• Development and Reproduction
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• v.24 no.3
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• pp.207-214
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• 2020

Primary cell culture is a sufficient method frequently used to study the cellular properties and mechanisms of isolated cells in a controlled environment. In this study, an embryonic cell line (FGBC8) derived from the blastula stages of embryos of olive flounder Paralichthys olivaceus was developed. Furthermore, conditions for optimal long-term maintenance of this primary embryonic cell culture were investigated. Morphologically, FGBC8 cells were composed primarily of epithelial-like cells. FGBC8 cells were subcultured for >160 passages over ~830 days. The doubling time of FGBC8 cells was 73.8 h, and the modal diploid chromosome number was 48. FGBC8 cells transfected with green fluorescence protein (GFP)-expression plasmid exhibited a strong signal 48 h after transfection. Consequently, we demonstrated that fish serum is a crucial supplement for the long-term survival and maintenance of comparable morphology in these primary embryonic cells. Our results can be used as a guide for primary embryonic cell cultures for other fish species and may be useful for cell biotechnological applications.

• Journal of dental hygiene science
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• v.19 no.1
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• pp.39-47
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• 2019

Background: Light-emitting diodes curing unit (LCU), which emit blue light, is used for polymerization of composite resins in many dentistry. Although the use of LCU for light-cured composite resin polymerization is considered safe, it is still controversial whether it can directly or indirectly have harmful biological influences on oral tissues. The aim of this study was to elucidate the biological effects of LCU in wavelengths ranging from 440 to 490 nm, on the cell viability and secretion of inflammatory cytokines in MDPC-23 odontoblastic cells and inflammatory-induced MDPC-23 cells by lipopolysaccharide (LPS). Methods: The MTT assay and observation using microscope were performed on MDPC-23 cells to investigate the cell viability and cytotoxic effects on LCU irradiation. Results: MDPC-23 cells and LPS stimulated MDPC-23 cells were found to have no effects on cell viability and cell morphology in the LCU irradiation. Nitric oxide (NO) and prostaglandin $E_2$ which are the pro-inflammatory mediators, and interleukin-$1{\beta}$ and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) which are the proinflammatory cytokines were significantly increased in MCPD-23 cells after LCU irradiation as time increased in comparison with the control. LCU irradiation has the potential to induce inflammation or biological damages in normal dental tissues, including MDPC-23 cells. Conclusion: Therefore, it is necessary to limit the use of LCU except for the appropriate dose and irradiation time. In addition, LCU irradiation of inflammatory-induced MDPC-23 cells by LPS was reduced the secretion of NO compared to the LPS alone treatment group and was significantly reduced the secretion of TNF-${\alpha}$ in all the time groups. Therefore, LCU application in LPS stimulated MDPC-23 odontoblastic cells has a photodynamic therapy like effect as well as inflammation relief.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.67-80
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• 2024

Background: Post-ovulatory aging (POA) of oocytes is related to a decrease in the quality and quantity of oocytes caused by aging. Previous studies on the characteristics of POA have investigated injury to early embryonic developmental ability, but no information is available on its effects on mitochondrial fission and mitophagy-related responses. In this study, we aimed to elucidate the molecular mechanisms underlying mitochondrial fission and mitophagy in in vitro maturation (IVM) oocytes and a POA model based on RNA sequencing analysis. Methods: The POA model was obtained through an additional 24 h culture following the IVM of matured oocytes. NMN treatment was administered at a concentration of 25 μM during the oocyte culture process. We conducted MitoTracker staining and Western blot experiments to confirm changes in mitochondrial function between the IVM and POA groups. Additionally, comparative transcriptome analysis was performed to identify differentially expressed genes and associated changes in mitochondrial dynamics between porcine IVM and POA model oocytes. Results: In total, 32 common genes of apoptosis and 42 mitochondrial fission and function uniquely expressed genes were detected (≥ 1.5-fold change) in POA and porcine metaphase II oocytes, respectively. Functional analyses of mitochondrial fission, oxidative stress, mitophagy, autophagy, and cellular apoptosis were observed as the major changes in regulated biological processes for oocyte quality and maturation ability compared with the POA model. Additionally, we revealed that the activation of NAD⁺ by nicotinamide mononucleotide not only partly improved oocyte quality but also mitochondrial fission and mitophagy activation in the POA porcine model. Conclusions: In summary, our data indicate that mitochondrial fission and function play roles in controlling oxidative stress, mitophagy, and apoptosis during maturation in POA porcine oocytes. Additionally, we found that NAD⁺ biosynthesis is an important pathway that mediates the effects of DRP1-derived mitochondrial morphology, dynamic balance, and mitophagy in the POA model.

