• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.379-384
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• 1996

This study was undertaken to investigate the sister chromatid exchange (SCE) frequency and embryonic development after exposure to cryoprotectants and vitrification of mouse zygotes. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). After mouse zygotes were exposed to EFS40 at $25^{\circ}C$ for 30 sec., they were immediately plunged into $LN_2$ or cultured for cryoprotectant toxicity test without freezing. The results obtained in these experiments were summarized as follows: After thawing, survival rates to the 2-cell stage of zygotes exposed to or vitrified in EFS 40 (98.5%, 95.2%) were not significantly difference compared with that of control (100%). However, the developmental rates upto blastocyst and hatching blastocyst in vitrified groups (66.7, 50.0%) were lower than those of control (93.9, 81.8%) or exposed group (94.0, 78.8%) (p<0.05). When the influence of vitrification and exposure to cryoprotectant on the in vitro development of mouse zygotes was assessed by the SCE frequency, the SCE frequency in exposed ($20.2{\pm}2.1$) to or vitrified embryos ($21.4{\pm}3.2$) was higher than that in control embryos ($16.8{\pm}1.5$). These results suggest that the frequency of SCE was increased after cryoprotectant exposure or Vitrification although developmental rates of zygotes upto blastocysts and /or hatching blastocysts were not afected by cryoprotectant.

• Proceedings of the Ginseng society Conference
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• 1984.09a
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• pp.27-36
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• 1984

It was previously reported that red ginseng extract inhibited carcinogenesis by urethan, DMBA and aflatoxin $B_1E (Cancer Detection and Prevention, 6: 515-525, 1983). In an attempt to investigate the mechanism of the anticarcinogenic effect of ginseng, we assayed natural killer (N.K) activity in mice treated with urethan and benzo(a)pyrene. In our experiment newly born Swiss Webster mice, less than 24 hrs. old, were given a single subcutaneous injection of lmg of ure-than and 40ug of benzo(a)pyrene. The mice had been administered with ginseng since weaning, and sacrificed at various intervals. Major organs were examined both, with the naked eye and microscopically. N.K. activity of spleen cells was analyzed in a 12-hour $^{51}Cr^-release$ assay against YAC-1 cells. Administration of ginseng resulted in an increase of N.K. activity by $18\%$ at 4 weeks, $20\%$ (P < 0.05) at 6, $29\%$ (P < 0.05) at 12, and $13\%$ at 24 following a single injection of urethan. At the same time, significantly lower incidences of lung adenoma were noted at 6 weeks $(50\%)$ and 12 weeks $(27\%)$ following the administration of ginseng to urethan-injected mice. This result indicates that the enhancement of N.K. activity by ginseng makes a contribution to its anticarcinogenic effect. On the hand, N.K. activity was suppressed by benzo(a)pyrene during the time span of this experiment and it almost returned to the level of controls following the adminsitration of ginseng. However, the lung adenoma induced by benzo(a)pyrene began to occur at 48 weeks in which N.K. activity had naturally declined to a very low level in all experimental mice, and administration of ginseng did not decrease the incidence. In explanation of this result, we might propose that the recovery of the N.K. activity by ginseng had little effect on the incidence of lung adenoma because of the long latent period of carcinogenesis by benzo(a)pyrene. In conclusion, these results suggest that the anticarcinogenic effect of ginseng in urethan-treated mice may be related to the augmentation of N.K. activity.

• Food Science and Preservation
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• v.23 no.6
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• pp.876-882
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• 2016

