• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.17 no.4
• /
• pp.34-42
• /
• 2016

In order to prepare for the type approval test for the United States Coast Guard (USCG) Phase-II of Ballast Water Treatment System (BWTS), a phytoplankton mass culture was conducted in a mesocosm enclosure. We evaluated the response of the phytoplankton community after nutrient addition (+N, +P, and +NP) and investigated the development of the species with increasing culture time. After nutrient dosing, the phytoplankton population significantly (p < 0.05) increased from day 1 to day 3, depending on the nutrient treatments In particular, the specific growth rate of the phytoplankton community in the case of +NP treatment and + N treatment were estimated to be $2.47d^{-1}$ and $1.98d^{-1}$, respectively. The phytoplankton population density in the case of + NP treatment was approximately 50 times higher than that of the control group, suggesting that these treatments could be useful for mass culturing phytoplankton (> 75% of natural community) for the approval regulation of USCG Phase-II. In the phytoplankton community of the mesocosm, Pseudo-nitzchia spp. dominated in the logarithmic growth phase. The cell density decreased significantly (p < 0.05) with increasing time, coinciding with the nutrient limitation. At that time, the dominance of Pseudo-nitzchia spp. shifted to that of Cylindrotheca closterium. Therefore, the optimum nutrient concentration ($N:30{\mu}M$, $P:3{\mu}M$) and reasonable harvesting time (after 3 days in summer) found in this study for the mass culturing of phytoplankton may be helpful to meet the USCG Phase-II biological criteria to be used in BWTS.

• Journal of Plant Biotechnology
• /
• v.29 no.4
• /
• pp.247-252
• /
• 2002

The combination of radiation technique with an in vitro culture system was initiated to develop salt tolerant rice. We established an in vitro culture system to select tolerant lines against salt stress. NaCl tolerant cell lines were selected from the callus irradiated with gamma ray on N$_{6}$ medium with 1.5% NaCl and 2 mg/L 2,4-D. Regenerants (M$_1$) were obtained from the tolerant callus which was cultured for 30 days auxin-free medium. The M$_2$seeds were harvested from M$_1$plants on an individual plant basis. Thirty seedlings from each 450 M$_2$lines were transplanted in a field and total 5,000 M$_3$lines were harvested with an average 90 percent of fertile grain. M$_3$lines were utilized for selection of salt tolerance. Salinity-tolerant lines (225) were selected among 5,000 M$_3$lines. Of the 225 lines tested, the morphological traits of two lines (120-10 and -11) were far superior to control (Donagjinbyeo) in agromomic traits such as plant height, root length and no. of roots. Control and tolerant lines were analyzed by RAPD markers. Three polymorphic bands were presented in only tolerant lines, demonstrating a genetic difference between control and the tolerant lines. Such tolerant lines could be used as genetic resources to improve salt tolerance.e.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.9
• /
• pp.1333-1339
• /
• 2011

After mulberry (Morus alba) leaves were fermented with Hericium erinaceum mycelium by solid-state culture to enhance physiological activity, fermented mulberry leaves (MA-HE) was extracted by hot-water (MA-HEHW) and ethanol (MA-HE-E). MA-HE-HW showed enhanced mitogenic and intestinal immune system modulating activities (1.41 and 1.52 fold of saline control, respectively) compared to hot-water extracts of non-fermented mulberry leaves (MA-HW) and H. erinaceum mycelium (HE-HW) at $100\;{\mu}g$/mL. Meanwhile, when we tested the inhibitory effects of extracts on nitric oxide (NO), tumor necrosis factor (TNF)-${\alpha}$, and interleukin (IL)-$1{\beta}$ and IL-6 production, MA-HE-E significantly inhibited these pro-inflammatory mediators in LPS-stimulated RAW 264.7 cells (45.1, 41.3, 70.2, and 55.7% inhibition of LPS control at $1,000\;{\mu}g$/mL). In addition, MA-HE-HW and MA-HE-E did not show any cytotoxicity on RAW 264.7 cells at $1,000\;{\mu}g$/mL whereas HE-E and MA-E indicated cytotoxicity (80.1 and 30.7% cell viability of saline control). These results suggest that mulberry leaves fermented with H. erinaceum by solid-state culture might have enhanced immunomodulatory and anti-inflammatory effects compared to non-fermented mulberry leaves, resulting in ingredients biotransformed for fermentation with H. erinaceum mycelium.

