tomato. Emergence of pepper and tomato seeds was markedly enhanced by osmotic priming or SMP treatment although the final emergence percentage was not significantly influenced. On the other hand, early growth was not significantly influenced by osmotic priming or SMP treatment of pepper and tomato seeds.

• Korean Journal of Environmental Biology
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• 제37권1호
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• pp.19-30
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• 2019

To investigate the characteristics of seasonal environment and phytoplankton community structure in the coastal area of Dokdo, a survey of Dokdo around waters was conducted during the four seasons. Phytoplankton of 4 phylum 72 species in four seasons were collected in Dokdo around water. The seasonal mean abundance of phytoplankton were $3.32{\times}10^4cells\;L^{-1}$ in winter, $1.04{\times}10^4cells\;L^{-1}$ in spring, $0.28{\times}10^4cells\;L^{-1}$ in summer, and $4.86{\times}10^4cells\;L^{-1}$ in autumn in Dokdo around water. During winter, the diatoms Chaetoceros spp. had dominated. During spring, when the nutrients in the euphotic layer were depleted, the nano-flagellates and Cryptomonas appeared at surface layer. In summer, the abundance of phytoplankton was relatively low, which lead to occurrence of diatoms such as genus of Chaetoceros, Rhizosolenia, and Skeletonema. In autumn, Pseudo-nitzschia spp. was the most dominant species and tropical species such as Amphisolenia sp. and Ornithocercus magnificus were observed, implying that they may have introduced within warm water current such as Kurosiwo Current. Therefore, although natural phytoplankton communities in the vicinity water of Dokdo are mainly influenced by Tsushima Warm Current branched Kurosiwo Current, their population dynamics was affected on the spatio-temporal change of physicochemical factors by short-term wind events, namely "island effect". Long-term survey research is needed to facilitate food-web response in marine ecosystem associated with phytoplankton biomass and physicochemical factors including the warm water current in oligotrophic offshore water of Dokdo, which may have significant role for sustainable use of Dokdo.

• Korean Journal of Ecology and Environment
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• 제37권1호통권106호
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• pp.26-35
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• 2004

We investigated the effect of physicochemical environmental factors on the community dynamics of phytoplanktons and bacteria at the Hoeya Dam Reservoir, a drinking water reservoir for Ulsan city. Water samples were collected and analyzed every two to four weeks at three sites along the reservoir from April to October, 2001. During the study period, the Secchi depths were between 0.4 and 3.5 m. At the surface layer of water column, temperature ranged 10.2 ~ $32.0^{\circ}C$, pH 7.3${\sim}$9.6, dissolved oxygen 5.5 ${\sim}$ 12.4 mg $L^{-1}$, $BOD_5$ 0.8 ${\sim}$ 5.0 mg $L^{-1}$, $COD_{Mn}$ 3.7 ${\sim}$ 10.0 mg $L^{-1}$, and Chl-a 8.9 ${\sim}$ 60.9 mg $m^{-3}$. At the bottom layer, temperature varied 7.2 ${\sim}$ $28.9^{\circ}C$, pH 7.1 ${\sim}$ 9.3, dissolved oxygen 0.6 ${\sim}$ 9.7 mg $L^{-1}$, $BOD_5$ 0.8 ${\sim}$ 4.5 mg $L^{-1}$, $COD_{Mn}$ 3.9 ${\sim}$ 10.0 mg $L^{-1}$, and Chl-a 4.3 ${\sim}$ 81.9 mg $m^{-3}$. The numbers of phytoplanktons were 7.4${\pm}10^2{\sim}2.6{\pm}10^5$ cells $mL^{-1}$ at surface and 2.5${\pm}10^2{\sim}2.4{\pm}10^4$ cells $mL^{-1}$ at bottom, and were positively correlated with water temperature and Chl- a concentration. Genus Stephanodiscus and genus Oscillatoria dominated on April and on May, respectively. Cyanobacterial blooms of Aphanizomenon, Microcystis, Anabaena were observed from June to early September, and thereafter Stephanodiscus and Aulacoseiral dominated again. Total microbial counts ranged 1.73${\pm}10^4{\sim}1.68{\pm}10^5$ cells $mL^{-1}$, and were positively correlated with water temperature and phytoplankton counts at surface water. Heterotrophic plate counts (HPCs) ranged 30${\sim}4.1{\pm}10^3$ CFU $mL^{-1}$, and were positively correlated with $BOD_5$ and $NO^3\;^-$-N concentration at bottom water. Unlike the total microbial counts, the numbers of fecal coliforms and fecal streptococci as well as HPCs were higher at the bottom than the surface layer and were highest at the upper a site among the three sampling sites. Since the concentrations of fecal coliforms and streptococci were still high at the bottom of site c, where intake for water treatment plant is located, it appeared that special management of water treatment processes may be needed especially after strong rainfall.

• Korean Journal of Animal Reproduction
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• 제21권2호
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• pp.167-176
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• 1997

This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

• Tuberculosis and Respiratory Diseases
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• 제49권1호
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• pp.37-48
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• 2000

Backgrounds : The pathophysiology of chronic airflow obstruction, such as bronchial asthma, is characterized by mucus hypersecretion, goblet cell hyperplasia(GCH), smooth muscle hypertrophy, and inflammatory cells infiltration. In fatal asthma patients, one distinct findings is mucus hypersecretion due to GCH. However, the mechanisms of GCH in these hypersecretory diseases remain still unknown. In this study, a rat model was rapidly induced with GCH by instillation of $TiO_2$, intratracheally. We intend to confirm GCH and association of concomitant inflammatory cells infiltration and to observe the effect of potent antiinflammatory agent, that is dexamethasone, on GCH with inflammatory cells. Methods : Twenty-one 8-weeks-old male Sprague-Dawley rats were divided into three groups. Endotoxinfree water was instilled intratracheally in group 1(control) ; $TiO_2$, was instilled in the group 2 ; and dexamethasone was injected intraperitoneally to group 3 before $TiO_2$ instillation. After 120 hours, all rats were sacrificed, and trachea, bronchi, and lungs were resected respectively. These tissues were made as paraffin blocks and stained as PAS for goblet cells and Luna stain for eosinophils. We calculated the ratio of goblet cell to respiratory epithelium and number of infiltrated eosinophils from each tissue. Results : (1) Fraction of goblet cells was significantly increased in group 2 than in group 1 in the trachea and in the main bronchus. (10.19$\pm$11.33% vs 4.09$\pm$8.28%, p<0.01 and 34.09$\pm$23.91% vs 3.61$\pm$4.84%, p<0.01, respectively). (2) Eosinophils were significantly increased in the airway of group 2 than that of group 1. (5.43$\pm$3.84% vs 0.17$\pm$0.47 in trachea and 47.71$\pm$16.91 vs 2.71$\pm$1.96 in main bronchi). (3) There was a positive correlation between goblet cells and eosinophils(r=0.719, p=0.001). (4) There was significant difference in the decrease of goblet cells after dexamethasone injection between group 2 and group 3 (p<0.01). Also, infiltration of eosinophils was suppressed by dexamethasone. Conclusion : We made an animal model of $TiO_2$-induced goblet cell hyperplasia. GCH was observed mainly in the main bronchi with concomitant eosinophilic infiltration. Both goblet cell hyperplasia and eosinophilic infiltration were suppressed by dexamethasone. This animal model may serve as a useful tool in understanding of the mechanism of GCH in chronic airway diseases.