The antioxidant and anti-inflammatory effects of Corni fructus extracts (CEF, EtOAc extraction; CBF, buthanol extraction; CWF, water extraction) were investigated. The total phenolics of CEF (173.3 mg TAE/g) were significantly higher than those of CWF (26.7 mg TAE/g) and CBF (94.8 mg TAE/g). DPPH and ABTS free radical scavenging activity of CEF (DPPH: $RH_{50}$; $25.1{\mu}g/mL$, ABTS: $RC_{50}$; $36.1{\mu}g/mL$) showed even higher than that of BHA and ${\alpha}-tocopherol$ used as positive control. All three Corni fructus extracts in the concentration of $1{\sim}100{\mu}g/mL$ were effective inhibitors of NO and prostaglandin $E_2$ ($PGE_2$). NO production was inhibited 71.3~92.2% by CEF, 76.8~85.5% by CBF and 74.4~96.9% by CWF, respectively. CEF, CBF and CWF ($1{\sim}100{\mu}g/mL$) inhibited also pro-inflammatory cytokines like $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 very effectively. $TNF-{\alpha}$ was inhibited up to 51.2% by CWF and $IL-1{\beta}$ was inhibited up to 67.1% by CEF. IL-6 was best inhibited by CEF up to 58.9%. This study suggested the potential of Corni fructus for use as an excellent antioxidant substance and inflammatory inhibiting mediators. Therefore CEF, CBF and CWF Corni fructus extracts may be used for therapeutic approach to various inflammatory diseases.

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.207-216
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• 2013

This study was conducted to establish the method for preserving PGCs that enables long-term storage in liquid nitrogen for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of freeze-thaw treatment on viability of PGCs in chickens. PGCs were collected separately from a germinal gonad of an early embryo of 5.5~6 day (stage 28) of Isa brown, Korean Oge (KO), White Leghorn and Commercial breeds. PGCs separated from a germinal gonad of an early embryo of 5.5~6 day (stage 28) are suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and glycerol). The PGCs were then purified using magnetic activated cell sorting (MACS) method. The viability of PGCs after thawing was $87.4{\pm}0.4%$ and $89.4{\pm}0.2%$ with the 10% EG treatments with no significant difference between the Isa brown and Commercial breeds. The viability of PGCs after freeze- thawing was significantly higher for Isa brown ($87.4{\pm}0.4%$) and Commercial breeds ($89.4{\pm}0.2%$) than Korean Oge (KO) ($77.6{\pm}1.1%$) and White Leghorn ($76.2{\pm}0.9%$)(p<0.05) using 10% EG cryoprotectant. This study established a method for pre- serving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at agermplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.5
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• pp.448-454
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• 2000

This study was to examine the physicochemical characteristics of coagulation reaction between ignited oyster shell powder (IOSP) and red tide organisms (RTO), and its feasibility, in developing a technology for the removal of RTO bloom in coastal sea,IOSP was made from oyster shell and its physicochemical characteristics were examined for particle size distribution, surface characteristic by scanning electron microscope, zeta potential, and alkalinity and pH variations in sea water. Two kinds of RTO that were used in this study, Cylindrotheca closterium and Skeletonema costatum, were sampled in Masan bay and were cultured in laboratory. Coagulation experiments were conducted using various c(Incentrations of IOSP, RTO, and a jar tester. The supernatant and RTO culture solution were analyzed for pH, alkalinity, RTO cell number, IOSP showed positive zeta potentials of $11.1{\~}50.1\;mV\;at\;pH\;6.2{\~}12.7$, A positive zeta potential of IOSP slowly decreased with decreasing pNa 4,0 to 2,0. When pNa reached zero, the zeta potential approached zero, When a pMg value was decreased, the positive zeta potential of IOSP increased until pMg 3.0 and decreased below pMg 3.0. IOSP showed 4.8 mV of positive zeta potential while RTO showed -9.2 mV of negative zeta potential in sea water. A positive-negative EDL (electrical double-layer) interaction occurred between $Mg(OH)_2$ adsorption layer of IOSP and RTO in sea water so that EDL attractive force always worked between them. Hence, their coagulation reaction occurred at primary minimum on which an extreme attractive force acted because of charge neutralization by $Mg(OH)_2$ adsorption layer of IOSP. As a result, the coagulation reaction was rapidly processed and was irreversible according to DLVO (Deriaguin-Landau-Verwey-Overbeek) theory. Removal rates of RTO were exponentially increased with increasing both IOSP concentration and G-value. The removal rates were steeply increased until 50 mg/l of IOSP and reached $100{\%}\;at\;400\;mg/l$ of IOSP. Removal rates of RTO were $70.5,\;70.5,\;81.7,\;85.3{\%}$ for G-values of $1,\;6,\;29,\;139\;sec^(-1)$at IOSP 100 mg/l, respectively. This indicated that mixing (i.e., collision among particles) was very important for a coagulation reaction.