• Journal of Plant Biotechnology
• /
• v.32 no.1
• /
• pp.51-55
• /
• 2005

Chinese leymus (Leymus chinensis Trin.) is a perennial grass that is widely distributed at high pH sodic and arid soil in the northeastern Asia. An efficient regeneration system was established through somatic embryogenesis of mature seeds to understand its high adaptability to harsh environmental conditions on the basis of molecular biology. The calli were efficiently induced (about $70\%$) from mature seeds on MS medium supplemented with $1.5\;\cal{mg/L}$ 2,4-D. Somatic embryos were formed from the surface of embryogenic callus on MS medium supplemented with $2.0\;\cal{mg/L}\;kinetin\;and\;0.5\;\cal{mg/L}$ NAA after 3 weeks of culture. Roots were induced from the shoot when transferred to MS medium without plant growth regulator for 1 week. Plant regeneration rate was $36\%$ and regenerated plantlets were grown to normal mature plants in pot. An efficient plant regeneration system in this study will be useful for molecular breeding of L. chinensis.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.4
• /
• pp.487-495
• /
• 2007

The objectives of the present study were to initiate cloning of Korean native goat by somatic cell nuclear transfer (NT) and to examine whether unovulated (follicular) oocytes can support the same developmental ability of NT embryos as ovulated (oviductal) oocytes after hCG injection in stimulated cycles of the goat. The in vivo-matured and immature oocytes were collected from the oviducts and follicles of superovulated does, respectively, and the immature oocytes were maturated in vitro. Ear skin fibroblasts derived from a 3-yr-old female Korean native goat were used as the donors of nuclei or karyoplasts. Following fusion, activation and in vitro culture to a 2- to 4-cell stage, 49 in vitro-derived and 105 in vivo-derived embryos were transferred to 6 and 17 recipient does, respectively. One doe and three does of the respective groups were identified as pregnant by ultrasonography on day 30 after embryo transfer. However, only one doe, which had received in vivo-derived embryos, delivered a normal female kid of 1.9 kg on d 149. The cloned kid gained more weight than her age-matched females as much as 87% during the first 4 mo after birth (17.7 vs. $9.4{\pm}0.8$ kg) and reached puberty at 6-mo age a few months earlier than normal female does. The telomere length of the kid, which was similar to that of the donor fibroblast at 2-mo age, decreased 8% between 2- and 7-mo ages. Moreover, at 7-mo age, she had 21% shorter telomere than her age-matched goats. To our knowledge, this is the first case in which a cloned animal born with a normal weight exhibited accelerated growth and development. The unusually rapid growth and development of the cloned goat may have resulted from SCNT-associated epigenetic reprogramming involving telomere shortening.

• Applied Microscopy
• /
• v.7 no.1
• /
• pp.21-43
• /
• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• KSBB Journal
• /
• v.17 no.1
• /
• pp.73-79
• /
• 2002

The method to optimize the microbial production of ethanol from CO using Clostridium ljungdahlii was developed. The kinetic parameter study on CO conversion with Clostridium ljungdahlii was carried out and maximum CO conversion rate of 37.14 mmol/L-hr-O.D. and $K_{m}$ / of 0.9516 atm were obtained. It was observed that method of two stage fermentation, which consists of cell growth stage and ethanol production stage, was effective to produce ethanol. When pH was shifted from 5.5 to 4.5 and ammonium solution was supplied to culture media as nitrogen source at ethanol production stage, the concentration of ethanol produced was increased 20 times higher than that without shift. Ethanol production from CO in a fermenter with Clostridium ljungdahlii was optimized and the concentration of ethanol produced was 45 g/L and maximun ethanol productivity was 0.75 g ethanol/L-hr.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.7
• /
• pp.933-941
• /
• 2021