• Journal of Aquaculture
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• v.21 no.4
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• pp.331-338
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• 2008

Monthly changes in the gonadosomatic index (GSI) and hepatosomatic index (HSI) of wild river puffer Takifugu obscurus, and water quality environment in spawning area during breeding season were investigated from March 1995 to February 1996. Monthly changes in GSI and HSI of T. obscurus, that was cultured in low salinity, were calculated. The external morphology of the gonads, germ cell differentiation during gametogenesis and the reproductive cycle with the gonad developmental phases were investigated by histological analysis. The optimum water quality environment in Ganggyung, Choongcheongnam-do, where is spawning ground of wild T. obscurus, was $15-20^{\circ}C$ (water temperature) and 0 psu (salinity). Monthly changes in the GSI in females and males reached a maximum in May, and then rapidly decreased. Therefore, it is assumed that in the natural condition the spawning period of wild T. obscurus is May to June. In females and males, it showed a negative correlationship between the GSI and HSI. The external morphology of the gonads in female and male T. obscurus, that was cultured in low salinity, is composed of a pair of saccular structure. Based on monthly changes in the GSI, it is assumed that in female T. obscurus, that was cultured in low salinity, spawn from March through May. Therefore, it showed a negative correlationship between changes in the GSI and HSI. On the whole, in females and males, it showed a similar pattern between wild and cultured T. obscurus. The reproductive cycle with the gonad developmental phases can be classified into successive five stages in females: the early growing stage, late growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In males, that can be divided into successive four stages: the growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In case of wild T. obscurus, the spawning period has once a year, however, those cultured in the high water temperature ($20-27^{\circ}C$) - low salinity (under 3.3 psu) condition have reproductive characteristics having possibilities of discharge of eggs and sperms year-round as a multiple spawner.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.1
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• pp.23-29
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• 1989

In order to investigate developmental habit and morphology of maize tillers, time and location of tiller development. number of tillers per plant, tiller angle, height and diameter of tillers and root systems of tillers were examined under field condition for maize with tillers. Materials used were mostly from Korean local lines and a few lines from other countries were also included for comparison. The time of the first tiller development was about 18 to 20 days after emergence when planted on May in Yusong. The second tiller appeared about 4 to 5 days after the first tiller appeared. The tiller number per plant varied with lines and hybrids and ranged from two to ten. The location of tiller development was usually basal nodes of the main stem. Each tiller appeared to have its own root system. The angle between tillers and main stem was variable depending upon the maizes and the tiller angle could be classified into three categories. The height of tillers was also variable and seemed to be under genetic control. The most productive tillers were found among the Korean local derivatives.

• Development and Reproduction
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• v.11 no.2
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• pp.127-135
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• 2007

In order to monitor the developmental features of embryos, larvae, and juveniles of Konoshiro Gizzard Shad Konosirus punctatus, the fertilized eggs were gotton using artificial insemination. Konosirus punctatus were caught in Mankyung-myeon, Kimjae, Jeollabuk-do at June of 2004, and experiments were carried out in Ichthyology laboratory in Chonnam National University. Konosirus punctatus spawned draft egg from March to June. The fertilized eggs were cultured in $19.0{\sim}23.0^{\circ}C$(mean, $21.2^{\circ}C)$. The eggs had spherical shape and the diameter is $1.14{\sim}1.34\;mm$(mean, 1.21 mm). The lens began to appear from 35 hr 53 min after fertilization. At the moment, the movement of larvae was more active, and the tail was separated completely from yolk, the heart had forms, and melanophore appeared. Hatching was observed from 37 hr 10 min after fertilization. The total length of the hatched larvae was $4.26{\sim}5.30\;mm$(mean, 4.96 mm), but the mouth and anus were not opened at the time when the larva had yolk sack, and had $22{\sim}27$ myometium, and the anus located just abdominal front of the tail fin, and melanophore accumulated in the eye. Post-larvae used yolk completely after 2 day of hatching, and the total length was $4.96{\sim}5.74\;mm$(mean, 5.24 mm). From 16 days after hatching, the tail had curved tail end, and appeared the stems for pectoral, dorsal, and caudal fins. At 53 days after hatching, the total length of post-larvae was $27.11{\sim}34.09\;mm$(mean, 30.11 mm), and the frontal part of head developed like an adult one. At this time, fins and body are transferred to those of adult Konosirus punctatus. Fishes have a different shape and location of melanophore even in the same family. This research was tried to elucidate the early developmental features of Konosirus punctatus, together with species-specific pattern of melanophore.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 1997.05a
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• pp.27-34
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• 1997