• Journal of Life Science
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• v.27 no.4
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• pp.383-389
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• 2017

This study intends to analyze the contents of rutin, procyanidin B3, quercetin, kaempferol, known to have antioxidant, anti-inflammatory and anti-carcinogenic effects, among the polyphenol type contained in the grape pruning stem extracts (GPSE), utilizing grape stems being discarded after harvest, measure the effects on the skin moisture, inhibition of skin cell proliferation, anti-inflammatory on the damaged skin of a HR-1 mice induced with UVB, and verify the applicability as a material for functional food and functional cosmetics. The results of verifying the photoprotection effects through the skin proliferation control through of GPSE showed similar result to suncream was achieved at the GPSE concentration of 2,000 mg/kg on the epidermis (p<0.05). The results showed anti-inflammatory effects on all groups applied with GPSE as compared to the control group irradiated with UVB, but at the GPSE concentration of 1,000 mg/kg, a lower COX-2 protein expression at 8%, lower than the 22% of suncream, was observed to achieve an excellent anti-inflammatory effect (p<0.05). The results of this study confirmed the existence of active polyphenol type, such as rutin, kaempferol, querocetin and procyanidin B3, within the GPSE, and GPSE has improvement effects on moisturizing effects, skin proliferation control effect, inflammatory control effect and improvement effects on the skin barrier function through UV ray damage. GPSE is a functional ingredient with a potential for skin protection effects, and has high utilization as an ingredient for functional food and functional cosmetics.

• The Korean Journal of Pesticide Science
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• v.18 no.3
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• pp.202-209
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• 2014

RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.611-616
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• 2001

The bacteriological and physiochemical analysis of seawater in Tongyeong harbor was conducted to evaluate sanitary conditions, The samples were collected at 6 stations established once a month from January to December, 2000. During the study period, the ranges of temperature, transparency, chemical oxygen demand, dissolved oxygen, dissolved nitrogen, phosphate and chlorophyll-a were $6.8\sim25.2^{\circ}C,\;1.0\sim2.5\;m,\;1.79\sim2.41\;mg/L,\;5.7\sim10.1\;mg/L,\;6.59\sim10.53{\mu}g-at/L,\;0.56\sim1.01{\mu}g-at/L\;and\;1.21\sim9.54\;mg/m^3$, respectively, The viable cell counts of seawater in Tongyeong harbor ranged from $3.0\times10^4CFU/mL\;to\;6.9\times10^6CFU/mL$. The coliform group and fecal coliform MPN's of the samples were ranged $23\~4,600\;MPN/100\;mL$ (means 540 MPN/100 mL) and $11\~1,600\;MPN/100\;mL$ (means 210 MPN/100 mL), respectively, The coliform group was classified with IMViC reactions and pathogenic vibrios were analyzed. Two hundred eighteen strains that were obtained from seawater samples in Tongyeong harbor represented Escherichia coli group, $66.1\%$; Citrobacter freundii group, $11.0\%$; Enterobacter aerogenes, $9.6\%$; and unknown, $13.3\%$, respectively. During the study period, infectious bacteria such as Vibrio cholerae O1, Salmonella sp. and Shigella sp. were not detected from the samples, but detection ratios of V. parahaemolyticus, V cholerae non-O1 and V. vulnificus were $10.0\sim30.1\%$.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.37-37
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• 2003