Ginsenoside Rg4 is a rare ginsenoside that is naturally found in ginseng, and exhibits a wide range of biological activities including antioxidant and anti-inflammatory properties in several cell types. The purpose of this study was to use an in vivo model of hair follicle (HF)-mimic based on a human dermal papilla (DP) spheroid system prepared by three-dimensional (3D) culture and to investigate the effect of Rg4 on the hair-inductive properties of DP cells. Treatment of the DP spheroids with Rg4 (20 to 50 ㎍/ml) significantly increased the viability and size of the DP spheres in a dose-dependent manner. Rg4 also increased the mRNA and protein expression of DP signature genes that are related to hair growth including ALP, BMP2, and VCAN in the DP spheres. Analysis of the signaling molecules and luciferase reporter assays further revealed that Rg4 induces the activation of phosphoinositide 3-kinase (PI3K)/AKT and the inhibitory phosphorylation of GSK3β, which activates the WNT/β-catenin signaling pathway. These results correlated with not only the increased nuclear translocation of β-catenin following the treatment of the DP spheres with Rg4 but also the significant elevation of mRNA expression of the downstream target genes of the WNT/β-catenin pathway including WNT5A, β-catenin, and LEF1. In conclusion, these results demonstrated that ginsenoside Rg4 promotes the hair-inductive properties of DP cells by activating the AKT/GSK3β/β-catenin signaling pathway in DP spheres, suggesting that Rg4 could be a potential natural therapy for hair growth.

• The Korean Journal of Mycology
• /
• v.46 no.2
• /
• pp.161-176
• /
• 2018

Fungal ${\beta}$-glucan, known to have immunostimulatory and antitumor activities, can be recognized by host immune cells as one of the pathogen-associated molecular patterns (PAMPs). Although there are several reports on the diverse immunostimulatory activities of ${\beta}$-glucan, little is known about the intracellular signal transduction of ${\beta}$-glucan. Stimulation of RAW264.7 macrophage cells with ${\beta}$-glucan from Ganoderma lucidum induced the expressions of dectin-1, toll-like receptor 2 (TLR2), TLR4, and TLR6 at the transcription stage. Treatment with ${\beta}$-glucan also induced inflammatory mediators such as macrophage inflammatory proteins (MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-$1{\gamma}$, interleukin (IL)-$1{\beta}$, and tumor necrosis factor (TNF)-${\alpha}$. Treatment of the cells with polymyxin B, an inhibitor of lipopolysaccharides (LPS), blocked the induction of inflammatory mediators in LPS- or ${\beta}$-glucan-stimulated systems. Pretreatment of the cells in our cell culture system with LY294002, a phosphoinositide 3-kinase (PI3K) inhibitor, or U0126, a mitogen-activated protein kinase/extracellular-signal-regulated kinase (MAPK/ERK) kinase (MEK)1/MEK2 inhibitor, led to a reduction in the induction of inflammatory mediators in a concentration-dependent manner. These results show that stimulation of the macrophage cells by ${\beta}$-glucan induced the expressions of both dectin-1 and TLRs. We also found that the PI3K/Akt and MEK pathways were involved in the induction of inflammatory mediators in macrophage cells during intracellular signal transduction of ${\beta}$-glucan.

• Journal of Life Science
• /
• v.28 no.11
• /
• pp.1347-1353
• /
• 2018

The fermentation of non-digestible soy meal can convert polysaccharides into many compounds that have a wide variety of biological functions. Bacillus strains are capable of hydrolyzing non-digestible saccharides, such as melibiose, raffinose, and stachyose, found in soy meal components. A highly active ${\alpha}$-galactosidase (${\alpha}$-d-galactoside galactohydrolase, EC 3.2.1.22) was isolated from a bacterium in a traditional Korean fermented medicinal herb preparation. The isolate, T2-16, was identified as Bacillus coagulans based on its 16S rRNA sequence and biochemical properties, and the strain was named Bacillus coagulans NRR-1207. When incubated in 10%(w/v) skim milk, Bacillus coagulans NRR1207 caused a decrease in the pH of the culture medium, as well as an increase in titratable acidity and viable cell counts. This strain also showed higher activities of ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, naphthol-AS-BO-phosphohydrolase, and acid phosphatase when compared to other enzymes. It hydrolyzed oligomeric substrates, such as raffinose and stachyose, and liberated galactose, indicating that the Bacillus coagulans NRR1207 ${\alpha}$-galactosidase hydrolyzed the ${\alpha}$-1,6 glycoside linkage. These results suggest that the decreased stachyose and raffinose contents observed in fermented soy meal are due to this ${\alpha}$-galactosidase activity. Bacillus coagulans NRR1207 therefore has potential probiotic activity and could be utilized in feed manufacturing, as well as for hydrolyzing non-digestible soy meal components.