Bovine oocytes surrounded with compact cumulus cells were cultured for 20 to 22 hours($38^{\circ}C$, 5% $CO_2$) in modified TCM-199 medium supplemented with 5% superovulated cow serum(SCS) and inseminated by in vitro capacitated spermatozoa. Day 7 to 8 embryos were equilibrated for 10 minutes in 1.3M methyl cellosolve(MC) <1.1M diethylene glycol(DEG), 1.8M ethylene glycol(EG), 1.6M propylene glycol(PG) and 1.1 M 1,3-butylene glycol(BG) solutions. They were then loaded into 0.25ml straws, placed into an alcohol bath freezer at $0^{\circ}C$, cooled from $0^{\circ}C$ to $-6^{\circ}C$ at $-1^{\circ}C$/minute, seeded, held for 10 minutes, and stored in liquid nitrogen. After thawing in $30^{\circ}C$ water, the embryos wee rehydrated in TCM-199 medium and then cultured for 48 hours in TCM-199 plus 5% SCS. Embryos were considered viable if they progressed to later developmental stages with a good morphology. Some of the embryos frozen in each cryoprotectant were thawed and transferred non-surgically without removing the cryoprotectant. Hatched embryos survived freezing and one-step dilution as follows : EG(50.0%), MC(53.6%), DEG(56.9%), PG(58.0%) and BG(11.5%). The survival rate of embryos cooled at -0.3^{\circ}C$ vs. $-0.5^{\circ}C$/minute was not significantly different(P<0.05), however, blastocysts hatched most often (P<0.01) in vitro when cooled at a rate of $0.3^{\circ}C$/minute(64.6%), 31/48) than at $-0.5^{\circ}C$/minute(22.6%, 12/53). Pregnancy rates resulting from embryos frozen in the different cryoprotectants were as follows : MC(48%, 10/21); DEG(30%, 3/10); EG(74%, 20/27); and PG(40%, 4/10). These results indicate that MC, DEG, EG and PG have utility as cryoprotectants for the freezing and thawing of IVF Bovine embryos.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.26 no.5
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• pp.458-464
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• 1993

From November 1992 to January 1993, the rockfish, Sebastes inermis was reared in the laboratory, and observed the developmental stage of the eggs and larvae squeezed from the parent fish. On December 4 1992, developing eggs at morula stage were obtained from female parent fishes($17.4{\sim}20.3mm$ in total lenght(TL)). The diameter of eggs ranged from 1.20 to 1.35mm(n=50), and oil globules in white color were found in the eggs. Hatching began about 55 hours after morula stage at the water temperature of $9.2{\sim}12.3^{\circ}C$. The newly hatched larvae measured from 2.90 to 3.35mm in TL with $8+12{\sim}13=20{\sim}21$ myomeres. Total lengths of the larvae will be from 5.75 to 6.15mm in 27 and 30 days after the hatching, respectively. Myomere number was $8+17{\sim}18=26$.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.229-234
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• 2008

An efficient micropropagation was established by using axillary bud explants from two-year-old tree(Echinosphorea koreensis Nakai), which has been known as a rare and endangered species. Among various basal media tested, DKW medium was shown to be the best for axillary shoot elongation. The addition of both BA and TDZ to the medium induced 6 to 10 shoots per explant during eight weeks of culture, without showing any abnormal morphology at the shoot proliferation stage. However, high concentration of TDZ(>0.05 mg/L) appeared to cause hyperhydration on either leaf or shoot at the later developmental stage. Approximately 20% of shoots produced roots by the addition of 1.0 mg/L NAA but not by IBA($0.2{\sim}1.0$ mg/L). Ex vitro micro-cuttings were better source for root induction; up to 58.6% of the micro-cuttings rooted when 100 mg/L IBA was applied to the soil(vermiculite). More than 90% of plantlets with roots were successfully acclimatized and grew normally in the field. Therefore, we suggest that this endangered tree species can be effectively micropropagated by axillary bud culture system developed in this study.

• Journal of Korean Society of Forest Science
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• v.30 no.1
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• pp.50-60
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• 1976

For finding out the proper method of developing Buxus koreana var. insularis, this study was carried out in Pokil-Do, Wan-Do, Chonnam, in 1975. The ecology, morphology and characteristics of the plant were mainly investigated and analyzed. The results are summarized as follows; 1. The small islands of the southern sea of Korea; Pokil-Do, Chin-Do and So-Huk-San-Do, turn out to be the site of the natural communities of B. koreana var. insularis. 2. The growth of the plant is fairly good in the acid soil of these islands. 3. This vegetation of composed as the three-layered communities, these communities consit of Camellia, Buxus and Cares. 4. The plant is a board leaved evergreen shrub that is tolerant to shade and native to the temperate zone. 5. The size of the leaves, fruits and seeds is slightly larger than those of B. koreans. 6. As fruiting and germination of the seeds are favorable. it is easy for us to make a number of seedlings. 7. The proper times for sowing and harvesting the seeds are around the middle of July and August, respectively. 8. The seedlings is difficult to grow in the naked ground because the germinated seeds are seriously damaged by frost. 9. The plant growth per annum is about 8 to 10cm in height, 1.5mm in diameter. It is graceful looking as the branches and leaves as of the dense foliage type 10. It is possible to develop the plant in many ways, such as; ornamental plant, bee plant, sculptural and industrial materials.