We have shown the $Ca^{2+}$-activated chloride current is present in cardiac myocyte in rabbit pulmonary vein (Kim et al., 2002). This current amplitude was increased as [N $a^{+}$]$_{i}$ was increased and we suggested this chloride current may be involve in the spontaneous action potential frequency change. Since this current is activated by the increase of intracellular $Ca^{2+}$, we would like to test what is the inducer of the increase of [C $a^{2+}$]$_{i}$ between a L-type $Ca^{2+}$-current or a reverse mode of N $a^{+}$-C $a^{2+}$ exchange current. White rabbit (1.5 kg) was used and anesthetized with Ketamin (100 mg/kg). Pulmonary vein (PV) was isolated and sleeve area between left atrium and PV was dissected. Using collagenase (Worthington 0.7 mg/cc), single cardiac myocytes were isolated. In the presence of 15 mM of N $a^{＋}$, three steps of voltage pulses were applied (holding potential : -40 ㎷, -80 ㎷ for 50 msec, 30 ㎷ for 5 msec, 10 ㎷ steps from -70 ㎷ to 60 ㎷). The inward and outward tail current was activated after brief 5 msec prepulse. The outward tail current was blocked by the removal of extracellular chloride substituted by glucuronic acid or by a chloride channel blocker, 5 mM 9-AC. But the inward tail current was still remained even though the amplitude was decreased. The reversal potentials were changed to the direction of the change of chloride equilibrium potential ( $E_{Cl}$ ) but the shift of equilibrium potential was not enough to match to the theoretical equilibrium potential shift. In the presence of L-type $Ca^{2+}$ channel blocker, nifedipine 1 uM, inward tail currents were greatly reduced but the outward current tail currents were still remained. In the presence of N $a^{+}$-C $a^{2+}$ exchange current blocker, 10 uM KB-R7943, the inward and outward tail currents were blocked almost completely. We tried to test the $Ca^{2+}$sensitivity of the chloride current with various [C $a^{2+}$]$_{i}$ in pipette solution from 100 nM to 1 uM but we failed to activate $Ca^{2＋}$-activated chloride currents even though the cell became contracted in the presence of 1 uM $Ca^{2+}$. From these results, we could conclude that the increase of [C $a^{2+}$]$_{i}$ to activate the outward $Ca^{2+}$-activated chloride current was mainly induced by the activation of the reverse mode of N $a^{+}$-C $a^{2+}$ exchanger, But for the increase of [C $a^{2+}$]$_{i}$ to activate the inward tail current, L-type $Ca^{2+}$ current may be the major provoking current. Since the cytosolic increase of [C $a^{2+}$]$_{i}$ through pipette solution have failed to activate $Ca^{2+}$-activated chloride current, this chloride current may have very low $Ca^{2+}$ sensitivity or a comparmental increase $Ca^{2+}$ such as in subsarcolemmal space may activate the chloride current. Since there are several reports and models that the increase of $Ca^{2+}$ in subsarcolemmal space would be over several to tens of uM, both possibility may be valid together.uM, both possibility may be valid together.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.3
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• pp.199-210
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• 2011

This study was conducted to develop functional sources of herbal cosmetics for treatment of skin aging and inflammatory disorders using volatile flavor extracts of four different herbal medicinal prescriptions including Cnidium officinale Makino (COM), Angelica gigas Nakai (AGN), Mentha arvense L. (MAL), Artemisiae argyi Folium (AAF), Paeonia lactiflora Pall (PLP), Rehmanniae Radix Preparata (RRP), Scutellaria baicalensis Georgi (SBG), Panax ginseng C.A. Meyer (PGM), Glycyrrhiza uralensis Fisch (GUF). The volatile flavor extracts of four different herbal medicinal prescriptions (HH-1: COM, AGN, PLP, RRP, HH-2: COM, AGN, PLP, RRP, SBG, PGM, GUF, HH-3: COM, AGN, MAL, AAF, HH-4: COM, AGN, MAL, AAF, SBG, PGM, GUF) were extracted using SDE and their antioxidant and anti-inflammatory effects were measured by using DPPH radical and SLO, respectively. As a result, HH-2 showed moderate DPPH radical scavenging activity (68.24 %) and the strongest SLO inhibitory activity (83.96 %) at 100 ${\mu}g$/mL. Moreover, HH-2 of four different prescriptions significantly inhibited NO production on LPS-stimulated RAW 264.7 cells in a dose-dependent manner without considerable cell cytotoxicity at range of 2.0 ~ 50 ${\mu}g$/mL. Additionally, HH-2 also effectively suppressed the production of $PGE_2$ and IL-6, which are responsible for promoting the inflammatory process. Major volatile components of HH-2 were identified as eugenol, paeonol, butyl phthalide, ${\beta}$-eudesmol and butylidene dihydrophthalide by GC-MS analysis. Thus, these results suggest that HH-2 may be useful as a potential source of anti-inflammatory agents in herbal medicinal cosmetics.